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CHAPTER 1

INTRODUCTION
Bangladesh is an agricultural country and its economic backbone is mainly
based on agriculture. Agriculture plays a vital role in national economy,
contributing about 16.33% of Gross Domestic Products (GDP) (Bangladesh
Economic Review, 2014). Livestock being one of the major components of
agriculture (crop, livestock, fisheries and forestry). The current contribution of
livestock sub-sector to overall GDP is about 1.78% (Bangladesh Economic Review,
2014). The export earnings from leather and leather goods is 3.21% of the total
export, 20% of the population is directly and 50% is partly dependent on this sector
(Bangladesh Economic Review, 2014). Goat was the first ruminant livestock that
was domesticated around 8000-11000BC (Luikart et al., 2006; Tapio et al., 2006).
Among the domestic animals valuable to men in the tropics, the goat is perhaps the
most important species. The largest number of goats is observed in Asia, followed
by Africa, representing about 59.7% and 33.8%, summing up to 93.5% out of the
total number of the world, respectively (FAOSTAT, 2012). Bangladesh possesses
25.611 million heads of goat (Bangladesh Economic Review, 2014).
The goats are mostly reared for meat purpose. Goats are very important group of
livestock in Bangladesh. It is known to all that the main product of goat is meat and
wool (Amin, 1994). Goats available in Bangladesh are of indigenous type having
small body size and producing hairy and course wool of varied colors. In our
country, the goat is primarily raised for meat and no attention is given for wool
production. Goat is much more resistant to rain and other environmental hazard
than other livestock animal. In rural areas of Bangladesh, most of the families rear
goat or goat for their economic consideration. The meat of goat and goat has high
demand in market and the milk of them is easily digestible. Their dung is used as
Agro fertilizer; hides, skin and wool is considered as a source of earning foreign
1

currency. Among the problems existing in Bangladesh hampering the livestock


development, parasitic diseases occupy prime position in Bangladesh. Asian
development bank estimated (1984) that the losses of animal production due to
parasitic diseases were 50% in Bangladesh.
Bangladesh leathers have good inherent quality because of warmer climatic
condition and food habits. Leathers made from hides and skins of animals
maintained in warmer climates are superior to those of animals raised in colder
climates, but such advantages are overshadowed by various defects (FAO, 2004).
Surprising leather defects due to skin disease, postmortem defects or curing defects
and industrial processing defects cut about 55.2% value of leathers resulting in
annual economic loss of approximately Taka 818 core (US$ 220.95 million).
Among such economic looses, US$ 24.1 million was due to Black Bengal goats
skin defects in Bangladesh (FAO, 2004).
Today, societal and scientific concerns regarding exclusive dependency on
chemicals have emphasized the need for the development and introduction of cheap
and environment-friendly pesticide alternatives. In a recent study conducted by
animal scientists from the University of the Philippines at Los Baos (UPLB), eight
medicinal plants were tested for their efficacy in controlling ectoparasites in poultry
and selected ruminants namely: carabao, goat, sheep, cattle, and dog. The selected
botanicals were: leaves of kakawate (Gliricidia sepium), akapulko (Cassia alata),
neem (Azadirachta indica), calabash (Crescentia cujete), alagaw (Premna odorata),
tobacco (Nicotiana tabacum), stems of makabuhay (Tinospora rumphii), leaves of
kutchai (Allium tuberosum), and roots of tubli (Derris philippinensis). The plant
parts were prepared either as a 100% crude powder and applied topically to the
whole body of the animal or as a decoction or crude water extract in various

concentrations. The decoction used oil or water as a carrier and it was applied on
the test animals as a spray, dip, sponge bath, or as a "pour-on" application.
Most of the farmers in Bangladesh cannot afford to buy modern medicine for the
treatment of the livestock because of high price of the drug and poor economic
condition of the farmers. So, if we can develop the traditional system of medicine
in Bangladesh it will be highly beneficial for the farmers and for the overall
improvement of the livestock. Recently much interest in the field of medicinal
plants has been grown throughout the world. Many countries have already come to
realize the medicinal plants as a potential means of therapeutic agent and also their
availability and cost effectiveness.
In Bangladesh many drugs are being used for long time to combat parasitic
infection in livestock. A large number of anthelmintics are now available in the
market. The indiscriminate use of anthelmintics made the parasites to be resistant
against the drug, which have been reported by experts throughout the world
including Bangladesh (Hannan, et al., 2001). So, we should have to back the
traditional uses of medicines.
Ectoparasites are distributed all over the world but occur mainly in tropical and
sub-tropical countries including Bangladesh. Ectoparasites result in a considerable
amount of blood loss, disturbances in feeding, retarded growth and decreased
production. The damages caused by ticks are irritation, inflammation, exudation of
lymph which coagulates to form crusts. Ectoparasites like ticks play an important
role as vector of deadly pathogens to animals and man. Mites cause dermatitis,
alopecia and sometimes animals are infected secondarily by bacteria causing
toxemia, septicemia and ultimately dead of the animals.

Among the

ectoparasiticides, Ivermectin, a new patent drug, this is effective against both endo
and ecto parasites and skin lesions of animals available in Bangladesh. Ivermectin

stimulates the release of GABA from nerve ending and enhance binding of GABA
to special receptors at nerve junctions.
Various types of herbal extracts showed the larvicidal and acaricidal effects
(Chungsamarnyart,et al. 1991). In fact, the preliminary study on the traditional
system of Veterinary Medicine by Food and Agriculture Organization (FAO, 1980),
revealed that indigenous system for the treatment of animal diseases is popular in
man in Asian countries including Bangladesh. There are several indigenous
medicinal plants (neem oil, Tobacco) have anthelmintics action (Nath, 1983,
Mostofa, 1983 and Hossain, 1994) and used against both ecto and endo parasites in
Bangladesh, (Mostofa, 1983, Safique 1983, Mannan, 1997). Neem has been used
historically to rid the body of several forms of ectoparasites and pests and the
action is rapid. Hormone mimicking activities of neem extracts causes interference
with the parasitic life cycle inhibiting their ability to feed as well as prevent the
hatching of eggs (Kumar and Navaratnam, 2013). Neem plant contents have been
reported to be an effective bioinsecticide (Chary, 2011) and found useful in the
control of many insect species of medical and veterinary importance. Research
work in this field is tilled now limited in our country.
At present there are no available effective drugs without toxicities. Although
ivermectin is safe and effective against both ecto and endo parasites of animals but
it is expensive. In contrast, the alternative cheapest and available source of drug is
herbal therapy. Experimental investigations, therefore, is imperative to assess the
therapeutic value of indigenous herbal plants and oil. From this point of view, this
research work has been designed to investigate the comparative efficacy of
indigenous medicinal plants (tobacco and neem oil) and ivermectin against
ectoparasites (ticks) of goats with their adverse effects, if any. Therefore, the
research work was undertaken with the following objectives:

i.

To investigate the efficacy of indigenous medicinal plant (Neem oil, Tobacco

and their combined) and Ivermectin against ectoparasites (ticks) in goats.


ii. To study the adverse effects of indigenous medicinal plant (Neem oil,
Tobacco and their combined) and Ivermectin against ectoparasites (ticks) in
goats.
iii. To study the effects of indigenous medicinal plant (Neem oil, Tobacco and
their combined) and Ivermectin on some hematological parameters against
ectoparasites (ticks) in goats.

CHAPTER 2
REVIEW OF LITERATURE
The main purpose of this chapter is to get up-to-date information regarding the
research works addressed here. Important information related to the present study
was represented below. The contributions made by numerous research workers are
compiled and the publications are documented in the references.
2.1 Effects of Tobacco against ectoparasites
Tobacco leaves contain more than 3.5% nicotine. They are widely distributed
throughout the world, mainly cultivates in Indonesia, Japan, China, India, Pakistan,
Afghanistan and Bangladesh. It originates in Argentina. Maximum tobacco
cultivate in America and their tobacco is best quality. Its main component is
nicotine (Nicotina tabacum) an alkaloid which is soluble in water and has an
antiparasitic activity. Once it was used as a drug to treat the ulcer and headaches.
Leaves and stems are used as a strong narcotics. Its extracted nicotine is used as
insecticides. Nicotine acts as an active anthelmintics.
Fajimi et al. (2012) Studied on the anti-louse activity of aqueous tobacco extract in
heavily parasitized West African dwarf goats. The goats were allotted into three
treatment groups of six animals each, viz: ivermectin treatment (subcutaneous
injection of 0.5mg/kg bodyweight ivermectin); tobacco extract treatment (topical
application of undiluted tobacco juice) and the untreated control. The assessment of
depth of infestation and cure was based on the geometric mean of the louse counts.
The geometric mean louse count decreased from 1690.71 to 2.62 (99.84% efficacy)
by day 14 in ivermectin treatment; while it decreased from 1627.39 to 0 (100%
efficacy) in the tobacco extract treatment. The geometric mean louse counts
continued to increase in the untreated control group. There were increases in live

weight gains in the ivermectin and tobacco treated groups against the
corresponding loss in weight recorded in the untreated control. At 78 days postchallenge employing two louse-infested goats, significant louse count could not be
registered in the tobacco-treated goats.
Aktar (2007) carried out an experiment to study the comparative efficacy of
ivermectin neem and tobacco. Thirty sheep of both sexes and aged upto 3 to 5 years
infested with ticks and lice were randomly divided into 6 equal groups. Group A
kept as infested control group. In group B, the efficacy of Invemectin (200 g/kg,
body weight) is 100% against ticks and lice after 7 days of treatment. In group C
and D, the efficacy of neem at 1% concentration is 56.25% and 63.71% against
ticks and lice respectively but at 2% concentration the efficacy is 64% and 71.73%
against ticks and lice respectively. In group E and F, the efficacy of tobacco at 1%
concentration is 60% and 65% against ticks and lice respectively but at 2%
concentration the efficacy against ticks and lice are 68.96% and 74.46%
respectively. Both neem and tobacco at 2% concentration shows higher efficacy
than at 1% concentration against ticks and lice. Tobacco shows higher efficacy than
neem in both 1% and 2% concentration against ticks and lice. TEC and Hb and
PCV level increased at 21st (p<0.01) and 28th (p<0.01) post treatment while ESR
values decreased significantly (p<0.01) at 21st and 28th post treatment.
Hossen et al. (2002) examined that tobacco vaseline, tobacco-termeric and Neemtobacco ointment were found to be 54.41%, 66.38% and 74.80% effective
respectively, whereas, ivermectin was found 100% effective against humpsor in
cattle.
Hossen and Mostofa (1999) evaluated the efficacy of ivermectin (ivomec) applied
at the dose rate of 200 g/kg body weight subcutaneously twice at 7-day intervals,
tobacco leaf extract (Nicotiana tabacum, 2%) four times at 2-day intervals topically
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and tobacco copper sulfate formulation (1% tobacco and 1% copper sulfate)
topically four times at 2-day intervals, against ticks and lice infestation in cattle.
The drugs were found 100% effective against the parasites.
Mannan et al. (1997) reported that the humpsore lesions were completely healed up
at 18th day with ivermectin and 15th day with Neguvon ointment but only 80%
lesions were healed up with Neem-tobacco ointment treatment at 45 th day of posttreatment in cattle. The author recommended that Ivermectin could be use against
humpsore in cattle in absence of Neguvon.
2.2 Effects of Neem (Azadirachta indica) against ectoparasites of goats
Muraleedharan et al. (2009) assessed the Acaricidal effect of certain
neem (Azadirachta indica) preparations against natural infestations of hard
ticks, Haemaphysalis and Hyalomma sp. on sheep and goats. These preparations
were applied on infested ear lobes and reduction of ticks was noted from the very
next day. Undiluted neem oil provided 100% reduction of ticks on day 3 to 5 on
sheep and 94.42% on goats on day 4. Aqueous extract of 25g neem kernel
processed in 100 ml water was reduced tick counts to 84.86% on day 10 on sheep.
One per cent aqueous emulsion of Neemrich I 80-EC recorded a reduction of 65.95
and 36.38% of ticks on sheep and goats, respectively on day 6.
Rahman et al. (2009) carried out an experiment to investigate the efficacy of
Ivermectin and Neem plants against ectoparasites in calves. They found that
therapeutic efficacy of Ivermectin was 100% against ectoparasites in calves on day
7, 14, 21 and 28 after the treatment of Ivermectin. Ivermectin and Neem showed
significant effectiveness at 7th, 14th, 21st and 28th day of post treatment. The effects
on TEC, Hb level was increased significantly at 7 th, 14th and 28th day of treatment
and PCV level was increased significantly on 28th day of treatment. On the other

hand indigenous medicinal plant Neem leaves shown the efficacy of 68% at day 28
and body weight of Ivermectin and Neem treated calves also increased. All the
calves after Ivermectin injection and Neem spray remained healthy, no adverse
effect and calves appetite increased, growth and coat color improved rapidly.
Khalid et al. (2005) studied the effect of medicinal plants (Neem and Pineapple)
Against Gastro-intestinal Nematodiasis in Sheep. The authors concluded that Neem
and Pineapple (10% water extract of leaves) may be used as moderate anthelmintics
against gastro-intestinal nematodiasis in sheep.
Haque et al. (2003) reported the leaves of the Neem Ganda, Mehedi and raw Halid
were applied as fresh aqueous pastes for the treatment of artificially produce wound
in guineapig. In comparison to their effect, Neem leaves paste showed the more
effectiveness where heating was completed on 15 days, halud paste and Ganda
leaves pastes showed the same results, where healting was completed on 17 days.
The Mehedi paste was the least effective in which heating occurred on 19 days.
This investigation suggested that Neem leaves fresh paste are more effective for
wound healing in guineapig.
Webb and David (2002) evaluated the use of Neem seed extract for controlling
common cattle ticks. Three bulls and three cows of the Tswana, Brahman and
Simmenlaler breeds were cleared of ticks using ether and hand picking techniques.
Each animal was then naturally infested with ticks. A 5% (w/v) water extract of
Neem seed kernel was applied at a rate of 5 g/kg body weight to various anatomical
sites on the animals selected for the experimental treatment, while the control
treatment animals were sprayed with tap water. Treated and control animals grazed
together and were mustered at weekly intervals for examination. Tick population
densities on animals treated with Neem seed extract were lower than on untreated
animals. Indigenous Tswana cattle harbored fewer ticks during periods of tick
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abundance than Brahman or Semmentaler cattle. It was concluded that Neem seed
extract is effective in controlling ectoparasites on livestock.
Chandrawathani et al., (2002) carried out a study on fresh leaves of the Neem tree
(Azadirachta indica), which were daily offered to groups of goat infected with the
nematode parasites, predominately Haemonchus contortus, for a 5 week period.
During this time, Neem leaf consumption, nematode faecal egg counts, and larval
development were regularly monitored. At the end of the trial, all animals were
slaughtered for the purposes of total worm recovery. The trial demonstrated that all
goat readily consumed Neem leaves. This had an effect of reducing both faecal egg
counts and larval recoveries, which was confirmed by a significant reduction
(p=0.00442) in the number of worms in the Neem fed animals.
Brelin (2002) observed that due to severe problems with anthelmintic resistance in
Malaysia and many other countries, there is need for alternative methods of treating
ruminants against gastrointestinal nematodes. This report is based on a trial where
the Neem tree (Azadirachta indica) has been tested and evaluated as a herbal
control agent against the Barbers poleworm (Haemonchus contortus). This report
contains a general literature section dealing with the effect of gastrointestinal
nematodes on grazing livestock, the importance of control, constraints to control
and alternative approaches to gastrointestinal nematode control. This is followed by
a section concerning the same issues but with specific emphasis on Malaysia,
which is followed by a chapter on the Neem tree. The trial was a pen trial, where
the effects of fresh Neem leaves on established worm burdens were studied. Twelve
goat were divided into three groups where one was control and the other two
received different amounts of fresh Neem leaves. It was found that the fresh Neem
leaves significantly reduced the numbers of H. contortus in the abomasums of the
treated goat. No dose-dependent effect was seen between the chosen doses.

10

Arunachal et al., (2002) experienced that Seventy-five lambs naturally infected


with gastrointestinal helminths were divided into five groups and given aqueous
extracts of Neem leaves, seeds and bark (GI-GIII) or standard dose of praziplus
(GIV). GV was used as control. Clinical efficacy was evaluated based on the
reduction in faecal egg count (epg) of Strongyle eggs by Stoll's method obtained
before the trial and every 10 days upon commencement of the experiment.
Anthelmintic efficacy of Neem extracts and praziplus were 53, 4938 and 87%,
respectively. Although reduction in epg was noticed in all the treatment groups, the
variation was statistically significant (P < 0.01) only on the 10th day posttreatment. The reduction in egg count indicates the anthelmintic properties of Neem
extracts.
Abdel-Shafy and Zayed (2002) studied the effects of the plant extract of Neem seed
(A. indica) on eggs, immature, and adult stages of H. anatolicum excavatum at
concentrations of 1.6%, 3.2%, 6.4% and 12.8%. The extract was found to have a
significant effect on the hatching rate of eggs. It significantly increased the hatching
rate during the first seven days post-treatment (DPT) giving incompletely
developed and dead larvae; however, it caused hatching failure at DPT 15, Neem
Azal F induced a significant increased in mortality rates of newly hatched larvae,
unfed with the extract concentrations. Although, it had no significant effect on the
moulting rates of fed nymphs, it caused malformation or deformities in 4% of
adults moulted. It was concluded that the concentrations of Neem Azal F which
may be used for commercial control of this tick species were 1.6 and 3.2.
Ravindra et al. (2000) assessed the efficacy of the herbal ectoparasiticidal
compound (AV/EPP/14) (containing the active ingredients of C. deodara, P.
glabra, A. indica, E. globulus and A. calamus) on 16 buffaloes and 8 cattle infested
with B. microplus, H. anatolicum anatolicum, H. marginatum isaaci and H.

11

drornedarii, by spraying the compound in 1: 4 dilution in tap water. Tick infested


buffaloes and cattle were sprayed five times at 6-day intervals (0, 6, 12, 18 and 24)
which resulted in elimination of 65.3%, 87.6%, 96.5%, 99.6% and 100% of the
ticks, respectively. The treated animals were free of ticks for a period of 30 days
after the last treatment.
Kalakumar et al. (2000) gave in vivo and in vitro trials to determine the acaricidal
activity of custard seed oil, Neem oil and pyrethrin against three tick species,
Boophilus microplus, Hyalomma anatolicum anatolicum and Rhipicephalus
haemaphysaloides. Custard seed oil and pyrethrin were found 100% efficacious,
whereas, Neem oil was only 60.75% effective in cattle and buffaloes infested with
ticks. Oviposition was inhibited in all tick species treated with custard seed oil
whereas females treated with Neem oil oviposited.
Pietrosemoli et al., (1999) investigated the antiparasitic effect of Neem
(Azadirachta indica) leaves when administered to grazing goat Neem leaves were
harvested, partially dried and mixed at various concentrations with maize flour,
wheat bran, salt, minerals, molasses and binding material to prepare blocks that
were offered ad libitum to 36 crossbred heifers, initial age 22.47 2.84 months and
weight 231.99 5.25 kg, for 98 days on a commercial farm in a tropical dry forest
area in Venezuela. Blocks containing 0, 10, 20 and 30% Neem leaves were
evaluated. Faecal samples were taken directly from the rectum 3 days before
treatment and 21, 28, 60 and 90 days after the initiation of treatment. Numbers of
nematode eggs per g of faeces (epg) were established using the McMaster
technique. Animals were weighed every 21 days. No differences between
treatments were found for epg before treatment and after 28 days. Statistical
differences between treatments were determined for epg on days 21, 60 and 90
(P<0.0002, P<0.0022 and P<0.0002, respectively) after the initiation of treatment.

12

There was no difference between blocks containing different concentrations of


Neem leaves. Treatment had no effect on weight gain. It was concluded that the
addition of Neem leaves to nutritional blocks can reduce the epg of grazing goat.
Mathew et al., (1998) carried out screening on crude extracts of leaves of Adhatoda
vasica [Justicia adhatoda] (collected from India) was carried out to determine
antifeedant and anthelmintic potential in comparison to that of seed kernels of
Azadirachta indica. All the extracts of Adhatoda vasica showed equi- or more
potent activities than Azadirachta indica. The formulation of the crude extract with
piperine exhibits enhancement in the anthelmintic activity confirming its
synergistic action.
Mannan (1997) reported that treatment of Stephamofilasial lesions of cattle by the
topical application with neem tobacco ointment (tobacco- 5%, neem 15% and
Vaseline 80%) twice daily for 45 days, reduced 80% of the stephanofilarial lesions.
Mostofa et al. (1996) stated that efficacy rate of imported anthelmintics between 80
and 100%. Thiophanate is reported to have reduced efficacy for nematodes,
particularly H. contortus. Studies on seeds of the indigenous medicinal plant neem
(Azadirachta indica) have shown an anthelmintic efficacy of 35 to 53% against
natural infections of gastro-intestinal nematodes.
Alam (1995) observed the therapeutic efficacy of Fenitrothion (Neguvon) and
Neem seeds against stephanofilariasis in cattle. Fourty clinical cases of
stephanofilarial dermatitis were selected to carried out the study. The animal were
divided into five groups (each group contain eight cattle). He treated the animal and
found the following results topically applied of fresh Neem seed ointment, Neem
seed ointment and 10% fenitrothion ointment were almost equally effective (100%)
against Stephanofilariasis in cattle. However the recovery was faster (within 18-

13

34days) with 10% Fenitrothion ointment followed by Neem seed kernel ointment
(27-42 days) and fresh Neem seed ointment (30-49 days). On the other hand 5%
Fenitrothion ointment was found almost ineffective against Stephanofilariasis in
cattle.
Hossain (1994) reported that sitaphal seeds and leaves, Supariseeds, Garlic bulb,
Palas seeds, Papaya seeds, male form rhizome and epical bulbs, Vawrang fruits,
leaves and barks, Karchi bark and Seeds, Neem leaves and fruits, Dried tobacco
leaves are commonly used as indigenous anthelmintics.
Hossain (1994) reported that pineapple, Acacia, Catachu, Neem, Palas, Batoshack
(Chenopodium album) Cherodendron infortunatum, Sefali leaves, Neasinde roots
can be used as anthelmintics.
Ahmed et al. (1994) used morental citrate (Banminth-II, Pfizer, at 10 mg/kg) and
Fenbendazole (Panacur Hoechst at 7.5 mg/kg body wt) orally with that of neem
(Azadirachta indica) seeds at 75 mg and 100 mg/kg body wt orally, were tested
against gastro-intestinal nematodiasis in sheep. The efficacy rates of Fenbendazole
(99.66%) and morental citrate (87.5%) were higher than neem seeds (35.2940.0%).
Rai et al. (1993) reported that Stephanofilarial dermatitis in cattle was cured if
smear topically with diethyl carbamazine citrate (DECC 20%, 2 applications 10
days apart), or daily with coumaphos ointment, tobacco liniment, sulfur ointment,
zinc oxide ointment or neem oil, or by i/m or s/c injection of 9-12 mg/kg
Levamisole hydrochlorides, or s/c injection of Ivermectin at 1 ml/50 kg body wt.
For the smaller sores (up to 2 cm diameter) Levamisole, DECC and Ivermectin
were satisfactory, and for the larger sores (20-30 cm diameter) Levamisole was
significantly better than all the other treatments.

14

Mostofa (1993) used an ointment made of 1% Neguvon 10% Neem leaves and
barks, 89% Vaseline to treat humpsore of twenty-five cattle. All the cattle were
cured within 30 days by topical application of the ointment twice daily.
Shilaskar and Parashar (1989) conducted an in vitro anthelmintic activity of 15
extracts of various indigenous plants (Andrographis paniculata, Azadirachta
indica, Butea frondosa, Caesalpinia crista, Piper betle, Psoralia corylifolia,
Swertia chirata, Vernenia anthelmintica) was evaluated against Ascaridia galli
collected from birds postmortem. Washed active worms were incubated with the
test materials at 410C and examined at 1, 2, 3 and 24 h for mortality, which was
indicated by loss of motility. Percentage mortality was determined using angular
transformation methods. Ether and alcohol extracts of seeds of B. frondosa and V.
anthelmintica, ether extract of seeds of C. crista, successive alcohol extracts of
seeds of P. corylifolia and essential oil from leaves of Piper betle had highly
significant anthelmintic activity, and alcohol extracts of whole plants of Swertia
chirata had significant activity.
Nath (1983) reported that water-washed neem seed kernel can be used as a cattle
feed for digestibility, feed consumption, weight gain and nutrition.
Baquar and Tasnif, (1975) stated the brief morphology that the Neem tree is 18-21
m in height, simple leaves (13-16) and lanceolate shape. The tree is found in
Bangladesh, India and in area of hot and humid climates. Chemical analysis of
Neem exact of different parts revealed a number of constituent among them the
major is azadiractin. The others are nimbidin, nimbasole nimbectin and acid. Neem
is famous for its medicinal properties. Azadirechtin is well known for its
anthelmintic effect. The seeds are emetic laxative and anthelmintic. The oil from
the seeds gives as an anthelmintic in doses of 30 minims to one drachm. Externally
the oil is applied as an antiseptic dressing in leprosy, suppurating glands, urticaria,
15

ringworm etc. and maggot-infested wounds. The bark is a bitter tonic, astringent,
antispasmodic and stimulant and anthelmintic. The leaf juice also acts as an
effective anthelmintic.
2.3 Effects of Ivermectin against ectoparasites
Ivermectin is a broad spectrum antiparasitic drug which is registered in over 60
countries. It is currently registered for use in horses, goats, swine, camels, reindeer,
bison, and dogs (Di Netta, 1989). Ivermectin is active against two major phyla of
animal parasites, the Nemathelminthes and the Arthropoda (Campbell et al., 1983).
Ivermectin has not been evaluated previously by the Joint FAO/WHO Expert
Committee on Food Additives.
2.3.1 Chemistry
Ivermectin (CAS-7-288-86-7) is a mixture of two compounds belonging to a class
of substances known as avermectins (See Figure 1). The chemical names are 5-0demethyl

22,

23

dihydroavermectin

A1a

and

5-0-demethyl-22,

23-

dihydroavermectin A1b. These are also known as 22, 23-dihydroavermectin B 1a and


22,23-dihydroavermectin B1b. Ivermectin contains at least 80% of 22,23dihydroavermectin B1a and less than 20% of 22,23-dihydroavermectin B 1b. The
avermectins are derivatives of pentacyclic sixteen-membered lactones. Within the
family of the avermectins, there exist two series, A and B, within which are two
structural subsets, designated 1 and 2, consisting of two homologs a and b.
Members of the A-series are methoxylated at the carbon atom in position five,
whereas the B-series compounds have an underivatized hydroxyl-group at this
position. Compounds of the 1-subset possess an olefinic bond between the two
carbon atoms C22 and C23; this double bond is hydrated in the 2-subset, resulting in
a hydroxyl group at position 23. This difference has a profound effect on the

16

conformation of the ring bearing these functionalities and causes subtle changes in
bioactivity (Chabala et al., 1980). The a- and b-homologs differ by their
substituents at position 25, with a-homologs having an isopropyl group, derived
from L-valine, and b-homologs possessing a sec-butyl group derived from L-isoleucine during biosynthesis. Avermectins are glycosides with a disaccharide
attached to the hydroxyl group at C 13. The two identical sugars have been identified
as L-oleandrose, a dideoxy-methyl-aldohexose.

Structure of 22, 23-dihydroavermectin B1a, the major component of ivermectin


Ivermectin (as its major component, 22, 23-dihydroavermectin B1a) is an off-white
powder that is highly lipophilic and hydrophobic. It dissolves in most organic
solvents but is poorly soluble in water. It is stable at room temperature in nonacidic solutions but is degraded by UV light.

17

2.3.2 Mode of action


It was originally thought that the macrolides endectocides increased the release of
-amino butyric acid (GABA) from synaptosomes of the nervous system. This, in
turn, opened GABA-gated chloride channels. It is now known that these
compounds open chloride channels in invertebrates via a specific binding site that
is glutamate-gated, although the binding site apparently occurs in close anatomic
proximity to GABA-gated sites, and the macrolide endectocides may potentiate
GABA-gated sites as well. About 50 percent of the effect of a macrolide
endectocide can be reversed with picro toxin, a GABA antagonist active at the
chloride channel. In nematodes, the synapse between inter-neurons and excitatory
motor neurons is the primary site of action, whereas the myoneural junction is the
primary site in arthropods. In either case the chloride ion influx lowers cell
membrane resistance and causes a slight hyper polarization of the resting potential
of postsynaptic cells. This makes neurotransmission more difficult so that
transmission of stimuli to muscles is prevented, resulting in a flaccid paralysis of
affected parasites followed by their death or expulsion.
2.3.3 Formulation
Ivermec injectable formulation is a solution containing 2% Ivermectin (w/v) in an
organic vehicle for topical administration to goat, goat and swine. A 0.27% (w/v)
injectable formulation is used in young pigs. Ivermec oral for goat contains 0.4%
ivermectin (w/v) in an aqueous solution for use as a drench. A more dilute
formulation containing 0.08% ivermectin (w/v) is used as a drench in goat and
calves (Ivermec).

18

2.3.4 Pharmacokinetics
Pharmacokinetic studies with ivermectin are summarized by Fink and Porras
(1989). The specific formulation used, the route to administration, and the animal
species to which it is administered affect the pharmacokinetics of ivermectin. The
biological half-life (t1/2) of ivermectin in plasma following IV administration of 300
g/kg to goat is 2.8 days. IV administration to goat gives a similar biological halflife (t1/2 =2.7 days) to that in goat but a lower plasma concentration due to a greater
volume of distribution in goat than in goat (1.9 vs. 4.61/kg). Ivermectin is
eliminated more rapidly in dogs (t1/2 =1.6-1.8 days)
Usual injectable administration at the dose rate of 400g/kg & subcutaneous
administration of the commercial formulation of ivermectin to goat, goat etc. at a
dose rate of 200 g/kg results in a longer biological half-life (t1/2 =8 days) than IV
administration due to slow absorption from the injection site. A peak plasma
concentration (CP) of 44 ng/ml occurs at 2 days after S/C injection. Clinically
significant anthelmintic efficacy persists for approximately 2 weeks after S/C
injection and more than 2 weeks for pour-on formulation depending upon parasite
species. Oral dosing in goat results in a t1/2 of 3-5 days.
Following administration, ivermectin residues are lowest in brain and highest in
liver, bile, and fat (Chiu and Lu, 1989). Depletion half-lives were 4.8 and 7.6 days
for liver and fat, respectively, in goat and goat. Tissue redistribution patterns are
similar for goat, swine and rats, but depletion half-lives for liver and fat are shorter
in goat and rats than in goat or swine. The parent drug is the major liver residue for
3, 5, 7 and 14 days after dosing in rats, goat, swine and goat, respectively.

19

Fecal excretion is the main route of elimination, accounting for 98% or more of
excreted ivermectin, with the remainder appearing in the urine, except upto 5% of
the dose may be excreted in the milk of lactating animals.
2.3.5 Anthelmintic spectrum in goats
The efficacy of Ivermec pour-on (Ivermectin 500 g/kg body wt) in goat, is
highly effective for control of arthropod ectoparasites which include groups
(Hypoderma ovis, H. lineatum, Oestrus ovis), mites (Sarcoptes bovis, Psoroptes
ovis), and sucking lice (Linognathus vituli, Hematopinus eurysternus and L.
Pedalis). It is slightly less effective in controlling chewing lice (Damilinia spp.)
and the goat ked (Melophagus ovis). Ivermectin administered at a dose of 0.2mg/kg
orally or subcutaneously and 400-500 g/kg body wt. Pour-on in goat results in
efficacies of 97-100% against adult and 4 th stage larval forms of respective species
of

Haemonchus, Ostertagia (including inhibited L4 in goat), Cooperia,

Trichostrongylus (including T. axei), Strogyloides, Bunostomum, Nematodirus,


Trichuris, Oesophagusomum, Dictyocaulus and ovine Chabertia ovina.
Aziz et al. (2012) studied the comparative efficacy of ivermectin and diazinon
against ectoparasites in sheep. For this purpose 25 sheep heavily infested with ticks
and lice were randomly divided into 5 equal groups, i.e. groups B, C, D, E and F.
Another group of 5 sheep free from parasitic infestation were also selected and kept
as uninfected control (group A) and group B was kept as infected control group.
The sheep of groups C and D were treated with recommended (200 g/kg b.w.) and
higher than recommended (300 g/kg b.w.) doses of ivermectin subcutaneously. The
sheep of groups E and F were treated with diazinon (spray) at recommended (0.1%
soln.) and higher than recommended (0.2% soln.) doses. The efficacy of ivermectin
was found 100% whereas diazinon was 82-85% effective against ectoparasitic
infestation in sheep. Ivermectin and diazinon increased the feeding efficiency as
20

compared to infected control groups. No adverse effects was observed following


both the dose of ivermectin and recommended dose of diazinon. In this study, the
hematological parameters, i.e. TEC and Hb% were increased significantly (p<0.01)
in all treated four groups to the extent of 35-70% and 10-21% respectively. On the
other hand, ESR values were decreased significantly (p<0.05).
Hassan et al. (2012) assessed the efficacy of selective anthelmintics against ecto
and endoparasites of Black Bengal goats. Goats were treated with cevamec-1%
(ivermectin), endex-1500 (triclabendazole along with levamisole), and a placebo
(untreated). A reduction of eggs per gram count was very significant from day 7
(91.3% reduction) through day 28 (100%) with the treatment of ivermectin. The
reduction rate of eggs per gram was also significant with the treatment of
triclabendazole along with levamisole (75.8%-94.7%). Both of the drugs were
equally significant against endoparasitic infections of goats in this study (P < 0.05;
t-test). The percentage efficacy of ivermectin was also recorded against ticks and
lice of goats from day 7 through day 28 of the trial period. The packed cell volume
and hemoglobin levels increased significantly (P < 0.05) in both of the treated
groups which was indicative of effective treatments of those trial drugs.
Islam et al. (2005) the ectoparasiticidal efficacy of ivermectin and biskatali in
calves on the basis of lice and tick count in a selected area (20 square cm). The
authors concluded that ivermectin showed efficacy 100% and 2% biskatali spray
showed 100% efficacy.
Hanif et al. (2005) suggested that therapeutic efficacy of ivermectin was 100%
against ectoparasites (lice and tick) after 7 days of treatment. The authors
concluded that ivermectin is a very much suitable broad spectrum anthelmintic for
sheep and its pour on formulation may be used effectively and conveniently to treat
ectoparasite infestations.
21

Khalid et al. (2004) suggested that Vermic may be used as a broad spectrum
anthelmintic against gastro-intestinal nematodiasis in sheep. After treatment with
Vermic, Total Erythrocyte Count (TEC), Hemoglobin (Hb) content and Packed
Cell Volume (PCV) were increased significantly (P<0.01 and P<0.05) in sheep.
Islam et al. (2003) studied the efficacy of ivermectin against gastro-intestinal
nematodes and ectoparasites in calves. The authors concluded that ivermectin is
100% effective against lice infestation 20 th, and 25th post-treatment day. Body
weight increased 6.63% in calves after the treatment with ivermectin.
Hossen et al. (2002) examined that tobacco vaseline, tobacco-termeric and Neemtobacco ointment were found to be 54.41%, 66.38% and 74.80% effective
respectively. Whereas, ivermectin was found 100% effective against humpsor in
cattle.
Hannan et al. (2001) tested the efficacy of ivermectin in goats infected with
gastrointestinal nematodes, lice and ticks. The authors suggested that ivermectin @
500 g/kg body weight was 1400% effective against these ectoparasites and
removed all nematode eggs even form the severely infected goats after 7 days of
treatment. All infected goats gained significantly (P<0.01) higher body weight with
compared their pretreatment body weight. Observation at different intervals after
treatment also caused significant (P<0.001) changes in different haematological
values (Hb%, PCV, ESR).
Yazwinski et al. (1997) reported 100% effectiveness of Ivermectin against parasitic
infection. No adverse reaction was observed in trial animals.
Ryan et al. (1997) conducted an experiment to determine the mean total weight
gain of Ivermectin treated goats naturally infected with helminth infection. The

22

mean total weight gain was 33.9 kg greater for the Ivermectin treated goats than
those of untreated control animals (p<0.02); a 34% increase. It was concluded that
the Ivermectin S/C influenced the weight gain by limiting the impact of subclinical
parasitism.
Imrul (1997) reported the efficacy of ivermectin against gastro-intestinal
ectoparasites and ectoparasites like ticks and lice was100% in goat. Body weight of
ivermectin treated goat was increased by 6.205% on day 28 post treatment. Hb,
PCV and ESR values were changed significantly after treatment with ivermectin in
goat.
Ballwebner et al. (1997) studied the average daily weight gain of goat treated with
ivermectin. The average daily weight gain of each individual was recorded as
0.125-272 kg compared to non treated controls. The difference was statistically
significant (p<0.05) in 3 of the 4 studies.
Vizzio and Caro (1995) studied the efficacy of ivermectin and lufenuron against
flea infection. Ivermectin was given subcutaneously at a dose rate 0.1 ml/kg b. w.
every 20 days (3 or 4 times) and lufenuron P.O at 1 tablet every 30 days (4-6 times)
and reported that no live fleas were observed on the animals after the first week of
treatments. The authors also concluded that a combination of ivermectin and
lufenuron is an effective treatment for flea infections in dogs.
Taylor et al. (1995) reported the effectiveness of ivermectin against Ostertagia
ostertagi and Cooperia oncophora in goat, during a comparative study of early and
mid grazing season suppressive anthelmintic treatment for first year grazing season
goats and their effects on natural and experimental infection. Five groups of goat
were taken and groups 2 and 3 were treated by ivermectin and groups 4 and 5 were

23

treated with morantel. Over the 2 years observation period groups 2 and 4 showed
significantly better weight gain than other groups.
Stacey et al. (1995) reported that the goats treated with Ivermectin had significantly
gained higher weight than untreated goats.
Sangwan et al. (1995) studied the efficacy of ivermectin Ivomec, 0.4 mg/kg b.w.)
on goat infection with Psoroptes ovis at karnal, India and reported that 3
subcutaneous injection of ivermectin cured the animal completely. The author also
reported that diazinon (Neocidol, Malathion, Cythion) also cured the animals
but was more laborious than the pour method of ivermectin.
Titcherer et al. (1994) studied the efficacy of Ivermectin (Ivomec injectable)
against sucking and biting lice of cattle. Calves were naturally infected with
Linognathus vituli and Bovicola bovis (Damalinia bovis). Ivermectin was given
topically. A louse count was performed on each animal at 7, 14, 21 and 28 days
after treatment and lice were enumerated by species. Ivermectin was highly
effective against Linognathus vituli and none of these lice were found on any of the
treated calves from 7 days after treatment. Ivermectin was also highly effective
against Bovicola bovis and no lice were found after treatment.
Thompson et al. (1994) studied the effectiveness of Ivermectin against lice
infestation in long wooled goat. In these cases, naturally occurring infestation of
lice, Bovicola ovis were successfully treated with Ivermectin Jetting fluid in Merino
and Corriedale goat.
Thompson et al. (1994) studied the effectiveness of ivermectin against lice
infection in goat. In this case naturally occurring infestation of lice, Bovicola ovis
were successfully treated with ivermectin jetting fluid in Merino and corriedole
goat.
24

Umur and Irmak (1993) studied the comparative efficacy of Ivermectin and phoxim
against naturally sarcoptic mange in goat. The author treated 15 clinically affected
goat with 2 subcutaneous injections of Ivermectin (Ivomec at 0.2 mg/kg body
weight) at a 21 days interval and watched another 15 animals twice in a 0.2%
phoxim emulsion (sebacial) at a 10 days interval and reported that after a single
treatment no Sarcoptes bovis (S. scabiei) were found in 12 (80%) of the 15
Ivermectin treated goat and in 13 (86.6%) of the 15 treated with phoxim. The
second treatment eliminated the mites, and none were found 28, 42 and 56 days
later.
Rossanigo and Silva (1993) treated one group of goats with ivermectin monthly
(G1), another group strategically for 2 months (G2) and left another group as
untreated control (G3) and observed live weight gain and G1 adults were 5.92 kg
heavier than G2 adults and 4.21 kg heavier than G3 adults.
Mottelib (1993) studied the efficacy of ivermectin (Ivomec ) against naturally
infected with sarcoptes in camels. The drug was injected subcutaneously at 200
mg/kg b. w., repeated after 15 days and reported that the treatment reduced the
number of mites present on the 18 th day of therapy. Blood values returned to normal
after the second treatment although anaemia, leukocytosis, eosinophilia and
monocytosis were present before treatment. The author also stated that ivermectin
gave good results against nematode infection in some camels.
Kar et al. (1993) evaluated the efficacy of ivermectin in 60 patients in Orissa, India
infested with bancroftian filariasis and microfilaraemia. Ivermectin was given as a
single oral dose at 4 dosages level (20, 50, 100 and 200 g/kg b. w) and reported
that blood micorfalariae were cleared in all patients at all dosages within 1 to 14
days.

25

Foreyt (1993) treated the experimentally infected goat Psoroptes sp. at a dose rate
of 1 mg/kg body weight in a pelleted ration daily for 7 consecutive days and
reported that mites could not found in the goat 1, 2 or 3 months after treatment and
lesions of the affected goat resolved within 6-8 weeks after treatment.
Brien et al. (1993) studied the efficacy of Ivermectin against goat scab and treated
30 ewes by subcutaneous injection of Ivermectin (1% ivomec ) @ 200 g/kg b. w.
on 2 occasions days part and reported 97% animals were cured by a single injection
and 100% recovery was reported by a second injection 10 days later.
Zajac et al. (1992) treated 4 groups of goat infected with nematodes @ 200 g/kg
b. w. through S/C injection at monthly interval (traditional) at O, 3, 6, 9 and 12
weeks after pasture turnout (strategic) or 0, 8, 16. 20 and 24 weeks after turnout
(Tactical) of with levamisole (Tramisol) orally at 8 mg/kg b. w. at 0, 3, 6, 9 and 12
weeks turnout. They reported that none of these 4 treatments provided satisfactory
control of parasites although traditional Ivomec treatment gained significantly more
weight than other 2.
Cosoroaba et al. (1992) compared the efficacy of some ectoparasiticides in goat.
The theraputic effectiveness of Ivomec (ivermectin), Bayticol 6% (flumethrin).
Bayticol injectable and Lindavet E 15 were studied. The Ivomec in a single dose of
200 g/kg body weight S/C, Neocidal 600 EC and bayticol 6% in a solution of
0.1% applied with a brush, 3 times at an interval of 10 days, assumed 100%
efficacy in Chorioptes ovis parasitism. Bayticol injectable at the dose rate of
1ml/10kg b. w. 2 treatments at an interval of 10 days and Ivomec 200 g/kg b. w. in
a single dose had 100% efficacy in Damalinia bovis (Bovicola bovis) parasitism.
Similar results were obtained in Linognathus vituli parasitism.

26

Yeruham et al. (1991) treated 20 goat infected with Chorioptic bovis with Ivomec
(at 0.2 mg/kg body weight) 10 Of Which were treated once and 10 twice (2 week
interval) and reported that Ivomec did not give satisfactory result.
Kennedy (1990) studied the effect of Ivermectin on gaining body weight of goats
in USA. Goats treated with Ivermectin injectable preparation (200 g/kg S/C)
significantly gained more weight than the control animals.
Guerrero (1989) reported the better effect of Ivermectin against nematodes
(Hypoderma SP) Lice and ticks (including boophilus microplus) in grazing cattle. A
total of 19 trials involving 5124 cattle of different ages and breeds treated with S.C
injection of Ivermectin performed better and were economically more productive
their untreated control or animal treated with other types of anthelmintics and
external parasiticides.
Laka et al. (1988) treated the goat for scabies with Ivomec (ivermectin) at a dose
of 1 ml/50 kg b. w. and reported a very good acaricidal effect against ectoparasites.
Yamov et al. (1987) investigated the efficacy of Ivomec against goat parasites and
reported 97% efficacy of Ivermectin in goat against Oestrus ovis infestation in goat
at recommended doses and 94.8% and 89.5% against Dictyocaulus and
Haemonchus respectively.
Sevcikova et al. (1987) carried out trials on the effectiveness of 1% Ivermectin
preparation in goat, horses, goat, pigs, dogs, cats, rabbits, foxes, deer and Zoo
animals in Czechoslovakia. The authors reported high efficacy of Ivermectin
against ecto and endo parasites.
Rastov et al. (1986) carried out a comparative study of therapeutic effects of some
preparations against Ectoparasite in goat and goat in Bulgaria and reported 68.8%,

27

63.6% and 72.7% cure by Neocidol 60 (Diazinon), Chlorophos (Trichlorofon)


and IvomecR (Ivermectin at 300 g/kg b. w. subcutaneous) respectively.
Ninkov and Savin (1986) conducted and experiment on the activity of Ivermectin
against goat scab in Yugoslavia and reported 100% efficacy against goat scab
following subcutaneous injection from 12 days after the administration with no
adverse effect of the drug.
Danek et al. (1985) studied the efficacy of Ivomec against sarcoptic mange in goat
and goat and psoroptic mange in goat and Haematopinus eurysternus infestation in
goat. The author stated the high efficacy of Ivomec R against gastrointestinal
helminths.
Kashai et al. (1984) carried out Ivermectin Pour-on trials in goat and pigs
infected with endo-and ecto-parasites in Hungary and reported that a S/C injection
of Ivermectin at 0.2 mg/kg b.w. cured 65.2% young goat mixed strongly infection
of 142 to 250 EPG, as do judged by fecal tests on day 19 after treatment. The
weight gain of the treated group exceeded those in untreated groups by 6.8 kg.
Ivermectin at the same doses reduce adult ostertagia population by 99.98% and
mucosal L4

population by 99.90%. Ivermectin reduced Ascaris suum infection

intensity by upto 99%. Ivermectin (as ivomec) was also effective against psoroptic
mange in goat.

28

CHAPTER 3
MATERIALS AND METHODS
3.1 Study area
The experiment was taken to determine the comparative efficacy of neem oil,
Tobacco and Ivermectin (Vermic) against ectoparasites in goat. The experiment
was conducted from January 2014 to June 2014, at Sylhet Government Goat Farm,
Sylhet and with the co-operation of Department of Pharmacology and toxicology of
Sylhet Agricultural University, Sylhet.
3.2 Collection of animals
25 Goat of both sexes aged between 12 to 24 months were primarily selected in this
study. All the goats were examined for the presence of ectoparasites and the
efficacy of Ivermectin and indigenous medicinal plant (Neem oil and Tobacco)
were studied against these naturally infested ectoparasites.
3.3 Collection of drugs
Injectable Ivermectin (Vermic , S/C formulation, Techno Drugs Limited,
Bangladesh) was selected for the experiment and purchased from local market at
Sylhet.
3.4 Collection and Processing of tobacco
The tobacco leaves were collected from a tobacco shop in Sylhet town and brought
to the laboratory of the Department of Pharmacology. After collection and bringing
to the laboratory, all leaves was washed in running tap water and cut into small
pieces. Firstly the leaves materials was dried in shade and then they were dried in
the oven at 55-600C for 48-72 hours to gain constant weight. Neem oil was

29

purchased from Modern Herbal Company, Sylhet and brought to the laboratory of
the Department of Pharmacology.
3.5 Preparation of tobacco dust
Dusts were prepared by pulverizing the dried leaves with the help of a manual
grinder, haman dista. A 25-mesh diameter sieve was used to obtain fine dust and
preserved them into airtight plastic container, till their use in extract preparation.
3.6 Preparation of tobacco solutions
Solutions of tobacco extract were prepared on the basis of gm/ml concentration.
From the dust the aquous extract was prepared. For this the dust of 20 g was added
with the 100 ml distilled water and the mixture was homogenized. The mixture was
then evaporated while boiling at water bath at 50 0C. Final volume of the mixture
was then reached at 10 ml/20 gm and stored at this concentration in refrigerator as a
stock solution until its used for preparation of solution. The solution of 1% was
prepared by adding required amount of water in the stock solution. All solution
prepared at the day of experiment.
3.7 Other chemicals
Other chemicals were saturated salt solution, normal saline (0.9%), anticoagulant
(sodium citrate 3.8%), Hayems solution, 0.14% Hydrochloric acid solution and
their were prepared in the Pharmacology Laboratory, Faculty of Pharmacology,
Sylhet Agricultural University, Sylhet.
3.8 Selection of goats and experimental schedule
Among twenty five goats, twenty of heavily infested with different ectoparasites
were selected and randomly divided into 5 equal groups (group A, B, C, D and E).
Each group consisting of 5 goats. Goat of group A was kept as infected control.
30

Group B was treated with Neem oil. Group C was treated with Tobacco solution.
Group D was treated with combined spray with Tobacco and Neem oil. Rest Group
E was treated with Ivermectin. Goat of different groups was treated with indigenous
medicinal plant (neem and tobacco) and ivermectin as per following schedule.
Group of sheep

Drug with dose and route

Group A

Infected without treatment (control)

Group B

4 ml of neem oil topically applied in the affected area of


body once daily for 28 days

Group C

Spray with 1% tobacco solution

Group D

Combined spray with 1% tobacco solution and 4 ml of


neem oil

Group E

Ivermctin @ 200 g/kg body weight subcutaneously on


day 0 and 14

3.9 Detection of ectoparasites


The ectoparasites (ticks) were detected by physical examination of the animals and
the presences of the ectoparasites were recorded.
Ticks infestation was examined physically on individual animal. The procedure of
examination was to count the adult ticks of paralumbar region with an area of 54
i.e. 20 square cm. The selected 20 square cm. were marked with a permanent colour
and ticks within this area were counted at pretreatment and post-treatment days.

EXPERIMENTAL LAYOUT

31

Total animals (goats) 25

Group A (5)
control infected

Group B (5) spray


with Neem oil

Group C (5) spray


with Tobacco
solution.

Group D (5) combined


spray with Tobacco
and Neem oil

Clinical parameters

Heamatological test

Severity of
infestation

Body
weight

Clinical signs

Group E (5)
Ivermectin @
200g/kg b.
wt. SC

TEC

Hb

PCV

After injection of ivermectin and topical application of herbal drugs (neem oil and
tobacco solution) goats of 4 groups with one control group were closely observed
for 28 days and 14 days of post treatment and following parameters were studied.
a) Clinical parameters (severity of infestation, feeding efficiency, conditions of
hair coat, body weight and adverse effects).
b) Hematological parameters (TEC, Hb and PCV)
3.10 Clinical parameters

32

3.10.1 Severity of infestation


The severity of infestation of ectoparasites (ticks) was observed by counting the
number of ectoparasites in a selected area of the individual goat. The ectoparasites
were detected by visual examination of the goats and the number of parasites was
recorded. The ticks within this area were counted at pre-treatment (0 day) and (7 th,
14th, 21st, 28th) treatment period.
3.10.2 Feeding efficiency
Feeding efficiency of treated and untreated (infected control) groups was compared
at pretreatment (0 day), treatment (7 th, 14th, 21st and 28th) and 14 days post treatment
period.
3.10.3 Hair coat
Hair coat of the treated goats as well as the untreated infected control group was
physically examined by visual examination before and after treatment as per abovementioned schedule i.e., 0, 7th, 14th, 21st and 28th day of treatment and also on 14
days of post treatment period.
3.10.4 Body weight
To observe the effects of the three drugs on this parameter body weight of each
goats of treated and non-treated control group was taken before treatment (day 0)
and 28 days of treatment and on 14 days of post treatment period.

3.11 Hematological parameters

33

Blood samples were collected from Jugular vein of the goats of treated and control
group in vials containing anticoagulant (Sodium Citrate 3.8%) at day 0, 7, 14, 21,
28 of treatment period and also on 14 days of post treatment to determine the
effects three patent drugs and two herbal preparations on the following
hematological parameters.
3.11.1 Total erythrocyte count (TEC)
Thomas red blood cell pipette was used. The pipette was filled up to 0.5 mark with
blood and diluting fluid (Hayems solution) was then drawn till it reached the 101
mark. The contents were thoroughly mixed for 2 minutes. The dilution of the
contents was 1:200. The neubauer-counting chamber and cover slip were cleaned
and dried properly. The cover slip was placed on the chamber in proper position.
The content of the pipette was again shaken and 2 or 3 drops of the fluid were
expelled. The chamber was then filled with the contents of the pipette at a angle of
about 400 to the chamber so that the fluid was flown under the cover slip by
capillary action. The ruled area at the chamber was filled completely, taking care
that excess did not run into the troughs and no air bubble appeared under the cover
slip, The counting chamber was then placed under microscope and examined first
with a low power objectives (10x) to ensure that there was and even distribution of
the cells. The cells were then counted with the aid of high power objectives (100x).
The central squares of the counting were counted in the far corner squares and one
central square of the chamber. The counting and calculation of red blood cells were
performed as per methods indicated by Coffin (1955). The number of RBC was
calculated as follows:
Number of RBC = No. of cell counted x 10,000 and the result was expressed in
million/cu.mm.

34

3.11.2 Determination of hemoglobin (gm %)


The hemoglobin estimation was performed by the acid Haematin method with the
Hellige Hemometer (Coffin, 1955) Hydrochloric acid (N/10) solution was taken in
the special graduated tube up to its 2 mark. The special Sahli pipette was filled with
well mixed oxalated blood up to 20 mark and blood on the side of the pipette was
wiped out by cotton. The content of the Sahli pipette was expelled into the special
graduated diluting tube and thoroughly mixed. The tube was then allowed to stand
for 10 minutes for development of acid haematin. Distilled water was then added
drop by drop and each time mixed with the solution with the help of stirring rod
until the colour of the solution matched with the standard colour of the comparator.
The result was read as per method described by Coffin (1955).
3.11.3 Packed cell volume (PCV)
The PCV was determined as per method described by Coffin (1955). The wintrobe
haematocrit tube was filled up with well-mixed blood by special loading pipette up
to 10 mark. Then the tube was centrifuged at 3,000 rpm for half an hour and
reading was taken.
3.12 Statistical analysis
Data obtained from the experiment on body weight, hematological parameters
(TEC, PCV, Hb) were analyzed statistically using students t test.

35

CHAPTER 4
RESULTS
A research work was conducted to evaluate the comparative efficacy of tobacco,
Neem oil and ivermectin (injectable formulation) against tick infestation in goatss
by observing the severity of parasitic infestation i.e. number of ectoparasites with
their death % after giving treatment. In addition, effects of patent and herbal drugs
on some clinical (severity of infestation, color of hair coat, body weight and
adverse effects) and hematological (TEC, Hb and PCV) parameters in goats were
also investigated.
4.1 Comparative efficacy of Ivermectin (injectable), neem oil and tobacco
leaves against tick infestation in goats
4.1.1 Clinical Parameters
4.1.1.1 Severity of infestation i.e. number of ectoparasites with death % after
giving treatment
Efficacy of patent and two herbal drugs against tick infestation in goats is shown in
the Table 1 and figure 1

36

Table 1

37

Figure 1. Efficacy neem oil, tobacco leaves, neem oil-tobacco leaves combined
and ivermectin against tick infestation in goats

38

In control group A, ticks were gradually increased on 7 th, 14th, 21th and 28th on 14
days of treatment. On the other hand, groups of B, C, D and E treated with Neem
oil, tobacco leaves, combined with neem oil-tobacco leaves and ivermectin showed
variable efficacy against ectoparasitic infestation by killing ticks i.e. Neem oil56.25%, tobacco leaves-60% and combined with neem oil-tobacco leaves-68.96%
whereas ivermectin showed 100% efficacy in group E within 7 th day of treatment
(shown in Table 1).
4.1.1.2 Rough hair coat
On day 0 (pre-treatment) the hair coat of the infected goats (group B, C, D and E)
was rough. After giving treatment with neem oil, tobacco leaves, combined with
neem oil-tobacco leaves and ivermectin, the hair coat started to become smooth and
shiny gradually upto14th day of post treatment period. Among them, patent drugs
showed better results than herbal drugs.
4.1.1.3 Body weight
In control group (A), body weight of goats was decreased up to 12.48% on 28 th day
of treatment. On the other hand, the body weight was increased in all groups (B, C,
D and E) to the extent of 1.60%-7.88%. So, body weight of goats increased
significantly in all treated groups on 28th day of treatment.

39

Table 2

40

Figure 2. Body weight (BWT)

41

4.1.1.4 Adverse effects


No visible adverse effects were observed in four treated groups.
4.1.2 Efficacy of neem oil against tick infestation in goats
Group B goats treated with neem oil with the dose rate at 4 ml topically applied in
the affected area of body once daily for 28 days was 56.25% effective against tick
infestation in goats. On day 0 the number of mean value of tick was 6.400.50
and on 7th, 14th, 21st and 28th day after the treatment, the number of mean value of
tick was 5.000.54, 4.000.54, 3.200.58 and 2.80.37 respectively.
4.1.3 Effect of tobacco leaves (spray) against tick infestation in goats
Group C goats treated with tobacco leaves with the dose rate at 1% concentration
were effective up to 60% only against tick infestation in goats. On day 0 the
number was 6.000.44 and on 7th, 14th, 21st and 28th day after the treatment the
number of mean value of tick was 5.000.44, 3.800.37, 3.200.20 and 2.400.24
respectively.
4.1.4 Effect of neem oil-tobacco leaves (spray) combined against tick
infestation in goats
Group D goats treated with neem oil-tobacco leaves (spray) combined with the
dose rate at 4ml neem oil and 1% tobacco leaves concentration was found to be
68.96% effective against tick infestation in goats. On day 0 the number was
5.800.48 and on 7th, 14th, 21st and 28th day after the treatment the number of mean
value of tick was 4.600.50, 3.600.24, 2.800.20 and 1.800.20 respectively.

42

4.1.5 Efficacy of ivermectin (Injectable, S/C) against tick infestation in goats


Ivermectin at recommended dose (200 ug/kg, S/C) was 100% effective in group E
against tick infestation and on day 0, the number of mean value of tick was
5.40.67 and on day 7th, 14th, 21st and 28th day after the treatment of ivermectin, no
tick were found within the selected area of goats.
4.2 Effects of neem oil, tobacco leaves and ivermectin on hematological
parameters
a) Total erythrocytes count (TEC million/cu. mm of blood)
The effects of neem oil, tobacco leaves and ivermectin on TEC of goats is shown in
Table 3 and Figure 3.

43

Table 3

44

Figure 3. Efficacy of neem oil, tobacco leaves, neem oil-tobacco leaves


combined and ivermectin on TEC (million/cu. mm) in goats

45

The mean values of TEC on 0 day in control group A was 7.550.07 which
decreased gradually and it was 6.850.01 on 28th day of treatment.
On the other hand, neem oil, tobacco leaves, neem oil-tobacco leaves combined and
ivermectin increased the TEC values in 4 groups of treated goats (B, C, D and E).
The pre-treatment mean values of erythrocytes (million/cu.mm of blood) were
6.810.04, 7.880.07, 7.130.06 and 7.280.03 in the treated groups of B, C, D
and E respectively. On the 28th day of the post-treatment, the mean values of TEC
were significantly (p<0.01) increased up to 8.630.06, 9.750.03, 9.320.03 and
10.500.06 in the group of B, C, D and E respectively.
b) Hemoglobin content
The effects of neem oil, tobacco leaves, neem oil-tobacco leaves combined and
ivermectin on Hb (gm %) of goats is shown in Table 4 and Figure 4.
The mean values of Hb (gm) on 0 day in control group A was 7.700.07 and
decreased to 7.140.06 on the 28th day of treatment.
Like TECs, Hb (gm%) increased significantly (p<0.01) in 4 groups (group B, C, D
and E) of neem oil, tobacco leaves, neem oil-tobacco leaves combined and
ivermectin treated goats. The pre-treatment (0 day) of Hb (gm %) were 7.510.09,
7.760.13, 7.260.15 and 7.280.09 in groups of B, C, D and E respectively. On
the 28th day of the post-treatment, the mean values of Hb (gm %) were significantly
(p<0.01) increased up to 8.210.09, 8.710.09, 8.420.06 and 8.720.10 in the
group of B, C, D and E respectively.

46

Table 4

47

Figure 4. Efficacy of neem oil, tobacco leaves, neem oil-tobacco leaves


combined and ivermectin on hemoglobin content (gm%) in goats

48

c) Packed cell volume (PCV %)


The effects of neem oil, tobacco leaves, neem oil-tobacco leaves combined and
ivermectin on PCV (%) of goats is shown in Table 5 and Figure 5.
The mean values of packed cell volume of infected control group A on 0 day was
30.740.05 which decreased to 29.800.06 on the 28th day of treatment.
The treated neem oil, tobacco leaves, neem oil-tobacco leaves combined and
ivermectin were significantly (p<0.01) increased the PCV value in 4 treated groups
(group B, C, D and E). The pre-treatment mean values of PCV were 30.570.09,
30.710.08, 30.530.08 and 30.340.07 in groups of B, C, D and E respectively.
On the 28th day of the post-treatment, the mean values of PCV were 31.270.09,
31.590.07, 31.660.09 and 32.40.09 in the group of B, C, D and E respectively.

49

Table 5

50

Figure 5. Efficacy of neem oil, tobacco leaves, neem oil-tobacco leaves


combined and ivermectin on PCV (%) in goats

51

CHAPTER 5
DISCUSSION
After the treatment of neem oil, tobacco leaves, neem oil-tobacco leaves combined
and ivermectin, all the five treated and control group of goats were kept for 28
days. The ectoparasites were detected by visual examination of the goats and the
number of ticks was recorded. The procedure of examination was to count the ticks
of abdominal region within an area of 5 cm 4 cm (20 square cm). The selected 20
square cm were marked with a permanent color and the ticks within this area were
counted at pre-treatment (0 day) and post-treatment (7 th, 14th, 21th and 28th) period.
The hematological parameters were investigated at 7 days interval (0 day, 7 th day,
14th day, 21st and 28th day of treatment).
The efficacy of neem oil was found to be 56.25% against tick infestation in goats.
Sanjib (2007) reported that efficacy of neem oil was 70% against tick infestation in
goats which is 19% lower than the result of the present study in goats.
Muraleedharan et al. (2009) reported that undiluted neem oil provided 94.42%
reduction of ticks on day 4 on goatss which is 39% lower than the result of the
present study on day 28 on goats. Kalakumar et al. (2000) reported that neem oil
was found 60.75% effective in cattle infested with ticks which are 7% lower than
the result of the present study in goats. However, Nahar (2004) reported that
aqueous extract of neem showed 86.67% efficacy in vitro on spray method using
2% concentration.
The efficacy of tobacco at 7th days interval was evaluated against tick infestation in
goats. Tobacco at 1% concentration showed 60% efficacy against tick infestation.
Due to lack of available literature compare and contrast cannot be made. However,
Hossen and Mostofa (1999) reported that the efficacy of tobacco-leaf extract (2%
solution) and tobacco-copper sulfate (1% tobacco and 1% copper sulfate)
52

formulation against ticks and lice were found to be 100% effective at 21 day post
treatment in cattle.
The efficacy of neem oil-tobacco leaves combined at 7 th days interval was
evaluated against tick infestation in goats. Combined spray with 1% tobacco
solution and 4 ml of neem oil showed 68.96% efficacy against tick infestation. Due
to lack of available literature compare and contrast cannot be made. However,
Hossen et al. (2002) examined that tobacco vaseline, tobacco-termeric and Neemtobacco ointment were found to be 54.41%, 66.38% and 74.80% effective
respectively against humpsor in cattle.
The efficacy of ivermectin (200 g/kg body weight) against ticks was found to be
100% effective at 7th post treatment day and remained effective up to 28 th day of
post treatment. This result was in agreement with the earlier record of Aziz et al.
(2012), Hassan et al. (2012), Fahima (2003), Yazwinski et al. (1997), Sangwan et
al., (1995), Vizzio and Caro (1995), Pedroso et al., (1994), Thomson et al., (1994).
But some other early workers found 72.7% (Kostov, 1986), 86.6%, (Unur and
Irmark, 1993), 97% (O Brien et al. 1993), and 90% (Imrul, 1997) efficacy.
On comparing the efficacy of neem oil, tobacco leaves, neem oil-tobacco leaves
combined and ivermectin on the basis of disappearing of ticks, ivermectin was
found to be best drug and was effective within 7 days of post treatment. On the
other hand, On the other hand, other drugs showed comparatively less efficacy
against tick infestation within 7 days of post treatment.
Hematological changes were determined pre and post treatment with neem oil,
tobacco leaves, neem oil-tobacco leaves combined and ivermectin. The Hb content
and TEC of treated group (B, C, D and E) significantly (p<0.01) increased to the
extent of 9.32-19.78% and 26.72-44.23% respectively. Significant (p<0.01)

53

changes of PCV values were observed in the treated group of goats and this might
be due to removal of parasites from the surface of the body. Changes in
hematological parameters were also observed by other researcher Aziz et al.
(2012), Hassan et al. (2012), (Hanif et al., 2005), (Fahima, 2003), (Imrul, 1997)
and (Kumar and Jashi, 1992).
The hair coat of all goats treated with neem oil, tobacco leaves, neem oil-tobacco
leaves combined and ivermectin were observed smooth and shiny at the end of
post-treatment period. Rough and discolored hair coat in control groups due to
severe parasitic burden on the skin surface. Due to removal of the ticks from the
skin surface on 28 day of treatment, hair coat of treated goats were almost alright.
Body weight changes were determined pre and post treatment with neem oil,
tobacco leaves, neem oil-tobacco leaves combined and ivermectin. The body
weight of treated group (B, C, D and E) significantly (p<0.01) increased to the
extent of 1.60-7.88% respectively. Ryan et al. (1997) found that the mean total
weight gain was 33.9 kg greater for the Ivermectin treated goats than those of
untreated control animals (p<0.02); a 34% increase. However, Ivermectin S/C
influenced the weight gain by limiting the impact of subclinical parasitism.

54

CHAPTER 6

SUMMARY AND CONCLUSION


The experiment was performed to determinei. The efficacy of indigenous medicinal plant (Neem oil and Tobacco) and
Ivermectin against ectoparasites (ticks) in goats.
ii. The adverse effects of indigenous medicinal plant (Neem Oil and Tobacco)
and Ivermectin against ectoparasites (ticks) in goats.
iii. The effects of indigenous medicinal plant (Neem Oil and Tobacco) and
Ivermectin on some hematological parameters against ectoparasites (ticks) in
goats.
Twenty five goats were selected and divided into 5 equal groups (A, B, C, D and E)
and each group consisting of 5 goats. The group of goats A was kept as infected
control group. The group B was treated with 4 ml of neem oil topically applied in
the affected area of body once daily for 28 days. The group C was treated with 1%
tobacco solution. The group D was treated with 1% tobacco solution and 4 ml of
neem oil combined. The group E was treated ivermectin (200 g/kg body weight,
S/C).
In group B, the efficacy of neem oil at 4 ml topically applied in the affected area of
body was found 56.25% respectively. In group C, efficacy of tobacco at 1%
concentration was found 60% against tick. In group D, the combined efficacy of
neem oil-tobacco leaves were found 68.96% against tick respectively. The goats of
group E, therapeutic efficacy of ivermectin observed 100% against tick. During the
study of hematological parameters, it was observed that TEC, Hb and PCV content
were increased significantly (P<0.01).

55

Rough hair coat become smooth and shiny on 28 day of treatment. The body weight
of treated group (B, C, D and E) significantly (p<0.01) increased to the extent of
1.60-7.88% respectively.
The efficacy of neem oil, tobacco leaves, neem oil-tobacco leaves combined and
ivermectin on the basis of disappearing of ticks, ivermectin was found to be best
drug and was effective within 7 days of post treatment. On the other hand, On the
other hand, other drugs showed comparatively less efficacy against tick infestation
within 7 days of post treatment.
Although ivermectin is 100% effective against ectoparasites but the price of
ivermectin is very high, application of the drug required skilled person and
unavailable in the rural area of Bangladesh. Neem and tobacco and their combined
are cost effective; application is very easy and found everywhere in Bangladesh.
So, it is concluded that neem and tobacco and their combined could be used as an
ectoparasiticide instead of ivermectin and other chemical substances.

56

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