2012
VI BIENNIAL
NETROPICA
MEETING
Final Program
Hosted by the Graduate Program in
Infectious and Zoonotic Diseases;
School of Microbiology, UNAH
Co-Sponsored by the Teasdale-Corti
Project, Honduras-Canadá
Designed and Edited by Gustavo Fontecha-Sandoval-UNAH
Welcome
Dear Colleagues:
Welcome to Copán Ruinas, Honduras, and to the 2012 Biennial NeTropica Meeting!
Our tagline this year, "Research without Borders”, conveys our belief that science can be
both a medium and a purpose to bring people together. And indeed we have set a new
record in NeTropica meetings’ attendance: we are proud of hosting more than 120
participants from eleven countries in Central America, North America, South America,
Europe, and Asia.
Congregated here in the mythical City of Copán, one of the most important scientific
centers of the Mayan culture, speakers from all these countries will present over 80
posters, seven International Conferences, two Symposia and two Round Tables on topics
such as Infectious Diseases Research, Research Policies, National Health Research Systems,
Nutrition, Global Health, and Research Partnerships.
By attending the various sessions and interacting with colleagues and friends, there will be
stimulating opportunities for everyone to catch up with scientific discoveries; learn of new
possibilities, and very importantly, hear and or pose new questions. But besides intellectual
and personal growth, an important goal this year is to spark formal and informal discussions
around research sustainability in the Central American region. This is certainly an ulterior
motive for the VI NeTropica Meeting.
Every scientist in Central America faces a dual challenge: first, to do high-quality science
that meets international quality standards and is relevant for the development of the
region; and second, to maintain robust and uninterrupted research agendas that
necessitate continuous research funding in an increasingly competitive world. Long term
success in meeting these challenges can only be achieved by establishing academic research
programs through which new generations of young scientists are formed.
The education and training of these future scientists is, however, a tall order; it is expensive
and requires resources, dedication and sacrifices from most of us who are here today. But
together we can. I honestly believe that only through the union and dynamic partnerships
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between Central Americans and the collaboration of colleagues from more developed parts
of the world, we will be able to secure the scientific advancement of the region. I truly hope
this meeting will mark a new era of renewed friendships and stronger professional
connections leading to lasting and productive collaborations.
Finally, I invite you to make this meeting more than just about science. Take your time to
experience first-hand the rich historical heritage of the Mayan culture and enjoy the natural
beauty of the surrounding area. Copan has many gifts waiting for you.
On behalf of the Organizing Committee, I would like to extend my warmest welcome and
say we are delighted you are here.
Best wishes,
Gustavo Fontecha, PhD.
President VI VBiennial NeTropica Meeting
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Congress Committees
Ana Lourdes Sánchez (Brock University, CA)
Scientific Program
Committee
Esteban Cháves-Olarte (Universidad de Costa Rica)
María Elena Bottazzi (Baylor College of Medicine, USA)
Gustavo Fontecha-Sandoval (UNAH) (President)
Organizer and Logistics
Committee
Lourdes Enríquez (UNAH)
Maritza Canales (UNAH)
Annabelle Ferrrera (UNAH)
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Brief Summary about NeTropica
Since 1998 and with the financial support from Sweden’s International Development
Agency (SIDA/SAREC), and the participation of Central American public universities, the
Central American Network for Research and Training in Tropical Diseases, NeTropica, has
promoted the integration of regional scientific consortia to address problems of regional
interest in the field of Tropical Medicine, Neglected Diseases and other Infectious Diseases.
NeTropica’s specific objective has been to support investigative endeavors of researchers
trained through the Karolinska Institute Research and Training Program in Central America
(KIRT/CA) that operated in the region from 1988 to 1999. In a decade, the KIRT/CA
graduated a total of 49 researchers: 26 with MSc and 23 with PhD degrees.
The core principle behind NeTropica’s work is that the sustainable control of tropical
diseases can only be achieved through the dynamic engagement of scientific groups
working to address and understand the key determinants of diseases affecting local
populations. For KIRT trainees to accomplish such tall order, NeTopica has worked diligently
in making sure that upon acquiring specific training, researchers have access to two
fundamental factors for scientific success: a network of peers with who collaborate, and the
necessary operating budget to conduct research.
In its life span NeTropica has issued 6 calls for grant applications awarding a total of 54
grants averaging US $ 35,000 to local consortia of researchers. The impact of NeTropicasponsored research is measurable both in terms of academic advancement and health
outcomes for Central America.
NeTropica has also been dedicated to provide spaces and opportunities for researchers to
share and disseminate their findings. To this end, NeTropica has organized five bi-annual
international scientific meetings, which have been held in Costa Rica, Guatemala, Honduras,
Panama and Nicaragua. The sixth meeting will take place in Copan, Honduras, on July 2012.
At the end of the Swedish cooperation cycle, NeTropica is committed to continuing its
essential work for the advancement of scientific research in the region, specifically
supporting the training and establishment of a new generation of young researchers who
will lead the way into better health and development of the region. Ensuring NeTropica’s
sustainability is crucial for the Central American research community, but more
importantly, for the people benefited by the research done under the auspices of this
network.
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VI Biennial NeTropica Meeting:
Research without Borders
The “VI Biennial NeTropica Meeting: Research Without Borders” will be organized by the
Master’s Program in Infectious and Zoonotic Diseases (MEIZ), School of Microbiology at the
National Autonomous University of Honduras.
MEIZ is a Canadian-sponsored graduate program operating since 2009, and is the first of its
kind in Central America. MEIZ’s guiding principle is that CA countries must establish their
own graduate programs in which to train the next generation of researchers. By tapping
into the pool of existing local talent generated by other initiatives such as the Karolinska
Institute Research and Training Program in Central America (KIRT/CA) and by relying on and
producing their own resources, local graduate programs are more likely to be sustainable
and therefore have a stronger and more lasting impact.
The intersection of NeTropica and the legacy of KIRT/CA with MEIZ in organizing the “VI
Biennial NeTropica Meeting: Research Without Borders” signals a new era of research and
collaboration in Central America. The meeting will strive to bring together researchers,
students and stakeholders interested in advancing tropical disease research in CA and
examining health issues from a broad perspective of proximal and distal determinants.
Tropical diseases issues will be addressed through discussions, workshops, symposia and
presentations that consider broader biomedical and societal frameworks such as:
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Relevant Infectious and Zoonotic Diseases
The Millennium Development Goals
Innovation and technology for development and poverty alleviation
Strengthening of research capacities and networks
Investment in future generation of researchers
One Health
Neglected tropical diseases
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Sponsors
Co-Hosts
NeTropica
Global Health Research Initiative, Canada
Proyecto Teasdale-Corti Honduras-Canada
Universidad Nacional Autónoma de Honduras
Collaborating Organizations
Baylor College of Medicine
Colegio de Microbiólogos de Honduras
Brock University
Prodylab
Agencia ARDE, S de RL
Distribuidora Comercial, S.A. DICOSA
DIMEX Médica
Analítica Hondureña, ANAHLISA
Diprolab de Honduras
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Participating Organizations
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Baylor College of Medicine, USA
Brock University, CAN
Canadian Coalition for Global Health
Centro de Investigación en Enfermedades Tropicales, CR
Clínica Santa Rosa, HN
Clínica El Buen Pastor, HN
COHRED
Complejo Hospitalario Metropolitano, PN
Hokkaido University, JP
Hospital Escuela de Honduras
INISA-UCR
Instituto Clodomiro Picado, CR
Instituto Conmemorativo Gorgas, PN
Instituto Tecnológico de Costa Rica
International and Maternal's and Children's Health, Upssala Univ. SU
Karolinska Institute, SU
Laboratory of Pathology of Infectious Diseases, Med School of São Paulo University, BR
Laboratorio Nacional de Malaria de Honduras
Maestría en Enfermedades Infecciosas y Zoonóticas-UNAH, HN
Mississippi State University, USA
NeTropica
PAHO-Honduras
PIET-UNA, CR
Programa Nacional de Chagas y Leishmania de Honduras
IHIMV-Secretaría de Agricultura y Ganadería de Honduras
ISGlobal. Universitat de Barcelona - Hospital Clinic, Barcelona, España
Secretaría de Salud de Honduras
Subsecretaría de riesgos poblacionales, Honduras
TEPHINET
Universidad de Costa Rica
Universidad de San Carlos de Guatemala
Universidad del Valle de Guatemala
Universidad Nacional Autónoma de Honduras
Universidad Nacional Autónoma de Nicaragua
Universidad Nacional de Costa Rica
University of Texas
Uppsala University, SU
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Program-at-a-Glance
Time
Wed. July 25
8:30-9:30
9:30-10:00
10:00-10:20
Arrivals
10:20-10:45
10:45-11:10
11:10-11:35
11:35-12:00
12:00-1:00
1:15-2:15
2:15-3:15
3:15-3:30
Onsite Registration
3:30-4:30
4:30-6:30
6:30-7:00
7:00-8:00
8:00-10:00
Welcome
(Dr. Gustavo Fontecha, MEIZ-UNAH)
Keynote Address
Facts and Fiction of Global Health
(Prof. Hans Rosling, Karolinska Institute)
Welcome Cocktail
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Program-at-a-Glance
Time
Thursday July 26
8:30-9:30
Conference 1
National Health Research Systems
(Dr. Francisco Becerra, COHRED)
9:30-10:00
Poster Session*
10:00-10:20
Coffee Break
10:20-10:45
10:45-11:10
11:10-11:35
11:35-12:00
12:00-1:00
1:15-2:15
2:15-3:15
** Borrelia turicatae adaptation within the arthropod vector. Job López
Phylogenetic analysis of Type A Influenza viruses isolated from waterfowl in Guatemala, Central America.
Ana Silvia Gonzales
Preliminary results of a study on Trypanosoma cruzi congenital transmission in two endemic regions of
Honduras. Jackeline Alger
Sand fly density reduction is less marked in precarious housing after insecticide residual spraying. Luis
Fernando Chaves
Lunch break
Round Table 1
Vector-Borne and Zoonotic Diseases in Guatemala
(Convener: Dr. Pamela Pennington, Universidad del Valle de Guatemala)
Conference 2
New Paradigms in HPV-Associated Cancers Prevention
(Dr. Rolando Herrero, International Agency for Cancer Research, WHO)
3:15-3:30
Break
3:30-4:30
Conference 3
Texas and Mexico: Sharing a Legacy of Poverty and Neglected Tropical Diseases
(Dr. Maria E. Bottazzi, Baylor College of Medicine, USA)
4:30-6:30
Symposium 1
Microbial Genetics
(Convener: Dr. Esteban-Chaves-Olarte, University of Costa Rica)
6:30-7:00
Networking
7:00-8:00
Dinner
8:00-10:00
Cultural event
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Program-at-a-Glance
Time
Friday July 27
8:30-9:30
Conference 4
Nutrition and Human Development
(Dr. Reynaldo Martorell, Emory University)
9:30-10:00
Poster Session*
10:00-10:20
Coffee Break
10:20-10:45
** Snakebite Envenomings in Latin America: Current Situation and Challenges. José Ma. Gutiérrez
10:45-11:10
11:10-11:35
11:35-12:00
12:00-1:00
1:15-2:15
2:15-3:15
Competence of Leishmania species to determine different clinical and immunological features in murine
infection. Marcia Laurenti
Health care workers’ knowledge, attitudes and reported practices related to health care-associated
infections in an Indian teaching hospital. Senia Rosales
Establishing a Proteomics Laboratory at Instituto Clodomiro Picado (UCR): first two-year's experience in
serving local and regional research. Bruno Lomonte
Lunch break
Conference 5
Research to Policy
(Dr. Luis Gabriel Cuervo, PAHO)
Conference 6
Assessing North-South Partnerships: The Canadian experience
(Dr. Vic Neufeld, CCGHR, Canada)
3:15-3:30
Break
3:30-4:30
Round Table 2
Research Skills for a Successful Scientific Career
(Convener: Dr. Ana Sanchez, Brock University)
4:30-6:30
Symposium 2
Strategies for Research Sustainability in Central America
(Convener: Dr. Gustavo Fontecha, MEIZ-UNAH)
6:30-7:00
Concluding Remarks
(Scientific Committee)
7:00-8:00
8:00-10:00
Gala Dinner
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General Information
Conference Venue
Clarión Hotel Copan Ruinas, Honduras
+504 2651 4480 / +504 2651 4481 / +504 2651 4482
Cell Phone: +504 3190 3487 / +504 9726 9613
Emergency Cell Phones
+504 33935443 / +504 99966160 / +504 94870165
Opening Hours of Registration and Information Desk
Tuesday, July 24, 2012
08:30 – 18:00
Wednesday, July 25, 2012
8:30 – 18:00
Registration Fees
Professional: USD 150.00
Student:
USD 100.00
Social Program:
The Welcome Cocktail will be held on Wednesday, July 25 from 20:00-22:00hrs at the Patio
of the Hotel Clarión.
A Mayan cultural event will be held on Thursday, July 26 from 20:00-22:00hrs at the Patio
of the Hotel Clarión, and a Gala Dinner will be held on Friday, July 27 from 20:00-22:00hrs
at the Restaurant of the Hotel Clarión.
Badges:
Please wear your name badge at all times during the conference in order to gain access to
the scientific program and all conferences.
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Congress Language
The language of the Meeting is English.
Speakers Center
An LCD projector and laptop for PowerPoint presentations are provided in the meeting
room. All speakers are requested to take their presentations on CD/USB stick to the
speakers’ center. All presentations must be received as early as possible, but no later than 1
hour before the presentation. Please use the meeting room laptop computer rather than
your personal computer for your presentation. A student will be available to assist the
speakers.
Meals and Coffee breaks
The hotel fee includes all the meals from dinner of Wednesday July 25 to dinner of Friday
July 27 as well as the coffee break during the meeting. Any other consumption will be
charged to your room.
Airport Taxes
You should be aware that any departure flight from Honduras is charged with a
Government tax of USD 38.00 that you must pay when you arrive to the San Pedro Sula
Airport before taking your flight back home.
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Plenary and Keynote Speakers
HANS ROSLING (Karolinska Institute)
Keynote Address: Facts and Fiction on Global Health
Professor of International Health, Karolinska Intitutet,
Director of Gapminder Foundation www.gapminder.org
Stockholm
Hans Rosling is Professor of International Health at Karolinska Intitutet
the medical university in Stockholm, Sweden. When working as a young
doctor in Mozambique he discovered a previously unrecognized paralytic disease that his
research team named Konzo. His 20 years of research in global health concerned the
character of the links between economy and health in Africa, Asia and Latin America
He has been adviser to WHO and UNICEF, co-founded Médecines sans Frontiers in Sweden
and started new courses and published a textbook on Global Health. He is a member of the
International Group of the Swedish Academy of Science and of the Global Agenda Network
of the World Economic Forum in Switzerland.
He co-founded Gapminder Foundation with his son and daughter-in-law. Gapminder
promotes a fact based world view by converting the international statistics into moving,
interactive, understandable and enjoyable graphics. This way first done by developing the
trendalyzer software that Google acquire in 2007.
Using animations of global trends, Hans Rosling lectures about past and contemporary
economic, social and environmental changes in the world and he produces thematic videos
using the same technique. His award-winning lectures on global trends have been labeled
"Humorous, yet deadly serious" and many in the audience realize their own world view is
lagging many decades.
Hans Rosling's 5 points on global trends are:
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1. There are no longer two types of countries in the world, the old division into
industrialized and developing countries has been replaced by 192 countries on a
continuum of socio-economic development.
2. Many Asian countries are now improving twice as fast as Europe ever did.
3. A new gap may form between 5 billion people moving towards healthy lives with
education, cell phones, electricity, washing machines and health service and more
than 1 billion people stuck in the vicious circle of absolute poverty and disease.
4. So far all progress towards health and wealth has been achieved at the price of
increased CO2 emission that drives the imminent climate crisis.
5. There are reasons for optimism regading the future of the world because the world
is so poorly governed at present. Hence we have enormous opportunities to
improve the life of all humans by turning our already converging world into an
equal, secure sustainable and free place to live in.
Plenary and Keynote Speakers
FRANCISCO BECERRA-POSADA (COHRED)
Conference 1: National Health Research Systems
A Mexican national, is a Medical Doctor from the National
Autonomous University of Mexico. And his MPH from The Johns
Hopkins University School of Public Health. He is currently enrolled in
the Doctor in Public Health Program at the National Institute of Public
Health (INSP) in Mexico. He has been an international consultant for PAHO, WHO, USAID,
CARE, CORAT-Africa and Save the Children-US, among others. In Mexico he has held diverse
positions, such as Director General for the State of Morelos Health Services, Director for
Academic Agreements for the Coordinating Office of the National Institutes of Health,
Mexico, Assistant Director for the Center for Health Systems Research, INSP. Before joining
COHRED, he was Joint Director General for Federal Hospitals in Mexico. In COHRED he is
developing the COHRED LAC unit and is Head of COHRED-LAC, he is also part of the
COHRED-TECH team work, and is the coordinator of the EU/FP7 funded project ‘MASCOT’,
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he participates in another EU/FP7 project: ‘EU-LAC HEALTH’. Email: [email protected]
Website: www.cohred.org Phone: +5255 4622 5441
Plenary and Keynote Speakers
ROLANDO HERRERO (International Agency for Cancer Research,
WHO)
Conference 2: New Paradigms in HPV-Associated Cancers
Prevention
A Costarican national, is a Specialist Medical Doctor from UCR (Oncologist). His
PhD degree from Karoliska Institue, Stockholm and obtained a post doctoral
position at National Cancer Institute, NIH, Rockville, MD. He is currently the Head of the Prevention
and Implementation Group, Early Detection and Prevention Section, International Agency for
Research on Cancer, Lyon, France. He has been an international Professor and Researcher in the
United States of America, Costa Rica and France. In Costa Rica he has held positions such as
Coordinator of the study Population-based natural history of cervical neoplasia in a high-indicence
area of Latin America, in collaboration with NCI. He has been Director of the National Cervical
Cancer Screening Program, Ministry of Health, CR; Epidemiologist, Unit of Field and Intervention
Studies, International Agency for Research on Cancer, Lyon, France. Principal Investigator of the
Proyecto Guanacaste "HPV Type 16-18 Vaccine Trial in CR", and Principal Investigator of the project
Determinants of HPV infection among women 45-70 years old. Dr. Herrero has received many
Honorifc Mentions such as the “Premio Institucional en Investigación Científica 1990, Caja
Costarricense de Seguro Social”, and “Fred L. Soper Award, 1991, for significant
contributions to the literature on Health in Latin America, Pan American Health Foundation
2002”.
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VIBM_1047_0106
New paradigms in HPV-associated cancer prevention
Rolando Herrero
Prevention and Implementation group. International Agency for Research on Cancer. World Health
Organization, Lyon France
A limited number of human papillomaviruses (HPV) are associated etiologically with cervical,
vaginal, vulvar, anal, penile and oropharyngeal cancer. Recent knowledge about the natural history
of infection and the development of new preventive interventions have opened the door to the
possibility of controlling an important proportion of cancers associated with these infections. The
most common cancer associated with HPV is cervical cancer, responsible for nearly 600,000 cases
and 300,000 deaths per year worldwide. Screening programs based on cytologic screening have
succeeded in controlling this disease in developed countries, but not in developing countries, where
such programs have been impossible to establish or have been ineffective. The development of
new screening tools based on detection of viral DNA offers a sensitive and reproducible test that
has been shown to perform better than cytology. In particular, its high sensitivity for detection of
precancerous cervical lesions gives the test a high negative predictive value. Thus, women who are
HPV negative are at very low risk of developing cervical disease for several years after the test. This
allows a better selection of the groups at risk and the extension of the screening intervals. The
current proposal for screening in many areas is to do primary screening with an HPV test and to
triage positive women with cytology, although new molecular methods are under development for
more accurate selection of women with precancer. The recent development of highly effective and
safe vaccines against the two main HPV types associated with most cancers, namely HPV 16 and 18
is expected to have a large impact in the reduction of cervical cancer. The vaccines have also
proven effective against vulvar, vaginal, penile and anal infections and lesions and are likely to
reduce incidence of tumors of those localizations. Vaccination of adolescent women has started in
many countries, and it is likely to be extended to boys. The main challenge remains to make the
currently expensive vaccines available in developing countries, where they are mostly needed.
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Plenary and Keynote Speakers
MARIA ELENA BOTTAZZI (Baylor College of Medicine, US)
Conference 3: Texas and Mexico: Sharing a Legacy of
Poverty and Neglected Tropical Diseases
Maria Elena Bottazzi, Ph.D. is the associate dean of the National School of
Tropical Medicine and a professor of pediatrics at Baylor College of Medicine.
She also directs research and administrative activities of BCM’s new Section of
Pediatric Tropical Medicine and she leads product development for the Sabin vaccine development
program at Texas Children's Hospital and BCM.
Bottazzi previously was associate professor and vice-chair for administration in the Department of
Microbiology, Immunology and Tropical Medicine at George Washington University.
Bottazzi is an internationally-recognized scientist with more than 10 years of experience in
translational research and vaccine development for neglected tropical diseases. In addition, her
major interest is the role of vaccines as control tools in international public health programs and
initiatives.
She is a native of Tegucigalpa, Honduras, where she obtained her degree in microbiology and clinical
chemistry in 1989, followed by her Ph.D. in molecular immunology and experimental pathology at
the University of Florida in Gainesville in 1995. Her post-doctoral training in cellular biology was
completed 1995-2001 at the University of Miami and the University of Pennsylvania. She became an
assistant professor in 2001 after moving to the George Washington University.
Bottazzi is an associate for the Public Library of Science (PLoS) Neglected Tropical Disease Journal
and is the author or co-author of more than 60 scientific and technical papers in molecular, cellular
biology, immunoparasitology, and vaccine development. She is the recipient of multiple awards and
was recently appointed as a member of the International Advisory Committee of the Carlos Slim
Health Institute in Mexico.
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Texas and Mexico: Sharing a Legacy of Poverty and Neglected
Tropical Diseases
Peter J. Hotez, Maria Elena Bottazzi, Eric Dumonteil, Jesus G. Valenzuela, Shaden Kamhawi, Jaime
Ortega, Samuel Ponce de Leon Rosales, Miguel Betancourt Cravioto, Roberto Tapia-Conyer
The NTDs are the most common infections of the poorest 120 million people in the Americas who
live on less than US$2 per day. They include ancient scourges such as hookworm and other soiltransmitted helminth infections, Chagas disease, amoebiasis, schistosomiasis, vivax malaria,
leishmaniasis, and dengue. Together, these NTDs produce a burden of disease in the western
hemisphere that in certain regions even exceeds HIV/AIDS, while simultaneously trapping Latin
America's “bottom 100 million” in poverty through their deleterious effects on child physical and
intellectual development, pregnancy outcome, and worker productivity. Most of the NTDs occur in
Mexico's poorest states, led by the contiguous southern states of Chiapas, Guerrero, and Oaxaca.
Overall, these three southern states, in addition to neighboring Campeche, Quintana Roo, Veracruz,
and Yucatan, exhibit the lowest human development indices in Mexico. Across the border, the state
of Texas is neither immune to poverty nor to the NTDs. Indeed, at a 17% poverty rate, Texas has a
significantly higher rate than the overall 14% poverty rate in the United States. Emerging evidence
over the last few years has revealed a hidden burden of NTDs and related neglected infections of
poverty in Texas. The high prevalence and incidence of the major NTDs in both Mexico and south
Texas afford an opportunity for joint cooperation to address the highest prevalence conditions,
especially Chagas disease, CL, dengue, and the soil-transmitted helminth infections. There is also an
urgent need to develop alternative control tools for the major NTDs. A perspective on the vision and
mission of the newly established National School of Tropical Medicine at Baylor College of Medicine
will be presented in order to achieve elimination of these ancient conditions in the coming decade.
Ref: PLoS Negl Trop Dis 6(3): e1497.
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Plenary and Keynote Speakers
REYNALDO MARTORELL (Emory University, US)
Conference 4: Nutrition and Human Development
Reynaldo Martorell's research interests include maternal and child
nutrition, child growth and development, the significance of early
childhood malnutrition for short and long term human function,
micronutrient malnutrition, and the emergence of obesity and chronic diseases in
developing countries. Dr. Martorell's policy interests include global health concerns,
particularly programs and policies in food and nutrition, issues dealing with hunger and
malnutrition, and the health implications of changes in diet and lifestyles in developing
countries. He is the Robert W. Woodruff professor of International Nutrition and Senior
Advisor of the Global Health Institute and until recently, chair of the Hubert Department of
Global Health, a position he held for 11 years. He also holds a faculty appointment at the
National Institute of Public Health of Mexico. He is a Director of the Nevin Scrinshaw
International Nutrition Foundation and is a Member of the Board of Trustees of Helen
Keller International.
He is an advisor to UNICEF, the World Health Organization, the Interamerican Development
Bank and the World Bank. Dr. Martorell is past president of the Society for International
Nutrition Research and a former associate editor of the Journal of Nutrition. Recent awards
include the Carlos Slim Lifetime Achievement Award in Health, the McCollum International
Lectureship, the International Nutrition Prize from Kellogg's and the Society for
International Nutrition Research, and the Marion Creekmore award from Emory University.
He was elected to the Institute of Medicine of the US National Academy of Sciences in
2003. Prior to his position at Emory, Dr. Martorell held positions at Cornell University,
Stanford University, and the Institute of Nutrition of Central America and Panama (INCAP)
in Guatemala. Dr. Martorell received a bachelor's degree in anthropology from St. Louis
University and a Ph.D. in biological anthropology from the University of Washington. He
was born in Honduras, Central America.
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VIBM_4458_0097
Nutrition and Human Development
Reynaldo Martorell, Ph.D., Robert W. Woodruff Professor of International Nutrition Rollins
School of Public Health, Emory University
Human development is very sensitive to nutritional influences particularly during the first
1000 days, that is, pregnancy and the first two years of life. Early life is a time of rapid brain
growth and maturation as well as rapid physical growth in general. This leads to very high
nutritional requirements. In developing countries, infectious diseases, and diarrheal
diseases diarrheal diseases in particularly, are important causes of malnutrition along with
diets of poor quality and quantity. Malnutrition in early life leads to immunodeficiency and
accounts for nearly a third of child deaths. Survivors suffer long term consequences in their
human development. A remarkable study and subsequent follow-up studies from
Guatemala carried out by the Institute of Nutrition of Central America and Panama (INCAP)
from 1969 to 1977 provides experimental data on the long terms effects of improving
nutrition in the first 1000 days on a variety of human capital outcomes. Substantial impact
on adult human capital and on economic productivity was found in follow up studies in
1988-89 and 2002-04. The 1988-89 study showed that adult body size and work capacity
were increased for those provided improved nutrition through age 3 y, while the 2002-04
follow-up showed that schooling was increased in women and that reading comprehension
and intelligence were increased in both men and women. Participants were 26 to 42 y of
age at the time of the 2002-04 follow-up, facilitating the assessment of economic
productivity. Wages of men were increased by 46% in those provided with improved
nutrition through age 2 y.
Findings for cardiovascular disease risk factors were heterogeneous; however, they suggest
that improved nutrition in early life is unlikely to increase cardiovascular disease risk later in
life and may indeed lower risk.
In conclusion, the substantial improvement in adult human capital and economic
productivity resulting from the nutrition intervention provides a powerful argument for
promoting improvements in nutrition in pregnant women and young children.
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Plenary and Keynote Speakers
LUIS GABRIEL CUERVO (PAHO/WHO)
Conference 5: Research to Policy
Résumé: www.linkedin.com/in/lgcuervo
Luis Gabriel Cuervo is a medical doctor and specialist in Family
Medicine with a MSc in Clinical Epidemiology & Biostatistics.
Throughout his professional career he has been an agent of change advancing initiatives
that integrate research for health, health policies and health care. He has championed the
strengthening of health research governance, knowledge translation and its use to
strengthen health systems. Dr. Cuervo has contributed to the development of milestone
statements and the development and coordination of policies and landmark consultations
on research for health at international, national and local settings. Examples of these
include the regional contributions from the Americas to the Bamako Ministerial Forum on
Research for Health (2008), WHO's Strategy on Research for Health (2010), and the policy
on research for health for the Americas approved by PAHO/WHO Member States in 2009.
Dr. Cuervo has firsthand experience working with local, national and international leading
experts, influencing public health policies, and building capacities to produce and use
research for health. He has worked also as a clinician in rural and urban settings, and
developed partnerships involving low, middle and high income settings. He worked as a
senior clinical editor for the BMJ Publishing Group and peer reviews for leading
international and local journals. Dr. Cuervo contributes to working groups aimed at
improving research reporting and publication standards, research management and health
research governance. He has extensive experience organizing teams to improve research
management and governance. More recently he has coordinated the development of the
Art for Research exhibitions that illustrates through art the returns of investment in
research for health to society, with an emphasis in economic and social development and
competitiveness (www.paho.org/artforresearch).
22
Plenary and Keynote Speakers
VIC NEUFELD (CCGHR)
Conference 6: Assessing North-South Partnerships: The
Canadian experience
Dr. Vic Neufeld is Professor Emeritus in the Faculty of Health Sciences at
McMaster University, Hamilton, Canada, where for 27 years he served
as a general internist, educator and researcher. He was the founding
director of McMaster’s Centre for International Health. With a growing interest in health
research for development, he left McMaster in 1997 to work as a senior advisor with the
Geneva-based Council on Health Research for Development (COHRED), focusing on health
research priority setting in low and middle-income countries. In 2003, he became the
founding national coordinator of the Canadian Coalition for Global Health Research
(CCGHR). Professor Neufeld continues to have a special interest in health research capacity
strengthening, including leadership development.
23
VIBM_BA70_0098
Assessing South-North
Experience
Research
Partnerships:
the
Canadian
Vic Neufeld, Canadian Coalition for Global Health Research
Among the major trends in the field of research for health development, the role of
collaboration and partnerships is becoming increasingly important. An example is the shift
in “south-north” research partnerships toward collaborations that are mutually-beneficial
and driven by the agendas of low and middle-income countries (LMICs).
This presentation will include the following issues:
•
The process and results of a study conducted by the Canadian Coalition for Global
Health Research (CCGHR) and supported by the International Development Research
Centre (IDRC). Entitled “Building Effective and Sustainable Partnerships for Global Health
Research”, the study focused on the “southern voice” and experience in research
partnerships. Included in the study was a consultation conducted in the Andean region of
Latin America. A key product of the study has been the “Partnership Assessment Tool”
(PAT), a resource that is available in both English and Spanish on: www.ccghr.ca .
•
Lessons from the Canadian experience in promoting and implementing best
practices in south-north research partnerships.
•
Some suggestions for NeTropica to consider in order to promote increased
effectiveness and “harmonization” of research collaboration in the Central American
region.
24
Round Tables
Round Table 1
Vector-Borne and Zoonotic Diseases in Guatemala
Convener: Dr. Pamela Pennington
•
•
Introduction. Lic. Celia Cordón-Rosales. (Director Centro de Estudios en Salud,
Universidad del Valle de Guatemala).
Eco-epidemiology of Chagas Disease in Guatemala. Pamela Pennington, PhD (Director
Departamento de Bioquímica y Microbiología, Centro de Estudios en Biotecnología afiliado
al Centro de Estudios en Salud, Universidad del Valle de Guatemala).
•
Diversity and origin of avian and swine influenza viruses in Guatemala. Lic. Ana Silvia
González Reiche. (Veterinary Medical Sciences PhD program, Department of Veterinary
Medicine, University of Maryland College Park, Maryland, USA).
•
Development of polymorphic, neutral markers to study the dynamics of Plasmodium vivax
infections. Lic. Renata Mendizábal, MSc. (Centro de Estudios en Salud, Universidad del Valle
de Guatemala).
•
Q/A Discussion.
VIBM_7789_0075
Eco-epidemiology of Chagas disease in Guatemala.
Pamela M. Pennington and Sandra De Urioste, Universidad del Valle de Guatemala.
Chagas disease is a zoonotic disease caused by Trypanosoma cruzi, a parasite that infects
mammalian reservoirs and is transmitted by hematophagous insects that inhabit animal nests. The
principal vector in Central America is Triatoma dimidiata. Found from Mexico to Colombia, it
colonizes human dwellings of rustic construction. A vector control initiative based on insecticide
treatment inside homes is ongoing in Central America. In Guatemala, a region bordering with El
Salvador in the department of Jutiapa shows low control efficacy despite multiple spraying
campaigns. A multidisciplinary study was conducted to evaluate the role of peridomestic animals in
triatomine infestation. Household surveys on knowledge, attitudes and practices related to
25
peridomestic animals were undertaken in 472 homes. These were followed by a study on canine
Chagas seroprevalence and rodent infestation and infection. Blood meal analysis was performed to
determine the primary sources of blood for T. dimidiata. Data were triangulated through
Participatory Action Research and a rapid ethnographic study to confirm findings related to animals
and the household economy. An explicative model was generated. The results indicate that having
egg-laying hens inside the household poses a major risk factor for triatomine infestation (Odds Ratio
2 ,1.2-3.3), being a primary source of blood for triatomines (64%). Dog ownership is also an
important risk factor (OR 2.3, 1.3-4.0), where having more dogs increases the likelihood of
infestation. Of 80 sampled dogs, 35% were found to be infected and 15% of triatomines had fed on
dogs alone or dogs and humans. Having Mus musculus and/or Rattus rattus infestations also poses
a risk factor for triatomine infestations (OR 4.0, 1.1-14.1) and for infection in the house, as 20% of
rodents collected in households having infected triatomines showed T. cruzi infections.
Environmental factors potentially related to rodent nesting habitats, such as dirt floors (OR 3.4, 1.96), grain storage in sleeping quarters (OR 1.6, 1-2.5) and vegetation such as coffee trees (OR 1.9,
1.2-3.2) showed associations with triatomine infestation. Animal management programs that
include rodent management may provide a novel method to reduce infestations and infection in
areas where traditional insecticides are not sufficient to reduce infestation.
VIBM_0117_0080
Phylogenetic analysis of Type A Influenza viruses isolated from
waterfowl in Guatemala, Central America
Ana S. González-Reiche (a,b)*, María E. Morales-Betoulle (a), Danilo Alvarez (a), Jean-Luc Betoulle (a,c), Maria
L. Müller (a), Silvia M. Sosa (a), Daniel R. Perez (b)
a) Laboratorio de Ecología de Arbovirus y Virus Zoonóticos, Centro de Estudios en Salud, Universidad del Valle
de Guatemala (CES-UVG). 18 avenida 11-95 zona 15, Vista Hermosa III, Guatemala, Guatemala 01015. b)
Department of Veterinary Medicine, University of Maryland College Park, and Virginia-Maryland Regional
College of Veterinary Medicine, 8075 Greenmead Drive, College Park, MD 20742, USA. c) Fundación Para el
Ecodesarrollo y la Conservación (FUNDAECO), Guatemala, Guatemala 01001.
Background/rationale/Introduction: The role wild bird species play in the transmission and ecology
of avian influenza virus (AIV) is well established; however, there are significant gaps in our
understanding of the worldwide distribution of these viruses. Although numerous wild avian species
migrate between North and Central American the potential for influenza viruses moving with them
currently is unknown and no information is currently available about the prevalence and/or
significance of AIV in Central America. Research Objectives: To study the the presence, diversity and
origin of influenza A viruses circulating in wild birds in Guatemala. Methodology: From February
2007 to Juanuary 2010, hunter-harvested waterforl and mistnet captured terrestrial birds were
26
sampled at the Pacific and Atlantic coast from Guatemala. Cloacal and tracheal swab samples were
tested by real time RT-PCR, and virus isolation. Isolates were full length sequenced and phylogenetic
analysis was perfomed for individual gene segments. Results: A total of 1913 samples were
collected, and virus was detected in swab samples from migratory waterfowl (Blue-winged Teals,
n=28). Virus isolation was attempted for the RT-PCR positive samples, and fifteen isolates were
obtained from samples collected in 2008, 2009, and 2010. Identified subtypes include H7N9, H11N2,
H3N8, H5N3, H8N4 and H5N4. Phylogenetic analysis of the viral genome sequences revealed that
AIV isolates are highly similar to viruses from the North American linage. Conclusions: Results
obtained suggest that bird migration from the North dictates the ecology of influenza viruses in the
Guatemalan bird population. Increased surveillance in the Central America is needed to address the
major determinants for AIV ecology, evolution and transmission in the region.
VIBM_0078_0112
Development of neutral SNP markers for low-cost, low-technology
follow-up of within host dynamic of P. vivax infections.
Mendizábal-Cabrera, R., P. Pennington, L. Ortiz, M.E. Castellanos and N. Padilla. Center for Health Studies,
Universidad del Valle de Guatemala.
Monitoring the effectiveness of antimalarial drugs is a requirement for the development of
treatment policies. Monitoring is commonly performed by in vivo assays that may be validated by
parasite molecular typing techniques to conclude if treatment failure is a true recrudescence, a
relapse or a new infection with a different parasite genotype. Several molecular marker panels have
been proposed to validate these results, but generally apply expensive instrumentation and
demand qualified technical expertise, hampering its application in endemic countries. Using a
strategy of in silico identification of neutral single nucleotide polymorphism (SNP) markers in
sequences of P. vivax from strains reported across endemic areas, six SNP markers were identified
in genes coding for β-tubulin, MSP1, MSP3α, MSP7, MSP9 and MSP10. Hemi-nested primers with
allele-specific 3´ ends were designed and PCR conditions were standardized with control samples.
Twenty seven samples from two endemic areas of Guatemala were analyzed. SNP markers
identified in β-tubulin and MSP10 were not polymorphic (allele frequency > 0.95), but have
potential to be useful in other regions with P. vivax malaria. Primers for MSP3α showed to be nonspecific and this marker was not included in the analysis. However, SNP markers designed on MSP1,
MSP7 and MSP9 genes showed to be polymorphic and specific to P. vivax. In addition to haplotype,
a size polymorphism was also identified in the MSP1 region, which in combination with haplotypes
of polymorphic SNPs produced different genotype profiles for 70% of the analyzed samples. Data
regarding neutrality and allele association of SNPs and MSP1 size polymorphism will be also
discussed. This PCR-based genotyping technology, being low cost and simple, would be an
27
affordable alternative to conventional automatic sequencing, making easier its adoption in
laboratories of developing countries.
Round Table 2
Research Skills for a Successful Scientific Career
Convener: Dr. Ana Sanchez
•
•
•
•
•
Introduction. Ana Sanchez, PhD. (Brock University).
It’s never too late to give up. Hans Rosling, PhD. (Karolinska Institute, Stockholm, Sweden)
Ten Simple Rules for Aspiring Scientists in a Low-Income Country. Edgardo Moreno, PhD.
(Tropical Disease Research Program, Veterinary School, National University, Heredia, Costa
Rica).
TBA. Reynaldo Martorell, PhD. (Robert W. Woodruff Professor of International Nutrition
Hubert Department of Global Health, Emory University).
Q/A Discussion.
Symposia
Symposium 1
Microbial Genetics
Convener: Dr. Esteban-Chaves-Olarte
•
•
•
•
•
Introduction. Esteban Chaves-Olarte. PhD. (School of Veterinary Medicine, University of
Costa Rica).
Carcinogenic properties of bacterial genotoxins. Dr. Teresa Frisan. (Department of Cell and
Molecular Biology, Karolinska Institute, Stockholm, Sweden).
Using whole genome sequences to understand global spread and evolution of Vibrio
cholerae and Shigella sonnei. Nicholas R. Thomson, PhD. (Senior Scientist Pathogen
Genomics. Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK).
Biology of Brucella ceti infection in stranded dolphins from Costa Rica. Caterina Guzman,
PhD. (School of Veterinary Medicine, University of Costa Rica)
Q/A Discussion
28
VIBM_6701_0063
Carcinogenic properties of the bacterial cytolethal distending toxin
1
1
1
2
3
4
Riccardo Guidi *, Lina Guerra *, Laura Levi , Bo Stenerlöw , James G. Fox , Christine Josenhans , Maria G.
1
1
Masucci , Teresa Frisan
1
2
Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden; Division of Biomedical
3
Radiation Sciences, Rudbeck Laboratory, Uppsala University, Sweden; Department of Biological Engineering,
Division of Comparative Medicine, Massachusetts Institute of Technology, Cambridge, Massachusetts, United
4
States of America; Institute for Medical Microbiology and Hospital Epidemiology, Hannover Medical School,
Hannover, Germany.
Chronic inflammation and infection is associated with an increased risk of cancer
development. While much is known about the role of chronic viral infections in
tumorigenesis, the mechanisms by which bacterial affliction contribute to carcinogenesis
are still poorly characterized. Several Gram-negative human pathogens produce a
genotoxin (the cytolethal distending toxin, CDT), which induces DNA damage in the target
cells. The effects of intoxication have been described as “radiation damage”, since they are
similar to those evoked by ionizing radiation, a well-characterized genotoxic stress that
causes alteration at genome levels, thus promoting tumour development. We are focusing
on how CDT intoxication alters processes involved in the regulation of genomic integrity,
cell cycle progression, cell survival as well as cytoskeleton dynamics and tissue remodelling
in in vitro 2D and 3D models of intestinal epithelial cells. We demonstrated that chronic
exposure to the genotoxin cytolethal distending toxin (CDT) of Gram-negative bacteria
promotes genomic instability and the acquisition of phenotypic properties of malignancy.
Cells grown for more than 30 weeks in the presence of sub-lethal doses of CDT showed
increased mutation frequency and accumulation of chromatin and chromosomal
aberrations in the absence of significant alterations of cell cycle distribution, decreased
viability or senescence. Cell survival was dependent on sustained activity of the p38 MAP
kinase signaling pathway. The ongoing genomic instability was associated with impaired
activation of the DNA damage response and failure to efficiently activate cell cycle
checkpoints upon exposure to genotoxic stress. Independently selected sub-lines showed
enhanced anchorage independent growth as assessed by the formation of large colonies in
semisolid agarose. These findings support the notion that chronic infection by CDT
producing bacteria may promote malignant transformation, and point to the impairment of
cellular control mechanisms associated with the detection and repair of DNA damage as
critical events in the process.
29
Investigating the population diversity and epidemiology of Shigella
sonnei at the local and global level
Nicholas Thomson. Pathogen Genomics. Wellcome Trust Sanger Institute, Hinxton, Cambridge, UK.
Diarrhoeal disease caused by members of the bacterial genus Shigella remains an ongoing
public health issue in many developing countries. Shigellosis is synonymous with profuse
bloody diarrhoea and can cause mortality in the young, elderly and otherwise susceptible. A
recent investigation in six Asian countries found that Shigella was the most common
bacterial form of gastro-intestinal infection in children. There are four Shigella species
causing bacillary dysentery in man; S. dysenteriae, S. flexneri, S. boydii and S. sonnei. In
1999, the WHO estimated 160 million episodes worldwide per annum and 1.1 million deaths
of which most are children under 5 years old (Kotloff 1999). Shigella sonnei has been
associated with wealthier Western countries. However, this species is now emerging as a
problem in low income economies, where it is replacing endemic dysentery agents such as
Shigella flexneri even as access to clean water and sanitation improves. We have focused
on examining a global collection of historic and contemporary isolates of S. sonnei. The
genome information has provided a comprehensive understanding of the population
structure and the unique epidemiological behaviour of S. sonnei. We show that the origins
of S. sonnei do not lie in the developing world, but that S. sonnei evolved in Europe ~500
years ago and has only recently moved out of this region via intercontinental spread as a
single, rapidly evolving lineage that has now established local clonal populations in several
developing regions. We have used population genomic methods to follow and understand
the rapid increase in antibiotic resistance.
VIBM_1076_0046
Biology of Brucella ceti infection in stranded dolphins from Costa
Rica.
1
1,2
1
3
1,4
Guzmán-Verri Caterina , Murillo Carla , Piche Ovares Martha , Archer Frederick I II , Slon Jose Luis ,
1
1,4
1,5
5,6
Barquero-Calvo Elías , Lizano Esteban , Hernández-Mora Gabriela , González-Barrientos Rocio , Morales
6
6
1
2
2
1,4
Juan Alberto , Alfaro Alejandro , Ruiz, Nazareth , Rivas Olga , Zúñiga Claudia , Chaves-Olarte Esteban ,
7
7
1
Lomonte Bruno , Sasa Mahmood , and Moreno E
1
2
PIET, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica. Centro de Investigación en
3
Biotecnología. Escuela de Biología, Instituto Tecnológico, Cartago, Costa Rica. National Oceanic and
Atmospheric Administration, National Marine Fisheries Service, Southwest Fisheries Science Center, La Jolla,
4
5
CA. USA. Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica. SENASA, Ministerio de
6
Agricultura y Ganadería, Heredia, Costa Rica. Laboratorio de Patología, Escuela de Medicina Veterinaria,
7
Instituto Clodomiro Picado, Facultad de Microbiología,
Universidad Nacional, Heredia, Costa Rica.
Universidad de Costa Rica, San José, Costa Rica.
30
Brucellosis caused by Brucella pinnipedialis and Brucella ceti is a disease prevalent in pinnipeds and
cetaceans, respectively. It is relevant in seals, walruses, dolphins and whales and it may constitute a
risk for other animal species, including humans. Stenella coeruleoalba, commonly known as striped
dolphins are resident of tropical and subtropical oceanic regions worldwide. During 2004 and 2011,
twenty five striped dolphins stranded on the Pacific coast of Costa Rica. Clinical manifestations
consistent with nervous system alterations were observed in most cases. All animals were
serologically positive against Brucella antigens. Pathological findings, differential diagnosis and
isolation of B. ceti from cerebrospinal fluid, determined the cause of death as neurobrucellosis. To
gain insight of the brucellae involved in the course of infection in this wild life dolphin host
population, bacteriological characterization was carried out using genotyping, proteomic and fatty
acid profile analyses. Results demonstrated that these isolates from the Eastern Tropical Pacific of
Costa Rica belonged to the species B. ceti dolphin type, but constitute a different cluster from the
strains isolated in cetaceans from the Atlantic or Mediterranean seas. Phylogenetic analysis of
genotypical and phenotypical features of striped dolphins from the Mediterranean Sea, Pacific and
Atlantic Oceans revealed that S.coeruleoalba is an intertwined species, with clusters of individuals
geographically distributed in different oceanic areas. Moreover, S. coeruleoalba individuals from the
Pacific Ocean show sexual dimorphism with larger females, not observed in the Atlantic Ocean and
Mediterranean Sea populations. These features, together with the distinction of B. ceti strains
isolated in stranded S.coeruleoalba in the Pacific shorelines of Costa Rica, suggest that striped
dolphins from the Pacific Ocean may have remained isolated since the closure of the Panama
Isthmus, close to 7 million years ago. Successful pathogen adaptation to its host must certainly takes
into account the host evolution itself. Marine fauna with no or little contact with humans provides a
good model to study host pathogen coevolution and gain insights on how terrestrial mammals and
pathogens adapt when humans intervene.
Symposium 2
Strategies for Research Sustainability in Central America
Convener: Dr. Gustavo Fontecha
•
•
•
Introduction. Gustavo Fontecha, PhD & Maritza Canales, MPH. (Graduate Program in
Infectious and Zoonotic Diseases, School of Microbiology, National Autonomous University
of Honduras)
TBA. Vinod Diwan, PhD. (Chairman Karolinska International Research and Training -KIRT.
Head of Division – International Heath. Karolinska Institute, Stockholm, Sweden).
The Barcelona Institute for Global Health (ISGlobal): Collaboration Opportunities for
Research and Training in Global Health in Central America. Nuria Casjmitana, PhD. (Director
31
•
•
of Education and Training, ISGlobal. Universitat de Barcelona - Hospital Clinic). Barcelona,
España.
Research Program of the third Central American Higher Education Integration Plan and
the Seventh European Union Framework. Juan Alfonso Fuentes Soria, (MSc. General
Secretary, Central American Council for Higher Education. Consejo Superior Universitario
Centroamericano).
Q/A Discussion.
The Barcelona Institute for Global Health (ISGlobal): Collaboration
Opportunities for Research and Training in Global Health in Central
America
Núria Casjmitana, PhD. Director of Education and Training, ISGlobal
The Barcelona Institute for Global Health (ISGlobal) is a non-profit organisation that tackles
health problems affecting the world’s most vulnerable populations through the creation, sharing
and application of knowledge. Our vision is a world in which every person can enjoy health.
ISGlobal is a public-private partnership with a longstanding commitment to global health that
builds on the existing research, training and health activities of its founding institutions and
strives to contribute to the global effort to improve health worldwide.
We create partnerships based on cooperation and trust, sharing knowledge and fostering
mutual learning, with respect of social and cultural contexts when developing and promoting
solutions.
ISGlobal carries out activity throughout the value chain of knowledge in global health:
1. Knowledge creation, doing research with a multidisciplinary and translational approach
2. Knowledge management, setting new standards for innovative action
3. Knowledge transmission, strengthening the capacities of professionals and stakeholders
4. Knowledge Application to improve the impact of actions in the field.
We focus on health problems affecting the world's most disadvantaged people and for more tan
fifteen years we have been collaborating with institutions in Africa and Latin America. More
recently we started working in joint research and training initiatives with countries in Central
America and we want to explore the great potential offered by academic institution in this
region to develop collaborative programs in the field of global health.
32
The Research Program of the Third Plan for the Integration of
Higher Education Central and Seventh Framework Programme of
the European Union
Juan Alfonso Fuentes Soria. Secretario Gral CSUCA
The universities in Central America have to play an effective role in generating knowledge relevant
to social problems related to the countries in particular and the region in general, it is essential to
assess both conceptually and in practice research support.
This implies, first, that universities should make a consistent and growing effort to allocate more
resources for its own budget to strengthen this vital role, as well promote in the respective states
do the same to encourage and support more resources in the development of science and
technology.
This statement implies the need for an institutional cultural change within the universities, state,
national and regional levels. Moreover, at the Central American region there is little use of the
resources offered by the international community for research. In order to promote a change of
greater support for research, there is a need to strengthen and revitalize existing research networks
and to promote the creation of new networks in order to promote greater exchange between
researchers and enable greater chances of success for the management and resourcing. It refers to
some of the actions that the Confederation of Universities has been promoting through its existing
programs and systems that support them. Finally, we will share information on the Seventh
Framework Programme of the European Union and the ENLACE program.
33
Oral Presentations
1. Borrelia turicatae adaptation within the arthropod vector. Job López
2. Phylogenetic analysis of Type A Influenza viruses isolated from waterfowl in
Guatemala, Central America. Ana Silvia Gonzales
3. Preliminary results of a study on Trypanosoma cruzi congenital transmission in two
endemic regions of Honduras. Jackeline Alger
4. Sand fly density reduction is less marked in precarious housing after insecticide
residual spraying. Luis Fernando Chaves
5. Snakebite Envenomings in Latin America: Current Situation and Challenges. José Ma.
Gutiérrez
6. Competence of Leishmania species to determine different clinical and
immunological features in murine infection. Marcia Laurenti
7. Health care workers’ knowledge, attitudes and reported practices related to health
care-associated infections in an Indian teaching hospital. Senia Rosales
8. Establishing a Proteomics Laboratory at Instituto Clodomiro Picado (UCR): first twoyear's experience in serving local and regional research. Bruno Lomonte
34
Oral Presentation 1
VIBM_TBD_0022
Borrelia turicatae adaptation within the arthropod vector
Hannah Wilder and Job Lopez
Department of Biological Sciences, Mississippi State University
Background: Understanding microbial adaption within different natural environments remains a gap
in knowledge in emerging infectious disease research. Tick-borne diseases continue to afflict
human health, and given the vectors high reproductive potential, durability, and the ability to
transmit parasitic, viral, and bacterial pathogens, understanding pathogen colonization within the
vector is particularly relevant to public health. One such pathogen is tick-borne relapsing fever (RF)
spirochetes. The global distribution and economic burden of RF spirochetes on human health is
underappreciated, especially considering the disease’s prevalence and impact on populations in
developing countries. A causative agent of tick-borne relapsing fever throughout the Americas is
Borrelia turicatae. B. turicatae is maintained in enzoonotic cycles, and a unique characteristic of the
vector is that the ticks are long-lived (over 20 years), can be starved upwards of 5 years, yet the
spirochetes remain viable and transmissible. Additionally, the rapidity of feeding and transmission
dynamics of the spirochetes differs significantly from other tick-borne diseases, yet RF spirochetes
are studied only by a few labs. Research Objectives, Methodology, Results, and Conclusions: One
limitation hindering the development of disease preventing therapeutics has been a poor
understanding of the molecular events contributing to RF spirochete colonization of the tick vector.
We have hypothesized that differential gene regulation within the vector and mammalian host is
essential for B. turicatae to adapt and survive two physiologically and immunologically different
environments. Towards this, we have conducted microarray analyses to determine differential
gene expression profiles of B. turicatae within the tick and mammal. We have identified a series of
B. turicatae genes with unknown function that are up-regulated within the tick. The genes encode
for putative outer membrane proteins predicted to have adhesive properties. To determine the
role of these genes during the tick-mammalian infectious cycle, we developed the first system to
inactivate genes in B. turicatae. We have deleted one of the genes, the Borrelia repeat protein A
(brpA), and have determined that the mutants are attenuated when compared to wild type
spirochetes. These findings are providing a better understanding of the molecular mechanisms
required for spirochete adaptation within the tick vector.
35
Oral Presentation 2
VIBM_0117_0080
Phylogenetic analysis of Type A Influenza viruses isolated from
waterfowl in Guatemala, Central America
Ana S. González-Reiche (a,b)*, María E. Morales-Betoulle (a), Danilo Alvarez (a), Jean-Luc Betoulle (a,c), Maria
L. Müller (a), Silvia M. Sosa (a), Daniel R. Perez (b)
a) Laboratorio de Ecología de Arbovirus y Virus Zoonóticos, Centro de Estudios en Salud, Universidad del Valle
de Guatemala (CES-UVG). 18 avenida 11-95 zona 15, Vista Hermosa III, Guatemala, Guatemala 01015. b)
Department of Veterinary Medicine, University of Maryland College Park, and Virginia-Maryland Regional
College of Veterinary Medicine, 8075 Greenmead Drive, College Park, MD 20742, USA. c) Fundación Para el
Ecodesarrollo y la Conservación (FUNDAECO), Guatemala, Guatemala 01001.
Background/rationale/Introduction: The role wild bird species play in the transmission and ecology
of avian influenza virus (AIV) is well established; however, there are significant gaps in our
understanding of the worldwide distribution of these viruses. Although numerous wild avian species
migrate between North and Central American the potential for influenza viruses moving with them
currently is unknown and no information is currently available about the prevalence and/or
significance of AIV in Central America. Research Objectives: To study the the presence, diversity and
origin of influenza A viruses circulating in wild birds in Guatemala. Methodology: From February
2007 to Juanuary 2010, hunter-harvested waterforl and mistnet captured terrestrial birds were
sampled at the Pacific and Atlantic coast from Guatemala. Cloacal and tracheal swab samples were
tested by real time RT-PCR, and virus isolation. Isolates were full length sequenced and phylogenetic
analysis was perfomed for individual gene segments. Results: A total of 1913 samples were
collected, and virus was detected in swab samples from migratory waterfowl (Blue-winged Teals,
n=28). Virus isolation was attempted for the RT-PCR positive samples, and fifteen isolates were
obtained from samples collected in 2008, 2009, and 2010. Identified subtypes include H7N9, H11N2,
H3N8, H5N3, H8N4 and H5N4. Phylogenetic analysis of the viral genome sequences revealed that
AIV isolates are highly similar to viruses from the North American linage. Conclusions: Results
obtained suggest that bird migration from the North dictates the ecology of influenza viruses in the
Guatemalan bird population. Increased surveillance in the Central America is needed to address the
major determinants for AIV ecology, evolution and transmission in the region.
36
Oral Presentation 3
VIBM_3011_0111b
Preliminary results of a study on Trypanosoma cruzi congenital
transmission in two endemic regions of Honduras
1-3
4
5
4
1,3
Jackeline Alger, Norma Bustamante, Jaime H. del Cid, Benjamín Lopez, Edna Maradiaga, Concepción
3,6
7
8
8
9
Zuniga, María Luisa Matute, María Luisa Cafferata, Alvaro Ciganda, Pierre Buekens.
1
2
Unidad de Investigación Científica, Facultad de Ciencias Médicas (FCM), UNAH; Servicio de Parasitología,
3
Departamento de Laboratorios Clínicos, Hospital Escuela, MOH; Instituto de Enfermedades Infecciosas y
4
5
Parasitología Antonio Vidal; Tegucigalpa; Región Departamental de Salud de Santa Bárbara; Región
6
Departamental de Salud de Intibucá; Programa Nacional de Prevención y Control de la Enfermedad de
7
8
Chagas y Leishmaniasis MOH; Laboratorio Nacional de Vigilancia de la Salud MOH; UNICEM, Montevideo,
9
Uruguay; Tulane University, New Orleans, USA.
Background. Trypanosoma cruzi genotypes have been divided into TcI and non-TcI lineages (TcIITcVI). Infected women living in regions where TcI is predominant, e.g., Mexico and Central America,
could differ in transmitting T. cruzi to their children. Honduras is participating in a multicentre study
on T. cruzi congenital transmission. Objectives. To determine the rate of congenital transmission of
Chagas disease, to compare T. cruzi infected mothers’ characteristics and to describe birth
outcomes of infected infants, according to parasite genotype. Methodology. Prospective study to
enroll at delivery 10,000 women in Mexico, 5,000 in Argentina and 5,000 in Honduras. Honduras
study sites include Hospital Enrique Aguilar Cerrato, La Esperanza, Intibuca, and Hospital Santa
Barbara Integrado, Santa Barbara. We are measuring transmitted maternal T. cruzi antibodies by
performing two rapid tests in cord blood and, if at least one of the results is positive, we are
identifying infants who are congenitally infected by performing parasitological examinations on cord
blood and at 4-8 weeks, and serological follow-up at 10 months. We will also perform standard PCR,
real-time quantitative PCR, and T. cruzi genotyping on maternal blood and on cord blood, and
serological examinations on siblings. Results. Study enrollment began in May 2011. As of April 2012,
there have been 1694 women recruited in Intibuca and 1842 in Santa Barbara. There were 139
(3.9%) births with at least one positive rapid test results on cord blood, 91 (5.4%) in Intibuca and 48
(2.6%) in Santa Barbara. For the 91 cases from Intibuca, 37 women (40.6%) were <25 years old, 30
(33.0%) were primiparous, 1 (1.1%) had a gestational age <37 weeks and 19 newborns (20.9%)
weighed <2,500 gr. For the 48 cases from Santa Barbara, 9 women (18.8%) were <25 years old, 4
(8.3%) were primiparous, 1 (2.1%) had a gestational age <37 weeks and no newborn weighed
<2,500 gr. Conclusions. Information on T. cruzi congenital transmission and better knowledge about
risk factors will allow the implementation of a screening program in Honduras.
37
Oral Presentation 4
VIBM_D944_0000 a
Sand fly density reduction is less marked in precarious housing
after insecticide residual spraying
1
2
2
2
2
Luis Fernando Chaves , Azael Saldaña , C. Rigg , R. Rojas , A. Valderrama , J.E. Calzada
2
1
Programa de Investigación en Enfermedades Tropicales, Universidad Nacional, Heredia, Costa Rica &
2
Graduate School of Environmental Sciences, Hokkaido University, Sapporo, Japan. Departmento de
Parasitología, Instituto Conmemorativo Gorgas de Estudios de la Sálud, Ciudad de Panamá, Panamá.
Background/Hypothesis: Insecticide residual spraying (IRS) is a tool to control vector borne
diseases. It is generally assumed that IRS reduces vector density independently of housing
conditions. Few studies have been focused on Sand Flies while also looking at housing
characteristics. Methods: We conducted a 15 month longitudinal study that included two
deltamethrin based IRS interventions in 12 of the 24 houses at Comunidad de Trinidad Las Minas,
Capira, Panamá, a hyperendemic cutaneous leishmaniasis transmission village. During the study we
followed sand fly (SF) abundance. Results: We found a 50 to 80% reduction in SF density at sprayed
houses when compared with control houses, while controlling for seasonal changes in SF abundance
associated with rainfall. We found some heterogeneities in the reductions, as abundance changed
according to SF species, with Lutzomyia gomezi, L. panamensis, L. dysponeta and L. triramula
reducing their density between 40% and 90% after IRS, in contrast to L. trapidoi whose density
increased 5% after the IRS. Differences in the impact of IRS were associated with housing quality,
the most precarious houses, i.e., those with features that ease insect entrance, had a
disproportionally larger SF abundance, in some cases with an increased domiciliary SF density
following the IRS. Conclusion: Our results suggest that IRS success potential to control SF density
and Leishmaniasis transmission could be dependent on housing quality.
38
Oral Presentation 5
VIBM_1046_0012
Snakebite Envenomings in Latin America: Current Situation and
Challenges
José María Gutiérrez
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica. E mail:
[email protected]
Snakebite envenomings constitute a relevant neglected tropical disease in Latin America. This
pathology affects predominantly the rural population and has a high impact in regions where the
provision of health services is insufficient. The majority of envenomings are inflicted by species of
the genera Bothrops and Crotalus, classified in the family Viperidae. There are several laboratories
in the region which manufacture antivenoms for the treatment of these envenomings, although the
volume of production in some cases does not fulfill the national demands. A significant body of
knowledge has been gained in the taxonomy of the snakes, the biochemistry, toxicology and
immunology of venoms, and the clinical manifestations of envenomings, and the preclinical and
clinical performance of antivenoms. In addition, important advances have been made in the
prevention and treatment of this neglected tropical disease in Latin America. Nevertheless, there
are pending tasks that should be promoted in the region to further reduce the impact of this
disease, such as: (a) Improvement of our knowledge on snakes and their venoms; (b) assessment of
the actual incidence and mortality of snakebite envenomings; (c) increment in the volume of
antivenom produced and, in some cases, in the quality of antivenoms; (d) strengthening of national
quality control laboratories; (e) development of more effective strategies of distribution of
antivenoms, especially to remote rural areas where snakebites are frequent; (f) fostering
permanent education programs for the health staff in charge of the treatment of these
envenomings; (g) provision of support to people that suffer physical or psychological sequelae as a
consequence of snakebites; and (h) strengthening community programs aimed at improving the
prevention and adequate management of these accidents. This conglomerate of tasks should be
approached with a philosophy of solidarity, integration and cooperation in the region, with the
involvement of multiple actors and institutions.
39
Oral Presentation 6
VIBM_1072_0029 a
Competence of Leishmania species to determine different clinical
and immunological features in murine infection
1
Carvalho AK,
1
2,3
1
1
1
1
Silveira FT, Passero LFD, Gomes CMC, Corbett CEP, Laurenti MD
2
Laboratory of Pathology of Infectious Diseases (LIM-50), Medical School, University of São Paulo (SP), Laboratory
3
of Leishmaniasis, Evandro Chagas Institute and University of Pará, Belém (PA), BRAZIL.
American cutaneous leishmaniasis (ACL) is a zoonotic protozoal disease caused by different species of
Leishmania. In Brazil, L.(V.)braziliensis and L.(L.)amazonensis are considered the main pathogenic
species causing human ACL. The disease caused by these two Leishmania species presents a clinical–
immunological spectrum which has been associated with antigenic differences of the parasite species,
but also the host background. In order to evaluated the competence of the parasite species on the
development of clinical and immunological features on experimental ACL, BALB/c mice were infected
subcutaneously into the hind footpad with 106 promastigote with L.(L.)amazonensis or L.(V.)braziliensis.
The hind footpad swelling was weekly evaluated till the 8th week PI. At the 4th and 8th week PI, draining
lymph nodes were collected aseptically and used to determination of the parasite load using the
quantitative limiting-dilution and quantification of CD3+, CD4+ and CD8+ cells by flow cytometry. The
supernatant of the lymph nodes cell cultures under specific stimuli were used to determination of IL-4,
IL10, IFN- using Capture ELISA and NO by Greiss method.L.(L.)amazonensis induced a progressive
growth of skin lesions since the 3rd week (p<0.05); while L.(V.)braziliensis showed a small swelling in the
skin lesion between the 6th and 7th week (p<0.05) with regression at the 8th week PI. At 4th and 8th
weeks, the parasite load, in the draining lymph nodes of mice infected with L.(L.) amazonensis was
higher (p<0.05) than that animals infected with L.(V.)braziliensis. Concerning to T cells, there was a
decrease on the percentage of CD3+ and CD4+ cells at 8th week mainly in L.(L.)amazonensis infection;
while CD8+ cells showed a increase in both parasite infection. IL-4 and IL-10 levels in the supernatant of
lymph nodes cells culture were higher (p<0.05) in the L.(L.)amazonensis than those in the
L.(V.)braziliensis infection; while IFN- and NO levels were higher p<0.05)
(
in the L.(V.)braziliensis
th
th
infection than in the L.(L.)amazonensis infection either at 4 or at 8 week PI. The results highlight the
role of Leishmania species in the determination of different clinical-immunological features in the
experimental infection.
40
Oral Presentation 7
VIBM_0801_0021b
Health care workers’ knowledge, attitudes and reported practices
related to health care-associated infections in a Indian teaching
hospital
*
*
Vishal Diwan (1, 2) , Senia Rosales-Klintz (1) , H Shah (3), Rita Joshi (3), Megha Sharma(1,5), Ashish Pathak(6),
Johan Struwe(7), A.J. Tamhankar(8, 9), Cecilia Stålsby Lundborg(1) * authors contributed equally
(1) Division of Global Health (IHCAR), Department of Public Health Sciences, Karolinska Insititutet, Stockholm, Sweden. (2) Department of
Public Health & Environment, R.D. Gardi Medical College, Ujjain, India (3) Department of Microbiology, R.D. Gardi Medical College, Ujjain,
India (4). Department of Community Medicine, R.D. Gardi Medical College, Ujjain, India (5) Department of Pharmacology, R.D. Gardi
Medical College, Ujjain, India (6) Department of Pediatrics, R.D. Gardi Medical College, Ujjain, India (7) Swedish Institute for
Communicable Disease Control, Solna, Sweden (8) Department of Environmental Medicine, R.D. Gardi Medical College, Ujjain, India (9)
Indian Initiative for Management of Antibiotic Resistance (IIMAR), Mumbai, India
Introduction: Health care-associated infections (HAI) threaten patient’s safety, especially in settings
with limited resources. The simplest and most effective measure to prevent HAI is hand hygiene.
Evidence-based data to develop infection control strategies to contain HAI spread are highly needed
in India. Research objective: To assess knowledge and to explore attitudes and reported practices
related to HAI among health care workers (HCW) in a teaching hospital in India. Methodology:
Cross-sectional study conducted in a 570-bed rural teaching hospital associated to the R.D. Gardi
Medical College Ujjain district, Madhya Pradesh, India (2010-2011) as part of the baseline
evaluation of an ongoing intervention study. Participants were HCW (n=282) involved in direct
patient care, including doctors (n=53), medical students (n=86), nursing instructors (n=14), staff
nurses (N=113) and other HCW (n=16). An interviewer-administrated questionnaire comprising
mainly close-ended questions was presented in English, Hindi or Malayalam. A knowledge score was
calculated as the sum of correct answers (n=53) in questions related to transmission and prevention
of HAI. Participation was voluntary and responses were confidential. Ethical approval was granted
by the institutional ethics committee. Results: Most respondents acknowledged the importance of
HAI prevention (94%) and recognized the negative consequences associated to HAI occurrence
(80%). Preventive measures such as hand washing, glove use and general cleanliness were largely
recommended (>90%). However, only 31% of the participants recognized the role of HCW´s hands
as main transmission route for HAI. Hand washing is “always” practiced by 60% of the respondents
(self-reported compliance). Overall, the mean knowledge score was 35 (SD=10; 95% confidence
Interval = 34-36). Staff nurses had a significantly lower score when compared to doctors and
medical students. Conclusion: Our findings indicate the need to improve knowledge on HAI among
HCW in this setting. Increased knowledge and awareness of HAI transmission and prevention might
41
influence positively hand hygiene practices as well. Therefore, context-relevant interventions
targeting different professional groups are needed.
Oral Presentation 8
VIBM_8005_0020
Establishing a Proteomics Laboratory at Instituto Clodomiro Picado
(UCR): first two-year's experience in serving local and regional
research
1
2
Bruno Lomonte and Juan J. Calvete
1
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica;
Instituto de Biomedicina de Valencia, CSIC, and Departamento de Biotecnología, Universidad Politécnica de
Valencia, Spain
2
Background: Through the financial support of CONARE and Vicerrectoría de Investigación (UCR), a
basic core of laboratory equipment to perform proteomic analyses was installed at the Instituto
Clodomiro Picado (ICP), and officially inaugurated in April 2010. Methods: Efforts towards this
endeavour began two years earlier, by the training of academic personnel through exchange visits
to the Laboratory of Structural Proteinomics of Instituto de Biomedicina de Valencia (IBV), Spain.
This stage was followed by the acquisition of an equipment core, consisting of two mass
spectrometers (a MALDI-TOT-TOF and a nano-ESI triple quadrupole/LIT hybrid instrument), an Nterminal Edman protein sequencer, an automated in-gel protein digester, two HPLC and one FPLC
chromatographs. After further training of staff in the specific use of the instrumentation installed, a
first stage of implementation began, which consisted in the initial acquisition of analytical
experience/know-how, using protein and peptide samples provided by "internal" researchers at the
ICP, with expert assisstance from the IBV lab. This period spanned for approximately one year, and
has been followed by a second stage (currently), where analytical services are offered to the
academic community of biomedical researchers in Costa Rica. Also, a few researchers from other
countries (Colombia, Chile, Mexico, Papua New Guinea) have utilized the analytical services offered
in this newly-established facility. Conclusions: This project may be of special value and usefulness to
researchers in the Central American region, by providing powerful analytical tools to their diverse
projects in biomedical sciences involving protein identification and characterization. Although initial
experiences in this laboratory have dealt mostly with the proteomic study of snake venoms and
42
their toxins, analyses are now being diversified to applications in areas such as bacteriology,
immunology, parasitology, experimental pathology, and others.
Poster Presentation List
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
Research Priority Setting: Experience at the Faculty of Medical Sciences UNAH and the health units
network in Honduras, 2008-2012. Jackeline Alger et al
In vivo and in vitro regulation of Brucella abortus Type IV secretion system VirB by quorum sensing.
Pamela Altamirano et al
Assessment of TB10.4 and cross reactivity with environmental antigens in the context of Tuberculosis
immune recognition. Nancy Alvarez et al
Extended-Spectrum β-Lactamase-Producing Escherichia coli from Hospital Wastewater and Well Water
in León, Nicaragua. Erick Amaya et al
Vigilance of serotypes of Streptococcus pneumoniae in Honduras. Marjorie Anariba et al
An Integral Intervention for Malaria Control in the Municipality of Wampusirpi, Gracias a Dios. Engels
Banegas et al
Phenotypic and molecular characterization of Salmonella spp isolated in domestic animals in
susceptibility to antibiotic. Claudia Barahona et al
The role of neutrophils in the control of adaptive immunity against intracellular pathogen Brucella
abortus. Elias Barquero-Calvo et al
Nutritional Status Children Under Five years, Municipality of San Juan Ojojona, Honduras. January-May
2011. Wendy Barrientos et al.
Porcine calicivirus infections in home raised pigs from rural villages of Nicaragua. Patricia Blandón et al
Vaccine-derived NSP2 segment in rotaviruses from vaccinated children with gastroenteritis in Nicaragua.
Filemón Bucardo et al
Isolation and Detection of Leishmania Species among Wild and Domestic Hosts in an Endemic Area for
Cutaneous Leishmaniasis in Capira, Panama. José Calzada et al
Structural and biological characterization of a PII metalloproteinase from the venom of the arboreal
snake Bothriechis lateralis. Erika Camacho et al
Develop and validate an ELISA for detecting antibodies to SLPs B. abortus in cattle. Jorge Carrasco et al
Quality Standars on Blood Culture Sampling in the Hospital Escuela, Tegucigalpa, M.D.C. Honduras,
January - April 2011. Roxana Castillo et al
Presence of pathogenic agents and microbial indicators of contamination in wastewater, Metropolitan
Area, Costa Rica. Luz Chacón et al
The use of green fluorescent protein as a marker for Brucella vaccines. Carlos Chacón-Díaz et al
Sand fly density reduction is less marked in precarious housing after insecticide residual spraying. Luis F
Cháves et al
Sand Fly species composition in a rural setting with hyperendemic cutaneous leishmaniasis transmission.
Luis F Cháves et al
Capacity building through a new masters program in infectious diseases in Honduras: a case study of
international collaboration. Lourdes Enríquez et al
43
21. Phenotypic and molecular characterization of clinical isolates of enterotoxigenic (ETEC) and
enteropathogenic (EPEC) Escherichia coli in fecal samples of children under three years of Tegucigalpa
and Comayagüela—Advances. Jessy Espinoza et al
22. Research Ethics Governance in Honduras. Vilma Espinoza et al
23. Genetic polymorphisms associated with drug resistance in Plasmodium falciparum collected in Honduras.
Gustavo Fontecha et al
24. Patterns of prevalence and parasitological density of geohelminths, malaria and nutritional status among
public school going children in Honduras. Nelly Franco et al
25. Carcinogenic properties of the bacterial cytolethal distending toxin. Teresa Frisan et al
26. Genotypic characterization of Histoplasma capsulatum isolated from histoplamosis patients in Honduras.
Carmen Galo et al
27. Characteristics of Plasmodium vivax infections on individuals with and without fever from three endemic
communities in Olancho, Honduras, 2010. Jorge García et al
28. IL-17 expression in lesions of patients with different clinical forms of American Cutaneous Leishmaniasis
caused by Leishmania (L.) amazonensis and L. (V.) braziliensis. Claudia Gomes et al
29. Association between dengue transmission and river level variability in Dhaka, Bangladesh. Masahiro
Hashizume et al
30. Active Cases of Leishmaniasis Detection. Experience in Amapala, Valle, Honduras. Jessica Henríquez et al
31. Identification and evaluation of pre-malignant lesions associated with Helicobacter pylori infection in
Guatemala. Elisa Hernández
32. Validation of a Molecular Characterization Protocol for Bacterial Isolates from the Genus Brucella by
Using a Multiplex Assay Based on Single Nucleotide Polymorphism Detection. César Jiménez et al
33. Molecular Characterization of Rotavirus Strains Isolated from Domestic Animals in Costa Rica. Carlos
Jiménez et al
34. An assessment of competencies and perspectives gained by Canadian students during a global health
course. Daniel Korpal et al
35. Saliva from natural vector of Leishmania (L.) amazonensis did not promote the enhancement of
experimental infection. Marcia Laurenti et al
36. Amoebiasis in Leon, Nicaragua: Overdiagnosis and Overtreatment. Byron Leiva et al
37. Bcsp31 is overexpressed in Brucella abortus mutants defective in the type IV secretion system. Esteban
Lizano et al
38. Genetic Diversity of Plasmodium falciparum in Honduras. Ana C López et al
39. Surveillance of Viral Respiratory Pathogens in Tegucigalpa, Honduras 2006-2009. Ivette Lorenzana et al
40. Susceptibility of Aedes aegypti (Diptera: Culicidae) Larvae to Temephos (Abate) from Central District,
Francisco Morazan, Honduras. David Martínez et al
41. The two-component system BvrR/BvrS regulates the expression of the type IV secretion system VirB
in Brucella abortus. Magda Martínez et al
42. Efficacy of chloroquine for the treatment of uncomplicated Plasmodium falciparum malaria in Honduras.
Jessia Matamoros et al
43. Brucella canis is a Naturally Rough Bacteria Capable of Reach Intracellular Niches. Ma Concepción
Medina et al
44. External Evaluation of the Performance of Microscopic Diagnosis for Malaria in the Countries of the
Americas. Rosa Mejía et al
44
45. Implementation of Routine Monitoring using Genetic Markers for Detection of Anti-malarial Drug
Resistance in Plasmodium falciparum in Honduras. Meisy Mendoza et al
46. Genotyping Chlamydia trachomatis in Sex Workers Attending Two STI Clinics in Tegucigalpa and San
Pedro Sula, Honduras between March - June 2011. Suyapa Mendoza et al
47. Molecular Epidemiology of HIV-1 in Honduras. Wendy Murillo et al
48. Ecosystem and seroprevalence of the Leptospirosis in domestic animal in Santa Rosa de Aguan-Colon,
Honduras. Brenda Orellana et al
49. Genetic Diversity of Plasmodium vivax in Honduras. Andrés Ortiz et al
50. Evidence of the etiological role of noroviruses and human astroviruses in childhood diarrhea in
Tegucigalpa, Honduras. Jafet Ortiz et al
51. HIV-1 Drug Resistance among Pregnant Women in Honduras. Leda Parham et al
52. Nicaragua's Millennium Project: measures to help reduce poverty and improve child survival. Wilton
Pérez et al
53. Characterization of Genetic Diversity of Brucella ceti Isolated on the Pacific Coast of Costa Rica, Through
Sequencing of Nine Selected Genes. Martha Piche et al
54. Molecular Characterization of Mycobacterium tuberculosis isolates from patients with tuberculosis in
Honduras. Lelany Pineda-García et al
55. Characterization of species of Lutzomyia and Leishmania in endemic area of Non-Ulcerative Cutaneous
Leishmaniasis, municipality of San Francisco de Coray, Valle, Honduras. Doris Quan et al
56. Assessment of pediatric Norovirus diarrhea in rotavirus vaccinated children. Yaoska Reyes et al
57. Competence of asymptomatic naturally infected dogs to transmit Leishmania (Leishmania) infantum
chagasi to the natural vector. Vania Riveiro et al
58. Chromosomal and Extrachromosomal Location of vanA in Vancomicyn-Resistant Enterococcus faecium
Strains Isolated in Costa Rica. Olga Rivas et al
59. Association of variants of receptor FcγRIIA-131 with severe dengue in Honduras. Cynthia Rodríguez et al
60. Is outer membrane lipoprotein NlpI connecting bacterial RNA degradation with cell wall synthesis in
Salmonella enterica serovar Typhimurium? César Rodríguez et al
61. Phenotypic and molecular characterization of strains of drug-resistant Staphylococcus aureus obtained
from patients attending the Hospital Escuela and the Instituto Hondureno de Seguridad Social during the
period from October 2010 to January 2011. Norman Rojas et al
62. Genetic characterization of multidrug-resistant Mycobacterium tuberculosis isolates from Honduras.
Senia Rosales et al
63. Kato-Katz method in the diagnosis of hookworm infections: the importance of quality control. Ma
Mercedes Rueda et al
64. Molecular characterization of Brucella isolates by MLVA-16. Nazaret Ruiz et al
65. Peridomestic Attalea butyracea palm trees as a risk factor for dwelling triatomine infestations in rural
Panama and an alternative control strategy. Azael Saldaña et al
66. Homogenates of skeletal muscle injected with snake venom inhibit myogenic differentiation in cell
culture. Patrica Saravia-Otten et al
67. An Integral Intervention for Malaria Control in the Municipality of Wampusirpi, Gracias a Dios. Prabhjot
Singh et al
68. Characterization of species of Lutzomyia and Leishmania in endemic area of Cutaneous Leishmaniasis,
municipality of Dulce Nombre de Culmí, Olancho, Honduras. Wilfredo Sosa et al
45
69. Prevalence of Helicobacter spp. and H. pylori Presence in Canines from Metropolitan Area in Costa Rica,
and its Relationship with Gastric Pathologies. Marcela Suárez-Esquivel et al
70. Congenital HCMV Infection in Neonates from Honduras and Costa Rica. Lizeth Taylor et al
71. Molecular characterization and follow-up of a Hepatitis C viral infection in a hematology-oncology unit at
Children´s Hospital in Nicaragua. Lizeth Taylor et al
72. Challenges in achieving the Millennium Developing Goals in Nicaragua: Factors contributing to the
spread of HIV/AIDS in Nicaragua. William Ugarte et al
73. Soil Transmitted Helminths and Nutritional Status of School-Age Children in a Honduran Endemic Area.
Mary-Theresa Usuanlele et al
74. Hospital outbreak by KPC producing Klebsiella pneumoniae in a hospital in Panamá. Silvio Vega et al
75. Enterohemorragic Escherichia coli in Fecal Samples of Cattle from Dairy Farms in León city, Nicaragua.
Samuel Vilchez et al
76. Helicobacter pylori clarithromycin resistance from Guatemalan strains. Carmen Villagrán et al
77. A Baseline Pilot Study using Multidisciplinary Interventions to Improve Mother-child Health in Rural
Regions of Honduras: Integration of Health Education and Nutrition Intervention with Deworming
Programs in Olancho. Concepción Zuniga et al
Poster Abstracts
VIBM_3011_0111
Research Priority Setting: Experience at the Faculty of Medical
Sciences UNAH and the Health Units Network in Honduras, 20082012
1-3
1,3
1
1
4
Jackeline Alger,
Edna Maradiaga, Elizabeth Paguada, Gustavo Moncada, Sonia Flores, Odessa
5 1
2
Henriquez. Unidad de Investigación Científica, Facultad de Ciencias Médicas (FCM), UNAH; Servicio de
3
Parasitología, Departamento de Laboratorios Clínicos, Hospital Escuela, MOH; Instituto de Enfermedades
4
5
Infecciosas y Parasitología Antonio Vidal; Postgrado de Rehabilitación, FCM UNAH; Instituto Nacional
Cardiopulmonar, MOH; Tegucigalpa, Honduras.
Background. Research is a powerful tool for health and development. Priority setting and
developing the capability for priority setting are minimum requirements for a successful research
system. Objective. To describe the experience on health research priority setting at the Faculty of
Medical Sciences (FCM) UNAH and the health units network in Honduras during the period 20082012. Methodology. On April 2008, our institutions participated at the First Latin American
Conference on Research and Innovation for Health, Brazil. On November 2008, with COHRED and
46
PAHO support, an Inter-institutional Committee for Health Research was constituted with the
purpose to generate the Honduras National Health Research System. As a complementary activity,
the Scientific Research Unit FCM made a call for research priority setting to departments and
academic units as well as the public/private health units network. Meetings were held to explain the
importance of priority setting, to share the FCM research policy, to describe the experience of other
countries, to illustrate the usage of priority setting tools such as the Combined Approach Matrix and
weighted criteria matrix, to identify research lines and topics/subtopics by group work, to elaborate
a research agenda and to identify priorities by list reduction and weighting. Results. Meetings have
been held with 27 units, 15 (55.6%) within the FCM (pre-grade and postgraduate careers, nursing
and medicine), 1 (3.7%) other faculties within UNAH, 8 (29.6%) MOH and 3 (11.1%) others. The
development of the initiative has included the explanation of the importance and priority setting
tools (66.7%), elaboration of research agenda (22.2%), and completion of a weighted research
agenda (11.1%). The FCM research lines include 1) Health Problems (non transmissible chronic
diseases, transmissible diseases, neglected diseases, mother-child health, and mental health), 2)
Policy and Health System (quality, promotion, information, access, environment, biotechnology,
alternative medicine, legal framework, and human resources), 3) Institutional Line (academic
management, ethics/bioethics, service and teaching integration, opportunity/curiosity drivenresearch). Conclusions. To reduce inequities, improve population health and enhance development,
research must be performed within an articulated system with defined roles for governance, legal
framework and priorities. Institutional priority setting represents a preparation stage for a national
priority setting initiative.
VIBM_1122_0077
In vivo and in vitro regulation of Brucella abortus Type IV secretion
system VirB by quorum sensing
1
2
2
Pamela Altamirano Silva , Caterina Guzmán-Verri , Edgardo Moreno and Esteban Chaves-Olarte
1,2
1 Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica,
1000 San José, Costa Rica and 2 Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina
Veterinaria, Universidad Nacional, P. O. Box 304, 3000 Heredia Costa Rica
The ability to multiply within eukaryotic cells is essential for the pathogenesis exerted by Brucella
abortus. The mechanisms used by Brucella to survive intracellular remain largely unknown. The
Type IV secretion system VirB determines the intracellular traffic of Brucella to the endoplasmic
reticulum (ER) where it replicates. The transcription of the virB operon is regulated by several
systems. One of these systems is associated with the quorum sensing through the transcriptional
47
regulator VjbR, a protein that positively regulates virB by direct binding to the promoter region. This
activity seems to be inhibited by a homoserine lactone which accumulates when a high bacterial
density is reached. The aim of this project is to understand the in vitro and in vivo regulation of the
type IV secretion system VirB by the quorum sensing influence and how this regulation allows the
molecular changes needed by B. abortus to replicate intracellularly. To achieve this aim, cells were
infected with B. abortus at different phases of the growth curve and the intracellular replication was
followed at 0, 4, 24 and 48 h by plate counting and fluorescence microscopy. In addition,
intracellular bacteria at early times of infection (0-4 hours) were purified to monitor expression of
VirB. When the inoculum was in the lag and log phase the bacteria replicated intracellularly at very
high levels whereas, bacteria in stationary phase achieved significantly lower levels of replication.
These degrees of replication correlated with the level of activation of VirB with a significant increase
in its intracellular expression when the inoculum was in the log phase and a lower degree of
activation when the inoculum was in the stationary phase. In addition, we noted that upon
intracellular replication, bacteria reached a state where they are no longer recognized by antibodies
indicating the existence of molecular changes at its surfaces during restricted points of its
intracellular cycle. Our hypothesis is that Brucella replicating in the endoplasmic reticulum secrete
high concentrations of homoserine lactone which in turn regulate, through VjbR, the induction of
different intracellular changes allowing the survival of this pathogen.
VIBM_0615_0068
Assessment of TB10.4 and cross reactivity with environmental
antigens in the context of Tuberculosis immune recognition
1,2
1
3
1,
2,4
Nancy Alvarez-Corrales , Lelany Pineda-Garcia , Raija K Ahmed , Carol A Rodriguez Markus Maeurer
1
Escuela de Microbiología, Universidad Nacional Autónoma de Honduras (UNAH). Tegucigalpa, Honduras.
2
Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet. Stockholm, Sweden.
3
4
Swedish Institute for Communicable Disease Control (SMI), Stockholm, Sweden, Center for allogeneic stem
cell transplantation (CAST), Karolinska University Hospital, Stockholm, Sweden.
Background: Tuberculosis is one of the greatest infectious diseases worldwide. It is highlighted the
need to discover novel biomarkers and correlates of protection. Secreted proteins like TB10.4 are
major targets of immune recognition by T cells inducing IFN-γ release. Past infections individualize
the memory T cell pool and private T cell receptor specificities of preexisting populations influence
disease severity and outcome of subsequent unrelated infections with close sequence similarity.
Objective: To define the recognition and activation by host T cell response toward TB10.4 (Rv0288)
and study the relationship between heterologous immunity and TB.
48
Methodology: Recombinant mimicry peptides of different strains of yeast, fungus and bacteria
were assessed in a Whole Blood Assay stimulating T cell activation; we determined IFN-γ (ELISA),
multiple cytokines and chemoquines (Luminex) and B cell recognition (chip microarray technology)
in the cell culture supernatant of TB- and TB+ Honduran population.
Results: Based on IFN-γ, TB+ and TB- population had broadly recognition over the entire TB10.4
protein. TB+ responders toward mimicry peptides ranks 8-60pg/ml including S. cerevisiae and R.
lactaris; TB- group reached up to 174pg/ml recognizing TB10.4, S. cerevisiae and A. fumigatus.
Multiple cytokines demonstrated ongoing inflammation and cellular trafficking; B cell recognition to
TB10.4 showed different responses compared to cellular signature.
Conclusions: The pattern recognition along the recombinant protein and its relationship with
memory T cells toward unrelated organisms may skew the host immune responses either providing
protection or pathology. The variances of the responses are very diverse along the protein and
promote antigenic variation; however the expositions to natural microorganisms generate T cell
cross-reactivity when shared sequences of unrelated pathogens can be target of pre-existing
memory T cells and diverge the course of the clinical disease by changing the pattern of T cell
immune dominance or leading the selection of epitope variants.
VIBM_c010_0069
Extended-Spectrum β-Lactamase-Producing Escherichia coli from
Hospital Wastewater and Well Water in León, Nicaragua
1,3
2,3
3
3
1
2
Erick Amaya , Daniel Reyes , Margarita Paniagua , Soledad Calderón , Mamun Rashid , Patricia Colque ,
2
2
1
1
Inger Kühn , Roland Möllby , Carl Erik Nord , Andrej Weintraub *
1
Division of Clinical Microbiology, Department of Laboratory Medicine, Karolinska Institutet, Karolinska
University Hospital, Huddinge, Sweden
2
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Sweden
3
Department of Microbiology, Faculty of Medical Sciences, National Autonomous University of Nicaragua
(UNAN), León, Nicaragua
Antibiotic resistant bacteria have emerged by the selective pressure of antimicrobial use in humans
and animals. Water plays an important role for dissemination of these organisms among humans,
animals and the environment. Thus, tracking the spread of antibiotic resistant bacteria in water
samples is a useful source of information that can be used to design strategies for the control of
these microorganisms. This study was aimed to investigate the antibiotic resistance patterns and
presence of bla-genes among 493 Escherichia coli isolates from different aquatic environmental
sources collected through October 2008 to May 2009 in León, Nicaragua. Antibiotic resistance
analyses were done according to the Clinical and Laboratory Standards Institute guidelines.
Screening of bla-genes on extended-spectrum β-lactamase (ESBL)-producing E. coli was done by
49
multiplex PCR assay using universal primers blaSHV, blaTEM, blaOXA, and blaCTX-M. Sequencing
analysis of the amplicons was performed to identify the specific bla genes detected in the PCR.
Epidemiological relationship between E. coli isolates, harbouring bla-genes, from the hospital
sewage water and from well water samples was analyzed by RAPD-PCR. High levels of antibiotic
resistance were found in E. coli isolates from hospital sewage water and in 8 of 87 well water
samples. Among the E. coli isolates from the hospital sewage, ampicillin, chloramphenicol,
ciprofloxacin, nalidixic acid and trimethoprim-sulfamethoxazole was the most common multiresistance profile (67%). Differently, ampicillin, ceftazidime, ceftriaxone, cefotaxime,
chloramphenicol, ciprofloxacin, gentamicin, nalidixic acid and trimethoprim-sulfamethoxazole was
the most common multi-resistance profile (19%) for the E. coli isolates from the wells. ESBLproducing E. coli were detected in 33% of the resistant isolates from the hospital sewage samples
and in 26% of the resistant isolates from the well water samples. It was showed that 100% of the
ESBL-producing E. coli isolates harbouring genes for blaCTX-M. Genes encoding for blaTEM and
blaSHV were only found in the hospital samples (44% and 53%, respectively) and the gene encoding
for blaOXA was only detected in the well water samples (57%). Even though we did not perform a
longitudinal study, our results suggest that ESBL-producing E. coli were widely spread in hospital
sewage water and community water samples.
VIBM_0703_007
Vigilance of serotypes of Streptococcus pneumoniae in Honduras
1.
2.
Microbiologist, National Bacteriology Laboratory.
XII Masters Cohort on Epidemiology CIES 2009-2011
Alertness of pneumonias and bacterial meningitides, (PBM) began in Honduras in 1999 with the
modality of sentinel site; in the Escuela Hospital .This vigilance directed to St. pneumoniae, H.
influenzae and Neisseria meningitidis as the responsible of high percentages of infantile morbimortality in the country. The pneumonias acquired in the community are the main reason of
hospitalization and death of 5-year-old minors in the developing countries. The lethalty the
neumococo is between 10 % and 30 %. Streptococcus pneumoniae is a diplococcal gram positive,
lanceolado, anaerobic optional. The capsular polysaccharide is the base for its diversity antigenic.
There are currently 91 serotypes described. From 2000 to 2005 Honduras reported 65 isolations of
Streptococcus pneumoniae of invading character. 2006 started the stereotyping of the
autochthonous isolations of St. pneumoniae With the objective of identifying the circulating
serotypes in Honduras with the purpose of the introduction of a vaccine. In April, 2011, the Health
State department and PAI introduced the vaccine 13 valente with the modality of two doses and a
reinforcement, directed to 5-year-old minor children and adults (60-year-old) , identified this as
50
susceptible population. Method. The vigilance of the pneumonias and bacterial meningitides is
based on the reference of the Streptococcus pneumoniae isolations of the National Network
Laboratories. The isolations of this pathogenic are those that come from sterile samples, when they
have been capable of producing invading process. Results. From 2005 to 2010 there have been
received in the Laboratory 29 isolations of pneumococo autochthonous. From the 29 isolations of
Streptococcus pneumoniae that were received, 25 were stereotyped. The serotypes 14, 6 and 19
were the most isolated ones. Conclusions. To support and strengthen this vigilance to prove on a
scientific bases that the vaccine 13 ° valente gives the awaited protection to the most sensitive
population in our country. To Improve and increase the collection of samples of suspicious patients
of NMB. Less of the 50 % of the suspected of pneumonia took blood culture samples and to 70 % of
the suspects of meningitis. To implement the new technologies to allow a better diagnosis of NMB
VIBM_0801_0072
An Integral Intervention for Malaria Control in the Municipality of
Wampusirpi, Gracias a Dios
1
2
3
4
5
6
Engels Ilich Banegas , Rosa Elena Mejia , Prabhjot Singh , Efrain Burgos , Jose Aguilar , Karen Calderon ,
7
Wilberto Montalvan
1
2
National Malaria Program, Ministry of Health, Honduras. National Parasitological Laboratory, Ministry of
3
4
Health, Honduras. Pan American Health Organization, Honduras. Regional Health Department of Gracias a
5
6
Dios, Ministry of Health, Honduras. PAWANKA, Wampusirpi, Honduras. Municipal Health Team of
7
Wampusirpi, Ministry of Health, Honduras. Department of Health Promotion, Ministry of Health, Honduras.
BACKGROUND: The municipality of Wampusirpi in Gracias a Dios had the highest incidence of
malaria (118 malaria case per thousand inhabitants) in the country in 2009; a high burden of
Plasmodium falciparum cases and a maternal death due to malaria was reported in the same year.
This high burden of malaria is due to multiple factors including difficult access to health services
owing to geographical, economic and cultural reasons, complicated by lack of knowledge and
empowerment of people. An urgent intervention for malaria control, overcoming these barriers,
was necessary. RESEARCH OBJECTIVE: Develop a comprehensive and integrated intervention with
community participation to reduce the incidence of malaria transmission in the municipality of
Wampusirpi. METHODOLOGY: An intervention was developed and carried out in all 26 localities of
Wampusirpi. Brigades of community volunteers were formed and trained to conduct house to
house visits carrying out direct installation of bednets with 100% coverage of all sleeping spaces,
active case detection, supervised treatment for 3 days targeted towards a reduction in P. falciparum
cases and comprehensive 14 day treatment for P. vivax and mixed infections, and giving health
51
education with emphasis on use and maintenance of the bednet. Surveillance was maintained in the
municipality following the intervention. RESULTS: Coverage of 100% of localities and houses was
achieved using 3380 bednets installed in 902 households with a population of 6161inhabitants; 403
malaria cases were detected and treated of which 50% (201) were due to P. falciparum. Prevalence
rates of up to 29% in localities were found with a global prevalence of 6.39% (95% CI: 5.78-6.99).
Malaria declined by 85.5% in 2011 from 2009 with one case of P. falciparum malaria reported. Two
cases of P. vivax malaria have been reported in the first three months of 2012. CONCLUSIONS: The
implementation of interventions in a comprehensive and integrated manner with community
participation, surveillance and corresponding technical monitoring led to a sustained impact and
drastic reduction in malaria transmission. This serves as a model intervention and is to be scaled up
to other endemic areas of Honduras, especially those with P. falciparum transmission.
VIBM_C645_0110
Phenotypic and molecular characterization of Salmonella spp
isolated in domestic animals in susceptibility to antibiotic.
1,2
1,2
1
2
1
2
Dias SY, RS Nuñes, Barahona C, Carrasco J. Section of Bacteriology, IHIMV, SENASA-SAG, Section of
Immunology, Department of Microbiology, Faculty of Science, UNAH.
Introduction: Salmonellosis is a zoonosis and public health problem (1). Of 2,500 Salmonella
spp serotypes are known. (1). Poultry are the most exposed to Salmonella spp and to control the
bacteria disinfection is the procedure of choice. However, the surveillance of antibiotic resistance is
of utmost importance. Objective: To characterize the molecular and phenotypic resistance of
Salmonella spp isolated from domestic animals.Methodology: a) Source of Salmonella
spp: 34 isolates of Salmonella spp were likely included in this study; these isolates are from a group
of strains of domestic animals kept IHIMV. b) Serological identification of Salmonella ssp: isolates
wereserotyped with polyvalent anti-Vi antiserum. c) Antibiogram to Salmonella ssp: it was
performed by method Kirby-Bauer d) Molecular analysis of Salmonella spp:d1-DNA extraction and
purification: Salmonella spp DNA was extracted and purified by the modified technique of CTAB. d2PCR for virulence genes and genepHCM: The purified DNA of Salmonella spp, were mixed with
primers to amplify different virulence genes (invA and SPVC) and the MDR gene plasmid
incompatibility (pHCM) and replicas were made by 35 cycles. The amplified products were analyzed
in agarose gels and registered the product by digitalcamera.Results: Of the 34 isolates of Salmonella
spp, 18 were detected in the invA gene,17 were detected spvC gene and 16 were
detected pHCM plasmid incompatibility. Of the 34 Salmonella spp strains that were asked to study
susceptibility and the results were:a) 6-beta-lactams, none of the strains were resistant to
2penicillins, 29.4% were resistant to ceftiofur and none was resistant to the other 3 cephalosporins,
52
b) 5 aminoglycosides study: none were resistant togentamicin.Discussion: Of the 34 strains
of Salmonella spp re-confirmed, 53% (18/34) was detected invA gene and 50% (17/34) the
gene spvC. Furthermore, it was found that 77.8% (14/18) of both strains carry virulence genes.
VIBM_1103_0055
The role of neutrophils in the control of adaptive immunity against
intracellular pathogen Brucella abortus
Elías Barquero-Calvo1, Anna Martirosyan2,3,4, Vilma Arce-Gorvel2,3,4, Diana Ordoñez-Rueda2,3,4, Alejandro
Alfaro-Alarcón5, Bernard Malissen2,3,4, Marie Malissen2,3,4, Jean-Pierre Gorvel2,3,4, Edgardo Moreno1,6
1 Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria, Universidad
Nacional, Heredia, Costa Rica, 2 Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille University,
UM2, Marseille, France, 3 Institut National de la Santé et de la Recherche Médicale (INSERM), U1104,
Marseille, France; 4 Centre National de la Recherche Scientifique (CNRS), UMR7280, Marseille, France; 5
Pathology Department, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica, 6 Instituto
Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica
Polymorphonuclear neutrophils (PMNs) are the first line of defense against microbial pathogens. In
addition to their role in innate immunity, PMNs may also regulate events related to adaptive
immunity. To investigate the influence of PMNs in the immune response during chronic bacterial
infections, we explored the course of brucellosis in antibody PMN-depleted C57BL/6 mice and in
neutropenic mutant Genista mouse model. We demonstrate that at later times of infection,
Brucella abortus is killed more efficiently in the absence of PMNs than in their presence. The higher
bacterial removal was concomitant to the: i) comparatively reduced spleen swelling; ii) increased
infiltration of epitheloid histiocytes depicted as macrophages/dendritic cells (DCs); iii) higher
recruitment of monocytes and DCs; iv) significant activation of B and T lymphocytes, and v)
increased levels of proinflammatory Th1 type cytokines. These results reveal that PMNs have an
unexpected influence in dampening the immune response against intracellular Brucella infection
and strengthen the notion that PMNs actively participate in regulatory circuits shaping both innate
and adaptive immunity.
VIBM_0801_0079
NUTRITIONAL STATUS CHILDREN UNDER FIVE YEARS, MUNICIPALITY
OF SAN JUAN OJOJONA, HONDURAS; JANUARY-MAY 2011
53
Barrientos W
* Epidemiologist, Dr of Medicine
Background: Honduras is second with 29% (2), studies show that WFP as one in three children
under five suffer from chronic malnutrition and even in rural area west of the country can be found
levels up to 88% (3) In Honduras, San Juan de Ojojona located south of the capital, is one of 80
municipalities that are considered high priority for the government of the Republic, which since
2005 with its Nutrition Policy and Strategy in line with other programs intervention of national and
international agencies have implemented strategies to support local. Objective: To determine the
nutritional status of children under five years in the municipality of San Juan de Ojojona, Honduras,
January-May 2011. Methodology: Descriptive observational study of January to May 2011. The
Universe is the CESAMO LINVI of the municipality, with 338 children under five years, calculating the
sample with finite sampling formula being 180, stratifying and selecting in 13 districts under the
systematic random sampling. Workshops was conducted to standardize filling surveys and making
weight / height, with instructions and proper-Making Manual Anthropometric Measurements, was
signed informed consent, field supervisors, review the complete filling checking instruments, was
classified, organized by sequence number and area of residence. Was digitized in Epi-Info 3.5.2 and
anthropometric data Epi-Nut with population benchmark NCHS / WHO / CDC. Results: According to
this study Undernutrition in children under five years is associated with low education of mothers,
low monthly family income and housing conditions unfavorable, with 46%. Global Malnutrition is
common 12 to 23 months and the Chronicle with 41% of 48 to 59 months, with most males. Rice,
tortilla, beans are the foods most consumed by reminder 24 hours and 100% of children under six
months of exclusive breastfeeding was more than half of respondents aware of the existence of
center programs leading weight control / size and food aid. Conclusions: Make all health care
centers and private clinics Ojojona INCAP Dietary Guidelines which are selected as potential food
and culture of the region to be implemented. Consider treatment for children under five who was
diagnosed chronic undenutrition.
VIBM_2810_0054
Porcine calicivirus infections in home raised pigs from rural villages
of Nicaragua
1
1
1
2
2
Filemón Bucardo , Patricia Blandon , Yahoska Reyes , Qiu-Hong Wang and Linda J. Saif . National
1
Autonomous University of Nicaragua, (UNAN-León) , Food Animal Health Research Program, Ohio Agricultural
Research and Development Center, Department of Veterinary Preventive Medicine, The Ohio State University,
2
USA .
54
Norovirus (NoV) and Sapovirus (SaV) belong to the Caliciviridae family. They are an important cause
of outbreak and sporadic gastroenteritis in humans. NoV and SaV are highly diverse genetically and
are classified into genogroups (G), genotypes and genetic variants. NoV of GII and SaV of GI are the
most common found in human. Recently, NoV of GII was found in pigs and it was antigenically and
genetically related to the human NoV GII, therefore, questions about its possible zoonotic potential
were raised. A surveillance study was carried out in rural villages of León, Nicaragua between 2010
and 2011 to investigate for the first time in a Central American country the incidence of NoV and
SaV in home raised pigs. We wanted to examine the genetic relationship between viruses recovered
in humans and pigs in this area, where humans and pigs closely contact. In total 200 fecal samples
from asymptomatic male and female pigs of all ages were tested for NoV and SaV. Using degenerate
primers (JV33 and SR80) originally designed to detect human SaV, a total of 22 (11%) were PCRpositive. After sequencing the PCR products (330bp in the RdRp, nt 4580 - 4910), only 3 (NIC47-10,
NIC18-11 and NIC60-11) contained SaV sequences. These Nicaraguan sequences showed the highest
nucleotide identity, with the porcine SaV detected in Korea (88%, strain GN12), United States (89%,
strain OH-LL26), and Finland (92%, strain 338-1-FI-07). NoV screening was performed by RT-PCR
with degenerate primers (p290 and NVp110) and only 7 (3.5%) of 200, tested positive. Finally NoV
of GII was examined in all samples by real-time RT-PCR with primers (NVGIIF and NVGIIR) specific for
human NoV of GII and 9 samples were tested positive. There are not specific primers for all known
NoV and SaV and designing of generic primers is limited by the extreme genetic diversity of these
viruses, therefore sequence analysis of PCR products is undergoing to confirm whether the PCRpositive found in this study are true NoV. Our preliminary finding revealed the first porcine SaV
detected in a Central American country and suggested that NoV of GII might be also circulating in
pigs.
VIBM_4890_0057
Vaccine-derived NSP2 segment in rotaviruses from vaccinated
children with gastroenteritis in Nicaragua
a
b
c
b
Filemón Bucardo , Christine M. Rippinger , Lennart Svensson , John T. Patton .
a
b
Department of Microbiology, University of León, UNAN-León, Nicaragua. Laboratory of Infectious Diseases,
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
c
Division of Molecular Virology, Department of Clinical and Experimental Medicine, Linköping University,
Sweden
Rotavirus (RV) vaccination programs have been established in several countries using the humanattenuated G1P[8] monovalent vaccine Rotarix™ (GlaxoSmithKline) and/or the human-bovine
55
reassortant G1, G2, G3, G4, P[8] pentavalent vaccine RotaTeq™ (Merck). The efficacy of both
vaccines is high (~90%) in developed countries, but can be remarkably lower in developing
countries. For example, a vaccine efficacy against severe diarrhea of only 58% was observed in a
2007–2009 Nicaraguan study using RotaTeq. To gain insight into the significant level of vaccine
failure in this country, we sequenced the genomes of RVs recovered from vaccinated Nicaraguan
children with gastroenteritis. The results revealed that all had genotype specificities typical for
human RVs (11 G1P[8], 1 G3P[8]) and that the sequences and antigenic epitopes of the outer capsid
proteins (VP4 and VP7) of these viruses were similar to those reported for RVs isolated elsewhere in
the world. As expected, nine of the G1P[8] viruses and the single G3P[8] virus had genome
constellations typical of human G1P[8] and G3P[8] RVs: G1/3–P[8]–I1–R1–C1–M1–A1–N1–T1–E1–
H1. However, two of the G1P[8] viruses had atypical constellations, G1–P[8]–I1–R1–C1–M1–A1–N2–
T1–E1–H1, due to the presence of a genotype-2 NSP2 (N2) gene. The sequence of the N2 NSP2 gene
was identical to the bovine N2 NSP2 gene of RotaTeq, indicating that the two atypical viruses
originated via reassortment of human G1P[8] RVs with RotaTeq viruses. Together, our data suggest
that the high level of vaccine failure in Nicaraguan is probably not due to antigenic drift of
commonly circulating virus strains nor the emergence of new antigenetically distinct virus strains.
Furthermore, our data suggest that the widespread use of the RotaTeq vaccine has led to the
introduction of vaccine genes into circulating human RVs.
VIBM_1466_0058
Isolation and Detection of Leishmania Species among Wild and
Domestic Hosts in an Endemic Area for Cutaneous Leishmaniasis in
Capira, Panama
1
1
1
1
2
3
K. Gonzalez , A. Saldaña , A. Miranda , I. Rodríguez , M. Laurenti , M. Baldi and J.E. Calzada
1
1- Gorgas Memorial Institute for Health Studies (ICGES), Panama. 2- Facultad de Medicina, Departamento de
Patología, Universidad de Sao Paulo, Brasil. 3- Facultad de Veterinaria, Universidad Nacional, Heredia, Costa
Rica
Introduction: Cutaneous leishmaniasis (CL) is a major zoonotic vector-borne disease in Panama.
Several domestic and wild forest mammals have been implicated as reservoir hosts of CL. Since
humans do not normally transmit parasites back to the vector sandflies, human infections are
strongly linked with the dynamics of infection on mammal reservoirs. The detection of Leishmania
spp. in domestic and wild animals present in human endemic areas is of fundamental importance
for incriminating them as potential reservoirs of CL infection. Research Objectives: To apply
56
conventional and PCR-based diagnostic techniques for Leishmania spp diagnosis and
characterization in domestic and wild mammal reservoirs in Trinidad de Las Minas, an area of
intense CL transmission in Panama. Methodology: Dogs were tested for the presence of antibodies
against Leishmania by IFAT and ELISA tests using crude antigenic preparation. Blood and tissue
samples from all animals were evaluated to detect Leishmania species using parasitological
methods and by conventional PCR and real time PCR (qPCR). Positive Leishmania species were
characterized by PCR-RFLP and DNA sequencing. Results: We evaluated 51 dogs, 3 horses, 4 spiny
rats, 4 sloths and 3 opossums from this community. Nineteen dogs (19/51; 37.3%) were shown to
be positive for Leishmania antibodies with at least one serological test (ELISA or IFAT). None of
these dogs had clinical signs suggestive of leishmaniasis. Leishmania DNA was detected in three
sloths and it was possible to isolate the parasite from two of them. Positive Leishmania isolates
were confirmed as L. Viannia panamensis. Conclusions: Molecular and parasitological results
suggest that sloths are important in maintaining the sylvatic cycle of Leismaniasis in the area of
Panama. Serological results suggest the dogs are often infected with, but their potential
epidemiological role in Leishmania transmission needs further evaluation.
VIBM_1120_0017
Structural and biological characterization of a PII metalloproteinase
from the venom of the arboreal snake Bothriechis lateralis
1
1
1
1
Erika Camacho , Eva Villalobos , Alexandra Rucavado , Teresa Escalante , José María Gutiérrez
1
1 Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica
Snake venoms are complex mixtures of biologically active peptides and proteins. A non-hemorrhagic
PII metalloproteinase was isolated from the venom of the arboreal viperid snake Bothriechis
lateralis, by a combination of ion-exchange chromatography on DEAE Sepharose and hydrophobic
interaction chromatography on Phenyl Sepharose. The proteinase has a molecular mass of 36,624
kDa, determined by mass spectrometry. It showed a weak glycosylation pattern, as evidenced by
Pro-Q Emerald300 staining, when compared with Blat-H1, a hemorrhagic PII metalloproteinase
isolated from the same venom. The amino acid sequence of peptides obtained by in-gel trypsin
digestion was determined by ESI-MS/MS. In addition, a partial amino acid sequence was deduced
from cDNA obtained from B. lateralis venom gland. Based on the sequences obtained, the protein
belongs to the PII group of metalloproteinases, comprising a metalloproteinase domain followed by
a disintegrin RGD domain. The metalloproteinase has a weak proteolytic activity on casein and
gelatin, but a moderated fibrinogenolytic activity assayed by SDS-PAGE. In addition, the
fibrinogenolytic activity is not inhibited by alfa2-macroglobulin. The enzyme does not cleave the bait
region sequence of alfa2-macroglobulin. This finding might be due to the weak proteolytic activity
57
of this enzyme or, alternatively, to structural characteristics of PII metalloproteinases, since the
hemorrhagic PII Blat-H1 is not inhibited by alfa2-macroglobulin either. The isolated enzyme is able
to degrade nidogen and type IV collagen of Matrigel, after six hours of incubation. The enzyme does
not induce platelet aggregation, but inhibits ADP and collagen-mediated aggregation. This effect is
not reversed by Batimastat, a synthetic metalloproteinase inhibitor, suggesting that it does not
depend on proteinase activity and, instead, is due to the action of the disintegrin RGD domain.
VIBM_0801_0102
Develop and validate an ELISA for detecting antibodies to SLPs B.
abortus in cattle
1.2 Nuñez DG, 2Agüero KM, 3Alvarado XC, 4Posas C, 1Carrasco J. 1Section of Immunology, Department of
Microbiology, Faculty of Science, UNAH. 2Section Antigens Production, 3Seccion of Virology, 4Seccion of
Serology, IHIMV-SAG-SENASA
Introduction: Brucellosis is a contagious zoonotic disease and causes economic impact on the
livestock sector in developing countries. The techniques currently used for serological diagnosis of
brucellosis in cattle are: a) rose Bengal test (RBT), buffered agglutination test plate (BAP), Rivanol
and kit ELISA-sLPS1. Objectives: To develop, standardize and validate an ELISA-sLPS home to
diagnose bovine Brucellosis. Methodology: 1) Origin of samples: a total of 100 sera were obtained
from Serology Section -IHIMV, of which 50 sera were positive and 50 negative sera by RBT, BAP and
Rivanol. 2) Production of sLPS antigen: strain 1119-3 was used for B. abortus and seeded in 4 days in
potato agar was obtained a bacterial paste was mixed with phenol-water, the organic phase was
precipitated with methanol saturated with sodium acetate and the supernatant was mixed with
trichloroacetic acid. This mixture was dialyzed, then lyophilized and stored at -20°C until use1. 3)
ELISA-sLPS in house: a- Microplate coated sLPS: The sLPS mixed with borate buffer, was adsorbed on
the microplate and blocked with skim milk in PBS-T. b- ELISA-sLPS Assay: The sera were diluted and
deposited in the microplate, were incubated and washed with PBS-T. After was added conjugate
(anti-IgG-peroxidase) incubated and washed with PBS-T. Finally adding the substrate (ABTS) and
quantified at 405 nm. Results: A) From 52 bottles seeded with B. abortus strain 1119-3, 92.3% of
harvested bottles were obtained 50.67 g of wet bacterial paste and this paste was obtained 7.65 mg
of sLPS. B) ELISA-sLPS in house (cut off 1: 64) was compared with the three reference serological
tests, and the ELISA-sLPS in house was 100% specificity, 100% sensitivity and 100% efficiency.
Conclusion: The ELISA-sLPS in house showed an efficiency of 100% compared with the 3 reference
tests.
58
VIBM_0801_0073
Quality Standars on Blood Culture Sampling in the Hospital Escuela
Tegucigalpa, M.D.C. Honduras, January - April 2011
Roxana E. Castillo
Microbiologist, NationalBacteriologyLaboratory. XII Masters Cohort on Epidemiology CIES 2009-2011
Introduction: Blood culture is considered a standard method in the diagnosis of sepsis and
pneumonia. It presents a low positivity, however is the isolation of the agent that helps to
determine the etiology. The most common bacterial agents of pneumonia are: Streptococcus
pneumoniae, Haemophilus influenzae and Neisseria meningitidis, In the Hospital Escuela, the
institution responsible of the surveillance of pneumonia and bacterial meningitis, were received a
total of 5.819 blood cultures from January 1 to December 31 of 2010. Only 25.3% (1.471) have a
bacterial isolation result. There were found Streptococcus pneumoniae (1) and Haemophilus
influenzae at 0.1% (2). Method: Study Area: Hospital Escuela. Type of Study: Qualitative. Evaluation
type. Unit of analysis: The health personnel who take the blood culture sampling. Data Collection
was made through the application of techniques such as Interview Guide, Direct Observation and
Hospital Record Review. Results: There were 38 blood cultures evaluated in four different rooms of
the Hospital. Preparation: Specialist (0) 0% , Medical Care (1) 2.6%, Resident (34) 89.5%, Bachelor
on Nursing (3) 7.9%. The rooms: 0% (emergency pediatric nursing, nutrition and infant) do not have
printed standards. Training: 10 people for a 26.3% do not have training in less than 6 months,
people with training in more than 6 months (22) 57.9%.
Aseptic measures: Wash hands 28.9% (11). 92.1% (35) used laboratory coat, masks and gloves.
100% (38) do not allowed acting disinfectant solution used to sterilize the area to be punctured. 1%
had received antibiotics before taking the sample and only 3 (7.9%) had not received antibiotics.
15.8% (6) had fever at the moment of the sample taking. Conclusions: (1) The majority of the
personnel affirmed to know the procedure on culture blood, but this is not put in practice all the
time. (2) The laboratory applies international standards of quality, nevertheless the training,
evaluation, constant is fundamental. (3) In none of the medical documents of the patient was
registered name of the person responsible of taking the culture blood, being this a legal document.
VIBM_7017_0085
Characterization of the Promoter Region of omp25
Brucella abortus
1
1,2
2,3
Gene in
1
Amanda Castillo Zeledón , Esteban Chaves-Olarte , Edgardo Moreno , Caterina Guzmán-Verri .
59
Programa de Investigación en Enfermedades Tropicales (PIET), Escuela de Medicina Veterinaria, Universidad
1
Nacional, 304-3000 Heredia, Costa Rica ; Centro de Investigación en Enfermedades Tropicales, Facultad de
2
Microbiología, Universidad de Costa Rica, 1000 San José, Costa Rica ; Instituto Clodomiro Picado, Universidad
3
de Costa Rica, San José, Costa Rica .
Brucellosis is a zoonotic disease caused by intracellular facultative cocobacili bacteria belonging to
the Brucella genus. The mechanisms used to invade and replicate within cells are not well
understood. However, its virulence is linked to its outer membrane properties as well as to
secretion and gene regulation systems. Omp25 is the outer membrane protein most studied.
Although its function is unknown its gene expression is under the control of the BvrR/BvrS two
component regulatory system, relevant for virulence. The aim of this work was to characterize the
omp25 promoter region as a prototype of promoter sequences that might constitute a virulence
regulon under the BvrR/BvrS control. To accomplish this, transcriptional fusions of the omp25
upstream region to the lacZ promotorless reporter gene were created. The first fusion had the
complete 392 bp region upstream omp25. Likewise, two shorter fragments were cloned containing
262 bp and 151 bp of the same region. The expression of the transcriptional fusions in the wild type
Brucella abortus 2308 strain was evaluated during its growth curve using β-galactosidase assays and
compared to B.abortus 2308 containing empty vector and B. abortus 2308 with a chromosomal
omp25 lacZ transcriptional fusion. These assays showed the presence of a growth phase dependent
promoter activity in two of the constructs with more than 151 bp upstream omp25, and more than
90% reduction in gene expression in the construct containing the 151 bp fragment. Therefore, the
upstream region between positions -396 bp and -262 bp does not influence omp25 promoter
activity. The 10% activity obtained in the last fragment determine a basal promoter activity which is
located within 151 bp upstream of omp25 and this activity is positively regulated by further
upstream sequences located among the 111 bp between the -262 and -151 bp. This
characterization could be used as a model for comparative studies with other regulatory regions in
important Brucella genes, and in this way create better tools to control their expression and
therefore this pathogen.
VIBM_1107_0047
The use of green fluorescent protein as a marker for Brucella
vaccines
1
2
3
3
Chacón-Díaz Carlos , Mancilla Marcos , Barquero-Calvo Elías , Guzmán-Verri Caterina , Chaves-Olarte
1
2
2
4
3,5
Esteban , Moriyón Ignacio , Iriarte Maite , Grilló María Jesús , Moreno Edgardo .
1. Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica,
San José, Costa Rica. 2. Departamento de Microbiología y Parasitología, Universidad de Navarra, Pamplona,
Spain. 3. Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria,
60
Universidad Nacional, Heredia, Costa Rica. 4. Instituto de Agrobiotecnología, CSIC-UPNA-Gobierno de Navarra,
Pamplona, Spain. 5. Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San
José, Costa Rica.
Brucellosis is an important malady of productive and wildlife animals and a worldwide zoonosis. The
use of effective vaccines and the corresponding diagnostic tests that allow differentiating infected
from vaccinated animals are essential tools to control the disease. For this purpose, a prototype of
Brucella abortus S19 vaccine chromosomally expressing green fluorescent protein (S19-GFP) was
constructed. The S19-GFP was readily identified under ultraviolet light by macroscopic and
microscopic examination and maintained all the biochemical characteristics of the parental strain
S19 vaccine. S19-GFP replicated ex vivo and in vivo, and protected mice against challenge with
virulent B. abortus to the same extent as the isogenic S19. An immunoenzymatic assay designed to
measure anti-GFP antibodies allowed the discrimination between mice vaccinated with S19-GFP and
those immunized with S19. Both vaccines raised antibodies against lipopolysaccharide molecule to
similar levels. The experimental model constitutes a “proof of concept” for the use of Brucella-GFP
vaccines and associated diagnostic test to distinguish vaccinated from naturally Brucella infected
animals.
VIBM_1 11_0001
Presence of pathogenic agents and microbial indicators of
contamination in wastewater, Metropolitan Area, Costa Rica
1
1
1
2
Luz María Chacón Jiménez , Dra. Rosario Achí Araya , Kenia Barrantes Jiménez , Laya HunOpfer , Carolina
1
1
2
1
Santamaría Ulloa , Melissa Solano Barquero , Lizeth Taylor Castillo , Luis Ocampo Van Patten , Carmen
3
Valiente Álvarez
1
2
INISA, Universidad de Costa Rica Facultad de Microbiología, Universidad de Costa Rica
Costarricense de Acueductos y Alcantarillados
3
Instituto
Background: One of the main problems worldwide is the contamination of the hydric resource. The
World Health Organization reports that 1.8 millions of deaths yearly are due to diarrheal problems,
with around 88% of cases of diarrhea associated to the consumption of non-potable water. In Costa
Rica, the wastewater management is poor in the Central Valley, for example, in Heredia, only 5,5%
of the population has a wastewater treatment system, the rest are discharged on rivers. During the
last years the burden of water-transmitted diseases has increased, one example, in Costa Rica, was
the stroke of Norwalk virus in July of 2010. At present no studies regarding circulation of enteric
61
virus or pathogenic bacteria in water are available in our country. Objetives: We propose to
determine the presence of pathogenic viral agents and bacteria in wastewater treatment plants in
Costa Rica's Metropolitan Area and to compare their presence versus contamination indicators such
us coliphages and fecal coliforms. Methods: The presence of microorganisms is determined both at
the input and output of five treatment plants from the Metropolitan Area, in different moments
during 12 months. Molecular techniques (RT-PCR) for detection for Hepatitis A and E Virus,
Rotavirus, Enterovirus, Norovirus and Norwalk virus are used and a molecular technique (MultiplexPCR) will be developed for the detection of virulence genes of Enteropathogenic, Enterotoxigenic,
Shiga-toxin producers, Enteroaggregative and Enteroinvasive Escherichia coli. Results: We expect to
implement standardized methods for detection of enteric viruses in wastewater and to obtain
information about their presence in Costa Rica. We will compare and evaluate the predictive value
of using indicator microorganisms as predictors of the presence of pathogens in wastewater.
Conclusion: This information will represent a first step in the improvement of the wastewater
treatment and will contribute to the public health policies regarding wastewater in the country.
VIBM_D944_0000b
Sand Fly species composition in a rural setting with hyperendemic
cutaneous leishmaniasis transmission
1
1
1
1
1
2
Azael Saldaña , J.E. Calzada , C. Rigg , R. Rojas , A. Valderrama , Luis Fernando Chaves
1
Departmento de Parasitología, Instituto Conmemorativo Gorgas de Estudios de la Sálud, Ciudad de Panamá,
2
Panamá. Programa de Investigación en Enfermedades Tropicales, Universidad Nacional, Heredia, Costa Rica
& Graduate School of Environmental Sciences, Hokkaido University, Sapporo, Japan.
Background: American Cutaneous Leishmaniasis, ACL, is a zoonotic disease with a large richness of
co-occurring vector species in transmission settings. Here, we describe the patterns of
phlebotomine sand fly (Diptera: Psychodidade) species diversity at Comunidad de Trinidad Las
Minas, Capira, Panamá, an hyperendemic focus of ACL transmission. Methods: Our study setting
consisted of 24 houses, 12 subjected to two rounds of indoor residual with and the other 12 kept as
control. During 14 months (April 2010 – June 2011) we monitored Sand Fly species composition and
abundance with CDC light traps inside the houses (domicile) and outside (peridomicile). Results: We
collected a total of 5628 Sand Flies, and we were able to identify 5617 of the samples into 24
species, a number of species close to 25 ± 1.6, the estimate from the Chao2 Index. The most
abundant species were Lutzomyia gomezi (20%), L. triramula (20%) and L. trapidoi (20%). Cluster
analyses showed that most of the 24 houses had high similarity in abundance patterns of the six
most common sand fly species, with only few peripheral houses not following the main cluster
62
pattern. We also found that species richness was decreased to 22 species in the sprayed houses, of
which only 19 were found in the domiciliary environment. Changes in species richness were
especially notorious at the end of the wet season. Conclusion: Our results suggest that species
richness can be decreased following residual spraying in the domiciliary environments, primarily
affecting the less common species.
VIBM_0801_0011
Capacity building through a new masters program in infectious
diseases in Honduras: a case study of international collaboration
Lourdes Enríquez, MSc1; Maritza Canales, MPH1; & Ana Sanchez, PhD2
1
National Autonomous University of Honduras; 2Brock University, Ontario, Canada.
Background: Honduras suffers of a serious deficit of highly trained personnel dedicated to scientific
research. Moreover, foreign-trained researchers returning to the country have traditionally faced
the lack of a research support system that hinders their full professional development and makes it
difficult for them to reach leadership and mentorship positions. Case description: This paper
describes the conceptualization and implementation of a new 2-year research-based graduate
program in infectious and zoonotic diseases (MEIZ) in Honduras. The two main institutional partners
for this initiative were the National Autonomous University of Honduras (UNAH), where the
program was implemented and Brock University, a Canadian University willing to provide
leadership, technical and administrative support. The two institutions were awarded funding by the
Global Health Research Initiative to undertake the “Honduras-Canada Teasdale-Corti Project 200720012: Increasing capacity to achieve millennium development goal # 6 in Honduras: Combating
Infectious Diseases”. MEIZ curriculum was defined in alignment with Honduras’ Country National
Strategic Plan and designed following UNAH’s policies of endorsing research programs relevant to
the Honduran society. Course contents and theses projects responded to academic, governmental
and stakeholders recommendations and entailed crosscutting themes of research methodology,
ethics, biosafety, leadership and service to society. The program structure followed Canadian
standards in terms of projects’ design, literature analysis, knowledge synthesis and dissemination,
technical writing and protocols for theses proposal and final defenses. Discussion: The recently
developed graduate program has been extremely successful in graduating in 2 years 7 new Masters
of Science with different areas of specialization in infectious diseases. Moreover, graduates are
endowed with not only sophisticated laboratory skills but also with solid understanding of how to
undertake high-quality, safe and ethical science. Thesis projects’ research findings are being
disseminated through peer-reviewed journals and conferences and some graduates are considering
further graduate degrees.
63
Conclusions: International cooperation focusing on capacity building to foster scientific
advancement provides a long term solution to the ongoing problem of researchers’ shortage in
developing countries. MEIZ is an innovative model of a graduate program that while responding to
national needs has been implemented with the standards of highly developed country.
VIBM_0801_0028
Phenotypic and molecular characterization of clinical isolates of
enterotoxigenic (ETEC) and enteropathogenic (EPEC) Escherichia
coli in fecal samples of children under three years of Tegucigalpa
and Comayaguela. Advances
1
2
2
2
1
2
Jessy Espinoza Galeas, Yessy Cabrera, Elizabeth Posadas, Ofelia Calidonio, Lourdes de Madrid, Annabelle
Ferrera.
1
2
Infectious and Zoonotic Diseases Master´s Program, School of Microbiology, Science Faculty, School of
Microbiology, National Autonomous University of Honduras, Tegucigalpa, Honduras.
Introduction: Diarrheal diseases are one of the most important causes of illness and death all over
the world. In Honduras, the etiology of diarrheagenic bacteria and the virulence of various
Escherichia coli pathotypes have not been well-studied. ETEC and EPEC are responsible for more
cases of human diarrhea than previously anticipated and require further study to clarify their
virulence mechanisms, the pathogenesis of the infection and their epidemiological significance.
Objective: The aim of the study was to investigate the prevalence of ETEC and EPEC and their
epidemiological association with childhood diarrhea in Tegucigalpa, Honduras, as well as to
characterize the identified strains by phenotypic and genotypic methods. Methods: Stools samples
from children under 3 years of age attending 3 health care centers in Tegucigalpa, and presenting
acute diarrhea, were collected between October 2010 and June 2011, and their parents interviewed
concerning social, economic and environmental factors related to the epidemiology of the disease.
All samples were examined by conventional methods for E. coli and the isolated strains were
subjected to multiplex PCR targeting genes for the thermostable and -labile toxins (STh, STp and LT)
of ETEC, and for the eae and bfpA genes of EPEC. Results: In the present survey, E. coli was isolated
from 69 (77.5%) of the 82 studied children with episodes of diarrhea. ETEC was detected in 8 (12%)
of the E. coli cases; of these, half produced heat-stable toxin (ST) while the other half produced
heat-labile toxin (LT). Five (7%) episodes of E. coli diarrhea were identified as EPEC; one isolate was
classified as typical EPEC while 4 isolates were classified as atypical. Vomiting (69%), fever (30%)
and dehydration (23%) were recorded for cases with ETEC and EPEC diarrhea; additionally, 23% of
the latter infected children consumed just boiled water, not completely purified. Conclusion: These
preliminary data are relevant in estimating the burden of disease due to ETEC and EPEC in children
64
under five years old in Tegucigalpa, Honduras. Data obtained emphasize the need for additional
research into the role of toxin genes and other putative factors affecting the virulence of
diarrheagenic E. coli in Honduras.
VIBM_0501_0108
Research Ethics Governance in Honduras
Vilma E. Espinoza* and Ana L. Sanchez**
*National Autonomous University of Honduras, Tegucigalpa, Honduras; **Brock University, St. Catharines,
Ontario. Canada.
INTRODUCTION: In accordance with universal principles and international standards, countries
enact laws or policies and endorse the creation of research ethics boards (REB) to ensure the ethical
conduct of research with human participants. Biomedical and health research is indeed undertaken
in Honduras but little is known about the status of these regulations in the country. Similarly, there
is a dearth of information regarding the nature, localization and function of research ethics boards
functioning in Honduras. RESEARCH OBJECTIVES: To describe the history and current situation of
research ethics in Honduras as well as to map and characterize the operation of health REBs in
Honduras. METHODS: To gather information, a comprehensive review of Honduras laws and health
regulations was conducted, key informants were interviewed and site visits carried out. RESULTS:
Initiatives for the implementation of research ethics review processes were started by individual
physicians in 1994. Thereafter, academic institutions, professional associations and other
organizations became supportive, such as the Faculty of Medical Sciences of National Autonomous
University of Honduras, the Honduran College of Physicians, the Pan American Health Organization.
More recently, the unit for Science and Research of the Ministry of Planning has become greatly
involved. To date, Honduras lacks of a national law or other form of nation-wide regulatory
framework. However, important aspects of research ethics are included in several laws such as the
Health Code, the Special Law on HIV/AIDS, the Regulations of the Special Law on HIV / AIDS, the
Code of Ethics of the Medical College of Honduras. As for REBs, it was found that as of 2010, 9 REBs
were operating in the country: 6 in the capital city, Tegucigalpa; 2 in San Pedro Sula and 1 in Santa
Rosa de Copan. Four REBs were directly associated with universities and 5 with hospitals and all
reviewed biomedical research. REBs operated in isolation from one another. CONCLUSIONS: Despite
the lack of national regulations, which are urgently needed, there is great interest in Honduras to
uphold the ethical conduct of research. Harmonization of principles and processes among the REBs
would strengthen their work and maximize their potential.
65
VIBM_0801_0008
Genetic polymorphisms associated with drug
Plasmodium falciparum collected in Honduras
1
1
1,3
1
2
2
resistance
in
1*
Ana Cecilia López , Sosa W. , Jovel I. , Varela A. , Mejía R.E. , Banegas E. , Fontecha G .
1
2
MEIZ-Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras; Laboratorio
3
Nacional de Malaria, Secretaría de Salud de Honduras; Malaria Research Laboratory, Infectious Diseases
Unit, Department of Medicine, Karolinska University Hospital/Karolinska Institutet.
*Corresponding Author: MEIZ-Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de
Honduras, Ciudad Universitaria, Boulevard Suyapa, Tegucigalpa, Honduras. Telephone: +504 33935443; email:
[email protected]
BACKGROUND AND OBJECTIVE. Malaria is one of the most important infectious diseases for many
tropical and subtropical countries. The increasing number of drug-resistant parasites is a major
problem in controlling malaria. Central America is the last region of the world where Plasmodium
falciparum remains susceptible to chloroquine (CQ) and sulphadoxine-pyrimethamine (SP). While in
neighbor countries ceased to be useful long time ago [1]. In spite of this, the risk to develop
resistance in Central America is high due to spontaneous mutations and high migratory flow of
people headed to North America from South America where Plasmodium is mostly resistant to
these drugs. The goal of this study was to determine the presence of genetic polymorphisms
associated with drug resistance in Plasmodium falciparum collected in Honduras. MATERIALS AND
METHODS. Febrile patients from 5 different provinces with Plasmodium falciparum diagnosed by
microscopy were included. Patients agreed to participate voluntarily in the study. A blood sample
was collected on paper filter by capillary puncture for PCR analysis. DNA was extracted using a
Chelex-100 (Bio-Rad) approach [2]. P. falciparum chloroquine resistant (Pfcrt) and P. falciparum
dihydropteroate synthase (pfdhps) genes were amplified using nested PCR as reported elsewhere
[3-4]. P. falciparum dihydrofolate reductase (pfdhfr) gene was amplified using unpublished
techniques developed by CDC-Atlanta. Amplification products were observed on 1% et-br agarose
gels. PCR products were purified with Wizard ®SV Gel and PCR Clean-Up System (Promega)
according to the manufacturer's instructions and sequenced by Macrogen. Chromas Pro and Mega5
softwares were used for sequence analysis. RESULTS. Twenty eight P. falciparum samples were
collected. All samples presented the pfcrt 72-76 “CVMNK” genotypes associated with CQ
susceptibility. Positions 540K/581A/613A from pfdhps; and N51/C59/S108/I164 from pfdhfr gene
revealed 100% of wild type genotypes associated with SP susceptibility. CONCLUSIONS. The results
indicate that P. falciparum remains sensitive to CQ and SP in Honduras. This suggests that both
drugs are efficacious for treatment of falciparum malaria. Continuous monitoring of circulating
genotypes in the region is a useful tool to evaluate the potential development of drug resistance in
Central America.
66
VIBM_0501_0089
Patterns of prevalence and parasitological density of geohelminths,
malaria and nutritional status among public school going children in
Honduras
1
1
2
1
Franco, N. , Mejía, R. E , Mancero T. & Paz, M. Y
1
2
Secretaría de Salud, Tegucigalpa-Honduras. Panamerican Health Organisation, Tegucigalpa-Honduras.
Introduction. Neglected diseases including parasitic infections are related to social, economic and
environmental determinants. The 54th World Health Assembly approved the WHA54.19 resolution
which considers these soil-transmitted helminthic infections as a public health concern. Under this
framework member states are encouraged to define prevalence baselines or monitor the impact of
deworming campaigns to establish prevention and control strategies. Material and methods.
A descriptive study was carried out in 2011 in Honduras following the WHO standardized protocol
for school surveys, with a representative sample of students registered in public schools in 2010. A
total of 2,554 students from 48 randomly selected schools of 42 municipalities were included,
representing all the 18 states. With previous informed consent, the following were collected: a)
stool samples processed by Kato-Katz method to determine prevalence and density of
geohelminthic infection; b) blood samples (finger prick) processed for malaria using PCR (RDT and
microscopy were used only for fever cases) and to determine hemoglobin levels; d) anthropometric
measurements (weight and height) to define body mass index (BMI) for the definition of nutritional
status and e)questionnaire on individual and environmental factors related to soil-transmitted
helminthic infections. Conclusions. Of the students studied 43.5% (95% CI 41.6–45.5) had one or
more parasitic infections. Prevalence of specific parasites was: Trichuris trichiura 34.0% (32.2-35.9),
Ascaris lumbricoides 22.3% (20.7-24.0); hookworm 0.9% (0.6-1.3); Hymenolepis nana 0.9%;
Strongyloides stercoralis 0.4%; Enterobius vermicularis 0.1%. Only 43 students had severe infection.
All students with fever tested negative for malaria using both RDTs and microscopy. By PCR testing
of all the students, only 5 were positive for P. vivax, all from the same municipality. Anaemia was
detected in 19.2% (17.7-20.8) of children of which 46.9% (42.2-51.5) had simultaneous parasitic
infections. Majority of children had a normal BMI (84.1%), 3% had less than normal BMI, and 12.9%
were either overweight, obese or had morbid obesity.
Results of this study establish areas of low and no transmission of malaria supporting its elimination
in Central America. The high proportion of overweight and obese children is alarming and requires
immediate attention countering the epidemic by education campaigns and other interventions.
67
VIBM_0890_0038
Genotypic characterization of Histoplasma capsulatum isolated
from histoplamosis patients in Honduras
1
2
1
Carmen María Galo, Ana Sánchez, & Gustavo Fontecha
1
2
Universidad Nacional Autónoma de Honduras; Brock University, Ontario, Canada.
Background. Histoplasma capsulatum is a dimorphic fungus frequently found in nitrogen-rich soils,
especially those containing guano from bat colonies or bird roosts. The fungus causes opportunistic
infections in immunocompromised people and is endemic to the Americas but its epidemiology,
biology and genetic diversity is poorly understood in Honduras. Research objective. To analyze the
genetic diversity of 26 isolates from patients with histoplasmosis and assess their phylogenetic
relationship with isolates from other countries. Methodology. Four DNA sequences of partial
protein-coding genes were examined: arf (ADP-ribosylation factor); H-anti (H antigen precursor); ole
(delta-9 fatty acid desaturase) and tub 1 (alpha-tubulin). The nucleotide sequences of internal
transcribed spacer (ITS) regions of rRNA and 2 non-coding polymorphic sequences 604U and 642U
were also examined. Sequences were amplified by polymerase chain reaction followed by
sequencing to analyze the phylogenetic variability using both Neighbor joining (NJ) and UPGMA
methods. Results. We found between 8 and 13 clusters in the 26 isolates of H. capsulatum. Three
groups were closely related with bootstrap values greater than 60. Comparison of the sequences
obtained in our study with sequences from several geographical areas deposited in the NIH genetic
sequence database, GenBank (http://www.ncbi.nlm.nih.gov/genbank), showed higher similarity
with isolates from Brazil and Mexico than from those from Argentina or Medellin, Colombia, or from
North America or Africa. These findings provide evidence towards the classification of Honduran
isolates as H. capsulatum var capsulatum belonging to the Latin America A (LAm A) class. The
sequences generated were deposited in the GenBank nucleotide sequence databases. Conclusion.
This is the first report in Honduras and Central America concerning the phylogeny and genetic
variability of H. capsulatum. These findings will serve as reference for future studies on the
biological diversity of this important fungus.
VIBM_0801_0099
Characteristics of Plasmodium vivax infections on individuals with
and without fever from three endemic communities in Olancho,
Honduras, 2010
1
1
2
2
3
4
Jackeline Alger, Jorge García, Ofelia Martínez, Martin Ramírez, Ricardo Aviles, Miguel Quintana, Eric
5 1
2
Garges. Servicio de Parasitología, Departamento de Laboratorios Clínicos, Hospital Escuela; Region
68
3
Departamental de Salud, Juticalpa, Olancho; Elemento Médico, Fuerza de Tarea Conjunta Bravo, Comayagua;
4
5
Honduras; US Army Public Health Command Region - South, San Antonio, Texas; Army Institute of Public
Health, US Army Public Health Command, Silver Spring, MD.
Background. Malaria active case detection (ACD) is effective in identifying subclinical cases, but in
Honduras it is not performed routinely. Previous studies suggest that subclinical malaria cases exist
in communities throughout the country. Objectives. To characterize Plasmodium spp. infection in
individuals with and without fever from three endemic communities in Olancho, Honduras.
Methodology. From August 29th to September 2nd 2010, 30 homes were visited in the communities
of Sosa Lobo, Chacon and Villa Antonia, Juticalpa District. After verbal informed consent was
obtained, all the participants were interviewed and a focused physical exam was carried out to
include body temperature. Blood samples were taken using finger prick and a rapid diagnostic test
(RDT) was performed during home visits. Additionally, thick film slides were prepared for
microscopy and filter paper blots were prepared for molecular studies. Parasite diversity was
assessed using P. vivax polymorphic markers for MSP1, CSP and GAM1 genes. Results. Seventy-one
individuals participated in the survey, 19 (26.7%) reported a fever episode within 30 days period
prior to the home visit. One (1.4%) P. vivax positive case was detected by RDT. Subsequent
microscopic evaluation of thick films detected four additional P. vivax cases for a total of 5 (7.0%);
four cases (80.0%) presented low parasite density and two (40.0%) were afebrile; two cases (40.0%)
were women, three (60.0%) were older than 15 years old; 4 cases (80.0%) were from Chacon and 1
(20.0%) from Sosa Lobo communities. Molecular analysis detected one genotype based on a
combination of three markers (MSP1 5/6, ~380 bp; CSP, ~600 bp; GAM1, ~500 bp). All cases were
treated according to MOH guidelines. Conclusions. Through ACD we identified 2 malaria subclinical
cases (2.8%). The microscopic diagnosis was 4 times more sensitive than RDT in presence of low
parasite density. Molecular analysis demonstrated genetic homogeneity and absence of polyclonal
infections. Subclinical cases might contribute to the persistence of malaria transmission in endemic
areas of Honduras. This study was partially funded by Global Fund Honduras, Malaria component
and US Army Center for Health Promotion, San Antonio, Texas, United States of America.
VIBM_FE37_0092
IL-17 expression in lesions of patients with different clinical forms
of American Cutaneous Leishmaniasis caused by Leishmania (L.)
amazonensis and L. (V.) braziliensis
1
1
1
2
1
1
3,4
1
Menezes JGPB , Rodrigues FM , Morais IB , Gama ME , Laurenti MD , Corbett CEP , Silveira FT , Gomes CMC
1-Laboratory of Pathology of Infectious Diseases, Medical School of São Paulo University, São Paulo, Brazil; 2Federal University of Maranhão, 3-Parasitology Department, Evandro Chagas Institute (Surveillance Secretary
69
of Health, Ministry of Health), 4-Tropical Medicine Institute, Federal University of Pará, Belém, Pará, Brasil. Email: [email protected]
The immunopathogenesis of Leishmania infection was explained primarily through the balance
Th1/Th2 immune response. However, recent studies point to the presence of Treg and Th17 cells
playing a role in the immune response against parasite infections. Th17 cells are characterized by
the production of IL-17, which constitutes a pro-inflammatory cytokine secreted primarily by
activated T CD4+ and CD8+ cells. This study aimed to evaluate the profile of cells expressing IL-17 in
skin lesions of patients presenting different clinical forms of American cutaneous leishmaniasis
(ACL) caused by L.(V.) braziliensis and L.(L.) amazonensis, addressing a better understanding on the
role of Th17 cells in the immunopathogenesis of ACL in Brazil. Thirty-one patients were examined:
anergic diffuse cutaneous leishmaniasis (ADCL): 6; borderline disseminated cutaneous leishmaniasis
(BDCL): 6, both by L.(L.) amazonensis (DTH–); localized cutaneous leishmaniasis (LCL) also due to
L.(L.) amazonensis with DTH– (8) and DTH+ (5) and, LCL due to L.(V.) braziliensis with DTH+ (6).
Paraffin-embedded biopsies were submitted to immunohistochemistry using as primary antibody
anti-IL-17 (SC-7927), 1:250 dilution. For amplification and visualization of the reaction Novolink max
polymer was used. The immunostained cells were counted in 5–10 fields (400x) in section by using
an image analysis system (Zeiss). The comparison of IL-17+ cells density in the clinicalimmmunological spectrum of ACL showed a progressive increase in IL-17+ expression from the
central LCL (DTH+) caused by L.(V.) braziliensis to the polar forms, ADCL and BDCL (DTH–) caused by
L.(L.)
amazonensis,
as
follows:
(ADCLDHT-[506]>BDCLDHT-[384]>LCL/LaDTH[318]>LCL/LaDTH+[251]>LCL/LbDTH+[289]). In conclusion, our results showed that cells expressing IL-17
seems to play an important role in the immunopathogenesis of these different clinical forms of ACL
L. (V.) braziliensis or L. (L.) amazonensis, characterized by a polarized immune response and
different pathological expression.
VIBM_TZ02_0037
Association between dengue transmission and river level variability
in Dhaka, Bangladesh
Masahiro Hashizume1, Ashraf M. Dewan2,3, Toshihiko Sunahara1 and Taro Yamamoto1
1 Institute of Tropical Medicine (NEKKEN) and the Global Center of Excellence program, Nagasaki University.
1-12-4 Sakamoto, Nagasaki 852-8523, Japan. 2 Department of Geography and Environment, University of
Dhaka, Ramna 1000, Dhaka, Bangladesh. 3 Department of Spatial Sciences, Curtin University of Technology,
Kent Street, Bentley 6102, Perth, Australia.
Background: Many studies have investigated the relationship between climate and dengue in
various locations. Floods also can potentially increase the transmission of dengue. However, few
70
studies reported the association of dengue epidemics with flooding. Objectives: We estimated the
effects of river levels and rainfall on the hospital admissions for dengue fever at 11 major hospitals
in Dhaka, Bangladesh. Methods: We examined time-series of the number of hospital admissions of
dengue fever in relation to river levels from 2005 to 2009 using generalized linear Poisson
regression models adjusting for seasonal, between-year variation, public holidays and temperature.
Results: There was strong evidence for an increase in dengue fever at high river levels.
Hospitalizations increased by 6.9% (95 % CI: 3.2, 10.7) for each 0.1 meter increase above a threshold
(3.9 meters) for the average river level over lags of 0–5 weeks. Conversely, the number of
hospitalizations increased by 29.6 % (95 % CI: 19.8, 40.2) for a 0.1 meter decrease below the same
threshold of the average river level over lags of
−19
0 weeks. Conclusions: Our findings provid e
evidence that factors associated with both high and low river levels increase the hospitalizations of
dengue fever cases in Dhaka.
VIBM_0703_0083
Active Cases of Leishmaniasis Detection. Experience in Amapala,
Valle, Honduras
3
Henríquez J.¹, Bueso M.², Orellana L.¹, Valladares C.¹, Zúniga C.², Sosa W , Mejía R.E.¹,
1
2
Laboratorio Nacional de Chagas y Leishmaniasis, Secretaría de Salud de Honduras. Programa Nacional para
3
el control y la prevención de la Enfermedad de Chagas y Leishmaniasis, Secretaría de Salud. Escuela de
Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras
Background: The Leishmaniasis is in the Group of neglected diseases in many countries, being the
particular case in Honduras. They are zoonotic diseases that affect more people living in arid
country and wooded areas as it is the case of the island of Amapala that by their climatic and tourist
conditions increase the probability of maintaining and introducing vectors and new cases. In
Honduras, the department’s that inform most of the cases are (Olancho, Choluteca, Valle, El Paraíso
and Francisco Morazán). Objetives: Active cases of leishmaniasis surveillance, through the pursuit of
cases. Methodology: Active cases of Leishmaniasis detection, in the municipality of Amapala, prior
to the activity was conducted promotion in the health center, visits to colleges, schools, and House
to House giving priority to the towns with a history of presence of cases. Patients are attended in
the morning in the health centre and evenings, the laboratory was installed at home of
collaborators who had generally suffered the disease. The mobilization of the laboratory was
conducted in order to attract greater numbers of patients. Results: were attended 42 patients, all
with characteristics of cutaneous leishmaniasis not ulcerated lesions, of which 100% presented
amastigotes in parasitological examination. The 52.4% were female and 47.6% of the male. 61.9%,
71
correspond to under 18 age, 19.1% at ages 19 - 29 years, 7.1%, between 30-40 years, and 11.9%
people older than 41 years. With this research demonstrates that there is more spontaneous cases
which are reported in health facilities by demand, indicating that patients by different
circumstances are not attending with health facilities to be diagnosed in a timely manner and
provide appropriate treatment. Conclusions and recommendations: Surveillance for the disease is
very weak, despite the presence of cases, information system, it is not appropriate in the area,
there is little community participation. Local capacity must be developed to improve the diagnosis
for case detection and monitoring.
VIBM_K007_0019
Identification and evaluation of pre-malignant lesions associated
with Helicobacter pylori infection in Guatemala
1
1
1
2
1
E. Hernández, C. De Tercero, F. Arana, G.I. Perez-Perez, B. Hernández.
1 Facultad de Ciencias Médicas, Universidad de San Carlos, Guatemala. 2 Departments of Medicine and
Microbiology, NYU School of Medicine, New York, USA.
H. pylori infection is associated with superficial chronic gastritis, peptic ulcer diseases and gastric
adenocarcinoma. The histopatological cascade of events related with the evolution of H. pylori
infection was originally described by Correa et al. in 1976. As pathology progress at the gastric
mucosa level, H. pylori colonization has a tendency to be lost. Gastric carcinoma (GC) is gradually
decreasing in developed countries. However, GC continues to be one of the main causes of death in
developing countries. Guatemala is a country where GC mortality is very high and where very little
is known about this disease or H. pylori prevalence. The aim of this study was to determine H. pylori
prevalence and identify and evaluate the presence of pre-malignant lesions associated with this
bacterium in Guatemala City. The objective was determined the prevalence of H. pylori and identify
and evaluate the presence of pre-malignant lesions associated with this bacterium in Guatemala.
We studied 353 dyspeptic patients (mean age 46+15.6) with a 3:1 F to M ratio. Presence of H. pylori
was determined by culture, RUT, and microscopy. Histopathological scores were obtained from an
experience pathologist. Methods: We conducted the study from December 2008 to march 2010. We
studied a total of 353 patients with symptoms that were referred to two major hospital in
Guatemala City (San Juan de Dios and Roosevelt). In addition of demographic information, gastric
biopsy specimens from antrum and corpus were obtained. H. pylori status was determined by
culture, rapid urease test PCR and histology. We found a high prevalence of H. pylori (56.6%) with
no difference by gender. As expected, prevalence of H. pylori increased gradually and significantly
with age. High prevalence of pre-malignant lesions was observed (71.9%), with a significant
72
difference between H. pylori positive and negative patients (77% vs. 65% p-=0.01). In addition,
patients with gastric atrophy were significantly older than those without atrophy.The conclusions
The risk to be H. pylori positive increases with age. Age of the host is more relevant than H. pylori
status for the risk of distal gastric cancer development. Based on this study we concluded that in
Guatemala antimicrobial therapy used to interrupt the progression of the pre-malignant lesions
associated with H. pylori is a necessary preventive intervention.
VIBM_2062_0065
Validation of a Molecular Characterization Protocol for Bacterial
Isolates from the Genus Brucella by Using a Multiplex Assay Based
on Single Nucleotide Polymorphism Detection
1
Jiménez César , Ruiz Nazareth
1
Guzmán- Verri Caterina
1
1,2
1,3
1
1,3
1
, Slon Jose Luis , Barquero-Calvo Elías , Lizano Esteban , Moreno, E ,
2
PIET, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica. Centro de Investigación en
3
Biotecnología. Escuela de Biología, Instituto Tecnológico, Cartago, Costa Rica. Facultad de Microbiología,
Universidad de Costa Rica, San José, Costa Rica
Bacteria from the Brucella genus are responsible of causing brucellosis, the most extended bacterial
zoonosis around the world. There are 10 described species within the genus, some of them able to
cause disease in humans. Therefore, identification at the species level is relevant to achieve
diagnosis accuracy and an effective epidemiologic surveillance. Identification of isolates at the
species level is difficult due to their high similarity at genome level. The aim of this work was to
validate a protocol that allows, under our lab conditions, a reliable identification of bacterial isolates
at species level using a multiplex assay based on single-nucleotide polymorphisms for SNPs
identification. The results were compared with results obtained by MLVA-16 genotyping and
biochemical characterization, both able to distinguish Brucella isolates at the species level. Twentyone Brucella clinical isolates were analyzed using classical biochemical typing techniques. Seventeen
isolates were identified as Brucella canis and three as Brucella abortus. Genomic DNA was extracted
and analyzed using a multiplex assay based on single-nucleotide polymorphisms for SNPs
identification. The obtained extended DNA fragments were processed on a genetic analyzer and the
obtained electrophoretic patterns were compared to the ones provided by Brucella reference
strains. The same DNA was analyzed using MLVA-16 genotyping. The results from the MLVA-16
agreed with those obtained using SNPs identification. Concomitantly, these results matched those
obtained by biochemical typing with the exception of one isolate. Single nucleotide polymorphism is
73
an effective technique for Brucella identification at the species level in a fast and easy way, which
could lead to an accurate epidemiological control.
VIBM_3022_0096
Molecular Characterization of Rotavirus Strains Isolated from
Domestic Animals in Costa Rica
1
1
1
2
1
María del Rocío Cortés , Cristina Solís W ., Lizbeth Ramírez , Magaly Caballero , and Carlos Jiménez S .
1
2
National University of Costa Rica (UNA) , ACOPSA Laboratory, Costa Rica .
Group A rotavirus (RV) play an important role in human gastroenteritis and they are also viral
pathogens associated with neonatal diarrhea in different animal species. At the Central American
region there is limited knowledge of the impact of RV in domestic animals and in most of the
countries it is unknown the G and P groups involved. On the other hand, previously has been
reported, the presence of RV in humans with characteristics typical of RV from animals, which
allows us to suggest that domestic animals could act as reservoirs for human infections and as a
potential source of new genotypes. This investigation was designed to determine the presence and
genetic classification (Groups G and P) of RV affecting bovines, porcines and dogs of Costa Rica and
their relationships with RV strains isolated previously from humans. In the period between 2010 and
2011 we collected 166 fecal samples from calves, piglets, and dogs diseased with diarrhea. All
samples were analyzed for RV using immunochromatographic or immunoenzymatic assays and a
total of 71 samples were scored positive. The RNA from positive samples was analyzed by PAGE,
revealed in all cases a long electrophoretic pattern. By nested PCR was set the groups G and P in 14
samples. In addition the study included a review of the genotypes of Human Rotavirus identified in
previous research (Bourdett et al., 2008, Bourdett et al., 2009, Bourdett et al., 2010, Bourdett et al.,
2011). The molecular analysis performed on samples of RV from animals indicates the presence in
cattle from groups G5, G6, P1 and P5: while in pigs were diagnosed groups G3, G4, G9, G10 and G11
and groups P4, P5, P6, P7 and P10. Meanwhile in dogs were found groups G3, G9, P3 and P7.
Rotavirus group G3, G9, P4 and P6 have been previously found in diarrheal stools of children in our
country (Bourdett et al., 2008; Bourdett et al., 2010, Bourdett et al., 2011). The possible relationship
between the groups G and P found in animals and humans was established by sequencing and
phylogenetic analysis.
74
VIBM_QG10_0100
An assessment of competencies and perspectives gained by
Canadian students during a global health course
Daniel Korpal, Stephanie Della Smirra, Shamiram Zendo and Ana Sanchez
Department of Community Health Sciences, Brock University. 500 Glenridge Avenue. St. Catharines, Ontario,
Canada. L2S 3A1
Background: There is an expansion of Global Health (GH) training initiatives in Canadian universities,
the majority in the form of short-term immersion experiences in low and middle-income countries.
Despite their upsurge, little is known about the competencies students gain through these
experiences. Research Objective: To understand the GH competencies gained by Brock University
undergraduate students that completed the CHSC 3F97, the “Global Health Field Course to
Honduras”. This course is offered every two years to health science students and other majors. The
course, while focused on infectious diseases, provides an opportunity for exploring health issues
pertinent to developing countries. Methodology: Five groups totaling 85 students have taken the
course since 2004. Of those, 77 were eligible and invited via email to participate in the study. A 30minute long online questionnaire was designed with questions grouped into 4 categories:
demographics; perspectives of GH; GH competencies, and whether or not the course had a lasting
impact on their lives and careers. Most questions required participants to answer statements with a
gradient selection from strongly disagree to strongly agree and the majority provided space to
elaborate on the answers and provide examples or make comments. Results: Of 77 eligible, 48
alumni were successfully contacted and of these, 19 have completed the survey (response rate
40%). Only one student (5%) reported having academic GH experience prior to the course and 94%
stated that the course increased their interest in learning about other cultures and languages. The
majority (89%) agreed that the course helped them to better understand the complexities of NorthSouth partnerships, 73% agreed that the course strengthened their research skills and 68% agreed
that the course helped them in defining their career goals. Conclusion: The survey shows that
undergraduate courses such as CHSC 3F97 might be the first opportunity for Canadian students to
be exposed to the realities of the Global South. In particular, this course helped students to gain
essential GH competencies and not only made a lasting impact at the personal level but was also
valuable in helping define career goals and strengthen research skills.
75
VIBM_1072_0029 b
Saliva from natural vector of Leishmania (L.) amazonensis did not
promote the enhancement of experimental infection
1
Francesquini FC,
1
2,3
1
1
1
2
1
1
Silveira FT, Carvalho AK, Tomokane TY, Passero LFD, Barata IR, Corbett CEP, Laurenti MD
2
Laboratory of Pathology of Infectious Diseases (LIM-50), Medical School, University of São Paulo (SP), Laboratory
3
of Leishmaniasis, Evandro Chagas Institute and University of Pará, Belém (PA), BRAZIL.
Introduction and Aim: The promastigote forms of Leishmania are transmitted by sand flies during blood
feeding. Some reports have shown that vector’s saliva plays an important role in the modulation of
immune response, helping the establishment of the infection. But the most of these reports used L.
longipalpis or P. papatasi laboratory-reared as a source of saliva to infect different models with
different species of parasite; and conflicting data have been described when the natural vector/parasite
binomium is used. Thus, the present work has the main aim evaluate, in the natural vector/parasite
binomium, the effects of wild-caught vector saliva of Lutzomyia flaviscutellata in L. (L.) amazonensis
promastigotes infection in BALB/c mice. Material and Methods: The animals were inoculated into
the hind footpads with 106 L. (L.) amazonensis promastigotes in the absence or presence of salivary
gland lysate (SGL) from wild-caught Lu. flaviscutellata. Animals infected in the presence of SGL from Lu.
longipalpis were used as positive control. The lesion size was evaluated weekly till 60th day PI when
biopsies of the skin lesion were collected for histological studies and parasite burden determination.
Results: Increase on the lesion size was observed in all groups with the evolution of the infection, but
the lesion of mice infected in the presence of SGL from Lu. longipalpis was higher than those of mice
infected in the presence or in the absence of SGL from Lu. flaviscutellata, which were similar (p<0.05).
The histological features were similar between the groups and it was characterized by infiltration of
macrophages heavily parasitized, few lymphocytes and polimorphonulcear cells, and focal areas of
necrosis. The number of viable parasites was similar in the mice infected in the presence and absence of
SGL from Lu. flaviscutellata, which were lower than those of mice infected in the presence of SGL from
Lu. Longipalpis. Conclusion: The results presented herein indicated that SGL from wild-caught Lu.
flaviscutellata does not promote the enhancement of the L. (L.) amazonensis infection in BALB/c mice.
76
VIBM_288-_0024
Amoebiasis in Leon, Nicaragua: Overdiagnosis and Overtreatment.
3
2
Byron Leiva , Jadwiga Winiecka-Krusnell and Ewert Linder,
1
1,2.
.
2
Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden, Swedish Institute for
3
Infectious Disease Control, Solna, Sweden, Universidad National Autonoma de Nicaragua –León, Nicaragua.
Introduction, we have previously shown that over-diagnosis of Entamoeba histolytica is a problem
in health centers in the León, Nicaragua. In a group of 134 diarrhea patients, the most common
finding was E. histolytica/ E. dispar (24%) at the health center laboratory level. In the University
laboratory E. histolytica/E. dispar was found only in 4.5%. By PCR, E. dispar was recognized in 10
(7.5%) and E. histolytica in two cases (1.5%). The aims of this study were to obtain information on
the consequences of over-diagnosis of Entamoeba histolytica infection in the León region of
Nicaragua by retrospectively studying patient journals, and further aim was to evaluate the
diagnostic routines, and the professional skills of laboratory technicians at the health centers, finally
to evaluate two ELISA Test. Methods, first, 100 individuals with intestinal symptoms such as:
abdominal pain, diarrhea, constipation, other symptoms as well as the results of stool examination
and anti-parasitic drug were recorded. Second, in the quality control assessment, a questionnaire
was filled to individuals involved in the diagnostic routines and a parasite panel with 10 stool
samples was sent to each laboratory to be analyzed by the laboratories technician. Results, out of
100 patients recorded, 32 were diagnosed with E. histolytica/E. dispar, 23 of them were treated
with metronizadole and 5 with Tinidazole. In total 56 were treated with metronizadole and 7 with
Tinidazole. 41 were treated with some anti-amoebic drug despite the fact that stool examination
was not tested. The results of quality control for stool parasites performed in 15 laboratories were
reported. The evaluation of two ELISA tests, from 50 samples positive by microscopy 6 were positive
for E. histolytica by TechLab II ELISA test and 34 were positive by Remel test. In this study was
showed low cases of positive E. histolytica. In conclusion, the results showed that over-diagnosis of
E. histolytica/E. dispar is a severe problem as the number of false positives exceeds the number of
true positives. In a retrospective study the majority was overtreated and could develop drug
resistance and emerge as an important problem in public health.
77
VIBM_2061_0052
Bcsp31 is overexpressed in Brucella abortus mutants defective in
the type IV secretion system
1-2
1-2
2
2
Esteban Lizano , Carlos Chacón-Díaz , Pamela Altamirano-Silva , Caterina Guzmán-Verri , Edgardo Moreno
3
1-2
and Esteban Chaves-Olarte
2-
1-Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica.
2-Programa de Investigación en Enfermedades Tropicales, Escuela de Medicina Veterinaria, Universidad
Nacional. 3-Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica
The type IV secretion system VirB is crucial for the virulence of Brucella abortus and mutants in this
system are unable to survive intracelullarly in professional and non professional phagocytic cells. In
order to analyze the impact of VirB mutations at the level of the outer membrane, a proteomic
approach was undertaken. Outer membrane vesicles from exponentially growing Brucella, wild
type 2308 strain and isogenic mutants in VirB, were prepared and compared by 1D and 2D
electrophoresis. A protein with a molecular weight of 31 kDa and isoelectric point of 4.0 was
markedly and consistently overexpressed in all the VirB mutants tested and was identified by mass
spectrometry as Bcsp31, an immunogenic protein without known function. Bcsp31 was cloned and
produced in Escherichia coli as a his-tagged fusion protein and was used to produce a polyclonal
antibody. Expression analysis of Bcsp31 by western blotting indicated that this protein is
overexpressed in the VirB mutants being that the reason for its accumulation in outer membrane
fragments. In order to analyze the role of this protein in B.abortus pathogenesis, an isogenic
mutant was constructed and characterized. The ability of intracellular multiplication and the
sensitivity to polymixin B was not altered by the mutation on Bcsp31. Analysis by western blotting
using antibodies against VirB8 evidenced qualitative and quantitative differences in the Bcsp31
mutant as compared to the wild type. Altogether these results suggest that Bcsp31 is involved in
the adequate function and assembly of the Brucella abortus VirB system.
VIBM_0405_0007
Genetic Diversity of Plasmodium falciparum in Honduras
1
1
2
2
1,3
1*
Ana Cecilia López , Sosa W. , Mejía R.E. , Banegas E. , Jovel I. , Fontecha G .
1
2
MEIZ-Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras. Laboratorio
3
Nacional de Malaria, Secretaría de Salud de Honduras. Malaria Research Laboratory, Infectious Diseases
Unit, Department of Medicine, Karolinska University Hospital/Karolinska Institutet
78
*Corresponding Author: MEIZ-Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de
Honduras, Ciudad Universitaria, Boulevard Suyapa, Tegucigalpa, Honduras. Telephone: +504 33935443; email:
[email protected]
BACKGROUND AND OBJECTIVES. Merozoite surface proteins (MSP) of Plasmodium falciparum are
candidates for malaria vaccine [1-2]. However, these proteins show high polymorphism among
parasites. MSP-1 has variables blocks interspersed with conserved blocks [3]. Three main allelic
forms have been described: K1, MAD20 and RO33. MSP-2 has two allelic families: FC-27 and 3D7.
Both genes reveal variations within the same allelic type. This allows detecting polymorphisms in
parasite populations, as well as estimating the number of co-infecting parasites. The analysis of local
genetic variation of P. falciparum genes is an important contribution to the eventual development
of a vaccine in that geographical region. The aim of this study was to determine the genetic
variation present in the genes MSP-1 and MSP-2 of Plasmodium falciparum in Honduras.
MATERIALS AND METHODS. Febrile patients with P. falciparum malaria diagnosed by microscopy
were included. Patients came from five Provinces of Honduras. Blood was obtained by capillary
puncture on paper filter (3 MM Whatman). DNA was extracted using Chelex-100 (Bio-Rad) [4]. MSP1 gen was amplified using 3 sets of primers specific for allelic family by nested-PCR [5]. Eleven
samples were used to amplify MSP-2 [6]. Amplification products were observed on 2% et-br agarose
gels. Specific band size was determined for MSP-1. MSP-2 amplification products were purified and
sequenced. RESULTS. Twenty nine samples were analyzed. Both genes were highly polymorphic
with low allele frequencies. A total of 35 MSP-1 alleles (20 K1-, 9 MAD20-, and 6 RO33-types) were
recorded. Nine infections with more than one allelic family were observed (1 K1/MAD20, 5
K1/RO33, 1 MAD20/RO33, and 21 patients with two K1 bands). MSP-2 sequences revealed 11 3D7and 1 FC27-types (mixed-clone infection). Tree clusters were obtained from the phylogenetic tree
of 3D7 sequences with very low estimates of divergence within 10 of 11 sequences. CONCLUSIONS.
There was extensive polymorphism in MSP-1 and MSP-2 genes. The level of polymorphisms within
those genes and the allelic family distribution are comparable with reported in other geographical
areas with higher levels of malaria transmission.
VIBM_0801_0049
Surveillance of Viral
Honduras 2006-2009
1
Respiratory
2
Pathogens
3
in
Tegucigalpa,
1
2
Ivette Lorenzana de Rivera , Victor A. Laguna-Torres , Maribel Rivera , Cynthia Rodríguez , Erick S. Halsey ,
2
Tadeusz J. Kochel .
79
1
2.
Universidad Nacional Autónoma de Honduras,Tegucigalpa, Honduras, US Naval Medical Research Center
3
Detachment, Lima, Perú. Instituto Hondureño de Seguridad Social.Tegucigalpa, Honduras.
Background: Acute respiratory infections (ARI) constitute one of the principal causes of morbidity
and mortality in developing countries. The disease burden is estimated to be 3.9 million deaths
worldwide [2002]. In Honduras ARI is the number one cause of morbidity in children less than 5
years old. Aims: The purpose of the study was to establish the viral etiological agents associated to
ARI in participants enrolled at Hospital de Espacialidades del Seguro Social in Tegucigalpa, collected
in a surveillance basis. Materials and Methods: From 2006 to 2009, pharyngeal swabs were
collected from consented participants. They were frozen at -80 C and sent off regularly on dry ice to
NMRCD(Research Center of USA Marine) in Lima, Peru; were they were cultured on cell monolayers,
and viral antigens were detected by immunofluorescence staining. Genotyping of the Influenza A
isolates was done by sequencing analysis of the Hemagglutinin gene. Results: A total of 719
analyzed sample; 271(37%) were positive by any viral respiratory agent. A 48% of the cases were in
children between 1-5 years of age. A third of the patients took antibiotics unnecessarily. The
positivity by year range from: 23% to 46%, with the highest incidence observed in 2009. The most
common viruses isolated was Influenza A (175; 67%) Adenovirus (33;12.5%),Parainfluenza
(22,8.3%), Influenza B (17; 6.5%), Herpes simplex (10; 3.8%), RSV (4; 1.5%) and Human
Metapneumovirus (2;0.7%). Dual infections were detected in 2.3% of the cases. Discussion: A
significantly higher positivity was found in age group 1-5 years old. The higher prevalence by viral
etiological agent found was for Influenza A, followed by Adenoviruses. The prevalence of RSV was
low. The higher incidence observed in 2009 was due to worldwide epidemic of Influenza A H1N1.
Genetic analysis of influenza isolates showed 15 different genotypes all coming from the Americans
region. Conclusion: The data demonstrates that surveillance studies such as this are important, to
be able to report that are several viral agents that are associated with IRA in Tegucigalpa, Honduras
and to show seasonality, yearly peaks of incidence as well its association with age group and
severity of the disease.
VIBM_0801_0025
Susceptibility of Aedes aegypti (Diptera: Culicidae) Larvae to
Temephos (Abate) from Central District, Francisco Morazan,
Honduras
1
David Martínez and Eduardo Fernández
2, 3
1
Graduate Program in Infectious and Zoonotic Diseases, National Autonomous University of Honduras
2
3
(UNAH). Department of Community Health Sciences, Brock University Canada. Clinical Epidemiology and
Statistics, McMaster University Canada
80
Background: The development of resistance to larvicide temephos in Aedes aegypti has been
reported in some countries from Central America like Nicaragua, El Salvador and Costa Rica. There´s
no data published about the status of the resistance to this larvicide in Honduras. Objectives: The
objectives of this study were determining the susceptibility level to temephos in Ae. aegypti larvae
collected in three localities (Manchén, Villanueva and Kennedy) from the Central District, Honduras
and determining the lethal dosis of temephos, LD50, LD90 values by using the Rockefeller strain as
control; and estimating the resistance ratio (RR). Methodology: Quantal response bioassays
methodology was performed to evaluate the response of a field population. Samples of Ae. aegypti
eggs were obtained using ovitraps. The eggs were reared to adults and used to generate F1 colonies
for the bioassays. Colonies were established at the laboratory and thereafter maintained at 28 ± 3°C
with a photoperiod of 12:12 hours. Five concentrations of temephos were tested including the
diagnostic concentration recommended by World Health Organization (0.012 mg/liter). Results: The
LD50 and LD90 values were 0.011 mg/liter, 0.012 mg/liter to Rockefeller strain respectively and
0.073 mg/liter and 0.203 mg/liter to field population with a value of RR = 6.63. The results showed
that the field population has already developed a moderate degree of resistance to temephos.
Conclusions: It is essential to continue monitoring the resistance of this vector to temephos in order
to ensure its efficacy. Before this study, no information was available about the existence of
resistance in natural populations of Ae. aegypti in the studied area.
VIBM_C785_0061
The two-component system BvrR/BvrS regulates the expression of
the type IV secretion system VirB in Brucella abortus
1
3
1
1
4
Martínez-Núñez C , Lelany Pineda , Moreno E , Guzmán-Verri C , Chaves-Olarte E .
Programa de Investigación en Enfermedades Tropicales (PIET), Escuela de Medicina Veterinaria, Universidad
1
Nacional, 304-3000 Heredia, Costa Rica . Facultad de Farmacia, Universidad de Costa Rica 1000 San José,
2
3
Costa Rica . Escuela de Microbiología, Universidad Nacional Autónoma de Honduras . Facultad de
4
Microbiología, Universidad de Costa Rica 1000 San José, Costa Rica .
Introduction: Bacteria of the genus Brucella are important zoonotic pathogens that affect several
domestic mammalian species. The pathogenesis of Brucella is strongly related to the ability of these
bacteria to multiply intracellularly in the endoplasmic reticulum, an event controlled by the two
component system (TCS) BvrR/BvrS and the type IV secretion machinery (T4SS) VirB. Hypothesis: In
this project we worked with the hypothesis that TCS BvrR/BvrS determinates the expression of VirB
T4SS and VjbR transcriptional regulator. Methodology: In order to determine the transcriptional
regulation of the VirB T4SS by the TCS BvrR/BvrS in B. abortus, were compared by Western blot the
81
levels of different VirB proteins in wild type strain 2308 and strains with mutations in genes bvrS
and bvrR. Results: The results show that levels of VirB1 and VirB5 significantly decreased in the
mutants bvrR- and bvrS- while VirB8 and VirB9 proteins showed a slight decrease. Interestingly, all
proteins are over expressed in the strain complemented with a plamid encoding the functional gene
bvrR (strain bvrR-pbvrR+). In order to evaluate the expression of VjbR was developed and validated
a rabbit polyclonal antibody against this protein. With this antibody it was determined that VjbR is
undetectable in the mutants bvrS- and bvrR- while it is reconstituted to normal levels in the strain
bvrR-pbvR+. Conclusions: These results together indicate that TCS BvrR/BvrS is involved, directly or
indirectly, in the transcriptional regulation of two key elements for the virulence of B. abortus, the
VirB T4SS and the transcriptional regulator VjbR.
VIBM_0801_0082
Efficacy of chloroquine for the treatment of uncomplicated
Plasmodium falciparum malaria in Honduras
1
1
1
2
Rosa Elena Mejia Torres , Engels Ilich Banegas , Meisy Mendoza , Cesar Diaz , Tamara Sandra Mancero
3
4
5, 6
6
6
Buchelli , Gustavo A Fontecha , Md Tauqeer Alam , Ira Goldman , Venkatachalam Udhayakumar , José
7
Orlinder Nicolas Zambrano
1
2
National Malaria Laboratory, Ministry of Health, Honduras. National Malaria Eradication Service (SNEM),
3
4
Ministry of Public Health, Ecuador. Pan American Health Organization, Honduras. MEIZ-Microbiology
5
School, National Autonomous University of Honduras (UNAH), Tegucigalpa, Honduras. Atlanta Research and
6
Education Foundation, Decatur, GA. Malaria Branch, Division of Parasitic Diseases and Malaria, Center for
7
Global Health, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA 30333.
Department of Health Promotion, Ministry of Health, Honduras.
Background: Central American countries, including Honduras, use chloroquine (CQ) as the first line
of treatment for Plasmodium falciparum although resistance to the antimalarial is prevalent
worldwide. However, no clinical evaluation of the anti-malarial had been done in the country.
Objective: Evaluate the therapeutic efficacy of chloroquine (CQ) for the treatment of
uncomplicated P. falciparum malaria in the municipality of Puerto Lempira, Gracias a Dios,
Honduras. Materials and methods: An in vivo study to evaluate the efficacy of CQ was conducted in
the municipality of Puerto Lempira, Gracias a Dios in febrile patients at the Regional Health
Department and the Regional Hospital using the Pan American Health Organization (PAHO)-World
Health Organization (WHO) protocol with a follow up of 28 days. Patient history, clinical
examination, body temperature, body weight, parasitemia at day 0 (before treatment) were
recorded for each patient and supervised treatment was administered. Clinical examinations and
parasitological tests were repeated at days 1, 2, 7, 14, 21 and 28. The PAHO-WHO criteria were used
82
to classify the clinical and parasitological responses to treatment. Sixty nine patients from 6 months
to 60 years of age diagnosed with uncomplicated P. falciparum malaria were included in the
study. Blood samples from each patient were tested using microscopy at enrolment. Subsequently,
PCR was used to confirm the diagnosis and the presence of genetic mutations associated with CQ
resistance in the P .falciparum CQ resistance transporter (pfcrt) gene was analyzed. Results: Results:
Among 69 patients enrolled, one patient was found to have only P. vivax infection on PCR, although
microscopically positive for uncomplicated P. falciparum malaria. All patients who were treated with
CQ (25 mg/kg over three days) cleared parasitemia by day 3 and acquired no new P. falciparum
infection within 28 days of follow-up. There were no early or late treatment failures. Consistent with
the in vivo results, all the sixty eight samples sequenced for pfcrt gene had only CQ sensitive
ancestral genotype. Conclusions: In the municipality of Puerto Lempira, Gracias a Dios, Honduras,
CQ remains highly efficacious for the treatment of uncomplicated P. falciparum malaria. Molecular
data confirms the absence of CQ resistance associated pfcrt mutations in the parasites tested.
VIBM_1801_0027
Brucella canis is a Naturally Rough Bacteria Capable of Reach
Intracellular Niches
1
María C. Medina , Esteban Lizano
2, 4
2, 3
Moreno and Esteban Chaves
2, 3
3
2
2
, Carlos Chacón , Elías Barquero , Caterina Guzmán-Verri , Edgardo
1. Programa de Maestría en Enfermedades Infecciosas y Zoonóticas, Universidad Nacional Autónoma de
Honduras, 2. Programa de Investigación de Enfermedades Tropicales (PIET), Universidad Nacional, Heredia,
Costa Rica, 3. Centro de Investigación de Enfermedades Tropicales (CIET), Universidad de Costa Rica, 4.
Instituto “Clodomiro Picado”, San José, Costa Rica.
Background: Bacteria of the genus Brucella are classified into two major groups, called smooth and
rough bacteria, characterized by the composition based on the presence or absence of the O
antigen in its lipopolysaccharide. B. abortus, B. melitensis and B. suis belong to the first group,
whereas B canis and B. ovis are naturally rough bacteria. Studies on the pathogenicity of brucellosis
have focused primarily in natural smooth bacteria and it has been assumed that rough bacteria are
less virulent. However, both B. canis and B. ovis are able to induce a clinical outcome typical of
brucellosis in their primary hosts. Research Objectives: This study aimed to analyze the interaction
mechanisms used by Brucella canis in eukaryotic cells, using ex vivo studies of infection of Hela cell
lines and monocytes - macrophages RAW 264.7 and J774A1; and in vivo murine models. Molecular
techniques were also used to clarify their behavior including quantification of proinflammatory
cytokines and the expression of the VirB8 protein. Results: B. canis adhered massively to the cell
83
surface of infected cells at early times and was detected by immunofluorescence up to 72 hours of
infection in approximately 60 % of the cells. The recuperation of CFU was, however, very low at late
stages of the infection prompting us to hypothesize that this bacterium reaches a viable-noncultivable intracellular state. In vivo experiments indicated that B. canis is able to reach an active
replication stage in the mouse model following a replication kinetic similar to smooth strains.
Analysis of the molecular virulence mechanisms indicated clear differences in their pattern of
expression of the type IV secretion system virB (T4SS) that appears to be active throughout the
extracellular growth curve B. canis, in contrast to their smooth counterparts B. abortus.
Conclusions: Brucella canis is a naturally rough strain capable of intracellular multiplication and
persistence in the mouse model but presents biological differences when compared to smooth
strains of the genus in terms of the expression of virulence factors, and intracellular behavior.
VIBM_0411_0074
External Evaluation of the Performance of Microscopic Diagnosis for
Malaria in the Countries of the Americas
1
2
3
4
Rosa Elena Mejia , María Paz Ade , Prabhjot Singh , Sonia Gutiérrez y Nancy Arrospide , Engels Banegas
1
5
2
National Parasitological Laboratory, Ministry of Health, Honduras. Regional Malaria Program, Pan American
3
4
Health Organization, Washington DC. Pan American Health Organization, Honduras. National Institute the
5
Health Peru. National Malaria Program, Ministry of Health, Honduras
Background. Access to diagnosis and appropriate and timely treatment is one of the components of
the global strategy for malaria control. Access to adequate treatment can be guaranteed based on
the existence of a healthcare system that offers timely access to quality diagnosis. The quality of
preparation and examination of the blood slide for malaria requires following of standard
procedures. External evaluation is necessary for standardizing these processes across countries and
improving the quality of diagnosis. Research Objective: Establish a process for external evaluation
of quality of microscopic diagnosis of national reference laboratories in the countries of the
Americas. Methodology: National Reference Laboratories of Honduras and Peru were selected as a
Supranational Reference Laboratories, and prepared panels to be sent to participating countries.
Twenty three countries were invited to participate in the evaluation. Panels were prepared using
wild parasites (Plasmodium sp) from blood samples collected from patients. Each panel consisted of
20 slides, a mix of negative slides and slides positive for P. falciparum, P. vivax and mixed infections,
with parasitemia varying in positive slides from low to high. Panels were sent to countries that
entered the results in a virtual reporting system –NETLAB, within 10 working days of receipt of the
panel. Slides were evaluated for diagnosis, identification of parasite species, sexual stage and
84
measuring parasitemia using standard criteria. Results: Of the 23 countries invited, 10 participated
in the evaluation process in 2011. Of these, 70% of the laboratories had an acceptable degree of
diagnostic agreement, 10% (1/10) were consistent in identifying parasite species and 20% (2/10)
identified the sexual stages correctly. None of the laboratories evaluated were consistent in
measuring parasitemia. Conclusions: These results have shown that the microscopic diagnosis of
malaria in the countries of the Americas is deficient and needs improvement. Especially, focus is
required in identification of species type, sexual stage and measuring parasitemia. Strengthening of
national networks of public health laboratories in the countries is of utmost importance.
Parasitemia is measured differently across countries which is the reason for low concordance rates.
The evaluation will serve in standardization of processes and protocols across countries in
microscopic diagnosis of malaria.
VIBM_0078_0112
Development of neutral SNP markers for low-cost, low-technology
follow-up of within host dynamic of P. vivax infections.
Mendizábal-Cabrera, R., P. Pennington, L. Ortiz, M.E. Castellanos and N. Padilla
Center for Health Studies, Universidad del Valle de Guatemala.
Monitoring the effectiveness of antimalarial drugs is a requirement for the development of
treatment policies. Monitoring is commonly performed by in vivo assays that may be validated by
parasite molecular typing techniques to conclude if treatment failure is a true recrudescence, a
relapse or a new infection with a different parasite genotype. Several molecular marker panels have
been proposed to validate these results, but generally apply expensive instrumentation and
demand qualified technical expertise, hampering its application in endemic countries. Using a
strategy of in silico identification of neutral single nucleotide polymorphism (SNP) markers in
sequences of P. vivax from strains reported across endemic areas, six SNP markers were identified
in genes coding for β-tubulin, MSP1, MSP3α, MSP7, MSP9 and MSP10. Hemi-nested primers with
allele-specific 3´ ends were designed and PCR conditions were standardized with control samples.
Twenty seven samples from two endemic areas of Guatemala were analyzed. SNP markers
identified in β-tubulin and MSP10 were not polymorphic (allele frequency > 0.95), but have
potential to be useful in other regions with P. vivax malaria. Primers for MSP3α showed to be nonspecific and this marker was not included in the analysis. However, SNP markers designed on MSP1,
MSP7 and MSP9 genes showed to be polymorphic and specific to P. vivax. In addition to haplotype,
a size polymorphism was also identified in the MSP1 region, which in combination with haplotypes
of polymorphic SNPs produced different genotype profiles for 70% of the analyzed samples. Data
regarding neutrality and allele association of SNPs and MSP1 size polymorphism will be also
discussed. This PCR-based genotyping technology, being low cost and simple, would be an
85
affordable alternative to conventional automatic sequencing, making easier its adoption in
laboratories of developing countries.
VIBM_0801_0067
Implementation of Routine Monitoring using Genetic Markers for
Detection of Anti-malarial Drug Resistance in Plasmodium
falciparum in Honduras
1
2
3
1
Meisy Mendoza , Gustavo Fontecha , Engels Ilich Banegas , Rosa Elena Mejía
1
2
National Malaria Laboratory, Ministry of Health, Honduras. MEIZ-Microbiology School, National
3
Autonomous University of Honduras (UNAH), Honduras. National Malaria Program, Ministry of Health,
Honduras
Background: Resistance to chloroquine in Plasmodium falciparum is widespread in the world yet
Honduras uses the anti-malarial drug as the first line of treatment as clinical studies have shown no
resistance exists in the country. Pan American Health Organization -World Health Organization
(PAHO-WHO) recommends a clinical study every five years as a standard for identification of
resistance. However, given the risk of importation of resistant parasites from other countries or
development of resistance due to mutation, a continuous monitoring system is required in the time
period between two clinical studies. Research Objective: Early detection of resistance to
antimalarial drugs used in Honduras as first-line therapy for P. falciparum malaria. Methodology: In
all health facilities with a functional laboratory, microscopists obtained a blood sample on filter
paper for all patients positive upon microscopic diagnosis for P. falciparum or mixed infections all
year round in 2011, following the national malaria guidelines. These filter papers were then sent to
the national reference laboratory where they were collected and analyzed using nested polymerase
chain reaction (PCR) technique. The pfcrt gene was sequenced as the locus is characteristic for
chloroquine resistance in P. falciparum genome. All patients received treatment according to
national malaria guidelines. Results: Total of 55 samples were collected in 2011 and processed.
These samples were from 6 different districts of 3 states with the highest transmission of malaria in
the country: Olancho, Colon and Gracias a Dios. All samples were positive on PCR for P. falciparum
and all 55 samples sequenced for the pfcrt gene had the CVMNK amino acid sequence. Conclusion:
Resistant genes were not detected in any of the samples studied. An earlier study done in 2009 in
Puerto Lempira, Gracias a Dios following PAHO-WHO protocol had shown that no clinical resistance
to chloroquine is present in Honduras. This routine evaluation further lends credence to this fact
also amplifying the area from which no resistance has been detected till date. The country can
maintain chloroquine as its first line of treatment for P. falciparum malaria and use routine
monitoring using genetic markers to complement clinical studies for anti-malarial resistance.
86
VIBM_0801_0042
Genotyping Chlamydia trachomatis in Sex Workers Attending Two
STI Clinics in Tegucigalpa and San Pedro Sula, Honduras between
March - June 2011
1
3
2
Suyapa Mendoza , Nelba Tábora PhD , María Elena Bottazzi PhD, Nicholas Thomson
1
4
2
STI National Laboratory, Honduras, National School of Tropical Medicine, Baylor College Of Medicine,
3
4
Houston University, National University of Honduras, Sanger Institute
Background: Infection due to Chlamydia trachomatis is the most common sexually transmitted
bacterial disease of global health significance. In Honduras, the prevalence of this infectious agent in
sex workers is approximately 11.6 % in San Pedro Sula and 6% in Tegucigalpa. There is no data
concerning genotyping of Chlamydia trachomatis from Honduran samples. Objectives: The
objectives of this study were to determine Chlamydia trachomatis genovars from sex workers
attending two STI clinics from Health Centers of Honduras and compare Chlamydia trachomatis
genovars between the clinics of San Pedro Sula and Tegucigalpa. Methodology: A total of 786
endocervical samples were collected from sex workers from two different cities in Honduras, the
city of San Pedro Sula (366) and Tegucigalpa (420). All samples were screened for Chlamydia
trachomatis using a molecular test, PACE 2, developed by Genprobe. A total of 61 endocervical
positive specimens were genotyped by amplifying the ompA gene encoding the chlamydial mayor
outer–membrane protein, which is genovar specific. Real Time PCR was performed using a
Rotorgene Q6000 apparatus (QIAGEN). Results: Seven different Chlamydia trachomatis genovars
were found amongst the Honduran samples in the following proportions: E (32.8 %), F (32.8 %), D
(13.1%), J (8.2 %), I (8.2%), G (3.3 %), H (1.6 %) y K (0). In San Pedro Sula, the genovar E (37.1 %) was
the most prevalent followed by genovars F (28.6%), D (14.3 %), J (11.4%), I (5.7%), G and H (2.9%). In
Tegucigalpa, genovar F (38.5%) was the most prevalent, followed by the genovars E (27 %), D and I
(11.5%), G and J (3.8%). Conclusion: We have completed the first survey of Chlamydia trachomatis
genovars in Honduras. Our data shows that the distribution of Chlamydia trachomatis genovars in
the two cities of Honduras was similar, and is in concordance with genovars found in some of the
American Continent regions in which genovars E, F, and D are the most prevalent.
87
VIBM_C710_0016
Molecular Epidemiology of HIV-1 in Honduras
1, 2
2, 3
1
4
4
Wendy Murillo *, Helena Skar , Ivette Lorenzana de Rivera , Gabriela Paz-Bailey , Sonia Morales-Miranda ,
2, 5
2, 6
Jan Albert , Mattias Mild
1
Microbiology Department, National Autonomous University of Honduras, Tegucigalpa, Honduras.
3
Department of Microbiology, Cell and Tumor Biology, Karolinska Institute, Stockholm, Sweden. Theoretical
4
Biology and Biophysics, MS K710, Los Alamos National Laboratory, Los Alamos, New Mexico, USA. Del Valle
5
University of Guatemala, Guatemala City, Guatemala. Department of Clinical Microbiology, Karolinska
6
University Hospital, Stockholm, Sweden. Swedish Institute for Communicable Diseases Control, Stockholm,
Sweden
2
Background: Little is known about the Honduran HIV-1 epidemic, even though more than 25 years
have passed since the first case of HIV was reported in the country. The aim of this study was to
survey in detail the molecular epidemiology of HIV-1 in Honduras using a published sampling
designed to obtain a representative sample of HIV-1 variants that circulate in different population
groups and regions of Honduras. Methods: A total of 257 Honduran HIV-1 pol sequences from the
general populations, female sex workers, men who have sex with men and the Garifunas ethnic
group were analyzed together with relevant control sequences. Maximum-likelihood (ML) and
Bayesian-phylogenetic analyses were used to explore HIV-1 subtype distribution, demographic
history and migration patterns of HIV-1 into and within Honduras. Results: All Honduran sequences,
except two, were HIV-1 subtype B. The phylogenetic analyses showed that there had been at least
six successful introductions of subtype B into Honduras. One cluster, which was introduced around
1967, contained 164 (64%) of the Honduran sequences. The ML tree showed that sequences from
Haiti were found close to the root, whereas US/pandemic sequences had an intermediate position
and a majority of the Honduras sequences were distal from the root and closely related with the
US/pandemic sequences, suggesting that HIV-1 subtype B reached Honduras soon after the US
epidemic. Migration analyses showed that HIV-1 subtype B in general was intermixed both
geographically and between transmission groups and we only found limited genetic flow between
specific geographical regions. Conclusions: The Honduran HIV-1 epidemic is dominated by subtype
B that was multiply introduced starting from around 1966, soon after the start of the US epidemic.
One of the early introduction accounts for a majority of the current cases. Migration analyses
showed that viral lineages were intermixed geographically and between transmission groups.
88
VIBM_0401_0103
Ecosystem and seroprevalence of the Leptospirosis in domestic
animal in Santa Rosa de Aguan-Colon, Honduras
1
2
1
Orellana Brenda, Valladares Manuel. Seccion de Leptospirosis, IHIMV-SENASA /SAG, Honduras,
Departamento of Zoonosis, SENASA-SAG, Honduras.
2
Introduction: Leptospirosis in Honduras is endemic and the first registry was in bovines in 1975
Comayagua-Honduras (Espinoza, 1975) and the first human cases of leptospirosis were registered in
1998 in Marcovia-Choluteca. In 1985-until now there was a program of monitoring serology in
domestic animal (bovine and pig) in all the country stays, and it has been that they circulate around
16 serovars. In the Department of Colon the serosurvey has detected six serovars in bovines, and
the seroprevalence has been increasing of the year in a year. Due to this abnormal behavior
epidemiologist of the Leptospirosis in this zone set out a study of “Ecosystem and the
seroprevalence of the Leptospirosis in domestic animal in Santa Rosa de Aguan, Colon”. Objectives:
1) to determine the seroprevalence of the Leptospirosis in domestic animal, 2) to relate the
ecosystem and the epidemiology of the Leptospirosis in Santa Rosa de Aguan-Colon. Methodology:
Place of study: A) They were sampled in Santa Rosa de Aguan-Colon, 10% of the population 6
different domestic animals in 14 communities to determine the seroprevalence of the leptospirosis
in the zone and B) water samples of the 14 communities coming from 14 villages and small villages
were collected. Processing of the samples: A) The collected serums were processed by the
technique of conventional MAT using 14 serovars of L.interrogans and B) the collected water
samples were processed for the isolation of Leptospira spp and later identification. Result and
Discussion: Of the 430 serum samples of the domestic animal process using conventional MAT the
74, 7% were reactive and the seroreactivity in the different animal domestic were: A) bovine 83, 3%
(233 animals), goat 25% (12 animals), canine 65, 4% (26 animals), horse 73, 7% (76 animals), bovines
58, 2% (67 animals) and pigs 75% (16 animals). Of the eight water samples processed for the
isolation of Leptospira spp 100% were positive, but they even are in the study for its identification.
These results demonstrate the high prevalence to us of the Leptospirosis in the animal in Santa Rosa
de Aguan-Colon. This can be due to the high risk to which these animals in contaminated waters
with are exposed Leptospira spp. Although we have not even identified these leptospiras as
pathogenic he suggests them to us waters are the transmitters of this disease.
VIBM_0706_0009
Genetic Diversity of Plasmodium vivax in Honduras
1
1
1
1
2
2
1,3
1*
Andrés Ortiz , López AC. , Coello J. , Sosa W. , Mejía R.E. , Banegas E. , Jovel I. , Fontecha G .
89
1
2
MEIZ-Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras. Laboratorio
3
Nacional de Malaria, Secretaría de Salud de Honduras. Malaria Research Laboratory, Infectious Diseases
Unit, Department of Medicine, Karolinska University Hospital/Karolinska Institutet.
*Corresponding Author: MEIZ-Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de
Honduras, Ciudad Universitaria, Boulevard Suyapa, Tegucigalpa, Honduras. Telephone: +504 33935443; email:
[email protected]
BACKGROUND AND OBJECTIVES. Studies on genetic diversity in Plasmodium spp. have focused
mostly in P. falciparum due to its epidemiological significance. There are less studies on genetic
diversity of P. vivax even though it causes nearly 80% of the malaria cases in the Americas. Most
common genes studied are those expressed on the surface of the parasite given their interest as
vaccine candidates. The present study analyzed the genetic diversity of three genes for Plasmodium
vivax in Honduras. MATERIALS AND METHODS. Patients from five Provinces of Honduras and
diagnosed with malaria vivax were included. A blood drop sample was collected on paper filter. DNA
was extracted using Chelex-100 (Bio-Rad). All samples were amplified for the Domain I of AMA-1;
the repetitive region of CSP, and the central region (F2) of MSP-1 according to methods described
elsewhere. Amplification products were observed on 1% et-br agarose gels, purified and sequenced.
Sequence analysis and phylogenetic inferences were performed using Chromas Pro and Mega5.
RESULTS. Te results showed that P. vivax populations in Honduras are highly diverse. A large
number of distinguishable genotypes were found. PvAMA-1 revealed 48 (62.3%) different
sequences for 77 strains. A phylogenetic tree showed 3 separated clusters for PvMSP-1 from 51
nucleotide sequences. For PvCSP 53 (63.1%) different types of sequences were obtained in 84
strains including from 17 to 20 motif repetitions. Individuals with 20 repetitive segments showed
the higher frequency (60.7%). Phylogenetic trees and estimates of evolutionary divergence revealed
that the F2 segment of PvMSP-1 is more polymorphic than the other two genes. PvAMA-1 is the
most conserved gene included in this study. Allelic types were randomly distributed among the five
Honduran Provinces. Therefore, no correlation was found between geographical location and
genotypes. CONCLUSIONS. These results indicate that the P. vivax parasite population is highly
diverse in Honduras, as expected for the moderate level of transmission. PvAMA-1 is the most
conserved gene analyzed followed by PvCSP and PvMSP-1.
VIBM_C369_0041
Evidence of the etiological role of noroviruses and human
astroviruses in childhood diarrhea in Tegucigalpa, Honduras.
1
2
Jafet Ortiz , Annabelle Ferrera
1
Master´s Program in Infectious and Zoonotic Diseases (MEIZ), School of Microbiology, National Autonomous
2
University of Honduras, Tegucigalpa, Honduras; Faculty of Science, School of Microbiology, National
Autonomous University of Honduras, Tegucigalpa, Honduras.
90
Background: Diarrheal diseases are a common disorder in young children, and represent an
unsolved health problem in developing countries. In children, group A rotavirus is the major
etiologic agent of viral gastroenteritis, but viruses such as norovirus (NoV) and human astrovirus
(HAstV) among others, have also been recognized as etiological agents of this disease. However, the
importance of these agents in childhood diarrhea is poorly understood in Honduras as well as in
other low-income countries. Objective: To investigate the prevalence, clinical role and genetic
diversity of NoV and HAstV in pre-school children with acute gastroenteritis. Methodology: Fecal
samples and their corresponding epidemiological data were collected between October 2010 and
July 2011 from children under five years of age with acute diarrhea in three health care centers in
Tegucigalpa, Honduras. An immunoenzymatic assay was applied to all samples to screen for HAstV.
NoV detection and HAstV positive sample confirmation was performed applying reverse
transcriptase polymerase chain reaction (RT-PCR) followed by sequencing in order to assign their
genotypes. Results: NoV and HAstV were detected in 13% and 5% of studied children, respectively.
Among NoV positive samples, 92% belonged to genogroup II, being GII.4 the most common
genotype detected. Interestingly, only variants 2006b and 2010 of NoV GII.4 were identified by
phylogenetic analysis in the present survey. Human astroviruses were clustered into three different
genotypes: HAstV-1, HAstV-2 and HAstV-8. In general, these viruses were mainly encountered in
children less than two years of age. Diarrhea, vomits and fever were observed in more than 80% of
the cases and 33% of them required hospitalization. Conclusions: This is the first report that
describes the molecular epidemiology of NoV and HAstV in Honduras. Our results demonstrate that
these viruses have an important role as etiological agents in childhood diarrhea in Tegucigalpa, and
suggest that more extensive studies are needed in infantile population of Honduras in order to
establish effective infection control and prevention measures.
VIBM_0801_0051
HIV-1 Drug Resistance among Pregnant Women in Honduras
1,2,3
Leda Parham
2,4
5,
3
, Annika C. Karlsson , Odalys García , Jan Albert , Ivette Lorenzana de Rivera
1
1
Department of Microbiology, National Autonomous University of Honduras, Tegucigalpa, Honduras,
3
Department of Virology, Swedish Institute for Infectious Disease Control, Solna, Sweden, Department of
4
Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden, Department of Laboratory
5
Medicine Division of Clinical Microbiology, Karolinska University Hospital, Huddinge, Sweden, STD/HIV/AIDS
Department, Ministry of Health Honduras.
2
Introduction. Huge efforts have been undertaken to provide access to antiretroviral therapy (ART)
for prevention of mother-to-child transmission (PMTCT) of HIV-1 in resource-limited settings. In
91
Honduras the current treatment strategy for PMTCT has change from monotherapy to combination
therapy (cART). Even with the prevention role of cART, there is a possibility of the antiretroviral
drugs (ARV) to induce HIV resistance and therefore its possible transmission to the child. Research
Objective. To evaluate the frequency of HIV-1 drug resistance in Honduran pregnant women, under
cART between 2007-2010. Study population and Methods. Fifty HIV-1 pregnant women were
recruited under the MTCT program; 29 ART-naïve and 21 already under ART. To determine the
presence of HIV-1 resistance mutations, plasma samples collected at pregnancy and after delivery
were analyzed by sequencing of pol gene. Sequences were edited by Sequencher software (Gene
Codes Corporation-USA), and then inspected with Stanford HIVDB-program; HIV susceptibility to
nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors
(NNRTIs) and protease inhibitors (PIs) were estimated with Agence Nationale de Reserches Sur le
SIDA (ANRS) algorithm. Results. Resistance was detected in 20% (10/50) women, 14% of them ARV
treatment-naïve and 29% in the ones already under ARV treatment. Out of ARV naïve-women, 75%
of them had resistant virus already on the first sample, before receiving ARV prophylactic
treatment, and one woman (25%) developed resistance afterwards. Mutations in all of ARVexperienced women were detected at first time point. Resistance mutations in both groups include
M184V for NRTIs; K103KN, Y181C for NNRTIs; no PIs were found. Overall, resistance to NRTI drugs
was observed in 20% of women, to NNRTIs in 40% and combined resistance to both drug classes in
40%. Out of the women with resistance 70% had high level of resistance to different drugs.
Discussion. This constitutes the first report of the frequency of drug resistance in Honduran
pregnant women. The degree of resistance is significantly high therefore this will impact on their
response to future or actual ART. It is Imperative to continue studying the role of ART in pregnant
women in Honduras.
VIBM_C092_0032
Nicaragua's Millennium Project: measures to help reduce poverty
and improve child survival
1
1,2
1
Wilton Pérez , Rodolfo Peña , Lars-Åke Persson , Carina Källestål
1
5. International and Maternal's and Children's Health, Uppsala University
6. The Centre for Research and Interventions in Health (CIS), León, Nicaragua
Background. Poverty often contributes to ill health, and social interventions are usually slow to
reach those who need it most. To break this vicious circle, the world has designed the most
visionary plan: The Millennium Development Goals (MDGs). Halving poverty (MDG1) and reducing
by two-thirds the child mortality (MDG4) are two of the eight MDGs that the world must achieve by
2015. Targeting socially disadvantaged populations with sustainable and low-cost effective
92
interventions strengthen the progress towards the MDGs. Nicaragua is the second poorest country
in the Americas. According to the United Nation's MDG Report 2011, Nicaragua is lagging behind to
attain the MDG1. However, there is higher chance to meet the MDG4. Combating poverty and
reducing any health-related inequality remain as the greatest challenges. This study has two aims:
1) analyze the poverty reduction as well as drivers for its alleviation and 2) examine the social and
regional inequality trends of child mortality using good quality data of Health and Demographic
Surveillance Sites. Methodology. We undertook this study in two provinces in Nicaragua, the first is
a remote rural area and the second a mixed urban-rural. During 2003-2009 we followed a
population cohort and collected household (water, sanitation, floor and wall), individual (deaths,
births and migration) and geographical data. Poverty and child mortality were the main outcomes.
Micro-credit, home gardening and technical education were intervention factors for poverty
alleviation. Mother's education was used to assess mortality inequality. Results. In the remote rural
province, poverty and extreme poverty fell 13% and 5% between 2004 and 2009 respectively.
Microcredit, home gardening and technical training were significantly associated with poverty
reduction. The annual rate reduction for child mortality varied between 4.3% - 6.8%. Faster
reductions of mortality were observed when social inequalities declined. Conclusions. In this
unequal society it is unlikely that the MDGs will be achieved by 2015. However the necessary
measures to make it possible do exist. Our findings identify some measures to help overcome
challenges to poverty reductions and improvements of child survival in low- income settings.
VIBM_4019_0087
Characterization of Genetic Diversity of Brucella ceti Isolated on
the Pacific Coast of Costa Rica, Through Sequencing of Nine
Selected Genes.
Piche Ovares Martha1, Murillo Carla 1,2 Slon Jose Luis1,3, Barquero-Calvo Elías1, Lizano Esteban1,3,
Hernández-Mora Gabriela1,4, González-Barrientos Rocío4,5, Alfaro Alejandro5, Rivas Olga2, Zúñiga Claudia2,
Moreno E1 and Guzmán- Verri Caterina1
1PIET, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica. 2Centro de Investigación
en Biotecnología. Escuela de Biología, Instituto Tecnológico, Cartago, Costa Rica. 3 Facultad de Microbiología,
Universidad de Costa Rica, San José, Costa Rica. 4 SENASA, Ministerio de Agricultura y Ganadería, Heredia,
Costa Rica. 5 Laboratorio de Patología, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa
Rica.
Recently a new species of Brucella isolated from cetaceans has been described and named Brucella
ceti (Foster et al, 2007). Sonh et al, 2003 reported neurobrucellosis caused by B. ceti in humans.
Therefore the potential risk of zoonotic infection is under investigation. Since May 2006 until June
93
2011, thirteen Stenella coeruleoalba (striped) dolphins stranded on the Pacific coast of Costa Rica.
The cause of death was related to neurological signs and accordingly, bacteria isolated from
cerebrospinal fluid were identified as B. ceti using biochemical typing techniques. Genotyping
techniques such as multilocus sequencing typing (MLST) are becoming resourceful tools for bacterial
evolution and identification studies. By these means, Whatmore et al, 2007 classified 160 Brucella isolates in
27 sequence types (ST). Most marine isolates were classified as ST 26. The marine isolates obtained from
humans were classified as ST 27. The aim of this project was to analyze by MLST fourteen B. ceti isolates
obtained from thirteen stranded dolphins on the Pacific coast of Costa Rica in order to establish their
evolution links with the rest of marine isolates around the world. Genomic DNA was extracted from the
isolates by conventional techniques and analyzed by MLST. Nine genomic fragments from each isolate
were sequenced. The obtained sequences were compared to previously published genome data
from marine Brucella. Concatenated sequence data were used to construct an unrooted neighbourjoining tree representing the relationships between STs.
MLST results show that the isolates belong to ST 26. The more variable sequenced genes show some
single nucleotide polymorphisms (SNPs) not described before, whereas housekeeping genes are
conserved. The results show the ability of genotyping techniques such as MLST to classify and
compare Brucella species worldwide. These tools will improve diagnosis and epidemiological
controls due to a better understanding of the bacterial species circulating in the country and their
zoonotic potential.
VIBM_0801_0040
Molecular Characterization of Mycobacterium tuberculosis isolates
from patients with tuberculosis in Honduras
1,2,3
1
3,4
5
3,4
Senia Rosales , Lelany Pineda-Garcia , Solomon Ghebremichael , Nalin Rastogi , Sven Hoffner .
1 Departamento de Microbiología, Universidad Nacional Autonoma de Honduras. 2 Division of Global Health
(IHCAR), Department of Public Health Sciences, Karolinska Institute, Stockholm, Sweden. 3 Department of
Bacteriology, Swedish Institute for Infectious Disease Control,
Solna, Sweden. 4 Department of Microbiology and Tumor Cell Biology (MTC), Karolinska Institute,
Stockholm, Sweden. 5 Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de la Guadeloupe.
Background: Tuberculosis persists as a public health in Honduras. Molecular typing methods have
been used in epidemiology studies of Mycobacterium tuberculosis Complex (MTC) isolates for a
better understanding the dynamic transmission of the disease in the country. The aim of this study
was to provide an insight of the genetic biodiversity of MTC clinical isolates collected in Honduras
between 1994 and 2002. Genotyping was performed using spoligotyping and RFLP. The spoligotypes
obtained were compared with the SITVIT2 proprietary database of the Pasteur Institute of
Guadeloupe. Methodology: Two hundred-six clinical Mtb isolates from Honduran TB patients were
94
collected: Group 1 with eight-seven strains and Group II with 119 strains collected in 1994 and 2002
respectively. Drug-susceptible profile using isoniazid, rifampicin, streptomycin and ethambutol was
determined using BACTEC 460 system and the proportion method. All isolates were genotyped with
a spoligotyping commercial kit. The standard RFLP protocol was used to further characterization in
those strains belongs to a single spoligotype cluster. Results: Spoligotyping grouped 84% of the
isolates into 27 clusters. Of the 44 shared international types (SITs) identified among the Honduran
stains, 8 SITs were newly identified either within the present study or after match with an orphan
type previously identified in the SITVIT2 database. In addition, 16 patterns corresponded to orphan,
previously unreported isolates. The Latin American Mediterranean (LAM) lineage was the most
common in this study; 55% of the strains belonged to this family. Other genotypes found were
Haarlem (16%), T (16%), X-clade (6%), Unknown signature (5%) and S (1%). Only one Beijing strain
was identified (0.5%). We observed a high degree of diversity after characterizing the 43 isolates
belonging to the main spoligotyping cluster (SIT 33, LAM3) with IS6110-RFLP. Conclusions: The
findings obtained in this study show that tuberculosis transmission in Honduras is due to modern M.
tuberculosis lineages with high level of biodiversity.
VIBM_0301_0118
Characterization of species of Lutzomyia and Leishmania in
endemic area of Non-Ulcerative Cutaneous Leishmaniasis,
municipality of San Francisco de Coray, Valle, Honduras
1
1
1
2
3
3
1
Quan D ., Sandoval CM , Valladares MA , Mejía RE , Mejía RR , Zuniga C . Sosa- Ochoa W *,
1
2
Escuela de Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras. Laboratorio
3
Nacional de Vigilancia, Secretaria de Salud de Honduras. Programa Nacional de Prevención y Control de la
Enfermedad de Chagas y Leishmaniasis, Secretaria de Salud de Honduras. *Corresponding Author: Escuela de
Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras, Ciudad Universitaria,
Boulevard Suyapa, Tegucigalpa, Honduras; email: [email protected]
BACKGROUND AND OBJECTIVE. Non-ulcerative cutaneous leishmaniasis is a public health problem
in Honduras. In the country, four clinical forms of the illness: Cutaneous Leishmaniasis (CL),
Mucocutaneous Leishmaniasis (MCL), Visceral Leishmaniasis (VL) and Non Ulcerated Cutaneous
Leishmaniasis (NUCL) have been identified (1). In the 2011 there were 369 cases positive for Nonulcerative cutaneous Leishmaniasis. NUCL clinical form is distributed in the south of the country.
Thus, the main objective of this study is to determine the diversity of Luztomyia and Leishmania
species circulating in the area de San Francisco de Coray. MATERIALS AND METHODS. Nine
95
hundred nineteen Lutzomyia were captured using CDC trap for their taxonomic identification and
natural infection determination (2). Thirty eigth skin scrapings were smeared onto a sterile glass
slide, stained with Giemsa, and microscopically examined for the presence of Leishmania
amastigote forms. DNA from skin scraping was extracted using the Qiagen Blood & Tissue according
to the manufacturer´s instructions. DNA from female sandflies was placed in 1.5 –mL tubes
containing 100 µl of Chelex 100 (BioRad). RESULTS. Three Lutzomyia species were identified, Lu.
longipalpis, Lu. chiapanensis and Lu. zeledoni. Lutzomyia longiplapis of major predominance.
Female sandflies were separated in 17 pools for PCR analysis. The overall natural infection rate with
Leishmania infantum in the 17 pools of sandflies was 5.88% (1/17). Preliminary result from skin
scrapings molecular analysis, we find a Leishmania infantun in 4/5 skin scraping. CONCLUSIONS.
We confirm to Lu. longipalpis as the main vector and Leishmania infantum as the etiologic agent of
Atypical Cutaneous Leishmaniasis in the area of San Francisco de Coray, Valle
VIBM_281-_0050
Assessment of pediatric Norovirus diarrhea in rotavirus vaccinated
children.
Filemón Bucardo
1,2
1
1
2
2
, Yaoska Reyes , Patricia Blandon , Johan Nordgren , and Lennart Svensson .
Department of Microbiology, Faculty of Medical Sciences, National Autonomous University of Leon Nicaragua,
1
2
(UNAN-León) .Division of Molecular Virology, Faculty of Medical Sciences, University of Linkoping, Sweden
Rotavirus (RV) and Norovirus (NoV) are the major cause of acute gastroenteritis worldwide. Studies
carried out following RV vaccine introduction in Nicaragua revealed reductions in terms of RV
environmental transmission (sewage), as well as, severe RV infections. However, these studies also
showed that the burden of diarrhea in Nicaragua remains high. In order to investigate if NoV
contributes to this burden of diarrhea, we carried out a community and hospital surveillance study
of RV and NoV infections between 2009 and 2010 in 2 different cities of Nicaragua (León and
Jinotega). A total of 366 samples of children≤ 5 years of age with acute diarrhea were analyzed by
Real-time PCR. Interestingly, while NoV was found in 22% (81/366) of the samples RV was only
found in 8% (28/366). The most common NoV Genogroup (G) was the globally dominating GII with
90% (73) followed by GI with 9% (7) and co-infection with both GI:GII in one case. Nucleotide
sequencing (n = 60) and phylogenetic analysis revealed; GII.4-2006b (57%), GII.4-2010 (17%), GII.4X
(7%), GII.6 (5%), GII.13 (5%), GII.9 (2%), GI.2 (3%), GI.3 (3%) and GI.4 (2%). Of note, the frequency of
NoV was 23% (67/289) in children vaccinated and 15% (6/38) in not vaccinated (OR=1.6, p=0.4). No
differences were observed regarding the number of vaccine doses and NoV frequency. Our findings
96
reveal that NoV is the most common viral pathogen associated with gastroenteritis in a Rotateq™
vaccinated population of Nicaragua and that NoV GII.4 remains as the most common genetic variant
circulating. This study extends previous knowledge about the influence of RV vaccine over ecology
of viruses associated with gastroenteritis.
VIBM_8410_0030
Competence of asymptomatic naturally infected dogs to transmit
Leishmania (Leishmania) infantum chagasi to the natural vector
1
1
2
3
1
1
4
4
Laurenti MD ; Matta VL ; Rossi CN ; Horna A ; Tomokane TY ; Corbett CEP ; Secundino NFC ; Pimenta PFP ;
2
Marcondes, M
1
Laboratory of Pathology of Infectious Disease, Medical School, University of São Paulo, São Paulo (SP),
Department of Clinics, Surgery and Animal Reproduction, College of Veterinary Medicine, São Paulo State
3
4
University, Araçatuba (SP), Universidad de Panamá, Panamá, Laboratory of Medical Entomology, Centro de
Pesquisas René Rachou, FIOCRUZ, Belo Horizonte (MG), Brazil.
2
The aim of the present study was to evaluate the ability of dogs naturally infected with Leishmania
(Leishmania) infantum chagasi in parasite transmissibility to the vector. Thirty eight dogs with
visceral leishmaniasis were grouped according to clinical signs into symptomatic (n=24) and
asymptomatic (n=14) animals. All dogs were submitted to xenodiagnosis with F1 laboratory-reared
Lu. longipalpis. After being sedated, the dogs were exposed to the bite of around 50 sand flies
placed in round plastic boxes for 60 minutes. On the fourth day after the blood meal sand flies
were dissected to search for promastigotes. After euthanasia, fragments of skin, lymph nodes and
spleen were collected and processed by immunohistochemistry to evaluate tissue parasitism. Skin
parasitism was observed in 58.3% of the symptomatic dogs and in 35.7% of the asymptomatic
ones, and parasites were visualized in the spleen of 66.7% and 71.4% of them, respectively. All
dogs from both groups presented amastigotes in lymph nodes. All asymptomatic dogs, except one
(93%), were competent to transmit Leishmania to the vector, including eight (61.5%) without skin
parasitism, against 67% of the symptomatic, six (37.5%) of them with no detectable amastigotes in
skin. Besides, vectors that fed on asymptomatic dogs presented a higher infection rate (p=0.0494).
There was no correlation between skin parasitism and dog infectiousness to sand flies. In fact, the
presence of Leishmania in lymph node was the most related finding to a positive xenodiagnosis.
Finally, our data emphasize the relevance of the asymptomatic dogs in the maintenance of L. (L.) i.
chagasi in endemic areas of visceral leishmaniasis, being also involved in spreading the disease in
such regions.
97
VIBM_3036_0084
Chromosomal and Extrachromosomal Location of vanA in
Vancomicyn-Resistant Enterococcus faecium Strains Isolated in
Costa Rica
1
Rivas-Solano Olga , García-Santamaría Fernando
2
1
Centro de Investigación en Biotecnología. Escuela de Biología, Instituto Tecnológico de Costa Rica, Cartago,
2
Costa Rica. Centro de Investigación en Enfemedades Tropicales. Facultad de Microbiología, Universidad de
Costa Rica, San José, Costa Rica.
Vancomycin-resistant Enterococcus (VRE) can disseminate clonally or through horizontal transfer of
resistance genes. vanA is associated with a high level of phenotypic resistance to glycopeptides. In
Europe, it is suspected that VRE originated because of the use of avoparcin as growth promoter to
feed farm animals for human consumption. This resulted in a selective increase of resistant strains,
which disseminated to humans through the food chain (Manson et al, 2003, Tomita et al, 2002;
Biavasco et al, 2007; Novais et al, 2008). Since this antibiotic was used in Costa Rica until 2000, the
aim of the project was to determine vanA location in Enterococcus faecium strains isolated from
fecal samples of human (n=7) and swine origin (n=14), taken prior to the withdrawal of avoparcin
from the market (Bustamante et al, 2003). Plasmid DNA was extracted according to Anderson &
McKay (1983), then analyzed by pulse-field gel electrophoresis (Barton et al, 1995; Turabelidze et al,
2000; van den Braak et al, 1998) and transferred to nylon membranes (Sambrook et al, 1989), which
were sequentially hybridized with digoxigenine-labeled vanA and 16S-rRNA probes. Results showed
that some of the strains of human origin had a chromosomal location of vanA, while most of the
strains of swine origin carried vanA-positive extrachromosomal elements of approximately the same
size (80 kb). Because of their high molecular weigth, those elements could be conjugative plasmids.
Interestingly, in some strains of both origins, vanA was located in both chromosomal and
extrachromosomal elements. Those results suggest that in Costa Rica, the epidemiology of VRE is
similar to what has been described in Europe, but different from the USA, where it is believed that
VRE originated at clinical environments, since avoparcin was never used as growth promoter
(Tomita et al, 2002, Stobberingh et al., 1999; Mundy et al., 2000; Wegener, 1998).
98
VIBM_0801_0031
Association of variants of receptor FcγRIIA-131 with severe dengue
in Honduras
1
1
2
Cynthia Rodríguez , Ivette Lorenzana de Rivera , Mark Loeb .
1. Universidad Nacional Autónoma de Honduras, Tegucigalpa, Honduras. 2. McMaster University at Hamilton,
Canada.
Background. Dengue Virus infection is a global emerging disease with particular importance in
tropical and subtropical areas. Antibody- dependent enhancement theory have dominated the
explanation for severe dengue pathogenesis, however not all severe cases occur in the context of
the secondary dengue infections, therefore there must be another reasons, therefore there is an
increased interest in looking to host genetic factors. Aims. In this study we have investigated if
Single Nucleotide Polymorphisms (SNP’s) in genes coding for DC-SIGN, Fc-receptor (FcγRIIA),
Vitamin D receptor (VDR), TNFα, and HPA1& HPA2 are determinants for severe dengue disease in
Honduran patients. Population and Methods. A total of 200 dengue patients were investigated
from Tegucigalpa, Honduras; 100 Dengue Fever cases (DF) confirmed by laboratory means and 100
Dengue Hemorrhagic (DH) cases classify as grade I and II (WHO classification 2005). Genotyping was
performed using !PLEX assay (SEQUENOM®Inc. USA). Results. Significant increased risk was
observed in carriers of genotypes TC of FcγRIIA, (Chi2: 5.29, P: 0.02, OR: 2.22). The analysis on
frequencies of the genotype CC of VDR, showed a tendency to increase in severe forms than in mild
ones of the disease (Chi2: 3.36, P: 0.06, OR: 0.35). The G allele of the variant DC-SING-336 was not
associated with severe dengue disease. There were no statistically significant differences for TNFα,
HPA1& HPA2 polymorphism frequencies between DF and DHF. Conclusions. FcγRIIA and VDR, both
have an immunoregulatory function; for instance FcγRIIA is widely distributed in all subclasses of
IgG, mediating antibody dependent enhancement. The polymorphism changes the IgG biding
affinity, associated with reduced opsonization of the IgG2 antibodies associated with the variant
(CC). The exact role VDR is not really known. Our preliminary data contributes with new knowledge
on the role of genetic factors in dengue disease, the increase number of define SNP will greatly
increase the potential for genetic studies in dengue and should provide insights into possible
mechanisms of pathogenesis.
99
VIBM_4016_0115
Is outer membrane lipoprotein NlpI connecting bacterial RNA
degradation with cell wall synthesis in Salmonella enterica serovar
Typhimurium?
1,2
2,3
2
César Rodríguez , Syed Fazle Rouf , Mikael Rhen
1
Faculty of Microbiology and Research Center for Tropical Diseases, Universidad de Costa Rica. Costa Rica.
2
3
Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet. Sweden. Department of Medical
Microbiology and Hospital Epidemiology, Medical School Hannover. Germany
Background and rationale. The RNA degradosome complex steers the turnover of various RNA
transcripts. In Enterobacteriaceae, Pasteurellae, and Vibrionaceae the genes of the RNA
degradosome components PNPase (pnp) and DeaD are separated by nlpI; a gene coding for an
outer membrane lipoprotein. Data at hand from Salmonella enterica serovar Typhimurium (ST)
reveal several overlapping phenotypes for nlpI and pnp mutants. On the other hand, since NlpIdeficiency in Escherichia coli leads to osmotic sensitivity and filament formation, this lipoprotein has
also been implied in cell division. We hypothesize that NlpI connects the outer membrane with the
peptidoglycan synthesis machinery and with cytoplasmic biochemical activities. Objectives: To gain
experimental evidence confirming a connection between NlpI, the cell wall synthesis machinery,
and the cytoplasmic RNA degradosome in ST. Methodology: Phenotypic tests included cold
acclimatization, motility on soft agar plates, induction of bacterial autolysis upon exposure to
pharmaceutical inhibitors of cell wall synthesis, and classical disk diffusion tests for antibiotic and
detergent sensitivity. Gene expression analyses involved qRT-PCR and Northern blotting.
Biochemical assays list protein-protein interaction analyses, as well as determinations of
peptidoglycan composition and degradation. Results: nlpI and pnp mutants revealed defective cold
acclimatization and flagellar based motility, as well as a substantial stabilization of the motilityrelated regulatory RNAs CsrB and CsrC. While ampicillin at 5 g ml-1 blocked replication of wild type
ST, the isogenic nlpI mutant replicated until the entry of the stationary phase of growth even at 10
g of ampicillin per ml. Moreover, nlpI mutant cells appeared somewhat filamentous, displayed
osmotic sensitivity in low salt liquid media, and had an altered representation of penicillin binding
proteins (PBP) and muropeptides. The periplasmic protease Tsp, known to process the septumspecific PBP3; MurE, an enzyme specifically involved in peptidoglycan synthesis; and GlmS, needed
to synthesize UDP-N-acetyl-glucosamine, interacted with NlpI. Conclusions: Our data implicate NlpI
in a supramolecular protein complex spanning from outer membrane lipoproteins to the bacterial
cytoskeleton that orchestrates peptidoglycan synthesis. Since the bacterial cytoskeleton anchors the
RNA degradosome to the inner leaflet of the cytoplasmic membrane, NlpI seems to functionally
connect cell wall synthesis with RNA degradation.
100
VIBM_3027_0045
Phenotypic and molecular characterization of strains of drugresistant Staphylococcus aureus obtained from patients attending
the Hospital Escuela and the Instituto Hondureno de Seguridad
Social during the period from October 2010 to January 2011.
Sosa L *, Rojas N**
* Master of Infectious and Zoonotic Diseases, UNAH. ** Research Center for Tropical Diseases, CIET, School of
Microbiology, University of Costa Rica, UCR
Staphylococcus aureus is a major cause of nosocomial infections, but in view of the occurrence of
such bacteria in the community over the past decade, a main concern has emerged. There are
multiple mechanisms of drug resistance, particularly beta-lactams (oxacillin and its analogues). One
of its major mechanisms of resistance is the acquisition of a gene called mecA, responsible for the
production of a protein with low affinity to these drugs, called PBP2a or penicillin binding protein
2a. This study was aimed to determine the level of resistance to beta-lactamics / vancomycin and
detection of the PBP2a protein and mecA gene in clinical isolates obtained in this study and
determine their correlation. 96 clinical isolates were obtained from the Hospital Escuela (HE) and
the Instituto Hondureno de Seguridad Social (IHSS), identified and examined for the resistance to
oxacillin, methicillin, cefoxitin.and vancomycin, by conventional and molecular tests, such as disk
diffusion, E-test and mecA-specific PCR. An overall rate of 15% resistance to beta-lactams was
found. There was a 100% concordance for latex agglutination test and PCR. None were resistant to
vancomycin, and their minimal inhibitory concentrations were below 2 µg/ml. The resistance rate
found in this study was relatively low compared to the percentage of MRSA in other Latin American
countries and the world, but it is very similar to those reported in Nicaragua and Venezuela. Other
studies are needed, such as genotyping of MRSA isolates to identify the type of chromosomal
cassette involved and their origin (pandemic clone).
101
VIBM_0801_0021
Genetic characterization of multidrug-resistant Mycobacterium
tuberculosis isolates from Honduras
Senia Rosales-Klintz
3
Sven E Hoffner
1,2,3
1
3
3
3
, Lelany Pineda-García , Solomon Ghebremichael , Jim Werngren , Pontus Jureen ,
1
Departmento de Microbiología, Universidad Nacional Autónoma de Honduras (UNAH), Tegucigalpa,
2
Honduras. Division of Global Health (IHCAR), Department of Public Health Sciences, Karolinska Institutet,
3
Stockholm, Sweden. Department of Preparedness, Swedish Institute for Communicable Disease Control
(SMI), Solna, Sweden
Introduction: Multidrug resistant tuberculosis (MDR-TB) represents a major challenge for TB
control. This type of tuberculosis (TB) is resistant to at least isoniazid (INH) and rifampicin (RIF), the
main first-line drugs used for TB treatment. Molecular-based methods have been developed for the
timely detection of MDR-TB cases, based on the detection of genetic biomarkers related to
phenotypic drug-resistance. Research objective: To increase knowledge on the mechanisms
underlying the occurrence of drug-resistant TB in Honduras through phenotypic and genetic
characterization of clinical MDR-TB isolates. Methodology: Twenty-five MDR-TB strains were
isolated on Lowenstein-Jensen (LJ) medium at the National TB Reference Hospital (1994-1995, n= 6)
and the National TB Reference Laboratory (2004-2009, n=19). Conventional biochemical tests were
used for identification as M. tuberculosis. Proportion method on LJ and BACTEC 460 TB were used
for drug susceptibility testing (DST) of first-line drugs. Minimum inhibitory concentrations (MICs) for
INH and RIF were determined using either the resarzurin microtitre assay or BACTEC 460 TB. The
MIC range for INH and RIF was 0.25-4 mg/L and 0.5-16 mg/L, respectively. Sanger DNA sequencing
was used to identify resistance-related mutations for RIF (rpoB gene) and INH (katG gene and
mabA-inhA promoter region). Spoligotyping and IS6110-RFLP were used to genotype the strains.
Results: The majority of the MDR-TB isolates had high MICs for INH ≥( 4mg/L) and RIF (≥16 mg/L).
The most frequent rpoB mutation was Ser531Leu. The katG mutation Ser315Thr was the most
prevalent. Two isolates, both without mutations in katG, had the C-15T mutation in the mabA-inhA
promoter region. The most prevalent spoligotyping clades were the Latin American Mediterranean
(LAM) and T families. Only two small spoligotyping clusters belonging to the LAM3 (n=2) and T3
(n=3) genotypes were identified and confirmed by IS6110-RFLP. Conclusion: The heterogeneous
distribution of mutations and genotypes among these isolates indicate that MDR-TB cases in
Honduras are caused by acquired resistance rather than ongoing transmission of MDR-TB strains.
These findings are of importance to the National TB control program and for the future
development and implementation of suitable molecular-based diagnostic algorithms for MDR-TB in
Honduras.
102
VIBM_0605_0035
Kato-Katz method in the diagnosis of hookworm infections: the
importance of quality control
1
2
1
Maria Mercedes Rueda , Jose Antonio Gabrie , Maritza Canales & Ana L. Sanchez
1
2
2
Department of Microbiology, National Autonomous University of Honduras; Department of Community
Health Sciences, Brock University, Canada.
Background. Soil-transmitted helminth (STH) infections are a group of neglected tropical diseases
mainly caused by three species of intestinal parasites: Ascaris lumbricoides, Trichuris trichiura and
hookworms (Ancylostoma duodenale and Necator americanus). The diagnostic method most
commonly used for STH detection is called Kato-Katz. The World Health Organization recommends
Kato-Katz as the method of choice for qualitative and quantitative diagnosis of human STH as well
as for monitoring and evaluation of control and mass-treatment programs. Objective. To
demonstrate how the quality of detection of hookworm infection by Kato-Katz method is affected
by excessive clearance and / or alteration of diagnostic forms in the sample. Methodology. An
epidemiological study for SHT was in several rural communities located in the department of
Olancho, Honduras. An initial survey conducted in 351 children yielded a very low hookworm
prevalence of 5.6%, which prompted the investigators to review the Kato-Katz procedure performed
in the laboratory. It was found that in 228 samples the clearing time exceeded 2 hours, which could
have negatively affected hookworm eggs detection. To overcome this problem, a second sampling
was performed in the same population and care was taken to complete the egg count within 90
minutes from the sample preparation. Results. Only 195 children of the 228 requested were able to
provide a second sample. Comparison of the paired samples showed a statistically significant
difference in the percentage of positivity for hookworms: 1% in the first sample y15.9% in the
second (p <0.001). No statistical difference was found for Ascaris (35.9 % vs. 32.8 %, sample 1 and 2,
respectively) or Trichuris (69.7 % vs. 72.8 %, sample 1 and 2, respectively) (p>0.05). Conclusion.
Underestimation of hookworm infections can occur if the time between preparation and analysis of
the samples is greater than 2 hours; this is due to excessive clearing of the sample and possibly to
alteration of diagnostic stages. Standardization and quality assurance of Kato-Katz method are
paramount, particularly when results provide evidence on STH burden in endemic communities and
inform the decision-making process of STH control and prevention programs.
103
VIBM_1139_0086
Molecular characterization of Brucella isolates by MLVA-16.
1,2
1,2
1
1,3
1
1,3
Ruiz Nazareth , Murillo Carla , Jiménez César , Slon Jose Luis , Barquero-Calvo Elías , Lizano Esteban ,
1,5
4,5
4
2
2
Hernández-Mora Gabriela , González-Barrientos Rocio , Alfaro Alejandro , Rivas Olga , Zúñiga Claudia ,
1
Guzmán-Verri Caterina
1
2
PIET, Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica. Centro de Investigación en
3
Biotecnología. Escuela de Biología, Instituto Tecnológico, Cartago, Costa Rica. Facultad de Microbiología,
4
Universidad de Costa Rica, San José, Costa Rica. Laboratorio de Patología, Escuela de Medicina Veterinaria,
5
Universidad Nacional, Heredia, Costa Rica. SENASA, Ministerio de Agricultura y Ganadería, Heredia, Costa
Rica.
Brucellosis in one of most important bacterial zoonoses worldwide spread. The disease is caused by
bacteria belonging to the Brucella genus. Based mainly on host specificity and biological behavior,
the genus is divided into ten species. At the genomic level, these species show nearly 98% similarity.
Therefore, identification at species level or beyond is cumbersome if only conventional
microbiological techniques are used. The multiloci variable number tandem repeats analysis (MLVA)
is up to date one of most used molecular techniques for bacterial typing. The aim of this work was
to determine genotypic variation within Brucella isolates obtained from terrestrial and marine hosts
and correlate these data with those obtained by other genotyping and biochemical techniques.
Thirty-six Brucella costarican isolates, twenty-one terrestrial and sixteen marine isolates were
identified at species level by MLVA-16, and subsequently compared with biochemical and single
nucleotide polymorphism (SNP) typing results. All marine isolates belong to B. ceti species. On the
other hand, the biochemistry and molecular identification by MLVA-16 of the terrestrial isolates
agreed in all cases but one. Clusters among isolates with similar pathological features were
observed. The variability among marine Brucella isolates was higher than in the terrestrial isolates.
In agreement with previous reports, four VNTR´s showed high diversity in terrestrial isolates: bruce
04, bruce 07 and bruce 16. In marine isolates the same VNTR´s as well as bruce 09 where the most
diverse ones. These results prove the ability of MLVA-16 for Brucella genotyping and the usefulness
as a tool for epidemiological monitoring.
104
VIBM_1832_0044
Peridomestic Attalea butyracea palm trees as a risk factor for
dwelling triatomine infestations in rural Panama and an alternative
control strategy
1
2
1
1
1
1
1
A. Saldaña , E. Risch , Z. Cedeño , L. Hurtado , K. Gonzalez , A.M. Santamaría , C. Wald ,
1
1
L. Cáceres and J.E. Calzada .
1- Gorgas Memorial Institute for Health Studies (ICGES), Panama. 2- Department of Environmental Sciences,
University of Virginia, USA.
Introduction: Chagas disease is transmitted in Panama by Rhodnius pallescens, a wild triatomine
that lives in the crowns of “royal palms” (Attalea butyracea ) and feeds on the blood of tree-living
mammals like opposums. R. pallescens, unlike other Trypanosoma cruzi vectors, usually does not
colonize the human dwelling. This behavior prevents the traditional control for Chagas disease
based primarily on the application of residual insecticides. Objectives: 1- To determine the risk that
“royal palms” represents for dwelling triatomine infestations. 2- To evaluate the efficacy of a sheet
metal “banding” strategy to reduce the triatomine abundance in “royal palms”. Methodology:
Around 650 royal palms and 57 houses from a Chagas disease endemic area were characterized and
geo-referenced. The software ArcGIS 9.3 was used to calculate the number of palms within a 25,
50, 100, and 200 meters radius of each triatomine infested or non-infested houses. Additionally,
after partial pruning and installation of metal band of a meter wide placed at the base of 49 infested
palm tree, the presence of triatomines was evaluated by bait traps (Noireau type) during one year.
Triatomines captured were analyzed according to the quantity, species, stage, infection with T. cruzi
and / or Trypanosoma rangeli (PCR tests) and presence-type of blood meal (Dot-blot analysis).
Results: Infested houses presented in average 1.0 palms in a perimeter of 25 meters, the average of
the negative houses was of only 0.1 palms. In the 50 meters perimeter, the average was 2.1 and 0.9
palms for the positive and negative houses respectively. R. pallescens populations in the
intervened palms were significantly reduced after one year intervention with the sheet metal
“banding” instalation: The percentage of infested palms varied from 71.0% to 42.6%; the average
number of bugs per palm varied from 1.8 to 1.1 and the percentage of bugs with opossum blood
ingestion decreased from 7.5% to 0.0%. Conclusions: The sheet metal banding could be an easily
implemented alternative for a partial control of triatomine populations found in risky palm trees
located <50 mts from houses in endemic Chagas disease areas from Panama.
105
VIBM_7227_0005
Homogenates of skeletal muscle injected with snake venom inhibit
myogenic differentiation in cell culture
1
1
2
2
1
Patricia Saravia-Otten , Rosario Hernández , Teresa Escalante , Llira Bonilla , Bárbara Robledo , Alexandra
2
2
3
2
4
Rucavado , Bruno Lomonte , Jan-Ingmar Flock , José María Gutiérrez , Stefano Gastaldello .
1
2
Facultad de Ciencias Químicas y Farmacia, Universidad de San Carlos de Guatemala, Guatemala; Instituto
3
Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica; MTC,
4
Karolinska Institutet, Stockholm, Sweden, CMB, Karolinska Institutet, Stockholm, Sweden
Background: Viperid snakebite envenomings are characterized by skeletal muscle necrosis which, in
many cases, is followed by a deficient regenerative response ending in permanent tissue damage.
Research objectives: To understand some of the mechanisms involved in such poor regenerative
outcome by testing on myogenic cells in culture the effect of the muscle tissue environment
generated after injecting the gastrocnemius muscles of mice with Bothrops asper venom or with
two prototype tissue-damaging toxins from this venom. Methodology: Homogenates from
gastrocnemius muscles of mice injected with crude venom or with two tissue-damaging toxins from
this venom (Mtx, a myotoxic phospholipase A2 (PLA2), and BaP1, a hemorrhagic metalloproteinase),
were added to cultures of C2C12 myogenic cells, and myoblast proliferation and fusion were
assessed. Venom was quantified by immunoassay in muscle during the first week after injection.
Expression and activation of the triad MMP-2, MMP-9 and MT1-MMP was assessed in muscle by
gelatin zymography and Western blotting followed by densitometric analysis, during 15 days after
venom or toxins injection. Results: Homogenates from muscles collected five days after venom
injection induced a drop in the number of proliferating myoblasts and a complete abrogation of
myotube formation, whereas homogenates from muscle injected with either the myotoxic PLA2
(Mtx) or the metalloproteinase (BaP1) inhibited myotube formation without affecting myoblast
proliferation. The inhibitory effect induced by muscle homogenates from venom-injected mice was
completely abrogated by preincubation of the homogenate with antivenom antibodies, but not with
control antibodies, evidencing that the inhibitory effect on myotube formation is due to the action
of venom present in the tissue. In agreement, venom was detected by immunoassay in mouse
muscle during the first week after injection. A characteristic pattern of expression of matrix
metalloproteinases MMP-9 and MMP-2, as well as of MT1-MMP, was observed in muscles treated
with venom or with the isolated toxins Mtx and BaP1 in a similar pattern to that described in other
models of regeneration. Conclusions: Our observations suggest that traces of venom present in
muscle tissue by the time critical steps in muscle regeneration are occurring might inhibit myotube
formation and preclude a successful regenerative response.
106
VIBM_H010_0013
An Integral Intervention for Malaria Control in the Municipality of
Wampusirpi, Gracias a Dios
1
2
3
4
5
6
Engels Banegas , Rosa Elena Mejia , Prabhjot Singh , Efrain Burgos , Karen Calderon , Wilberto Montalvan ,
7
Melesio Prado Talabera
1
2
National Malaria Program, Ministry of Health, Honduras. National Parasitological Laboratory, Ministry of
3
4
Health, Honduras. Pan American Health Organization, Honduras. Regional Health Department of Gracias a
5
6
Dios, Ministry of Health, Honduras. Municipal Health Team of Wampusirpi, Ministry of Health, Honduras.
7
Department of Health Promotion, Ministry of Health, Honduras. Mayoral Office of Wampusirpi, Honduras.
BACKGROUND: The municipality of Wampusirpi in Gracias a Dios had the highest incidence of
malaria (118 malaria case per thousand inhabitants) in the country in 2009; a high burden of
Plasmodium falciparum cases and a maternal death due to malaria was reported in the same year.
This high burden of malaria is due to multiple factors including difficult access to health services
owing to geographical, economic and cultural reasons, complicated by lack of knowledge and
empowerment of people. An urgent intervention for malaria control, overcoming these barriers,
was necessary. RESEARCH OBJECTIVE: Develop a comprehensive and integrated intervention with
community participation to reduce the incidence of malaria transmission in the municipality of
Wampusirpi. METHODOLOGY: An intervention was developed and carried out in all 26 localities of
Wampusirpi. Brigades of community volunteers were formed and trained to conduct house to
house visits carrying out direct installation of bednets with 100% coverage of all sleeping spaces,
active case detection, supervised treatment for 3 days targeted towards a reduction in P. falciparum
cases and comprehensive 14 day treatment for P. vivax and mixed infections, and giving health
education with emphasis on use and maintenance of the bednet. Surveillance was maintained in the
municipality following the intervention. RESULTS: Coverage of 100% of localities and houses was
achieved using 3380 bednets installed in 902 households with a population of 6161inhabitants; 403
malaria cases were detected and treated of which 50% (201) were due to P. falciparum. Prevalence
rates of up to 29% in localities were found with a global prevalence of 6.39% (95% CI: 5.78-6.99).
Malaria declined by 85.5% in 2011 from 2009 with one case of P. falciparum malaria reported. Two
cases of P. vivax malaria have been reported in the first three months of 2012. CONCLUSIONS: The
implementation of interventions in a comprehensive and integrated manner with community
participation, surveillance and corresponding technical monitoring led to a sustained impact and
drastic reduction in malaria transmission. This serves as a model intervention and is to be scaled up
to other endemic areas of Honduras, especially those with P. falciparum transmission.
107
VIBM_C352_0034
Characterization of species of Lutzomyia and Leishmania in
endemic area of Cutaneous Leishmaniasis, municipality of Dulce
Nombre de Culmí, Olancho, Honduras
1
1
2
2
3
Sosa- Ochoa W *, Sandoval CM , Avila G , Carrasco B ., Figueroa M .
1
2
Escuela de Microbiología, Facultad de Ciencias, Universidad Nacional Autónoma de Honduras. Universidad
3
Nacional de Agricultura, Honduras. Organización No Gubernamental (ONG) El Buen Pastor, Catacamas,
Olancho. *Corresponding Author: Escuela de Microbiología, Facultad de Ciencias, Universidad Nacional
Autónoma de Honduras, Ciudad Universitaria, Boulevard Suyapa, Tegucigalpa, Honduras; email:
[email protected]
BACKGROUND AND OBJECTIVE. The Leishmaniasis is an infectious disease caused by protozoans of
the gender Leishmania; it constitutes a severe public health problem. The parasite infects human
beings by the sting of a Lutzomyia sandfly that previously has fed in an infected reservoir (1). In
Honduras it is a major cause of mortality, in the year 2010 there were 1394 cases positive for
Leishmaniasis. In the country, four clinical forms of the illness: Cutaneous Leishmaniasis (CL),
Mucocutaneous Leishmaniasis (MCL), Visceral Leishmaniasis (VL) and Non Ulcerated Cutaneous
Leishmaniasis (NUCL) have been identified (2). Thus, the main objective of this study is to determe
the diversity of Lutzomyia and leishmania species circulating in the municipality of Dulce Nombre of
Culmí. MATERIALS AND METHODS. One hundred sixty four Lutzomyia were captured using CDC
trap for their taxonomic identification and natural infection determination (3). Thirty skin scrapings
were smeared onto a sterile glass slide, stained with Giemsa, and microscopically examined for the
presence of Leishmania amastigote forms. For molecular analysis, the skin scraping was placed in
1.5 –mL tubes containing 100 µl of Chelex 100 (BioRad). RESULTS. Thirteen Lutzomyia species were
identified, Lu. panamensis, Lu. ylephiletor and Lu. carrerai thula were of major predominance.
Female sandflies were separated in 7 pools for PCR analysis. The overall natural infection rate with
Leishmania (Viannia) braziliensis in the 7 pools of sandflies was 14% (1/7). Twenty eight skin
scrapings were positive for Leishmania Viannia subgenus. Leishmania (Viannia) braziliensis was
detected in twenty four skin scrapings and Leishmania (Viannia) panamensis in four skin scraping.
CONCLUSIONS. We reported for the first time in Honduras three new species of Lutzomyia: Lu.
geniculata, Lu. carrerai thula and Lu. aragaoi. Leishmania braziliensis and Leishmania panamensis
are confirmed as the etiologic agents for Cutaneous Leishmaniasis and Mucocutaneous
Leishmaniasis in the area of Dulce Nombre de Culmí, Olancho.
108
VIBM_1122_0062
Prevalence of Helicobacter spp. and H. pylori Presence in Canines
from Metropolitan Area in Costa Rica, and its Relationship with
Gastric Pathologies
1
2
3
2
Marcela Suárez-Esquivel , Caterina Guzmán-Verri , Alejandro Alfaro-Alarcón , Elías Barquero-Calvo .
1. Physiology Department, Veterinary Medicine School, National University of Costa Rica. 2. Tropical Diseases
Investigation Program (PIET), Veterinary Medicine School, National University of Costa Rica. 3. Pathology
Department, Veterinary Medicine School, National University of Costa Rica.
There is a lot of information regarding H. pylori infection in humans and its association with gastric
malignancies, however, even when the presence of Helicobacter non-pylori has been described in
animals and humans, it's role in pathogenesis has not been elucidated. Canines warrant major
relevance because of their inclusion in humans' home. The proximity with their owners places them
in a possible zoonotic role, and pets’ well being is a matter of concern that justifies investment in
proper veterinary care. In the Small Animal Hospital of the National University, chronic vomiting is
the second cause of canine hospitalization for gastric problems. Considering the high prevalence of
H. pyori in human population and that Costa Rica is second place in gastric cancer prevalence, the
determination of the prevalence and presence of Helicobacter spp. and H. pylori, and it's association
with alterations in canine gastric mucosa is an initial step in comprehending a possible causative
role of these microorganisms. Gastric biopsies by endoscopy were taken from 80 canines from the
Metropolitan Area of Costa Rica. The samples were analyzed for rapid urease test, polymerase chain
reactions and histopathology, with hematoxylin- eosin, and Warthin-Starry stains. Our results show
a 90% prevalence of Helicobacter spp. in the sampled population. Histopathology revealed that 25%
of canines had mild subclinical gastritis, 26% of them had moderate to severe gastritis, and 45%
were categorized as healthy individuals, in spite of some mild gastric alterations. Up to 41% of the
individuals showed an increased mononuclear inflammatory infiltrate in lamina propia, 67% showed
edema, and 68% had hyperemia. None of them were positive to H. pylori. According to our findings,
the presence of Helicobacter spp. in stomach mucosa can indeed trigger inflammatory responses
capable of inducing different degrees of mucosal damage; however, it should be considered that
injury development is probably a result of several factors, similar to H. pylori- induced gastric
cancer.
109
VIBM_1-54_0109 b
Congenital HCMV Infection in Neonates from Honduras and Costa
Rica
1
1
2
2
2
Taylor Lizeth ,Baltodano Jose , Ortiz Jafet , Cabrera Jessy , Ferrera Annabelle
2
1Virology, Microbiology School, University of Costa Rica; Microbiology School, National Autonomous
University of Honduras
Introduction: Cytomegalovirus (CMV) is the most common cause of congenital and perinatal
infections; however, few data are available from developing countries where the overall burden of
infectious diseases is frequently high. Objective: The aim of the study was to determine the
prevalence of congenital CMV infections in a defined population in Honduras and Costa Rica and to
assess the seroprevalence and activity of the virus in the mothers. Methods: During 2010-2011,
urine samples were collected and tested by PCR for the presence of HCMV DNA in newborns;
sequencing was done in all positive samples to determine the genotype. At the same time,
mothers´ blood samples were tested for the virus by PCR and by ELISA for IgG/IgM antibodies; the
IgG avidity test was performed to determined IgG maturation and evolution of infection and
antigenemia for CMV pp65 to show viral replication. Results: In Costa Rica, from 145 collected
samples, 6.2 % babies were infected: 2 from 2010 asymptomatic children and 7 from 2011
symptomatic children. All mothers´ samples were antigenemia negative; 97 % were IgG Anti CMV
positive and 83 % presented past infections. Only 16 % were IgM Anti CMV negative and 2 mothers
presented a primary infection (IgM positive, IgG negative), but her babies were negative. Genotype
gB2 and gB3 was established only in 2 samples. In Honduras, 253 samples were collected. Of 100
samples analyzed up to now by PCR, 10% of babies and 31% of mothers presented CMV infection.
Of the 253 women tested by serology, 98% were IgG positive and 65% had both IgG and IgM
antibodies at delivery. Three (1.2%) women presented low avidity IgG antibodies with positive IgM,
indicating primary infection; 51.4% showed IgM antibodies with a high avidity IgG index, implying
recurrent infections; 11% presented positive pp65 antigenemia. Conclusion: Based on the present
data, in Costa Rica congenital CMV infection was low. On the contrary, in Honduras congenital
CMV infection was relatively high. Given the low awareness of the infection in the general public,
there is a need to promote knowledge of congenital CMV and support research initiatives into this
important infection.
110
VIBM_1-54_0109
Molecular characterization and follow-up of a Hepatitis C viral
infection in a hematology-oncology unit at Children´s Hospital in
Nicaragua
1
1
2
Taylor Lizeth , Vargas David , Somarriba María Mercedes .
1.Virolgy, School of Micribiology, University of Costa Rica. 2. Hemato-Oncology Unit, Children Hospital,
Managua, Nicaragua
Introduction: A “silent” outbreak of viral hepatitis in a hematology/oncology unit at the public
Children’s Hospital in Nicaragua showed 53,3 % HCV infection and 29,4 % HBV in 1999. HBV vaccine
was applied and improvements were performed in the Hospital infrastructure and the use of
disposable material was implemented. However in control groups analyzed in 2003 and 2006 it was
shown that HCV is still an important problem in this unit (unpublished data). The aim of this project
is to perform the molecular characterization of HCV associated to this outbreak and to identify, if it
is possible, the origin of the infection, to recommend appropriate measures to stop the outbreak.
Methods: A total of 600 samples were collected during 2009-2010. Screening was done by HCV EIA
combo to the first 414 samples. All samples were analyzed by RT-PCR to the 5’ no coding region.
Viral load and RT-PCR of hyper-variable region and sequencing were done in the positive samples
(60 samples from 1998-2006 were included). Phylogenetic analyze is in process. General
information and disease status were obtained for each patient. Results: During the ten year of
infection study, the population conserves their demographics characteristics, near 50 % boys and
girls, around 60 % are under 9 years old and 35 % live in Managua. HBV down from 10 % to 2.4 %
because of partial vaccination implementation, but HCV only change from 53 % to 26 %(EIA combo
and RT-PCR showed a good correlation). The viral load were over 105 copies/mL in 50 % of cases
during 2006 compare to 23 % during 2009.The most affected group was always children with
leukemia. Preliminary studies showed most frequency or genotype 1a and a common cluster by
phylogenetic studies. There was increased risk between become positive, most hospitalization time
and to be immunosuppressed. Conclusion:The results showed a possible persistent nosocomial
infection by HCV and good control of HBV by vaccination. To avoid new cases and to give the
correct therapeutic treatment to the infected children is necessary to implement corrective
measures, principally to give separate attention to positive and negative children, and to obtain
financial support to maintain this strategy.
111
VIBM_C101_0070
Challenges in achieving the Millennium Developing Goals in
Nicaragua: Factors contributing to the spread of HIV/AIDS in
Nicaragua
William J. Ugarte Guevara,
1,3
2
3
Eliette Valladares Cardoza, Birgitta Essén, Ulf Högberg,
3
1
Center for Demography and Health Research, National Autonomous University of León Nicaragua.
3
Department of Obstetrics and Gynecology, National Autonomous University of León, Nicaragua. Department
of Women’s and Children’s Health, International Maternal and Child Health, Uppsala University, Sweden
2
Background: UNAIDS estimates that over 7000 people infected with HIV live in Nicaragua by
December 2010. Despite has one of the lowest HIV prevalence rates in Latin America, conditions as
the increasing number of HIV cases in Central America, high levels of poverty and migration rates,
and incomplete and outdated epidemiological data suggest the need to identify the factors causing
the slow of progress in achieving the Millennium Developing Goals (MDGs) on reducing gender
inequality (MDG 3) and combating HIV/AIDS (MDG6). Research Objectives: Determine contributing
factors in the transmission of HIV/AIDS in the Nicaraguan context. Methodology: We undertook a
systematic literature review through multiple search engines (PubMed, Web of Science, Google
ScholarTM, and relevant bibliographies) using Nicaragua as a required keyword. We searched for any
articles for the years 2000 to the current using the keyword as well as the terms 'HIV' and/or 'AIDS',
'prevalence', 'knowledge', 'attitudes', 'behaviors', 'perceptions', 'prevention', 'intervention',
'gender', drugs, alcohol, 'vulnerability', 'education', and/or 'health care infrastructure'. Results:
Current literature indicates that two groups of factors contribute to the spread of HIV/AIDS in
Nicaragua. Structural factors enlarge the vulnerability of people to HIV infection, especially Poverty,
low education and gender power imbalance remain as the main obstacles for safe sexual practices;
and behavioral factors determine the chance that individuals become infected, for instance
inconsistent condom use. A conceptual framework displays the factors driving the epidemic and the
evidence-based interventions to implement in the context of Nicaragua. Conclusions: The challenge
of combating HIV/AIDS and reducing gender inequalities in Nicaragua are far from being achieved.
Developing synergies among the MDGs agenda and actors, and addressing the structural and
behavioral barriers are needed to ensure the success in achieving the MDGs.
112
VIBM_A406_0059
Soil Transmitted Helminths and Nutritional Status of School-Age
Children in a Honduran Endemic Area
a
a
b
b
c
Mary-Theresa Usuanlele , Jose-Antonio Gabrie , Maria-Mercedes Rueda , Maritza Canales , Theresa Gyorkos
a
and Ana Sanchez
a
b
Faculty of Applied Health Sciences, Brock University, St. Catharines, ON. Canada; School of Microbiology,
c
National Autonomous University of Honduras, Tegucigalpa, Honduras; Division of Clinical Epidemiology,
McGill University, Montreal QC, Canada.
Background: Soil transmitted helminth (STH) infections and malnutrition often share common
geographical locations, mostly in resource-poor communities. Although STH infections can affect all
age groups, school-age children are the most vulnerable and experience detrimental effects on their
health, physical and cognitive development. Dearth of information on the association between STH
infections and malnutrition in Honduras necessitates urgent investigations into this problem.
Research Objectives: To evaluate the prevalence of STH infections among school-age children living
in rural communities in Honduras and to assess their association with children’s nutritional and
growth status. Methodology: A cross-sectional study was done among school-age children residing
several rural communities of the department of Olancho, Honduras in 2011. Demographic and
anthropometric measures (weight and height) were obtained and stool samples collected for STH
determination by the Kato-Katz method. Z-scores of height for age (HAZ) and body-mass-index
(BMI) for age were calculated using the WHO’s 2007 Anthroplus software. Stunting and thinness
were defined as HAZ and BMI-for-age z-score less than -2 SD of the growth reference. Results: In
total 320 children (7–15 years old) completed the study. The overall STH prevalence was 72.5% for a
specific prevalence of 30.31% for Ascaris lumbricoides; 66.87% for Trichuris trichiura; and 15.94% for
hookworms. Children aged 7-10 were 2.14 times more likely to be infected by any STH and girls in
general were less likely (OR: 0.482) to be infected with hookworms than boys. The odds of having
hookworms were almost three times higher in stunted children (OR: 2.856). Of the infected
children, 25 (7.8%) were malnourished, the majority categorized as stunted but no statistical
association was found between infection and malnutrition. Conclusion: There is a high prevalence
of STH infections in the study population but a statistical association cannot be proved between
infection and overt impairment of the children’s nutritional status. Notwithstanding, due to the
chronic nature of STH infections, intervention efforts are warranted in the study communities to
decrease parasites’ transmission.
113
VIBM_4102_0056
Hospital outbreak by KPC producing Klebsiella pneumoniae in a
hospital in Panamá
Silvio Vega, MD, MSc; Marcela de Paredes, MT; Rudy Kant, MD; Ribana Molino, MD.
Complejo Hospitalario Metropolitano, CSS. Panamá
Background. Carbapenem resistance in Klebsiella pneumoniae does not occur naturally and is due
mainly to the presence of acquired carbapenem hydrolyzing betalactamases. Carbapenemases
(KPC) type enzymes in carbapenem resistant K. pneumoniae strains were first reported in 2001 in
North Carolina, and until 2005, the geographical distribution of these enzymes in the family
Enterobacteriaceae in general and in K. pneumoniae specifically was limited to the eastern United
States, where KPC producing K. pneumoniae became a frequent nosocomial pathogen. Outside of
the United States, KPC producing K. pneumoniae has been reported in several European countries;
in China and in Latin America in Argentina, Uruguay, Chile and Colombia.
We describe the emergence and spread of KPC producing Klebsiella pneumoniae at a Panamanian
hospital; this is the first report in Central America. Methods. Isolates of Klebsiella pneumoniae with
a carbapenem minimum inhibitory concentration >1 µg/mL identified by Vitek 2 Compact (V2C,
BioMerieux) and a negative EDTA-imipenem disk synergy test result were submitted to boronic acid
disk test and to polymerase chain reaction (PCR) for KPC gene and sequencing. Records from
patients who had KPC producing K. pneumoniae isolated were retrospectively reviewed. Results.
From February through August 2011, 71 patients (27 in the intensive care unit [ICU]) were colonized
(n=21) or infected (n=50) by KPC producing K. pneumoniae. PCR performed analysis with specific
primer for blaKPC demonstrated genetic related to KPC-1. All the strains were phenotypically
positive for carbapenemase activity, but negative by metallo-betalactamase production. Multidrug
resistance characterized the studied isolates, with polimyxin, tigecycline, gentamicin, and
fosfomycin being the most active agents. The outbreak strain was introduced in February and
disseminated rapidly by cross-transmission. Twenty nine cases of respiratory tract infections, 14
bacteremia, 13 surgical site infections, and 11 urinary tract infections were identified as the most
relevant infections. Most patients received polimixin b, tigecycline or gentamicin-containing
combination treatment. Crude mortality was 54.9% among ICU patients and 37.5% among non-ICU
patients, but attributable mortality was 20.2% and 30.3%, respectively.
Conclusions. The emergence of KPC–producing K. pneumoniae in a Central American hospital
creates an important challenge for clinicians and hospital epidemiologists, because it is added to the
already high burden of antimicrobial resistance.
114
VIBM_C093_0071
Enterohemorragic Escherichia coli in Fecal Samples of Cattle from
Dairy Farms in León city, Nicaragua.
1*
1
2
Samuel Vilchez BSc PhD , Daniel Reyes MD MSc PhD , Sylvia Beckerdreps MD MPH , Margarita Paniagua BSc
1
1
MSc & Félix Espinoza MD MSc PhD
1
Centro de Enfermedades Infecciosas, Departamento de Microbiología y Parasitología, Facultad de Ciencias
2
Médicas, Universidad Nacional Autónoma de Nicaragua, León (UNAN-León). Department of Family Medicine,
University of North Carolina at Chapel Hill, North Carolina.*Corresponding author
Diarrhea is still a leading cause of childhood morbidity and mortality in children from developing
countries. Enterohemorragic Escherichia coli (EHEC) is an emerging pathogen associated with
enteric foodborne diseases related with products of animal origin. In order to determine the
occurrence, virulence markers, serogroups and phenotypes of EHEC strains, 188 fecal samples of
cattle randomly selected among 14 dairy artisanal farms surrounding León city in Nicaragua, were
investigated. For E. coli screening, feces were streaked onto Sorbitol MacConkey Agar and
incubated at 37°C overnight. EHEC identification was performed by amplification of Shiga toxin (stx)
and intimin (eae) genes through conventional PCR. Serogroups were investigated in samples
positives for stx by slide agglutination with eight (O26, O55, O103, O111, O121, O128, O145 and
O157) different antisera. Then, a subset of 195 E. coli isolates from 29 EHEC positives arbitrarily
selected samples were submitted to biochemical fingerprinting analysis using a PhP-RE system.
EHEC was identified in 45.7% (86/188) of the assayed samples.The genotypic virulence
characteristics of positives EHEC were distributed as follow: st1 (4.6%), st2 (46.5%), st1-st2 (9.3%),
eae-st1 (4.6%), eae-st2 (25.6%) and eae-st1-st2 (9.3%). Serogroups were identified in 24.4% (21/86)
of the EHEC samples. Serogroup O157 was only found in 1.2% of the samples. The rest were
distributed as follow: serogroup O26 (8.1%), O103 (3.5%), O111 (4.7%), and O121, O128 and O145
(2.3%) respectively. Biochemical fingerprinting analysis revealed the presence of 12 common
phenotypes, grouping 94.9% (185/195) of the isolates. Suggesting the occurrence of pathogenic
clonal groups circulating. Additionally, the bovines phenotypes 01, 02 and 11, were identical to
human phenotypes 06, 14 and 18 previously identified in a study of children with diarrhea in León
city. This is the first report of EHEC in dairy cattle in Nicaragua. One interesting fact is we found
seven out of the eight serogroups investigated, including O157. This data might suggest a role of
bovines in the epidemiology of EHEC associated diseases in León.
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VIBM_0047_0026
Helicobacter pylori clarithromycin resistance from Guatemalan
strains
1
1
1
Carmen de Tercero , Elisa Hernández de Rodas , Norma Marroquín , Guillermo Pérez-Pérez
2
2
1
Centro de Investigaciones Biomédicas, Facultad de Ciencias Médicas, USAC, New York University, Langone
Medical Center.
Helicobacter pylori is a Gram negative bacterium associated to duodenal or gastric ulcers, gastric
cancer and gastric mucosa-associated lymphoid-tissue. Antibiotic treatment should be established
according to clinical and microbiological criteria. Different therapeutic regimens have been used.
One of the most included among regimens is Clarithromycin even though resistance to have been
reported due to several punctual mutation in V domain of ribosomal RNA 23 s sub unit. The main
objective of this study was evaluating the resistance to clarithromycin in H. pylori strains isolated
from Guatemalan patients suffered erosive chronic gastritis or pre malignant lesions. Methods: 82
strains from patients with erosive gastritis or pre malignant lesions from Hospital Roosevelt and
Hospital San Juan de Dios from Guatemala City were obtained by gastric biopsies and isolated in
blood agar in anaerobic conditions at 37oC. Secondary culture was made in brucella brought and
then inoculated again in blood agar at 0.5 Mac Farland standard concentration simultaneously we
applied an impregnated with different concentrations of Clarithromycin strep (E –test) and reading
according to manufacture recommendations. Results: we found 21 strains presenting resistance to
Clarithromycin it is around 26 %, there is not difference between sex, age or clinical diagnosis.
Conclusions: The resistance to Clarithromycin is high, and similar to other studies made in Latin
America. We recommend making cultures and clarithromycin resistance test in samples from
patients before starts with treatment and making a surveillance system of resistance.
VIBM_0801_0078
A Baseline Pilot Study using Multidisciplinary Interventions to
Improve Mother-child Health in Rural Regions of Honduras:
Integration of Health Education and Nutrition Intervention with
Deworming Programs in Olancho
Mejía R.E.¹, Zuniga C.², Bottazzi M.E.
3
116
1
2
Laboratorio Nacional de Parasitología, Secretaría de Salud de Honduras. Programa Nacional para el control
3
y la prevención de la Enfermedad de Chagas y Leishmaniasis, Secretaría de Salud. Baylor College of Medicine,
Houston, TX. USA.
Background: With the setting of poverty and malnutrition, Honduras is plagued by some of the
highest rates of tropical infectious and parasitic diseases in the Western Hemisphere. Among the
endemic NTD infections in the region are classic tropical diseases such as soil-transmitted helminth
infections with high prevalence and intensity rates. Overall Goal: To assess the impact and
efficiency of incorporating STH assessments during the laboratory analysis to an already established
Honduran health program for pregnant women and their newborn children. Materials and
methods: The study was performed in mother/child clinics located in six municipalities of the
Department of Olancho. The proposed sample size was 500 pregnant women, to be enrolled
preferably in the first and second trimester of pregnancy where their offspring would be evaluated
during the first year. The methods and laboratory procedures included assessment of disease
burden of STH using the Kato-Katz method, and an assessment of the prevalence of anemia. Briefly,
samples from participants were examined for the presence and intensity of intestinal helminths
through the analytical and quantitative fecal examinations and the level of anemia with hemoglobin
/ hematocrit assessments. Results. The project recruited a total of 462 (92.4%) pregnant women to
be included in the study. The age distribution included 26.68% of teenage mothers, of women
between the ages of 10-19, and 5% of women over the age of 35. With the following order of
frequency and parasites found: a) Trichuris trichiura b) Ascaris lumbricoides, and c) Hookworm. As
per Honduran guidelines, all pregnant women with STH infections were treated with Mebendazole
after their third trimester. Women that also had evidence of anemia were treated with iron and
folic acid. Of all pregnant women with anemia, all recovered satisfactorily by treatment with iron
and folic acid, the treatment with Mebendazole 500mg single dose was 100% effective for Ascaris
and Trichuris trichiura Uncinaria but 73% was effective and in 27% the parasite was not eliminated
but reduced parasite density. Conclusions. With the results obtained in this study it was shown that
STH assessments are recommended to be incorporated into already established Honduran health
program for pregnant women and their newborn children during each prenatal visit and to their
offspring.
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