Khater et al.,
THE LOUSICIDAL, OVICIDAL, AND REPELLENT EFFICACY OF SOME
ESSENTIAL OILS AGAINST LICE AND FLIES INFESTING
WATER BUFFALOES IN EGYPT
HANEM F. KHATER*, MOHAMED Y. RAMADAN , REHAM S. EL- MADAWY
Department of Parasitology
Faculty of Veterinary Medicine, Benha University, Moshtohor – Toukh, 13736, Egypt.
*Correspondence: Hanem F. Khater, telephone: 002-013-2461411, Fax: 002- 2013- 2463074
e-mail: [email protected]
1- ABSTRACT
The lethal and repellent effects of five essential oils were estimated for the first time
against the buffalo louse, Haematopinus tuberculatus, and flies infesting water buffaloes in
Qalyubia Governorate, Egypt. For the in vitro studies, filter paper contact bioassays were used
to test the oils and their lethal activities and compared with that of d- phenothrin. Four minutes
post-treatment, the LC50 values obtained were 2.74, 7.28, 12.35, 18.67, and 22.79% for
camphor (Cinnamomum camphora), onion (Allium cepa), peppermint (Mentha piperita),
chamomile (Matricaria chamomilla), and rosemary oils (Rosmarinus officinalis), respectively,
whereas for d- phenothrin, it was 1.17%. The LT50 values were 0.89, 2.75, 15.39, 21.32,
11.60, and 1.94 min post-treatment with 7.5% camphor, onion, peppermint, chamomile,
rosemary, and d- phenothrin, respectively. All applied materials, except rosemary, were lethal
to the eggs of H. tuberculatus. Despite the results of the in vitro assays, the in vivo treatments
revealed that the pediculicidal activity was more pronounced with oils. All treated lice were
killed after 0.5- 2 min, whereas with d- phenothrin, 100% mortality was reached only after 120
min. The number of lice infesting buffaloes was significantly reduced 3, 6, 4, 6, and 9 days
post-treatment with camphor, peppermint, chamomile, onion, and d- phenothrin, respectively.
Moreover, the oils and d- phenothrin significantly repelled flies for 6 and 3 days posttreatment, respectively. No adverse effects were noted on either animals or pour-on operators
after exposure to the applied materials. Consequently, some Egyptian essential oils show
potential for the development of new, speedy, and safe lousicides and insect repellents for
controlling lice and flies which infest water buffaloes.
Key words: Haematopinus tuberculatus, buffalo, lice, essential oils, pediculicide, ovicidal, fly
repellent.
and Leite, 2005; and Lemcke, 2006). H.
tuberculatus has been recorded where
buffalo have been introduced and
domesticated, Egypt, the Philippines,
Australia, Madagascar, China, and
Myanmar (Lancaster and Meisch, 1986). It
has also been found on cattle, in close
association with buffalo (Lancaster and
2- INTRODUCTION
The buffalo louse, Haematopinus
tuberculatus,
Burmeister,
1839,
(Phthiraptera:
Haematopinidae)
is
principally an ectoparasite of Carabao,
water buffalo, Bubalas bubalis L.
(Lancaster and Meisch, 1986; Bastianetto
1
Khater et al.,
Meisch, 1986), on camel (Lawal et al.,
2007), and on wild ruminants (Marley and
Conder, 2002).
Buffalo lice cause anemia and loss
of body condition. They need to be
controlled,
particularly if
animals’
condition is affected (Lemcke, 2006).
Damage caused by blood sucking lice
involves low milk and meat productivity
(Bastianetto and Leite, 2005), blood loss,
and in serious cases, abortion and death
(Lancaster and Meisch, 1986). Moreover,
small calves can build up high numbers of
lice, making control necessary (Lemcke,
2006). The severity of the infestations and
the potential for transmission of rinderpest
may make such lice an important pest for
control measures (Woodworth, 1922).
Additionally, flies infesting animals
cause great economic losses and transmit
many diseases (Roberts and Janovy, 2005;
Taylor et al., 2007).
Among lousicides, most cattle lice
drug formulations are effective against the
buffalo louse (Colwell, 2002; Marley and
Conder, 2002; Hussain et al., 2006; and
Lemcke, 2006). The need for novel
solutions to control pediculosis has been
intensified due to the emergence of
resistance (Chosidow et al., 1994; Levot,
2000), environmental pollution (Gassner et
al., 1997), and insecticidal residues in milk
(Gottschall et al., 2005).
Essential oils have been used for
centuries as insecticides and insect
repellents for treating and preventing
infestations by lice (Priestley et al., 2006;
Williamson, 2007; and Williamson et al.,
2007). The constituents of plant volatile
oils have long been known to affect the
behavioral responses of pests; their
monoterpenoid components appear to be
the most useful as insecticides or antifeedants (Veal, 1996).
The aims of this study were to
determine the in vitro and in vivo lousicidal
efficacy of some essential oils in
comparison to d-phenothrin, as well as
their ovicidal and repellent effects against
flies when applied as pour-on solutions on
to water buffaloes.
3- MATERIALS AND METHODS
3.1. Lice
The buffalo louse H. tuberculatus
and their eggs were collected from infested
water buffaloes at the farms of Faculties of
Agriculture and Veterinary Medicine at
Moshtohor, Benha University, Qalyubia
Governorate, Egypt.
3.2. Tested substances
Five essential oils were tested,
namely, camphor (Cinnamomum camphora),
onion (Allium cepa), peppermint (Mentha
piperita), chamomile
(Matricaria
chamomilla), and rosemary (Rosmarinus
officinalis). All oils were obtained from ElKaptain Company, Egypt.
The use of 0.4% d-phenothrin
(Item®, Mash pharmaceutical company,
Egypt) as an anti-lice shampoo is
authorized in Coryne, Monaco, France. It is
also authorized by the ministries of health
in France and Egypt for the treatment of
adults and eggs of human head lice,
Pediculus humanus var capitis.
3.3. Testing for in vitro pediculicidal
activity
The filter paper contact bioassay
was chosen because it is more
representative of what could occur in
nature. Lice would directly contact
compounds, as they do in the filter paper
imitating
field
circumstances.
Consequently, in vitro assays were useful
for pre-screening of the efficacy of
materials before field application. The
method used to assess the pediculicidal
activity was adapted from World Health
Organization, WHO (1981) and according
to Priestley et al. (2006).
Preliminary experiments were
conducted
to
determine
suitable
experimental parameters, such as dilution
factors for tested substances and the
duration of exposure to lice. Bioassays
were performed at 27±2ºC and 75±5%
relative humidity (RH).
The finalized direct contact assay
was carried out as follows. Each test
substance was diluted in water to different
2
Khater et al.,
concentrations from 0.23 to 60%), and a
few drops of Tween 80 were added as an
emulsifier. A volume of 600 µl of the
diluted sample was distributed evenly over
a 9 cm diameter filter paper held in the
lower half of a 9 cm glass Petri dish. After
15 min, the liquid had spread out, the filter
paper was fully impregnated and no excess
moisture was left in the dish. Ten buffalo
lice, males and females, were placed on the
top of each filter paper disc. The control
groups were treated with distilled water
and Tween 80. Ten replicates were used for
each concentration.
Lice were examined under a
dissecting microscope at nine different
time intervals (1, 2, 4, 8, 10, 15, 20, 30, and
60 min). Death was defined as the lack of
limb and gut movement, and the failure to
respond when the legs were stroked with a
forceps (Priestley et al., 2006). The number
of fatalities was recorded, and the lethal
concentrations, LC50, LC90, and LC95,
were subsequently calculated.
mortality was subsequently calculated. All
hatched nymphs were classified as having
survived the treatment, and those failing to
hatch or only partially hatching as having
been killed.
3.5. In vivo control of Haematopinus
tuberculatus
3.5.1. Tested substances
The efficacy of the essential oils
and anti-louse shampoo was evaluated
under field conditions as pour-on solutions
on infested buffaloes during the period
from April to May 2008. Rosemary was
excluded because it was the least toxic oil.
3.5.2. Animals
Forty-eight water buffaloes, with an
average body weight of 400 kg and body
surface area was 4.37 m2, that appeared to
be healthy except for their natural lice
infestations with H. tuberculatus, were
kept in shaded areas at the previously
mentioned farms. Body surface area was
computed from body weight using the
following formula:
Body surface area (m2) = 0.12 body
weight (kg)0.06 as indicated by Hurnik and
Lewis (1991).
3.4. In vitro assessment of the ovicidal
effect
The discriminating doses (DD) of
the in vitro bioassays were calculated
according to the methods of Kristensen et
al. (2006) and were approximately twice
the lethal dose that kills 95% of insects, i.e.
the LC95, Table 1.
The ovicidal activity was assessed
according to the methods of Priestley et al.
(2006). The discriminating doses were
prepared and used to fill 20 ml glass
bottles. Twenty-five (non- hatched) eggs
were immersed in each test substance for
10 min. After this time, the eggs were
removed and blotted on a medical wipe
tissue. Eggs in the control group were
exposed to distilled water and a few drops
of Tween 80. After treatment, eggs were
incubated in separate glass Petri dishes, at
28±1ºC and 75% RH. Such treatment was
repeated four times. Therefore, the total
number of eggs examined was 100 for each
dose.
Hatchability of the eggs was
checked on a daily basis for 20 days. Egg
3.5.3. Experimental design
The pour-on application method
was chosen because it is easy to carry out,
environmental pollution is reduced (in the
case of using insecticides), and also
because it is a very practical method,
especially where no dip tanks are available
or when just a few animals need to be
treated (Anon, 2008).
Pour-on treatment required the
application of the used material (2.5 L of
each DD) along the backline of the animal
using graduated squeeze bottle, where the
liquid was dispersed over the animal's body
surface, with the exception of the head, to
contact lice.
Buffaloes were grouped into six
groups (8 animals per group) and doses of
each discriminating dose of the compounds
were poured on the animals. The
experiment was carried out as follows:
3
Khater et al.,
Group 1: animals were treated with
camphor, 1.4 ml/kg b.w.
Group 2: animals were treated with onion,
2.9 ml/kg b.w.
Group 3: animals were treated with
peppermint, 3.6 ml/kg b.w.
Group 4: animals were treated with
chamomile, 3.4 ml/kg b.w.
Group 5: animals were treated with dphenothrin, 0.6 ml/kg b.w.
Group 6: animals were treated with
distilled water and few drops of
tween 80, serving as the
untreated control group.
The inspection was conducted as
follows. Twenty lice were collected from
each animal and examined 0.5, 1, 1.5, 2, 5,
25, 75, 90, and 120 min post-treatment,
until 100% mortality was observed.
The numbers of lice infesting both
sides of the animals, on neck, shoulder,
trunk, abdomen, limbs, and tail, were
counted daily for 10 days post-treatment in
order to calculate the reduction rates.
Animals and pour-on operators
were observed daily for any abnormal
health observation and skin irritation.
data
were
subjected
to
Probit
transformation followed by regression
analysis to determine the lethal values
(LC50, LC90, and LC95) as well as the
slope of the regression lines by computer,
using POLO-PCO according to the method
of Finney (1971). Moreover, the lethal time
(LT) estimates, LT50 and LT90, were
calculated.
For
statistical
analysis,
Duncan´s multiple range test was used with
the SPSS program (SPSS v10, SPSS Inc.,
Chicago, IL, UDA).
The reduction of egg hatchability
and lice percentages as well as repellency
indices were calculated
Hatchability reduction % =
[(Control – Treated) / Control]*100
Lice Reduction % = [(Pre- treatment countPost- treatment count) / Pretreatment count] *100
Repellency index (RI) =
[(Nc-Nt) /Nc ]* 100
Where Nc is the number of flies
infesting buffaloes (in the pre- treatment
day), and Nt is number of flies in the
treatment.
3.6. The repellent effect of the applied
materials
Flies infesting buffaloes were
collected through fly nets and then
identified according to Furman and Catts
(1986). While doing the in vivo treatments,
the repellent effect and protection time of
the applied materials toward flies, Musca
domestica,
Stomoxys
calcitrans,
Haematobia irritance, and Hippobosca
equina, were investigated daily for 10 days
post-treatment, by counting the total
number of flies present on the neck,
shoulder, trunk, abdomen, limbs, and tail of
animals in the treated and control groups.
Flies at both sides of the animal were
counted from a distance of 2 m away from
the animals, a proper distance for not
disturbing flies that came to infest animals.
4- RESULTS
The direct contact, in vitro,
bioassays revealed that the lousicidal
efficacy of the used materials increased as
the concentration and the exposure time
increased (Fig 1- 6). Just one minute after
treatment, 100% lousicidal efficacy was
achieved by 30% camphor, chamomile,
onion, and peppermint oils. It was
surprising to attain such a rapid lousicidal
effect.
The sensitivity of H. tuberculatus to
the materials used (Table 1), was
demonstrated by the LC50 values, obtained
after treatment for 4 min, of 2.74, 7.28,
12.35, 18.66, and 23.47 % for camphor,
onion, peppermint, chamomile, and
rosemary oils, respectively; the LC50 value
obtained for d-phenothrin was 1.17%.
Based on LC50 values of the tested
3.7. Data analysis
Live and dead lice were counted to
determine the mortality rates. The mortality
4
Khater et al.,
materials and that of rosemary, as a
reference substance, the relative potency
indicated that camphor, onion, peppermint,
chamomile, and d- phenothrin were 8.57,
3.22, 1.90, 1.26, and 20.06 times,
respectively, more effective than rosemary.
With regard to the time response
observations, the LT50 values were 0.89,
2.75, 15.39, 21.32, 11.24, and 1.94 min
after treatment with 7.5% camphor, onion,
peppermint, chamomile, rosemary, and dphenothrin, respectively (Table 2).
All
tested
materials
except
rosemary (not applied) showed an ovicidal
effect; the hatchability rates for treated
eggs were 10, 4, 14, 18, and 32, whereas
the reduction of the hatchability
percentages were 88.24, 95.29, 83.53,
78.82, and 62.35% for camphor, onion,
peppermint, chamomile, and d- phenothrin,
respectively (Table 3).
Concerning
pour-ons,
100%
mortality of H. tuberculatus was observed
0.5, 1, 1.5, 2, and 120 min after treatment
with camphor, peppermint, chamomile,
onion, and d- phenothrin, respectively
(Table 4). Additionally, the number of lice
infesting treated buffaloes was significantly
(P < 0.05) reduced from the pre- treatment
count up till 3, 6, 4, 6, and 9 days posttreatment, respectively (Fig 7).
The flies found to be infesting
buffaloes before treatment were M.
domestica, S. calcitrans, H. irritance, and
H. equina. The essential oils and dphenothrin repelled flies significantly (P <
0.05) for 6 and 3 days post-treatments,
respectively (Fig 8).
There is a long tradition of using
aromatic plants as insecticides and
repellents around the home and in animal
bedding (Williamson et al., 2007). The
data of our study demonstrate that the
applied oils were highly toxic to H.
tuberculatus. One minute after the in vitro
treatments, 100% of lice were killed by the
application of all oils except rosemary.
Therefore, the oils used were highly
effective as pour-on solutions for louseinfested water buffaloes, and their effect
persisted 3- 6 days post-treatment.
d-phenothrin was the most effective
material in case of the in vitro assays. It
significantly reduced the number of lice
infesting buffaloes for 9 days posttreatment. In contrast, it had the least
ovicidal and repellent efficacy.
In the present study, the control
group showed an increasing lice infestation
trend during the period of study. A similar
result was observed by Hussain et al.,
2006. Control of buffalo louse must be
carried out over short intervals to interrupt
the natural life cycle of the parasite
(Bastianetto and Leite, 2005; and Lemcke,
2006).
Concerning the treatment of buffalo
lice, most cattle-lice formulations including
single dose types were effective as pourons or sprays, and two doses of a synthetic
pyrethrin-based pour-on were effective in
eradicating
lice
(Lemcke,
2006).
Cypermethrin showed 94.7 % control of
lice infesting buffaloes, after 28 days posttreatment (Hussain et al., 2006).
Macrocyclic lactones have been
used to control parasites of domesticated
wild ruminants, including H. tuberculatus,
and Hypoderma spp. (Marley and Conder,
2002). Moreover, the efficacy of a single
S/C administration of ivermectin, 200
ug/kg, to buffaloes was reported to be
100% on the 28th day of medication
(Colwell, 2002 and Hussain et al., 2006).
In general, the repeated use of lousicides
results in the development of marked levels
of resistance (Levot, 2000). Additionally,
the resistance of P. h. capitis towards dphenothrin has previously been reported
5- DISCUSSION
Pediculosis is one of the notorious
diseases affecting livestock production and
efficiency at global level (Lancaster and
Meisch, 1986). The louse, H. tuberculatus,
is the main ectoparasite that attacks water
buffalo and the itch caused by it is
responsible for the low productivity of milk
and meat of the animals (Bastianetto and
Leite, 2005; and Lemcke, 2006).
5
Khater et al.,
(Chosidow et al., 1994). Chlorpyrifosbased lice treatments should be avoided
because they have been found to be toxic to
some buffalo, particularly in hot weather
(Lemcke, 2006). Insecticides also pollute
the environment around animals (Gassner
et al., 1997).
The use of ivermectin has
deleterious effects on male fertility of cattle
(Avery
and Schmidt, 1995),
goat
(Tanyildizi and Bozkurt, 2002), and rats
(El-Nahas and
El-Ashmawy, 2008).
Moreover, adverse reproductive reports
include abortion, stillbirth, and infertility
following use of ivermectin in animals,
such as cattle, horse, pig, dog, and sheep,
were
reported
(Greene,
1991).
Additionally, ivermectin causes neonatal
toxicity in rats (Lankas et al. 1989).
Consequently, health-care providers
now face a serious lack of new commercial
pediculicides.
It is very important to safely
delouse animals to avoid many problems,
such as abortion and death (Lancaster and
Meisch, 1986), infestation of small calves
as they can build up high numbers of lice
(Lemcke, 2006), insecticide residues in
milk and especially milk fat (Gottschall et
al., 2005), and environmental pollution
(Gassner et al., 1997). Thus, new
alternative insecticides are being sought
after for safer louse control.
The rapid killing of lice (as shown
by the applied oils) is very important in
order to avoid the delayed mortality (28
days)
caused
by
currently
used
conventional insecticides (Colwell, 2002
and Hussain et al., 2006).
Plant essential oils are highly
acceptable to the public as they are natural
and pleasant smelling (Williamson et al.,
2007). They are widely used in traditional
medicine for their insecticidal and repellent
activity against many species of insects,
including lice.
Regarding the prospective of
employing botanical extracts as lousicides,
tobacco (Nicotiana tobaccum), tubli
(Derris
philippinensis),
makabuhay
(Tinosphora
rumphi),
and
neem
(Azadirachta indica) at concentrations of
10, 20, and 40% in oil emulsion induced
more than 90% mortality in carabao louse
in vitro, whereas in vivo experimentation
showed that only tobacco and makabuhay
induced 45.91 and 79.67% reduction in
louse infestations, respectively (Robles,
2004).
Pestoban®, an Indian herbal
preparation, (unknown constituents) was
used by several authors to control lice
infesting cattle and buffaloes. Pestoban®,
induced 100 and 70% mortality of lice after
75 min of in vitro treatments with 1:20 and
1:30 v/v solutions, respectively (Prasad et
al., 1989). Furthermore, 10% Pestoban®
showed
100%
efficacy
against
Haematopinus and Linognathus spp. on
naturally-infected cattle and buffaloes in
India (Srivastava and Sinha, 1990).
Several
studies
have
been
conducted to evaluate the efficacy of the
herbal aerosol spray, Ectozee® (extracts of
Cedrus deodara, Azadirachta indica, and
Embelia ribes). Ectozee® at 25 and 100%
was reported to not only kill cattleinfesting lice, Linognathus spp., after 1 and
3 minutes, respectively in vitro, but also
Hippobosca spp. after 3 and 6 min,
respectively (Maske and Bhilegaonkar,
1996). In addition, Das et al. (2003) proved
the efficacy of the same product on dogs
with ectoparasitic dermatitis, for 97.7 and
100% of dogs were found to be completely
cured and free from louse and fly
infestations, respectively. A dog infested
with Haematopinus piliferus, Burmeister,
was covered with a cloth on which oil from
the eucalyptus, Eucalyptus globulus, was
sprinkled drop by drop, and all lice were
found to be dead after 48 hours (Sergent
and Foley, 1915).
Furthermore,
Tobacco
extract
treatment was found to be a better
therapeutic procedure than ivermectin to
cure Linognathus vituli infestations of
West African Dwarf goats (Fajimi et al.,
2003).
In recent years, several studies have
demonstrated the in vitro pediculicidal
efficacy of some essential oils towards
6
Khater et al.,
female head lice. Eucalyptus, rosemary,
and pennyroyal, Mentha pulegium which is
a member of the mint genus, oils were
found to be at least, if not more, effective
(against P. h. capitis) than d-phenothrin
and pyrethrum, two commonly used
pediculicides (Yang et al., 2004a).
Essential oils, in particular, pennyroyal, tea
tree, and anise, have potent insecticidal
activity for killing head lice and their eggs
(Williamson, 2007). E. globulus leaf oilderived monoterpenoids were found to be
highly toxic to eggs and females of the
human head louse (Yang et al., 2004b).
Furthermore, essential oils contain
monoterpenoids which have lousicidal and
ovicidal effects against clothing lice, P.
humanus (Priestley et al., 2006).
Additionally, peppermint and rosemary oils
were reported to control P. humanus
(Veal, 1996).
A study comparing the lethal
activity of oils using both a filter paper
contact bioassay and a fumigation assay
found that potency differed depending on
which method of testing was used. For
example, eucalyptus, pennyroyal, and
rosemary oils were more effective in closed
containers than in open ones, indicating
that the effect of these oils was largely a
result of action in the vapor phase, thus
giving them a higher fumigant toxicity
(Yang et al., 2004a). This observation
explained why rosemary was the least toxic
oil in our study as we used a filter paper
contact bioassay.
Volatile oils reduce egg hatchability
due either to the toxicity of the oil vapors
to eggs (Schmidt et al., 1991) or to some
chemical ingredients present in the
volatiles of tested oils which probably
diffused into eggs, thus affecting vital
processes associated with embryonic
development
(Gurusubramanian
and
Krishna, 1996).
Regarding the insecticidal activities
of the oils used against insects other than
lice, onion (Khater, 2003) and rosemary
oils (Shalaby and Khater, 2005) are highly
effective as larvicides against Culex
pipiens. Chamomile and rosemary oils are
toxic to Lucilia sericata larvae (Khater,
2008). The activity of the previously
mentioned oils extends beyond larval
stages, as their sublethal concentrations
reportedly led to serious morphological
abnormalities that inhibit metamorphosis
and adult emergence. Therefore, such oils
are classified as insect growth regulators
(IGRs) (Khater, 2003; Shalaby and Khater,
2005; and Khater, 2008). Moreover,
peppermint and spearmint (Mentha viridis)
are highly effective against fed females of
the cattle tick Boophilus annulatus (AbdelShafy and Soliman, 2004). Furthermore,
rosemary oil reduces the hatchability of
eggs (62.65%) and adversely affects some
biological aspects of the potato tuber moth,
Phthorimaea operculella (Moawad and
Ebadah, 2007)
With regard to flies infesting
animals, great economic losses, disease
transmission (Roberts and Janovy, 2005),
and anaphylactic reaction after Hippobosca
equina bite in human (Quercia et al. 2005)
are major reasons for the use of insect
repellents.
The applied essential oils not only
killed lice very quickly (0.5- 2 min), but
also repelled flies (for 6 days) at the same
time. Similar result was reported for some
herbal preparations such as Pestoban®,
which lasted about 2 h (shorter protection
time than ours) when applied on surfaces in
dwellings (Prasad et al., 1989) as well as
Ectozee® (Das et al., 2003). Furthermore,
onion (Guarrera, 1999), peppermint (Erler
et al., 2006), and rosemary (Prajapati et al.,
2005) were reported to be repellents.
Pennyroyal and its benzyl component were
effective repellents against P. h. capitis
(Toloza et al., 2006).
Self-application methods, such as
dust bags and back rubbers used principally
for horn fly, Haematobia sp, have also
been used to reduce louse infestations
(Anon, 2008). Furthermore, insecticidal
ear-tags are effective deterrents to buffalo
fly, H. irritans (Lemcke, 2006). On the
other hand, such uses produce resistant
horn fly populations within a few weeks
(Sheppard and Joyce, 1992). The Influence
7
Khater et al.,
of permethrin, diazinon and ivermectin
treatments on insecticide resistance in the
horn fly was reported by Byford et al.
(1999). Additionally, permethrin, which is
usually used for ear tags applied to cattle,
was found on all surfaces analyzed,
including the cattle, the bark of trees in
their pasture, on a fence pole, and on grass.
Moreover, some residues were found three
months after the ear tags were applied
(Gassner et al., 1997).
The present data revealed that the
herbal pour-ons were highly effective as
insecticides and fly repellents. Their safety
needs to be fully evaluated, but there were
no abnormal health observations or skin
irritations related to treatment were
observed on the treated animals and pouron operators during the study. A similar
observation was also recorded for some
commercial herbal preparations, such as
Pestoban® (Srivastava and Sinha, 1990)
and Ectozee® (Maske and Bhilegaonkar,
1996 and Das et al., 2003).
Plant volatile oils consist of
numerous different, mostly volatile low
molecular weight (LMW) terpenoids
(Dewich, 2002; Priestley et al., 2006;
Williamson, 2007; and Williamson et al.,
2007). Such oils have long been known to
affect the behavioral responses of pests,
with the monoterpenoid components
appearing most useful as insecticides or
antifeedants, (Palevitch and Craker, 1994).
LMW terpenoids may be too lipophilic to
be soluble in the haemolymph after
crossing the cuticle, and proposed a route
of entry through the tracheae (Veal, 1996).
In addition, Priestley et al. (2006)
explained that most insecticides bind to
receptor proteins in the insect and, in doing
so;
they
interrupted
normal
neurotransmission, which lead to paralysis
and subsequently, death. Additionally,
recent evidence suggests that LMW
terpenoids may also bind to target sites on
receptors that modulate nervous activity.
Ionotropic γ- aminobutyric acid, GAPA
receptors, the targets of organochlorine
insecticides lindane and dieldrin, are
modulated by LMW terpenoids with vastly
different structures (Priestley et al. 2006).
6- CONCLUSION
The control of lice presents many
great research challenges and prospects for
the identification of new, safe and
environmentally acceptable insecticides.
The present study revealed the lousicidal,
ovicidal, and repellent activity, at low
concentrations and short exposure time, of
some Egyptian herbs. These plant products
have potential for the development of new
and safe control products for carabao louse
and fly infestations. Treatment is usually
effective and is best carried out 14- 18 days
apart to ensure that the life cycle is
completely broken. Moreover, all buffaloes
on the property should be treated at the
same time. All new buffaloes should be
treated on arrival and not mixed with
resident buffaloes until deloused.
FUTURE STUDIES
The applied plant oils, like all
chemically-based medicinal herbal plants,
should also undergo a battery of
experimental procedures to determine their
total
pharmacological
profile.
Enhancement of the potency of oils by
adding synergists and stabilizers will
prolong their effectiveness. Furthermore,
they may lead to future development of
potential sources of natural insect control
agents which recommended for field
evaluation and integrated into other pest
management programs for control of
insects of medical and veterinary
importance.
ACKNOWLEDGMENTS
The authors would like to thank Dr.
Azza Moustafa, Research Institute of
Medical Entomology, Egypt, as well as Dr.
Nagwa Ahmed, Parasitology Department,
Faculty of Veterinary Medicine, Benha
University, Egypt, for their support and
suggestions.
8
Khater et al.,
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Khater et al.,
Mortality %
Fig 1. In vitro treatment of buffalo lice
with various concentrations of camphor oil
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
0.90%
1.40%
1.80%
7.50%
30.00
%
control
1
2
4
8
10 15 20 30
Post-treatment time (min)
60
80
Mortality %
Fig 2. In vitro treatment of buffalo lice
with various concentrations of onion oil
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
1.80%
4.70%
7.50%
1
2
4
8 10 15 20 30
Post-treatment time (min)
12
60
80
15.00
%
30.00
%
control
Khater et al.,
Mortality %
Fig 3. In vitro treatment of buffalo lice
with various concentrations of peppermint oil
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
3.80%
7.50%
15.00%
22.50%
30.00%
control
1
2
4
8
10 15 20 30
Post-treatment time (min)
60
80
Mortality %
Fig 4. In vitro treatment of buffalo lice
with various concentrations of chamomile oil
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
7.50%
15.00%
19.00%
22.50%
30.00%
control
1
2
4
8 10 15 20 30
Post-treatment time (min)
13
60
80
Khater et al.,
Mortality %
Fig 5. In vitro treatment of buffalo lice
with various concentrations of rosemary oil
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
3.80%
15.00%
22.50%
30.00%
60.00%
control
1
2
4
8 10 15 20 30
Post-treatment time (min)
60
80
Mortality %
Fig 6. In vitro treatment of buffalo lice
with various concentrations of d- phenothrin
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
0.23%
0.45%
0.90%
2.70%
1
2
4
8
10 15 20 30
Post-treatment time (min)
14
60
80
30.00
%
control
Khater et al.,
Fig 7. Reduction of lice after various treatments
100.00
80.00
Camphor
60.00
Peppermint
40.00
Reduction %
Camomile
20.00
Onion
0.00
dphenothrin
Untreated
control
-20.00
-40.00
-60.00
-80.00
-100.00
-120.00
1
2
3
4
5
6
7
8
9
Post-treatment time (day)
10
Repellency index
Fig 8. Repellent effect of various materials
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
-10.00
-20.00
-30.00
Camphor
Peppermint
Camomile
Onion
d-phenothrin
Untreated
control
1
2
3
4
5
6
7
8
Post-treatment time (day)
15
9
10
‫‪Khater et al.,‬‬
‫التأثير السمي لبعض الزيوت األساسية القاتل للقمل و بيضة والطارد للذباب‬
‫الذي يصيب الجاموس في محافظة القليوبية ‪ -‬مصر‬
‫هانم فتحي خاطر‪ ,‬محمد يوسف رمضان‪ ,‬ريهام سمير المعداوي‬
‫قسم الطفيليات‪ ،‬كلية الطب البيطري‪ ,‬جامعه بنها‬
‫أجريت هذه الدراسة لمعرفة التأثير السمي والطارد لخمس أنواع من الزيوت األساسية علي القمل‬
‫والحشرات الطائرة التي تصيب الجاموس بمحافظة القليوبية بمصر‪ .‬لقد أجري اختبار ورقة الترشيح في‬
‫المعمل بهدف معرفة مدي كفاءة تلك الزيوت ومقارنتها بدلتا فينوثرين‪ .‬لقد تبين من الدراسة أن التأثير‬
‫النصفي القاتل (بعد ‪ 4‬دقائق من التعرض للمواد المختبرة) ‪،2.74‬‬
‫‪%22.79‬‬
‫‪،7.28‬‬
‫‪،12.35‬‬
‫‪،18.86‬‬
‫لزيت الكافور والبصل والنعناع والكاموميل والحصالبان علي التوالي بينما كانت النسبة‬
‫‪ %1.17‬لمادة بدلتافينوثرين‪ .‬أما التأثير الزمني النصفي القاتل فقد كان علي التوالي‪،2.75 ,0.89 ،‬‬
‫‪ 10.94 ،11.60 ,21.32 ,15.39‬دقيقة‪ ,‬ولقد أثبتت كل المواد المختبرة كفاءة عالية في قتل بيض القمل‪,‬‬
‫ولقد مات كل القمل المعالج بالزيوت بعد فترة زمنية قصيرة (من نصف الي دقيقتين) بينما مات القمل‬
‫المعالج بدلتافينوثرين بعد ‪120‬دقيقة‪ .‬وعندما تم اختيار هذه الزيوت علي الجاموس لدراسة مدي تأثيرها‬
‫علي القمل المتطفل عليه لوحظ انخفاض أعداد القمل بعد ‪ 9 ،6 ،4 ،6 ،3‬أيام من معالجة القمل بزيت‬
‫الكافور والنعناع والكاموميل والبصل وكذلك مادة الدلتافينوثرين‪ ،‬علي التوالي‪ ،‬هذا باإلضافة إلي أن كل‬
‫المواد المستخدمة حقليا أثبتت كفاءة في طرد الذباب بعيدا عن جسم الحيوان وبالتالي تكون الزيوت‬
‫األساسية قد أظهرت كفاءة عالية كمواد فعاله وآمنة وسريعة في قتل القمل وبيضه وكذلك طرد الحشرات‬
‫الطائرة التي تصيب الجاموس‪.‬‬
‫‪16‬‬