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In vitro anthelmintic efficacy of some indegeneous medicinal plants against gastrointestinal nematodes

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Abstract

The prevalence of natural gastrointestinal nematodes was observed in cattle during the period from June, 2004 to May, 2005 in Sadar upazila of Mymensingh district. The prevalence of gastrointestinal nematodes was 84.1% (rainy seasons-97%, summer-85.5% and winter seasons-69.8%). The prevalence of strongyles (Haemonchus sp., Trichostrongylus sp., Oesophagostomum sp. and Mecistocirrus sp.), Bunostomum sp., Strongyloides sp., Trichuris sp. and Capillaria sp. were 63.9%, 26.3%, 21.5%, 17.3% and 24.5%, respectively. Water extracts of 20 indigenous plants(neem, tobacco plant, barbados lilac, betel leaf, pineapple, jute, turmeric, garlic, devil's tree, papaya, lime tree, dodder, white teak, conessi tree, bitter gourd, sweet basil, white verticillia, pomegranate, sage, chaste tree) showed potential in vitro activities against adult parasites. Out of these, 20 plant extracts, 10 plants (neem, tobacco, barbados lilac, betel leaf, pineapple, jute, turmeric, garlic, dodder and bitter gourd) showed 100% efficacy against adult worms, 4 plants (devil's tree, papaya, white verticillia and chaste tree) showed 90- 98% and others (lime tree, white teak, conessi tree, sweet basil, pomegranate and sage) showed below 90%.
J. Bangladesh Agril. Univ. 7(1): 57–61, 2009 ISSN 1810-3030
In vitro anthelmintic efficacy of some indigenous medicinal plants
against gastrointestinal nematodes of cattle
M. R. Amin, M. Mostofa, M. E. Hoque and M. A. Sayed
Department of Pharmacology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh
Abstract
The prevalence of natural gastrointestinal nematodes was observed in cattle during the period from June, 2004 to
May, 2005 in Sadar upazila of Mymensingh district. The prevalence of gastrointestinal nematodes was 84.1%
(rainy seasons-97%, summer-85.5% and winter seasons-69.8%). The prevalence of strongyles (Haemonchus
sp., Trichostrongylus sp., Oesophagostomum sp. and Mecistocirrus sp.), Bunostomum sp., Strongyloides sp.,
Trichuris sp. and Capillaria sp. were 63.9%, 26.3%, 21.5%, 17.3% and 24.5%, respectively. Water extracts of 20
indigenous plants(neem, tobacco plant, barbados lilac, betel leaf, pineapple, jute, turmeric, garlic, devil's tree,
papaya, lime tree, dodder, white teak, conessi tree, bitter gourd, sweet basil, white verticillia, pomegranate, sage,
chaste tree) showed potential in vitro activities against adult parasites. Out of these, 20 plant extracts, 10 plants
(neem, tobacco, barbados lilac, betel leaf, pineapple, jute, turmeric, garlic, dodder and bitter gourd) showed
100% efficacy against adult worms, 4 plants (devil's tree, papaya, white verticillia and chaste tree) showed 90-
98% and others (lime tree, white teak, conessi tree, sweet basil, pomegranate and sage) showed below 90%.
Keywords: Anthelmintic, Gastrointestinal nematodes, In vitro, Prevalence, Medicinal plants
Introduction
Parasitism is an important limiting factor that responsible for deteriorating the health and productivity
of livestock. The agro-ecological and geo-climatic conditions of Bangladesh are highly favorable for
the growth and multiplication of parasites. As a result about 50% apparently healthy cattle populations
are affected with different species of parasites (Garrels, 1975). Infections by gastrointestinal helminth
parasites of livestock are among the most common, which are considered as economically important
diseases of grazing livestock (Perry et al., 2002). They are characterized by lower outputs of animal
products (meat, milk, hides and skins), manure and traction, which all have impact of the livelihood of
small holder farmers (Perry and Randolph, 1999). The production performances of these cattle are
low, because of wide spread occurrence of pathogenic parasites. Parasitic diseases are considered
important causing enormous economic losses through morbidity and mortality in livestock. Among the
parasitic diseases, gastrointestinal nematodes such as Haemonchus spp., Trichostrongylus spp.,
Cooperia spp., Oesophagostomum spp., Trichuris spp. and Strongyloides spp. are most common
(Qadir, 1981; Rahman and Mondal, 1983). Imported synthetic anthelmintics are considered the only
effective way of controlling parasitic infection. However, as these are expensive and unavailable,
livestock producers are not interested to use these anthelmintics. Furthermore, some serious
disadvantages of using those anthelmintics, notably the development of resistance to helminth
parasites (Waller and Prichard, 1985) against various anthelmintic compounds and classes, as well as
their residues and toxicity problems (Kaemmerer and Butenkotter, 1973) poses hazards to livestock
development and public health. For these reasons, interest in the screening of medicinal plants for
their anthelmintic activity has remained of great scientific interest despite extensive use of synthetic
chemicals in clinical practices (Akhtar et al., 2000). Until today very little works are performed in our
country to investigate the anthelmintic properties of indigenous medicinal plants in cattle. Considering
all of these constraints, this work was undertaken with following objectives: i) screening of medicinal
plants having in vitro anthelmintic activity, ii) prevalence of gastrointestinal nematodes in cattle of
Sadar Upazila of Mymensingh district.
Materials and Methods
Collection of fecal samples
Fecal samples were collected from 1200 randomly selected cattle of Sadar upazila of Mymensingh
district (100 month) were examined at various seasons (rainy seasons, summer and winter). Fecal
samples were collected from the rectum of cattle by hand and kept in polythene bags and examined
58 Gastrointestinal nematodes of cattle
by Floatation method (Rahman et al., 1996). The simple test tube flotation method is a qualitative test
for the detection of nematode and cestode eggs and coccidia oocysts in the faeces. It is based on the
separating of eggs from faecal material and concentrating them by means of a flotation fluid with an
appropriate specific gravity.
Collection of indigenous medicinal plants
Indigenous medicinal plants (Table-3) from different location of Bangladesh were collected and the
plants material were dried in shade and then dried off in the hot air oven at 55-60
0
C to gain constant
weight.
Preparation of plants powder
Powders were prepared by pulverizing the dried indigenous medicinal plants with the help of electric
grinder. A 25-mesh diameter sieve was used to obtain fine dust and preserved them into airtight
plastic container, till their use for extract preparation. Previously prepared plants powders were used
for preparation of plants extract. Ten grams of each powder were taken in a 500 ml beaker and
separately mixed with 100 ml of distilled water. Then the mixtures were stirred for 30 min by a
magnetic stirrer (6000 rpm) and left as such for next 24 hrs. The extracts were then filtered through a
fine cloth and again through filter paper (Whatman No. 1). The filtered material was taken into round
bottom flask and then concentrated by evaporation of water from filtrate in a water bath at 50
0
C till it
reached the final volume of 10 ml. After the evaporation of water from filtrate, the condensed extracts
were preserved in tightly corked-labelled bottle and stored in a refrigerator until used for screening of
anthelmintic activity.
Preparation of stock solution
Stock solutions of plant extracts were prepared by diluting the condensed extracts with water.
Different concentrations of each category of plant extracts were prepared by dissolving them in the
water prior to anthelmintic screening.
Collection and maintenance of adult gastrointestinal nematodes in the laboratory
Collection of parasites from abomasum were done by following standard procedure as described by
Taylor (1934) and Bell (1957). Viscera of cattle were collected from local market .The abomasums
were opened through its lesser curvature with the help of scissors. The contents were emptied in
glass jars containing normal saline. The abomasi were thoroughly washed and cleaned off ingesta
and put in a different jar containing normal saline and left for an hour or two to release the
attachments of parasites from the wall of the abomasum. The mucosal surface of the abomasum were
rubbed carefully between the fingers to remove any remaining worms adhering to the abomasal wall.
Finally the abomasal mucosa was examined with the help of a magnifying glass for any remaining
parasites still adhering to the mucous lining of abomasum. The contents were washed several times
with water and continued till the worms were free from debrises. The final wash was made with the
normal saline. The sediments were examined in large petri-dishes over a black background. The
parasites were collected with the help of curved needle and kept in normal saline. The parasites were
cleared off debris by brushing with camel hairbrush or shaking in normal saline and identified
according to the keys described by Rahman et al., (1996) and Yamaguti (1958). They were kept in
petri-dishes containing normal saline incubated at 37
0
C.
In vitro screening of plant extracts for anthelmintic activity
Screening of water extracts of plant at various concentration viz. 25 mg/ml, 50 mg/ml and 100 mg/ml
were performed in the petri-dishes containing adult live stomach worms of cattle collected from
slaughter house in Phosphate buffer saline (PBS). PBS (100ml) containing 50 adult worms (both male
and female) were pipetted in 3 petri-dishes at ratio 0.1 ml, 0.05 ml and 0.025 ml water extracts were
then added, respectively. The drug-parasite petri-dishes were incubated for three hours at room
temperature and the efficacy was observed by counting the dead parasites and expressed in
percentages (%).
Amin et al. 59
Results and Discussion
The prevalence of gastrointestinal nematodes was highest in the month of June and July. The
prevalence of gastrointestinal nematodes in cattle are shown in the Table 1 and Table 2. Highest
prevalence of gastrointestinal nematodes were found in rainy seasons (July-October) followed by
summer (March-June) and winter seasons (November-February). Prevalence of strongyles
(Haemonchus sp., Trichostrongylus sp., Oesophagostomum sp. and Mecistocirrus sp.) infection was
highest followed by Bunostomum sp., Strongyloides sp., Trichuris sp. and Capillaria sp. Waruiru et al.
(2001) observed the epidemiology of gastrointestinal nematodes of dairy cattle in Central Kenya. The
total worm burden in the animals was highest during the rainy season (March-June and October-
December) and lowest during the dry seasons (July-September and January-February). This result is
in conformity with earlier reports made by Keyyu et al. (2006),
Jager et al. (2005), Soca et al. (2003)
and Yildirim et al.
(2000).
Table 1. Prevalence of gastrointestinal nematodes in cattle in Kotoally thana of Mymensingh
district during different seasons
Prevalence percentage
Name of Parasites
Whole year
Rainy
seasons
Summer
Winter
seasons
Gastrointestinal nematodes 84.08% 97% 85.5% 69.75%
Strongyles (Haemonchus sp., Trichostrongylus sp.,
Oesophagostomum sp. and Mecistocirrus sp.)
63.92% 71% 66.25% 52%
Bunostomum sp. 26.33% 29% 27.25 % 22.75%
Strongyloides sp. 21.5% 24.25% 23.25% 17%
Trichuris sp. 17.25% 19.5% 18% 14.25%
Capillaria sp. 24.5% 27% 26% 20%
Table 2. Prevalence of Gastrointestinal nematodes in cattle in Kotoally thana of Mymensingh
district on monthly basis
Month
Gastrointestinal
nematodes
Strongyles Bunostomum Strongyloides Trichuris Capillaria
January 62% 46% 20% 14% 12% 18%
February 73% 55% 25% 16% 14% 20%
March 74% 57% 22% 18% 13% 22%
April 82% 64% 26% 24% 17% 24%
May 86% 68% 29% 26% 20% 28%
June 100% 76% 32% 25% 22% 30%
July 100% 72% 33% 28% 20% 28%
August 96% 75% 30% 22% 23% 30%
September 97% 70% 28% 25% 18% 26%
October 95% 67% 25% 22% 17% 24%
November 76% 58% 24% 21% 15% 23%
December 68% 49% 22% 17% 16% 19%
60 Gastrointestinal nematodes of cattle
The in vitro efficacy of different concentrations of water extracts of 20 plants against adult
gastrointestinal nematodes are shown in the Table 3. Water extract of 20 indigenous plants showed
potential in vitro activities against adult parasites. The efficacy of water extract of these plants at the
concentrations of 25 mg/ml and 50 mg/ml was much lower than that of concentration of 100 mg/ml
except tobacco plants. Tobacco plants (25 mg/ml and 50 mg/ml) showed 100% in vitro efficacy
against gastrointestinal nematodes of cattle. Neem, barbados lilac, betel leaf, papaya, dodder, bitter
gourd and white verticillia (50 mg/ml) showed 70-80% in vitro efficacy against gastrointestinal
nematodes of cattle. Turmeric (50 mg/ml) was 88% effective in vitro against gastrointestinal
nematodes of cattle. The plants (100mg/ml) had highly significant activity (90-100%) against adult
gastrointestinal nematodes in vitro were : neem (leaves and bark), tobacco plant (leaves), barbados
lilac (leaves and bark), betel leaf (leaves), pineapple (leaves), jute (leaves), turmeric (rhizome), garlic
(bulbs), devil's tree (leaves), papaya (leaves), dodder (whole plant), bitter gourd (leaves and seeds),
white verticillia (leaves) and chaste tree (leaves). Iqbal et al. (2006) observed in vitro anthelmintic
activity of Nicotiana tabacum leaves against gastrointestinal nematodes of sheep. The in vitro
inhibitory effect was evident from the paralysis and/or mortality of worms noted at 6-hours post-
exposure. Raje and Jangde (2003) found in vitro anthelmintic activity of decoction of Nicotiana
tabacum against Haemonchus contortus of goats.
Table 3. In vitro anthelmintic efficacy of different concentrations of water extracts of 20
indigenous medicinal plants against adult gastrointestinal nematodes of cattle
Indigenous plants
Percent non-motile adult worms at
different concentrations of water extracts
of plant
Sl. No.
Local name Scientific name English name Part used 25 mg/ml 50 mg/ml 100 mg/ml
1. Nim
Azadirachta indica
Neem Leaves 22 74 100*
Nim
Azadirachta indica
Neem Bark 32 76 100*
2. Tamak
Nicotiana tabacum
Tobacco plant Leaves 100* 100* 100*
3. Ghora Nim
Melia azedarach
Barbados lilac Leaves 26 78 100*
Ghora Nim
Melia azedarach
Barbados lilac Bark 28 74 98*
4. Paan
Piper betle
Betel leaf Leaves 22 72 100*
5. Anaras
Ananas comosus
Pineapple Leaves 52 66 100*
Anaras
Ananas comosus
Pineapple Fruit 18 40 64
6. Deshi pat
Corchorus capsularis
Jute Leaves 24 58 100*
7. Halud
Curcuma longa
Turmeric Rhizome 42 88 100*
8 Rashun
Allium sativum
Garlic Bulbs 50 80 100*
9. Chhatim
A
lstonia scholaris
Devil's tree Leaves 36 62 94*
10. Pepe
Carica papaya
Papaya Leaves 46 78 90*
Pepe
Carica papaya
Papaya Seeds 14 28 68
11. Lebu
Citrus aurantifolia
Lime Tree Leaves 20 38 56
12. Swarnalata
Cuscuta reflexa
Dodder Whole plant 32 72 100*
13. Gamari
Gmelina arborea
White Teak Leaves 26 44 74
14.
Kurchi
Holarrhena
antidysenterica
Conessi tree Leaves 22 48 82
15. Karalla
Momordica charantia
Bitter Gourd Leaves 24 80 100*
Karalla
Momordica charantia
Bitter Gourd Seeds 20 60 98*
16. Tulsi
O
cimum basilicum
Sweet basil Leaves 16 56 84
17. Dondakalos
Leucas aspera
White verticillia Leaves 42 70 94*
18. Dalim
Punica granatum
Pomegranate Leaves 30 52 86
19. Bhui-Tulsi
Salvia plebeja
Sage Leaves 36 64 88
20. Nishinda
Vitex negundo
Chaste Tree Leaves 26 56 92*
*Considered strong wormicidal
Amin et al. 61
It may be concluded that the medicinal plants and/or its extracts may be used against gastrointestinal
nematodes of cattle in Bangladesh. The pharmacokinetics together with its toxic effects need further
studies.
Acknowledgements
The financial support of BAURES, Bangladesh Agricultural University, Mymensingh for conducting the
experiment is gratefully acknowledged.
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... Then cut into small pieces and taken a plastic jar. Then perform air-dried and finally sun dried for 3 days on the roof by covering a piece of cloth as prevention oxidation such as antioxidants and others chemical components (Amin et al., 2009). Clove was cleaned and be prepared for use. ...
... Dried bitter gourd and clove dust was prepared with the help of a blender (Sujan et al., 2008). A 25-mesh diameter seize was prepared to obtain fine dust and were preserved them into air-tight plastic container until being used (Amin et al., 2009). c) Preparation of Crude methanol extract (CME) Crude methanol extract (CME) was prepared from the selected three medicinal herbs according to the standard herb extraction methods (Gilani et al., 2004). ...
... f) Herbal anthelmintic dose Herbal anthelmintic dose was prepared for in vivo screening by adding required amount of distilled water after weighting stock solution (Amin et al., 2009). For in vivo screening combine herbal anthelmintic dose was given 1 ml/kg (100mg/ml) body weight for this study. ...
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This study aimed to evaluate the efficacy of garlic and turmeric against the prevalence of gastrointestinal parasites in cattle. Selected 25 cattle of different ages infected with parasites were brought under the study. These animals were treated with three different types of herbal tablets made of turmeric, garlic, and mixed (turmeric-garlic), respectively, and a commercial anthelmintic, LT-Vet. Fecal egg count determined parasite prevalence through EPG count. Fecal samples and body weight were examined before treatment and on the 7 th , 14 th, and 28 th day of the treatment for each animal. Prevalence of parasites was found at 37.588% in adults and 62.412% in yearlings before treatment. Most of the Farmers that were 83.33% used local herbs as a primary medication for their cattle. According to the research output, garlic had an elevated inhibitory effect against parasitic growth between turmeric and garlic. A significant reduction in EPG count was found on 7 th , 14 th , and 28 th day of treated cattle with garlic, turmeric, mixed and LT-Vet, respectively. Bodyweight increased by 6.96kg, 5.38kg, 4.98kg, and 1.242kg on average for mixed, turmeric, garlic, and LT-Vet treated cattle. The present study reveals that garlic and mixed had remarkable effects while turmeric has a moderate impact against gastrointestinal parasites in cattle.
... Recently, herbaceous plants have been reported against several ectoparasitic arthropods. Curcumin is known for having antiparasitic and antimicrobial properties, it was an effective compound against gastro-intestinal nematodes when dosing cattle at 100 mg/mL, whereas when dosing curcumin to goat from 0.2% to 0.6% of the daily ration for 60 days, a remarkable decrease in fecal egg shedding of gastro-intestinal nematodes were recorded (Amin et al. 2009;Afrin et al. 2016;Wuest et al. 2017). So, the main objectives of this study are to evaluate the insecticidal effects of curcumin against G. intestinalis in donkeys as well as enzyme assay, comet assay and scanning electron microscopy of larvae exposed to six different concentrations of curcumin. ...
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... Parasitism has been ranked high among factors that threaten livestock production worldwide (Perry et al., 2002). Moreover, drugs needed for the treatment of endoparasites of humans or livestock are usually expensive and often out of reach for the populations of developing countries (Amin et al., 2009). ...
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The aim of the present study was to determine the in vitro effects of selected plants extracts used by small stock farmers in the Blouberg district, Limpopo Province against helminth parasites. Plant materials were collected and dried at room temperature in the laboratory; thereafter hot and cold water extracts were prepared from Cassia abbreviata, Schotia brachypetala, Senna italica, Pappea capensis and Peltophorum africanum and tested on egg hatch, larval development larval mortality bio-assays against Haemonchus contortus. At the lowest concentration of 2.5 mg/mL Peltophorum africanum inhibited the highest percentage of 25 and 22% eggs from hatching for hot and cold water respectively; P. africanum was also responsible for the highest inhibition percentage of larval development with 86 and 80% larval development inhibition for hot and cold water plant extracts respectively. The highest rates of mortality were also recorded for Peltophorum africanum and Senna italica hot water extracts, with 68% and 65% mortality rates within two hours at the lowest concentration and 63% and 64% for cold water extracts respectively. It was evident that the tested medicinal plant extract showed significant in vitro anthelmintic activity at different concentrations; therefore the extracts can be considered both ovicidal and larvicidal. Keywords: Aqueous extracts, Ethnoveterinary medicine, Haemonchus contortus
... The plant Artemisia herba-alba (Chukun in Amharic), which belongs to the Asteraceae family, is used as an anthelmintic agent as well as for other common applications in folk medicine [12,13]. From the Lythraceae family, Punica granatum (Roman) is used against GI nematodes [14]. P. granatum has been used in natural and holistic medicine to expel tapeworms and treat various ailments [15,16]. ...
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... The plant Artemisia herba-alba (Chukun in Amharic), which belongs to the Asteraceae family, is used as an anthelmintic agent as well as for other common applications in folk medicine [12,13]. From the Lythraceae family, Punica granatum (Roman) is used against GI nematodes [14]. P. granatum has been used in natural and holistic medicine to expel tapeworms and treat various ailments [15,16]. ...
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Gastrointestinal nematodes (GINs) are the major limiting factor for the successfulness of livestock production throughout the world. Emergence of resistance strains as well as scarcity and high cost of the currently available drugs has led to the evaluation of other alternative helminth control options, mainly from plants. The current study is aimed at investigating the in vitro anthelmintic efficacy of crude methanolic extracts of two traditionally important medicinal plants, Artemisia herba-alba and Punica granatum, against Haemonchus contortus using adult motility assay (AMA) and egg hatch inhibition assay (EHIA). Four graded concentrations of the extracts were tested for both the AMA (10, 5, 2.5, and 1.25 mg/mg) and EHIA (0.1, 0.25, 0.5, and 1 mg/mL) in replicates. Albendazole and phosphate-buffered saline (AMA) or distilled water (EHIA) were used as the positive and negative controls, respectively. The crude extracts of A. herba-alba and P. granatum exhibited a potential anthelmintic activity at all dose levels in a concentration- and time-dependent fashion. The highest concentration (10 mg/mL) of all the extracts caused a significantly (p
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