Muhammad Nisar Khan

Anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) against trichostrongylid nematodes of sheep.

ETHNOPHARMACOLOGICAL RELEVANCE Ziziphus nummularia (Rhamnaceae) and Acacia nilotica (Fabaceae) are being used as anthelmintics in ethnoveterinary medicinal system of Pakistan. AIM OF THE STUDY Present study was conducted to determine the anthelmintic activity of Ziziphus nummularia (bark) and Acacia nilotica (fruit) in order to justify their traditional use in veterinary medicine. MATERIALS AND METHODS In vitro anthelmintic activity of crude methanolic extract (CME) of both the plants was determined against Haemonchus contortus by the adult motility assay, the egg hatch test and the larval development assay. In vivo anthelmintic activity was evaluated in sheep naturally infected with gastrointestinal nematodes by administering increasing doses of crude powder (CP) and CME (1.0-3.0 g/kg). RESULTS Both the plants exhibited dose- and time-dependent anthelmintic effects by causing mortality of worms, and inhibiting egg hatching and larval development. Acacia nilotica (LC(50)=512.86 and 194.98 microg/ml) was found to be more potent than Ziziphus nummularia (LC(50)=676.08 and 398.11 microg/ml) in egg hatch test and larval development assay, respectively. In vivo, maximum fecal egg count reduction (84.7%) was recorded on day 13 post-treatment in sheep treated with Ziziphus nummularia CME (3.0 g/kg) followed by 78.5% on same day with Acacia nilotica CME (3.0 g/kg). CONCLUSIONS These data show that both Ziziphus nummularia and Acacia nilotica possess anthelmintic activity in vitro and in vivo, justifying their use in traditional veterinary medicine in Pakistan.

Gastrointestinal helminthiasis: prevalence and associated determinants in domestic ruminants of district Toba Tek Singh, Punjab, Pakistan

The objective of the present study was to determine the prevalence and associated determinants (e.g., sex, age, on-farm management and husbandry) of gastrointestinal (GI) helminths in the domestic animals of district Toba Tek Singh, Punjab, Pakistan. For this purpose, 1,140 cattle, 1,140 buffaloes, 660 goats, 840 sheep, and 156 camels were randomly selected and their fecal samples were screened every other week for a year using a modified floatation technique. The samples positive for strongyle-type eggs had the parasite species identified using coproculture. It was found that the prevalence of GI helminths was significantly higher (P < 0.05) in sheep (44.17%; 371/840) than in other livestock. Sheep were followed in order by goats (40.15%; 265/660), buffaloes (39.82%; 454/1,140), and cattle (33.68%; 384/1,140). The important helminth species identified were Fasciola (F.) gigantica, Fasciola hepatica, Haemonchus contortus, Toxocara vitulorum, Trichostrongylus spp., Oesophagostomum spp., Ostertagia spp., Cooperia spp., Strongyloides spp., Moniezia spp., and Trichuris spp. The prevalence of GI helminths except F. hepatica and F. gigantica was significantly higher in grazing animals, females (P < 0.05) and young (P < 0.05) of all the host species when compared with stall-fed animals, males and adults, respectively. Using ponds and rivers/canals as drinking water were found to have significant influence (P < 0.05) on the prevalence of GI helminths. The results provide a baseline data for planning future research and control strategies against GI helminthes.

Bovine fasciolosis: prevalence, effects of treatment on productivity and cost benefit analysis in five districts of Punjab, Pakistan.

The present study reports the prevalence, effects of treatment and cost benefit analysis of bovine fasciolosis in five districts of Punjab Province viz Sargodha, Jhang, Muzaffargarh, Lodhran and Layyah. From each of the five districts, 80 animals were selected and fortnightly screened through standard coprological procedures for a period of one year for the presence of eggs of Fasciola species. Of 4800 faecal samples analyzed, 1222 (25.46%) were found positive for fasciolosis. The occurrence of Fasciola (F.) gigantica (22.40%) was higher (P<0.05) than F. hepatica (3.06%). Highest month-wise prevalence (P<0.05) of fasciolosis was found in winter (39.08%) followed in decreasing order by spring (29.50%), autumn (20.33%) and summer (12.92%). District-wise prevalence of fasciolosis was highest (P<0.05) in Sargodha (40.31%) and lowest in Layyah (11.77%) while other districts were having intermediate values of prevalence of fasciolosis. Species-wise prevalence of fasciolosis was found higher (P<0.05) in buffaloes (30.50%) as compared to cattle (20.42%). However, there were no age and sex-related differences (P>0.05) in prevalence of fasciolosis. A strong positive association of grazing (OR=1.81), mixed farming of small and large ruminants (OR=1.39), stagnant pond bathing (OR=2.24) and river/canal bathing (OR=2.06) was found with the prevalence of fasciolosis as compared to stall feeding, separate farming of small and large ruminants and rivers/canal/tap water bathing, respectively. Post-treatment average milk increase of 0.62 L per animal per day with 0.35% higher fat was observed in fasciolicide-treated animals with the cost benefit ratio of 3.9. The results provided significant data on the epidemiology of five districts of Punjab province which may be helpful for the planners and small holder dairy farmers for control of fasciolosis in the study districts.

An account of the botanical anthelmintics used in traditional veterinary practices in Sahiwal district of Punjab, Pakistan.

AIM OF THE STUDY The present study was aimed at documentation of botanical anthelmintics used in the traditional veterinary practices in Sahiwal district of Punjab, Pakistan. MATERIALS AND METHODS In rapid rural appraisal, 331 traditional veterinary healers (TVH) were identified as key respondents in the study area followed by participatory rural appraisal for data collection using a well-structured questionnaire. Information was collected through interviews, focused group discussions and field visits over a period of 2 years. RESULTS A total of 49 traditional recipes, with 41 plant species representing 39 genera and 27 families, were recorded for the treatment of helminthosis in animals. Most frequently used plants (> or = 5 times) were Brassica campestris L. and Mallotus philippinensis (Lam.) Muell.-Arg. and most frequently used families (> or = 5 times) were Brassicaceae, Euphorbiaceae and Solanaceae. Most frequently used part of the plant was leaves (n=10) followed in order by seeds (n=9), whole fruit (n=5), aerial parts and whole plant (n=4), fruit (n=3), bulb (n=2) and bark, rhizome, stem, stem plus root and twigs (n=1). Five recipes out of 49 (10.2%) contained more than one plant species and rest 44 (89.8%) contained single plant species. CONCLUSIONS Twenty out of 41 plants (48.78%) are reported for the first time for their traditional use as anthelmintics in Pakistan. Further studies on pharmacokinetics using scientific procedures may prove these plants as promising candidates for their future use as anthelmintics.

Anticoccidial drug resistance in fowl coccidia: the state of play revisited

The development of drug resistance in Eimeria is common because of extensive use of anticoccidial drugs for the control of avian coccidiosis. The significance of chemotherapy is evident from the fact that, in spite of advancement in the field of immunological, biotechnological and genetic methods, prophylactic chemotherapy with anticoccidial drugs is still widely used for the control of coccidiosis. In such situations, new drugs should be available to replace the older ones against which resistance has developed, however it takes a long time to develop any new compounds. It is therefore currently necessary to develop strategies to minimise the emergence of resistance in Eimeria strains and to prolong the effect of available anticoccidial drugs. This paper summarises the resistance status of Eimeria species in different parts of the world and reviews different types of resistance, mechanism of resistance development, factors involved in the development and spread of resistance, management of resistant strains and strategies to preserve the efficacy of the available anticoccidial drugs. Use of vaccines, synthetic and botanical anticoccidials and educating farmers about recommended coccidiosis control practices are discussed in this review, along with the integration of currently available options for the management of drug resistance and, ultimately, the control of coccidiosis.

Epidemiology of Eimeria and associated risk factors in cattle of district Toba Tek Singh, Pakistan

A cross-sectional study was carried out to determine the prevalence, species characterization, and associated risk factors with Eimeria of cattle of district Toba Tek Singh from April, 2009 to March, 2010. Of the total 584 fecal samples examined for Eimeria, 275 (47.09%) were found infected with six species of Eimeria. Among the identified species of Eimeria, Eimeria bovis was found to be the highest prevalent species (52.36%), followed in order by Eimeria zuernii, Eimeria canadensis, Eimeria ellipsoidalis, Eimeria alabamensis, and Eimeria cylindrica with prevalence of 48.27%, 34.83%, 29.31%, 24.14%, and 8.62% respectively. Peak prevalence was observed in August. Cattle were infected more frequently during rainy (60.32%) and post-rainy seasons (59.25%). Calves had significantly higher prevalence (P < 0.05) of Eimeria than adults while higher prevalence of Eimeria was observed in female cattle. Among management and husbandry practices, feeding system, watering system, housing system, floor type, and herd size strongly influenced the prevalence of Eimeria in cattle. Coccidiosis was more prevalent in ground feeding system, pond-watered animals, closed housing system, and non-cemented floor type (P < 0.05) as compared to trough feeding system, tap watered animals, open housing system, and partially cemented floor types, respectively. Breed and body condition of animals were not found risk factors (P > 0.05) influencing prevalence of Eimeria.

Anticoccidial activity of herbal complex in broiler chickens challenged with Eimeria tenella

The objective of the present study was to evaluate the anticoccidial effect of different concentrations of the herbal complex of 4 plants (leaves of Azadirachta indica and Nicotiana tabacum, flowers of Calotropis procera and seeds of Trachyspermum ammi) in broiler chickens in comparison with amprolium anticoccidial. Three concentrations (2 g, 4 g and 6 g) of herbal complex were given to the experimental groups once a day and amprolium (at the dose rate of 125 ppm) was given orally in drinking water from the 14th to the 21st days of age. One group was kept as infected, non-medicated control and one as non-infected, non-medicated control. All groups were inoculated orally with 75,000 sporulated oocysts on the 14th day of age except the non-infected, non-medicated control. Among herbal complex medicated groups, the maximum anticoccidial effect was seen in the group medicated with 6 g herbal complex followed by 4 g and 2 g herbal complex medicated groups. Treatment with 6 g of the herbal complex significantly reduced the negative performance and pathogenic effects associated with Eimeria tenella challenge at a level that was comparable with amprolium when using a largely susceptible recent field isolate. In summary, concentration-dependent anticoccidial activity of the studied herbal complex suggests its use as an alternative anticoccidial agent to chemotherapeutic drugs for Eimeria tenella control.

In vitro and in vivo acaricidal activity of a herbal extract.

The anti-tick efficacy of combined aqueous herbal extracts of Azadirachta indica leaves, Nicotiana tabacum leaves, Calotropis procera flowers and Trachyspermum ammi seeds was evaluated using adult immersion test, larval packet test and ear bag method. The extract exhibited lethal effects on egg laying (index of egg laying=0.371404±0.00435), hatching (22.35%) and total larval mortality at 50 mg ml(-1) and reduced tick intensity on the infested calves (18 detached out of 35 at 45% (w/w) suspension, topically applied). The herbal extract exerted dose- and time-dependent response against all the developmental stages of Rhipicephalus (Boophilus) microplus considered in this study, thus justified their use in the traditional system of Pakistan.

Anticoccidial activity of Curcuma longa L. in broilers

Comparative efficacy of turmeric (Curcuma longa L.) crude powder and salinomycin sodium on the occurrence of coccidiosis and growth performance of broiler was evaluated. A total of 90, day-old chicks were randomly divided into six groups. From first day onward, ration was supplemented with 1, 2 and 3 % turmeric powder in groups 2, 3 and 4, respectively, group 1 received salinomycin sodium @ 12 g 50 kg-1 feed while groups 5 and 6 were kept as infected un-medicated and uninfected un-medicated controls. First five groups were infected with Eimeria tenella sporulated oocysts @ 1,00,000/chick at the age of 20 days. Body weight gain, feed consumption, feed conversion ratio were investigated throughout the experimental period, and bloody diarrhea and oocysts excretions were investigated at the first and the second week after infection. Maximum coccidiostatic effect was observed with turmeric (3%) showing mild bloody diarrhea as compared to other infected groups receiving turmeric containing rations. This effect was comparable with a standard coccidiostat i.e., salinomycin sodium. Similarly, the weight gain in the groups treated with salinomycin sodium (2280g) and 3% turmeric (2293g) were also significantly higher (p < 0.05) than that of infected control group (1955g). In the groups treated with ration supplemented with 3% turmeric powder and salinomycin sodium, the peak excretion of oocysts was delayed about 1 or 2 days relative to the control infected group. Concentration-dependent coccidiostatic effect of turmeric suggested that further studies should be carried out to determine the possible maximum safe levels of turmeric with least toxic effects to be used as coccidiostat.

Anthelmintic activity of Trianthema portulacastrum L. and Musa paradisiaca L. against gastrointestinal nematodes of sheep.

Evaluation of anthelmintic effects of Trianthema (T.) portulacastrum L. (Aizoaceae) whole plant and Musa (M.) paradisiaca L. (Musaceae) leaves against prevalent gastrointestinal worms of sheep was done that may justify their traditional use in veterinary clinical medicine. In vitro anthelmintic activity of the crude aqueous methanolic extract (CAME) of both the plants was determined using mature female Haemonchus (H.) contortus and their eggs in adult motility assay (AMA) and egg hatch test (EHT), respectively. In vivo anthelmintic activity of crude powder (CP) and CAME in increasing doses (1.0-8.0 g kg(-1)) was determined in sheep naturally infected with mixed species of nematodes using fecal egg count reduction test (FECRT) and larval counts. The study design also included untreated as well as treated controls. Fecal egg count reduction and larval counts from coprocultures were performed pre- and post-treatments to assess the anthelmintic activity of the plants. CAME of T. portulacastrum and M. paradisiaca showed a strong in vitro anthelmintic activity and pronounced inhibitory effects on H. contortus egg hatching as observed through AMA and EHT, respectively. Both plants exhibited dose and time dependent anthelmintic effects on live worms as well as egg hatching. M. paradisiaca (LC(50)=2.13 μg mL(-1)) was found to be more potent than T. portulacastrum (LC(50)=2.41 μg mL(-1)) in EHT. However, in vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 85.6% and 80.7% with CAME of T. portulacastrum and M. paradisiaca at 8.0 g kg(-1) on 15th day post-treatment, respectively as compared to that of Levamisole (7.5 mg kg(-1)) that caused 97.0% reduction in EPG. All the species of gastrointestinal nematodes (GINs), i.e. Haemonchus contortus, Trichostronglyus spp., Oesophagostomum columbianum and Trichuris ovis which were prevalent, found susceptible (P<0.01) to the different doses of CP and CAME of both plants. The data showed that both T. portulacastrum and M. paradisiaca possess strong anthelmintic activity in vitro and in vivo, thus, justifying their use in the traditional medicine system of Pakistan.