April 2007, Volume 12, Issue 1 The Food Safety Division of Alberta Agriculture and Food publishes the Animal Health Forum quarterly to inform readers of division activities and bring attention to agri-food issues with food safety and market access implications. Editors: Joan St. Amand, Food Safety Division, Airdrie AB Phone: (403) 948-8575; E-mail: [email protected] Jagdish Patel, Food Safety Division, Edmonton AB Phone: (780) 644-5093; E-mail: [email protected] Please direct all inquiries to Joan or Jagdish. Chief Provincial Veterinarian Update other situation which would cause numerous livestock mortalities. The project involves working with four pilot municipalities to develop and implement a toolkit that will provide guidelines for planning for emergency carcass disposal. Alberta’s Emergency Carcass Disposal Pilot Project Lisa Morin, Office of the Chief Provincial Veterinarian, Edmonton, AB An inter-departmental team, led by the Office of the Chief Provincial Veterinarian (OCPV) at Alberta Agriculture and Food (AF), is implementing a pilot project that will help to prepare the province for the safe disposal of large numbers of livestock carcasses in cases of fire, disease, flood or any A general mail-out to all of Alberta’s municipalities was sent in late January and twenty two letters of interest were received. Four municipalities (County of Lethbridge, County of Lacombe, County of Vermilion River and County of Barrhead No. 11) were selected by the Alberta Emergency Carcass Disposal planning team. It was a difficult decision to choose only four municipalities due to the interest and support for this project. The selection criteria used by the planning team included: In This Issue • The number and types of livestock present. Alberta’s Emergency Carcass Disposal Pilot Project ....................................................... 1 Accidental Monensin Toxicity in Reindeer .......... 2 Zoonosis Update: Ovine Abortion ........................ 3 Bovine Trichomoniasis .......................................... 4 Alberta Veterinary Surveillance Network: Update on Livestock Pathology Consultation Activities ............................................................. 5 An Automated High Throughput Screening Enzyme Linked Immunosorbent Assay for Johne’s Disease Antibodies in Bovine Serum ..................................................... 6 Continuing Education Opportunities .................. 7 Who’s Who in the Division .................................... 8 • The potential for identifying suitable land for burial (based on the Natural Suitability Index, a geologicallybased document showing potentially suitable burial land in Alberta). • The human population present. • The location in the province. Municipalities with varied profiles were chosen so that the toolkit will represent different ends of the spectrum. For example, municipalities with a high likelihood of identifying suitable burial land, and those with a poor likelihood were selected. Similarly, municipalities with varied animal and human population densities were chosen. Regional representation was also a consideration. Website: “Ropin’ the Web”: http://www.agric.gov.ab.ca/surveillance/ A consultant has been hired to work with the pilot municipalities to develop the toolkit, which will include processes and guidelines about: Post mortems performed by an LPCP pathologist on 9 of the females showed that the animals were in good nutritional condition. There was no evidence of an infectious disease process. Four of the reindeer had severe pulmonary congestion and edema, which was compatible with death due to acute heart failure. Histological examination of a comprehensive set of formalin-fixed tissues collected from the nine reindeer examined revealed the common lesion of acute multi-focal myocardial degeneration and necrosis. • How to include emergency carcass disposal in existing Municipal Emergency Plans. • How to identify carcass disposal needs and profile current disposal capacities. • How to secure available resources for an emergency disposal situation, including how to identify suitable burial sites. During a farm visit, the owner realized that the wrong dairy cattle premix might have been fed to the reindeer, during the past month. The owner had been advised by a nutritionist to feed a dairy dry-cow premix that did not contain Rumensin free choice to the herd as a mineral supplement. The producer had done so for several years without experiencing problems. Legal carcass disposal methods in Alberta include composting, burial, rendering, incineration and natural disposal. Burial site identification involves adhering to provincial regulations such as the Destruction and Disposal of Dead Animals Regulation and various environmental Codes of Practice. The toolkit will also provide guidelines for obtaining land if municipal land is unsuitable or unavailable. During the farm visit, 5 more animals were found dead in pastures. The carcasses were frozen and there was blood at the nostrils and blood spray on the snow indicating terminal epistaxis. There was no evidence of struggle at the sites of death. The primary outcome of this project is that Alberta will be better prepared to deal with emergency carcass disposal, in the event that it is necessary. Once the toolkit is complete, it will be distributed to the remaining municipalities in Alberta to assist them with establishing their own emergency carcass disposal plans. Samples of premix were collected at the farm for monensin analysis from a bucket and two separate bins containing lactating cow premix containing sodium monensin (920 mg/kg) and dry cow premix containing no monensin. The premix that was in the bucket was discarded and the troughs in the animal pens used to feed the premix were examined to ensure they were empty. The owner was advised that more deaths might occur over the next few weeks, depending on the amount of the premix that had been offered to and consumed by the reindeer. One month after the farm visit, the owner reported that no more deaths had occurred. The Emergency Carcass Disposal Pilot Project is being led by the OCPV in the Food Safety Division of AF in partnership with Alberta Environment (AE), Alberta Health and Wellness (AHW), Emergency Management Alberta (EMA), Canadian Food Inspection Agency (CFIA) and the Alberta Association of Municipal Districts and Counties (AAMDC). Disease Impact Analysis of the premix collected from the bucket for monensin matched the monensin level detected in the lactating cow premix collected from the bin (771 mg/kg). The manufacturer’s instructions for the lactating cow premix clearly indicated that the premix should not be fed undiluted. Trace amounts of monensin were detected in liver samples collected from the 9 reindeer necropsied. Accidental Monensin Toxicity in Reindeer Brian Miller, Veterinary Pathologist and Jagdish Patel, Surveillance Veterinarian, Alberta Veterinary Surveillance Network (AVSN), Food Safety Division, Edmonton, AB Monensin is an antibiotic ionophore used to promote growth and as a coccidiostat in target species. Increase use of ionophores has led to occurrences of toxicosis in target and non-target species. In dairy cattle, monensin is used to prevent the development of ketosis. At the cellular level, the biological activity of ionophores has been associated with changes in concentration of primary ions. Theses changes can directly or indirectly alter the intracellular pH ultimately causing damage to cells or cellular organelles, particularly within cardiac and smooth muscle tissue. In a dosing study in cattle, evidence of QT interval prolongation, QRS complex prolongation, first degree heart block, infrequent premature atrial beats, and increased T wave amplitude were noted in a few animals after 2 to 7 days of exposure. The LD50 levels quoted in the literature for various species are (mg/kg body weight): horse 3mg/kg , sheep 12 mg/kg, goat 26 mg/kg, and cattle 50 mg/kg. A reindeer producer, who was experiencing an increase in animals dying suddenly, was referred to the Livestock Pathology Consultation Program (LPCP) of the AVSN. Fourteen reindeer out of a herd of 70 (13 females and 1 male) had died suddenly. April 2007 2 Cases of monensin intoxication in companion animals, particularly in dogs that have eaten cattle feed, have also been reported in the literature. distinguish pathologically but there are a few tips that may suggest which agent to consider most strongly until laboratory confirmation is received. To the authors knowledge, no cases of monensin intoxication in reindeer have been reported and the LD50 level for monensin in reindeer is unknown. This outbreak however, suggests that reindeer may be highly sensitive to monensin toxicity. Generally, monensin toxicity has been associated with errors in feed mixing that result in non-target species becoming exposed. This outbreak illustrates that feed related toxicities should be considered during investigations of sudden death in cattle, and other domestic species, when signs of infectious disease are not apparent on post mortem examinations. If there is placentitis with: • Necrotic, hemorrhagic cotyledons and intercotyledonary (IC) areas of leathery thickening and edema, consider Chlamydophila abortus. • Intercotyledonary necrosis only, consider Coxiella burnetti. • Several small, yellowish, necrotic foci spread over the surface of cotyledons, consider Toxoplasma gondii. If the placenta has been retained, the ewes have metritis and there are necrotic foci in the fetal liver: • If necrotic foci are large, consider Campylobacter spp. For further information please contact Dr. Brian Miller at (780) 427-8201,[email protected] or Dr. Jagdish Patel at (780) 644-5093, [email protected] • If necrotic foci are small and numerous, consider Listeria monocytogenes or Salmonella spp. Regardless of the above, there is much overlap in pathological findings in ovine abortion and lack of grossly visible lesions does not rule out any of the above agents. It is difficult to confirm the cause of ovine abortion without confirmatory laboratory tests. Sending one of more whole fetuses with placenta to a laboratory is optimal. Serological tests are also available for many of these agents at the herd level. If you wish assistance with appropriate testing, contact a laboratory diagnostician for advice. Diagnostic Overview Zoonosis Update: Ovine Abortion Jan Bystrom, Veterinary Pathologist, Zoonoses Surveillance Priority Team, Food Safety Division, Airdrie, AB With respect to human health, it is prudent to advise clients about the risk to human health when handling ewes that are lambing. Warnings should include: • Handle all uterine and fetal fluids and tissues including the placenta with rubber gloves and avoid contamination of clothing with fluids. • All of the above agents transmit by the oral route so frequent hand washing and avoidance of hand to mouth contact should be emphasized. • Destroy all fluids and tissues from lambing beds and do not allow scavengers access. This time of year there are requests for information regarding diagnosis of ovine abortion. Questions typically arise when abortions occur in clusters or “storms” that concern producers sufficiently enough to call for veterinary assistance. From the perspective of human health, it is critical to remember that many of the pathogens involved have potential to cause serious disease in humans. • Humans in specific risk groups should not be in contact with lambing ewes, including the old, the young, the immunocompromised and pregnant women. For further information regarding diagnosis of ovine abortion or other diagnostic laboratory inquiries, please contact Dr. Jan Bystrom at (403) 912-3309, jan.bystrom@ gov.ab.ca or Dr. Brian Miller at (780) 427-8201, brian. [email protected]. There are several potentially zoonotic agents that can cause ovine abortion. These include Chlamydophila abortus (formerly Chlamydia psittici), Coxiella burnetti, Toxoplasma gondii, Campylobacter jejuni and C. fetus, Listeria monocytogenes, and Salmonella spp. These can be difficult to 3 April 2007 Bovine Trichomoniasis end of the breeding period. When breeding period is not closely supervised, the first indication of a problem is often an increased number of cows that are cycling and not pregnant at the end of the breeding period. In operations with the infection, majority of calvings are in the later half of the calving season. In contrast, in herds without infection, 80% of the cows will calve within the first two heat cycles of the calving period provided that the cows are in good body condition and there are no other problems. Dr. Jagdish Patel, Surveillance Veterinarian, Food Safety Division, Edmonton, AB Diagnosis Testing herd bulls rather than cows for trichomoniasis is more rewarding and economical because there are fewer bulls to test and the disease is self-limiting in cows. Collecting and culturing and laboratory examination of the smegma from bulls is the diagnostic technique for field use. Smegma can be collected using a bovine uterine infusion pipette attached to a 12 cc syringe. Pass the pipette to proximal prepuce fornix area and with negative suction move the pipette around the fornix area. Slight hemorrhage in aspirated smegma is accepted to carry out an effective sample collection. Data submitted to Alberta Veterinary Surveillance Network show that during fall of 2006 trichomoniasis was confirmed in bulls from some farms in southern Alberta. Older bulls will remain carriers and can carry the infection from one year to the next. On rare occasions a pregnant cow can be a carrier and be a source of infection. Impact on pregnancy rate and calf crop in operations that use natural service in their breeding programs can be devastating if the infection is introduced to a naïve herd. The collected sample should be placed immediately in tubes/ pouches (InPouchR) containing culture medium that is at 20 to 25oC. These tubes/pouches should be stored in container away from direct sunlight and during transport to the laboratory, these samples should not be allowed to cool to single digit temperatures or freeze. We are therefore recommending that in herds where a potential for introduction of the disease exists, testing of bulls and prevention measures mentioned below should be undertaken prior to the breeding season. The sensitivity of culture method is 80 to 90% based on three tests done at weekly intervals. We highly recommend doing three successive tests at weekly intervals just prior to the breeding season. The chance of detecting the parasite is increased if the bull is separated from cows for at least 2 weeks before testing. Transmission The parasite Tritrichomonas fetus is transmitted when an infected bull breeds with a susceptible heifer or cow. Infected cows can transmit the disease to virgin uninfected bulls during breeding. Although young bulls (<3 years) may clear the infection, infected bulls become long-term carriers and thereby carry the infection from one breeding season to the next. Some infected cows can deliver a normal calf (<1%). Fortunately these carrier cows are rare but may present a problem when trying to control the disease in a herd. Bulls from multiple sources on community pastures, sharing of bulls by multiple herds and purchasing and using untested older bulls in emergency situations during the breeding season pose the greatest threat of contracting the disease. Prevention Preventing introduction of this disease in the herd should be the main emphasis in any management plan. Prevention measures are effective in closed herds and are important in reducing the risk of acquiring and spreading the disease in community pastures. Many producers already conduct some of the following measures: • Only use virgin bulls and heifers as replacements; use as many home-raised heifers as possible. • Only buy confirmed pregnant heifers if outside replacements are needed. Clinical Presentation The parasite usually causes death of the developing foetus between 40 to 70 days of pregnancy with some cows developing post-coital pyometra. Some cows may abort as late as five months into pregnancy. Often the first indication of a disease problem in a herd is when a producer notices the cows he thought were pregnant resume cycling towards the April 2007 • Maintain a limited tight breeding season. • Keep the average bull age as young as possible. • Test all mature bulls at least three times at weekly intervals before introducing them into your herd. • For community pastures test all bulls at least three times before beginning of the breeding season. 4 • Monitor the breeding period to detect signs of excessive repeat breeding. • Veterinary practitioners have made appropriate diagnostic efforts, generally having done a farm visit to clearly define the problem and performed necropsies or other diagnostic tests with samples submitted to private diagnostic laboratories without a diagnosis or resolution of the problem. If the problem is severe enough, this requirement may be waived, e.g. suspicion of foreign animal disease. • Control other reproductive diseases like campylobacteriosis with appropriate vaccinations and perform breeding soundness exam of all bulls before turnout. • Cull open cows at pregnancy check and test cows that have recently aborted. • Necessary diagnostic services are not available otherwise. • Do not purchase open cows from auction market. • Suspected oilfield related concerns might qualify by referral using a specific process. • Avoid using and or sharing bulls from unknown herd history. • Maintain proper fencing of pastures to avoid mixing of animals from herds with unknown status. The following summary of LPCP cases from the past several months are divided into two categories. Targeted surveillance submissions are solicited under specific programs or projects. Non-targeted submissions arise when practitioners contact a member of the LPCP team or other AF veterinarians for assistance. If above recommendations cannot be adhered to, vaccinating cows in the herd is an option however it is expensive and not 100% efficacious. Treatment Currently there are no approved treatments for trichomoniasis in either the bull or cow. Targeted surveillance submissions: • Practitioners are encouraged to submit liver samples from cases of suspect bovine bacillary hemoglobinuria (BH) as part of a surveillance project designed to look at incidence and distribution of BH and association between BH and liver fluke infection in Alberta. For further information please contact Dr. Jagdish Patel at (780) 644-5093, [email protected] • Practitioners are also encouraged to submit bovine brains sent under Canada-Alberta BSE Surveillance Program (CABSESP) for diagnostic workup further to the BSE test. Cows showing neurological signs that are not BSE are of particular interest. Surveillance/Research Update • A surveillance veterinarian analyzes Veterinary Practice Surveillance (VPS) data and clinical syndromes that appear unusual in some way or that suggest diseases of interest to AF, are flagged and practitioners may receive a telephone call with an offer of assistance by AF diagnosticians. Alberta Veterinary Surveillance Network: Update on Livestock Pathology Consultation Activities Jan Bystrom, Veterinary Pathologist, Food Safety Division, Airdrie, AB Alberta Agriculture and Food’s (AF) Livestock Pathology Consultation Program (LPCP) assists veterinary practitioners with disease diagnosis. The program is not intended to take the place of private diagnostic laboratory services. It is an adjunct to those services under certain conditions described below. Non-targeted submissions from practitioners: • Goat kid from a herd with a problem the producer suspected was due to sour gas exposure. Diagnosis of severe malnutrition and anemia due to sucking lice and coccidiosis was made. • Unusual incidence of congenital cataracts in dairy calves. This problem was referred to AF from a private laboratory for more extensive workup. The problem has not resolved and ongoing investigation involves the Livestock Disease Investigation Network (LDIN) and LPCP diagnostic services. Gross necropsy and histology services are offered in cases meeting the following criteria: • Significant or unusual herd or flock problem. • Unusually high mortality in calves due to neonatal diarrhea investigated by LDIN/LPCP. Viral etiology was confirmed with ongoing further diagnostic efforts referred to Prairie Diagnostic Services (PDS). • Signs suggesting a new syndrome, an emerging disease, foreign/notifiable animal disease, or food safety, market access or public health concerns. • Unusually high mortality in adult cows on pasture; confirmed as moldy sweet clover poisoning. 5 April 2007 • Unusual syndrome of P3 necrosis/lameness in feedlot steers investigated by LDIN/LPCP. Diagnosis of systemic vasculitis associated with Bovine Viral Diarrhea virus (BVDv) by immunohistochemistry was made in these particular steers. Complexity of the working steps is kept to a minimum by using one unique operation program. The complete robotic workstation with its peripheral equipment is simple to operate, robust and easily maintained. These characteristics enable the elimination of forced downtime due to human and/or machine failure. A 5 to 10 fold improvement in cost, test capacity and productivity was achieved (Table 1). With this system, one diagnostician is able to carry out over 1,000 antibody-screening tests a day. Based on the automated nature of the test, there is ample walk away time (WAT) between ELISA tests, enabling the technologist to multitask – performing other important duties such as sample receiving and preparation, test validation, result interpretation, information dissemination and other related LIM work. • Unusually high mortality with sudden death due to monensin toxicity in reindeer; see report elsewhere in this issue. • Unusual syndrome of recumbency, death and abortion in adult beef cows investigated as possible emerging disease. Diagnosis of severe selenium deficiency was made based on serum mineral analysis. Practitioners are encouraged to contact AF with questions or concerns about diagnostic services. In certain instances, AF may be able to provide the service or can assist with obtaining service through private laboratories. The performance characteristics – such as repeatability, reproducibility, optimal cutoff values, identity score, test sensitivity and specificity were evaluated. These test validation results (Table 2) were based on data accumulated in an 8-year proficiency testing with the US National Veterinary Services Laboratory (NVSL). Our laboratory test has received certification from the United States Department of Agriculture to perform serologic testing for Johne’s disease since 1998. In 2000-2001, the Standard Council of Canada accredited this test scope and in 2004, this a-ELISA was validated according to the requirements of the ISO/IEC 17025 standard. Please contact Dr. Jan Bystrom at (403) 912-3309, [email protected] or Dr. Brian Miller at (780) 427-8201,[email protected] An Automated High Throughput Screening Enzyme Linked Immunosorbent Assay for Johne’s Disease Antibodies in Bovine Serum John T. Y. Wu1*, Lester S. Y. Wong1 and Evelyn E. Bowlby1 A manual Mycobacterium avium ssp. paratuberculosis antibody enzyme linked immunosorbent assay (ELISA) (IDEXX Inc.) was fully automated using an Automation Workstation System (AWS). The IDEXX commercial kit is used to screen for exposure to the Johne’s disease bacterium in cattle on a herd basis. Due to the large number of sera involved in this type of surveillance project, there is a need to increase the cost effectiveness and decrease the turn around time (TAT). The purpose of the work was to develop an automated ELISA (a-ELISA) for Johne’s disease antibodies screening and examine the productivity advantages, quality assured performance characteristics, laboratory information management (LIM) work and multi-tasking provisions under this format. Figure 1. Biomek FX workstation Table 1. Comparison of manual and automated ELISA testing capacity per technologist The AWS was assembled using a modular approach design (Figure 1). The high throughput screening configuration consists of a Beckman Coulter hybrid Biomek® FX liquid handling system integrated with a Molecular Devices Spectramax® microplate reader, a Biotek® ELx405 microplate washer, a Kendro Cytomat® 6000 hotel and an Axis PTZ Network camera. The integrated system function is programmed with an IBM computer using SAMI® software. Housed in a Class 2 Biosafety Cabinet, this fully automated system provides flexibility to accommodate the multi-step requirements of the immunoassay procedure. April 2007 6 Manual Automated Specimen/project No. of projects/year 9 55 5,000 No. of projects/year 4 28 10,000 No. of projects/year 3 18 15,000 Duration of project 5.56 weeks 0.93 weeks 5,000 Duration of project 11.11 weeks 1.85 weeks 10,000 Duration of project 16.67 weeks 2.78 weeks 15,000 Table 2. Testing Proficiency evaluation between AF laboratory and results from NVSL (1998 - 2004: Partially Automated; 2005: Fully Automated) IDEXX JD ELISA Biostatistics values 1998 1999 2000 2001 2002 2003 2004 2005 1998-2005 Kappa 0.71 1.00 0.91 1.00 1.00 1.00 0.86 1.00 0.95 Identity score 92.00% 100.00% 96.00% 100.00% 100.00% 100.00% 96.00% 100.00% 97.99% Test sensitivity 90.91% 100.00% 100.00% 100.00% 100.00% 100.00% 100.00% 100.00% 98.56% Test specificity 100.00% 100.00% 88.89% 100.00% 100.00% 100.00% 80.00% 100.00% 96.67% Positive predictive value 100.00% 100.00% 94.12% 100.00% 100.00% 100.00% 95.24% 100.00% 98.56% 100.00% 100.00% 100.00% 100.00% 100.00% 100.00% 96.67% 25/25 24/25 24/24 25/25 25/25 25/25 195/199 Negative predictive value 1 60.000% Identity 23/25 False neg/false pos 2 FN 1 FP 100.00 24/25 1 FP 2 FN, 2 FP NVSL reference serum pos 20 20 17 18 16 17 21 10 139 NVSL reference serum neg 5 5 8 6 9 8 4 15 60 Immunology Virology Unit, Biology Section, Agri-Food Laboratories Branch, Food Safety Division, Alberta Agriculture and Food, Edmonton, Alberta, Canada. * Corresponding Author: John T. Y. Wu, Immunology and Virology Unit, Biology Section, Agri-Food Laboratories Branch, Food Safety Division, Alberta Agriculture and Food. 6909 - 116 Street, Edmonton, Alberta, Canada T6H 4P2. Phone: (780) 427-8324 Fax :(780) 415-4527 e-mail:[email protected] Food Safety Division Highlights Continuing Education Opportunities Jan Bystrom, Veterinary Pathologist, Food Safety Division, Airdrie, AB presentations and interactive discussion on swine diseases from a pathological and diagnostic perspective. The session built upon a similar session held in April 2006 that also included a wet lab portion. The format for the day was very practical and informal, allowing for a frank and excellent information exchange. Extensive discussion occurred with practitioners providing as much information regarding the state of current swine practice and diagnostics as the pathologists provided information to them. These types of sessions are available on request for small to large groups of practitioners within regions, groups in similar practice types, groups with similar interests, etc. On January 16, 2007, a continuing education (CE) session was held at the Calgary Animal Referral and Emergency (C.A.R.E.) Centre in Calgary at the request of specialty swine practitioners in Alberta. Jointly presented by Dr.’s Jan Bystrom and Ted Clark, this session consisted of If you have ideas for CE sessions or particular formats you would like, please contact Dr. Jan Bystrom at (403) 912-3309, [email protected] or Dr. Brian Miller at (780) 427-8201, [email protected]. 7 April 2007 Who’s Who in the Division Delores Peters is a disease surveillance veterinarian, currently working to adapt the Alberta Veterinary Surveillance Network (AVSN) beef model to the unique needs of poultry industry. She grew up on a mixed farm at Didsbury, Alberta. After graduating from WCVM she worked as a veterinarian in Pincher Creek and managed clinical research trials for beef cattle and pigs where she was the liaison to the computer software developer for collecting chute side health information for the feedlot. Delores completed her MSc (Animal Biochemistry, University of Alberta) in 1994 and is in the process of completing a Master’s Degree in Veterinary Public Health Management through the University of Sydney, Australia. After 11 years in China with family she is thrilled to be back in Alberta. E-mail: [email protected] Sarah Turner received a BSc in Agriculture, majoring in Animal Sciences and specializing in swine nutrition from the Nova Scotia Agricultural College (NSAC) in 1998. After graduation, she worked in a 500-sow farrow–finish PIC multiplier unit in Nova Scotia. She then worked for the NSAC for two years before moving to Alberta and working six years for Alberta Quality Pork. While with Alberta Quality Pork, Sarah managed the delivery and administration of the Canadian Quality Assurance (CQA®) Program and the Animal Care Assessment (ACA) Tool for the Alberta pork industry. She is currently working at Alberta Agriculture and Food as the AVSN Coordinator. E-mail: [email protected] April 2007 8
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