The use of PCR technique in the identification of Mycobacterium species responsible for bovine tuberculosis in cattle and buffaloes in Pakistan.

Bovine tuberculosis is one of the important diseases of dairy and wild animals. The disease is prevalent all over the world, though developed countries have tremendously reduced the prevalence through eradication campaigns. The prevalence of disease in Pakistan on the basis of tuberculin testing or culture isolation of the organism has been reported previously. It is, however, important to use the latest diagnostic tools, i.e. PCR to confirm the type of Mycobacterium infecting the animals in Pakistan. Therefore, the present study was carried out to assess the utility of direct PCR on milk samples and nasal swabs to confirm the type of Mycobacterium infecting the animals. This study was carried out on 215 cattle and buffaloes of more than 2 years of age present at two livestock farms. The tuberculin results showed 22.5% prevalence at one farm and 25.9% at the other with an overall prevalence of 24.7%. The 92.5% of milk samples and/or nasal swabs showed positive PCR for Mycobacterium genus, 86.8% for Mycobacterium tuberculosis complex and 77.4% for Mycobacterium bovis. The M. bovis by PCR was detected in 13.2% of milk samples, 24.5% of nasal swabs and 39.6% of both milk samples + nasal swabs. The results suggested that there are 60% higher chance for a nasal swab to yield a positive PCR for M. bovis than the milk sample. It can be concluded from the present study that tuberculin testing is a useful method in studying the prevalence of disease as the PCR for Mycobacterium genus was positive in 92.5%, M. tuberculosis complex in 86.8% and Mycobacterium bovis in 77.4% cases.

Frequency distribution of hard ticks (Acari: Ixodidae) infesting bubaline population of district Toba Tek Singh, Punjab, Pakistan.

The current research was conducted to define the epidemiological parameters related to the prevalence and associated risk factors of tick infestation in buffaloes in the Toba Tek Singh District of central Punjab, Pakistan. The prevalence of ticks on buffaloes was 31.21 % (352/1,128). Among the species of ticks, the prevalence of Hyalomma marginatum (75.56 %; 266/352) was higher (P < 0.05; odd's ratios (OR) = 3.09) than Rhipicephalus microplus (24.44 %; 86/352). Female buffaloes (69.60 %; 245/352) and younger animals (59.09 %; 208/352) were more heavily infested than males (30.40 %; 107/352) and adult animals (40.91 %; 144/352), respectively, whereas breed was not a determinant (P > 0.05). With regard to management and husbandry practices, the prevalence of ticks was higher in animals kept on uncemented flooring (54.55 %; 192/352; OR = 1.90) followed in order by partially cemented (28.69 %; 101/352; OR = 1.71) and fully cemented flooring (16.76 %; 59/352). With regard to feeding systems, grazing animals (64.20 %; 226/352) were more burdened compared to stall-fed animals (35.80 %; 126/352). The highest tick prevalence was recorded in closed housing systems (52.27 %; 184/352), followed by semi-closed (34.09 %; 120/352; OR = 1.53), and open housing systems (13.64 %; 48/352). Rope-tied animals (70.73 %; 249/352) were more parasitized (P > 0.05) than open (29.27 %; 103/352). Prevalence in the study district was highest in tehsil Kamalia followed in order by T.T. Singh and Gojra. The primary body area of infestation by ticks (head, neck, ear, dewlap, back, abdomen, foreleg, shoulder, hind leg, congenital areas, and tail) ranged from highest at inside thigh (17 %) to lowest at rump. In the present survey, the highest prevalence was recorded in July and lowest in December. Comparison of hematological changes showed remarkable differences between infested and non-infested animals, in the form of low values of infested animals, whereas an increment in biochemical parameter values was observed in tick-infested animals. The present study provides significant data to enhance planning for tick control program in the study area.

The global burden of fasciolosis in domestic animals with an outlook on the contribution of new approaches for diagnosis and control.

Fasciolosis is an economically important disease for livestock, as well as being zoonotic. Recent figures on the prevalence of this disease have caused alarm concerning its potential for an increased prevalence in the future. The prevalence of fascioliosis has been documented from different regions of the world, helping us identify areas where future research needs to be focused. This manuscript is a review of the current status of the disease, the pathogenic species involved, diagnostic techniques (with new modifications and comparative specificity, sensitivity, and rapidity of these tests), chemotherapy, and vaccination. This also encompasses inaccurate reports on vaccination and drug development as well as the latest technologies to find promising candidates for drugs and vaccines. Drugs with lower efficacy have been used on some farms which lead to exacerbation of the clinical disease, presumably due to the development of drug resistance. Future studies should be focused on (1) the use of the most reliable diagnostic tests for periodic monitoring of the disease, (2) insights of the ecobiology and transmission dynamics of the snail intermediate host and the best possible methods of their control, (3) in vitro and in vivo testing of chemotherapeutic compounds using sensitive methods, and (4) the identification of novel drug and vaccine candidates using modern molecular markers. This approach may help increase the reliability of chemotherapeutic agents and control nuisance, ultimately reducing the economic losses attributable to the livestock industry around the world.

Aerobic Bacterial Contamination In Platelet Concentrates At The Regional Blood Centre, Peshawar.

BACKGROUND: Platelet concentrates play a crucial role in transfusion medicine, aiding in the management of various medical conditions, including haemorrhage, thrombocytopenia, and platelet dysfunction. However, their storage conditions at 22° C present an optimal environment for bacterial growth, making them susceptible to contamination. Of particular concern is the transmission of microorganisms from the skin flora during the phlebotomy process, which can lead to the transfusion of contaminated platelet concentrates. Such contamination poses significant risks to patients, potentially resulting in morbidity and mortality. Determining the frequency and identifying causative organisms of bacterial contamination in platelet concentrates. METHODS: It was a descriptive cross-sectional study conducted at the Institute of Pathology and Diagnostic Medicine, Khyber Medical University, and the Regional Blood Center in Peshawar from May to October 2021, spanning a duration of six months. The study included 500 participants aged between 18 and 50 years (mean: 28.13±7.67 years. A simple convenient sampling technique was employed. Blood products underwent screening for Hepatitis B, Hepatitis C, HIV, Syphilis, and Malaria. Leaked units were excluded from the study. Platelets were prepared using a Cryofuge and subsequently subjected to culture media. RESULTS: The mean age of the participants included in the study was 28.13±7.67 years, with an age range of 18 to 50 years. Out of the total sample size of 500, there were 483 (96.6%) male participants and 17 (3.4%) female participants. Among the collected samples, bacterial growth was observed in only 11 (2.2%) platelet concentrates. The isolated organisms were Staphylococcus epidermidis, found in 7 (1.4%) platelet concentrates, and Staphylococcus aureus, found in 4 (0.8%) platelet concentrates. CONCLUSIONS: Bacterial contamination of platelet bags is higher compared to developed countries. Therefore, implementing quality control procedures is necessary to reduce the risk of bacterial contamination in platelet concentrates. Additionally, employing enhanced skin disinfection techniques at the phlebotomy site can significantly minimize bacterial contamination.

Seroprevalence of Sars-Cov-2 antibodies among eligible blood donors of Peshawar, Pakistan.

BACKGROUND AND OBJECTIVES: To determine the seroprevalence of SARS-CoV-2 antibodies and the associated risk factors among healthy blood donors from Peshawar Pakistan, during the second and third waves of the COVID-19 pandemic. METHODS: The study was conducted on 4047 healthy (with no history or symptoms of COVID-19) blood donors attending regional blood center Peshawar between Nov 2020 and June 2021. Demographic data was collected and donors were screened for the presence of anti-SARS-CoV-2 antibodies using electrochemiluminescence immunoassay (ECLIA). RESULTS: The mean age of the participants was 27.27±7.13 and the majority (99%) were males. Overall, 59% (2391/4047) of the blood donors were reactive for SARS-CoV-2 antibodies. An increasing trend in seropositivity was observed from 45.5% to 64.8% corresponding to the second and third wave of the pandemic in Pakistan. Logistic regression analysis revealed significantly higher odds of seropositivity among male donors compared to females. Similarly, in multivariable analysis, the odds ratio for seropositivity among blood types AB, A, and B were, 1.6, 1.4, and 1.3 (CI 95%) times higher compared to blood group O (P-value ≤0.0001). CONCLUSIONS: Seropositivity of SARS-CoV-2 antibodies among blood donors gradually increased during the second and third wave of the pandemic in Pakistan indicating a widespread prevalence of Covid-19 in the general population. Susceptibility to SARS-CoV-2 varies with ABO blood types, with blood group O associated with low risk of infection.

Prevalence, associated determinants, and in vivo chemotherapeutic control of hard ticks (Acari: Ixodidae) infesting domestic goats (Capra hircus) of lower Punjab, Pakistan.

A total of 800 goats of various breeds, age, and sex were randomly selected from Muzaffargarh (M. garh) and Layyah districts of lower Punjab, Pakistan. The selected goats were visited twice a month to collect information about determinants influencing goat tick infestation prevalence. For acaricidal efficacy, 360 tick-infested adult goats were subjected to an acaricidal treatment and post-treatment quantitative assessment of tick burden. Quantification of adult tick detachment 24 h post-treatment and the duration of treatment efficacy were calculated. Overall prevalence of goat tick infestation in both study districts was 60.1% (481/800). The prevalence was higher in district M. garh than in district Layyah. Tehsil-wise prevalence in district Layyah was highest in tehsil Layyah followed in order by Chaubara and Karor. In district M. garh, highest prevalence was found in tehsil M. garh followed by Kot Addu, Alipur, and Jatoi. Hyalomma a. anatolicum (75.9%; 365/481) and Rhipicephalus sanguineus (24.1%; 116/481) were the predominant species in both the districts. The highest month-wise prevalence was 56.9% and 62.7% in Layyah and M. garh districts, respectively, during July 2008, and the minimum (0%) prevalence was reported in November and December, respectively. Regarding host determinants, female goats were more heavily infested (72.8%) than males (47.5%), and younger animals were (63.5%) more burdened than older ones (56.7%). Teddy goats were the most susceptible breeds followed in order by Beetal, cross-bred, Nachi, and Dera Din Pannah. The preferred sites of attachment were inside and outside of the ear. Both the ivermectin (IVM)- and cypermethrin (CYM)-treated groups resulted in significantly lower (P < 0.05) tick counts relative to controls on all post-treatment counting days. The lowest tick burden in the IVM-treated group was significantly higher (P < 0.05) as compared to the CYM-treated group, the latter being close to zero. Hence, the in vivo efficacy trials of injectable IVM vs CYM pour-on revealed better results for the latter. These observations provided the first insights into what determinants impact goat tick infestation, and laid a foundation for planning of future control programs in the lower Punjab, Pakistan.

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.

Comparative anthelmintic efficacy of Arundo donax, Areca catechu, and Ferula assa-foetida against Haemonchus contortus.

In the present study, anthelmintic activities of Arundo (A.) donax L., Areca (Ar.) catechu L., and Ferula (F.) assa-foetida L. were determined. Leaves of A. donax L., latex of F. assa-foetida L. and seeds of Ar. catechu L. in different solvent fractions were subjected to in vitro (egg hatch assay; EHA, and adult motility assay; AMA) and in vivo (faecal egg count reduction test; FECRT) tests of anthelmintic activity using Haemonchus contortus model. In the AMA, crude aqueous methanol extracts (CAME) and ethyl acetate fractions of F. assa-foetida at 10 hr post-treatment showed maximum mortality of H. contortus at 12.5-50 mg mL-1. In the EHA, CAME of F. assa-foetida was identified as a potent ovicide based on its low LC50 (16.9 µg mL-1), followed in order by Ar. catechu and A. donax. Results from the FECRT also showed the extract of F. assa-foetida L. to be more effective than those of Ar. catechu L. and A. donax L., against the gastrointestinal parasitic nematodes. Chloroform and ethyl acetate fractions showed better anthelmintic activities against the adult worms in vitro, while CAME of these plants were better than their crude powders in vivo. It is recommended to document and investigate indigenous knowledge of possible medicinal plants to plan scientific trials that may justify their endorsement.

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 hypodermosis--a global aspect.

Cattle hypodermosis (warble fly infestation) is a notorious veterinary problem throughout the world. Larvae of Hypoderma species cause a subcutaneous myiasis of domesticated and wild ruminants. This disease is caused by, Hypoderma bovis, Hypoderma lineatum in cattle whereas, Hypoderma diana, Hypoderma actaeon, and Hypoderma tarandi, affect roe deer, red deer, and reindeer, respectively. Adults of the cattle grub are commonly known as heel flies, warble flies, bomb flies or gad flies. The biology of hypodermosis is complex because it passes through ecto- as well as endoparasitic stages in the life cycle. The parasitic stage of hypodermosis lasts about 1 year in domesticated as well as in the wild animals, while in the adult stage, a free-living fly lasts only for few days. The diagnosis of hypodermosis is of prime importance for planning treatment and the eradication program. Generally, there are two methods that are routinely used for diagnosis of hypodermosis, i.e., the direct clinical examination and immuno diagnosis by the use of pooled serum and/or milk sample. For the control of hypodermosis, different preparations are available and their use in most of the countries is limited to an individual level but never cover the whole cattle population of a country. Re-infestation in the herd occurs due to the untreated animals that remain the reservoir of the disease. The disease causes huge economic losses in animal production due to the effect of this disease on meat, milk, and the leather industry. It can also affect the general health status as well as the immune system of the body of the diseased animals. As regards the control measures of the disease, different methods have been efficiently practiced and consequently this disease is controlled at national level in many European countries.

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.

Effect of ivermectin on the cellular and humoral immune responses of rabbits.

The objective of this paper is to determine the effect of ivermectin administration on cell mediated (CMI) and humoral immunity (HI) of rabbits. CMI against dinitrochlorobenzene (DNCB) and sheep red blood cells (SRBC) in rabbits was determined by delayed-type hypersensitivity and macrophage engulfment assay (MEA), respectively; whereas, HI to Pasteurella multocida B2 vaccine and SRBC was determined by indirect haemagglutination assay (IHA) and Jerne hemolytic plaque formation assay (JHPFA), respectively. The rabbits were divided into four major groups (A through D) each subdivided into four sub-groups (1 through 4). Rabbits of group A served as vehicle control while those of groups B, C and D were treated with ivermectin at the dose rates of 200 microg/kg, 400 microg/kg and 600 microg/kg b.w., respectively. Cellular immunity was determined in sub-groups 1 and 2 through DNCB and MEA, respectively while HI was determined in sub-groups 3 and 4 through IHA and JHPFA, respectively. The skin sensitivity to DNCB at 24 and 48 h and macrophage engulfment of SRBC were highest (P>0.05) in rabbits administered with 600 microg/kg b.w. The highest geometric mean titers (14.00+/-0.31) and number of plaque forming units (1860+/-0.75) were found in rabbits that received ivermectin at a dose of 600 microg/kg b.w. followed, in order by the groups that received 400 microg/kg, 200 microg/kg b.w. and controls. Leukocyte counts were significantly higher in ivermectin-treated groups (C and D) than group A (vehicle control) and B (ivermectin at the rate of 200 microg/kg). A graded dose immune response suggested an immunopotentiating effect of ivermectin at higher doses.

Direct and indirect anthelmintic effects of condensed tannins in sheep.

Anthelmintic activity of condensed tannins (CT) was evaluated both in vitro and in vivo. In vitro tests included egg hatch test and paralysis/mortality assay on adult Haemonchus contortus. In vivo anthelmintic effect was determined by faecal egg count reduction test in lambs. To this end, 18 lambs were divided into three groups (low tannin, high tannin and control). The lambs of low and high tannin groups were fed diets containing 2 and 3% CT while the control group was fed on diets without CT. In vitro trials showed a dose-dependent inhibition of nematode egg hatching; whereas, there was no effect of CT on adult H. contortus. In vivo trials indicated reduction in faecal egg counts in lambs fed diets containing CT. Feed intake and nutrient digestibility of CT-fed sheep was lower and nitrogen balance was higher as compared to control. Maximum weight gain was observed in animals fed diets containing 3% CT. The direct anthelmintic effect of CT, therefore, was evidenced by inhibited egg hatching; whereas, faecal egg counts reduction in sheep was through improved nutrient utilization.

Prevalence, economic analysis and chemotherapeutic control of small ruminant fasciolosis in the Sargodha district of Punjab, Pakistan.

This study describes the epidemiology, the economic significance of small ruminant fasciolosis in animals slaughtered in the abattoirs of the Sargodha district, Punjab, Pakistan between January and June 2012. In vivo fasciolicidal efficacy of commercially available compounds was examined using a randomised complete block design. Microscopically screened faecal samples revealed 40.51% positive animals for fasciolosis. The prevalent species included Fasciola hepatica (35.64%) and Fasciola gigantica (8.21%). Mixed infections were noted in 3.33% subjects. Prevalence rates were significantly higher in females (42.25%) than in males (39.52%), and in adults (51.20%) compared to younger animals (33.98%). The disease was recorded more often in emaciated animals (63.63%) followed in order by average (43.45%), thin (43.22%), and fat (32.12%) animals. Between January and June 2012, fasciolosis in Sargodha district, Punjab, Pakistan, was estimated to incur US$0.036 million and US$0.177 million direct (liver condemnation) and indirect (carcass depreciation) economic losses, respectively. In vivo fasciolicidal efficacy of oxyclozanide proved to be the most effective method of control, compared to triclabendazole, and levamisole. Results provide useful information on the frequency distribution of fasciolosis and its economic significance. Finally, data on in vivo fasciolicidal trials show that oxyclozanide is the most efficacious compound for the treatment of the disease in the district Sargodha, Punjab, Pakistan.

Prevalence and associated risk factors of haemoparasites, and their effects on hematological profile in domesticated chickens in District Layyah, Punjab, Pakistan.

Several haemoparasites commonly infect avian species, including chicken that significantly decline productivity and ultimately lead to high mortality rate. The current study was designed to assess the prevalence and associated risk factors of haemoparasites in domesticated chickens in and around District Layyah, Punjab Province, Pakistan. For this purpose, blood samples from a total of 384 backyard chickens were collected from wing vein using sterile insulin syringe. The parasites were identified from Giemsa stained thin blood smears based on morphological features using standard keys. Results demonstrated that a total of 265 backyard chickens (69%) were infected with haemoparasites in District Layyah. According to genus-wise distribution, 31.5%, 24.4% and 13% prevalence of genera Plasmodium/Haemoproteus, Leucocytozoon and mixed species were recorded, respectively. Among associated risk factors, the prevalence was relatively higher in females, chicks, naked neck breeds, scavengers feeding patterns and chickens reared at fully open coops type. The information given in the study could be of much importance in planning of an effective haemoparasites control program at District and Provincial level.

An inventory of the ethnobotanicals used as anthelmintics in the southern Punjab (Pakistan).

A survey was conducted in southern Punjab, Pakistan, in order to document existing ethnobotanical knowledge by the herdsmen/key respondents about anthelmintics in ruminants. A 3-satge process was used to document the plants being used to treat and/or control helminthes. This paper describes 29 plants to treat helminthosis in ruminants. The main plants used were Lamium amplexicaule L., Mallotus philippinensis Muell., Withania somnifera (L.) Dunal., Azadirachta indica A. Juss., and Citrullus colocynthis (L.) Schrad. A few of these plants have been scientifically validated for their claim by herdsmen on modern lines while majority of them still needs investigations. This documentation could provide a foundation for the scientific study and verification of those plants which merit such study.

Anthelmintic activity of Swertia chirata against gastrointestinal nematodes of sheep.

This paper describes in vitro and in vivo anthelmintic activities of Swertia chirata. In vitro studies revealed that at 25 mg/ml (P>/=0.05) the crude aqueous (CAE) and methanolic extracts (CME) of S. chirata whole plant showed an anthelmintic effect on live Haemonchus contortus. Moreover, in the in vivo study, the whole plant of S. chirata administered as crude powder (CP), CAE and CME at the dose of 3 g/kg to sheep naturally infected with mixed species of gastrointestinal nematodes, showed a significant reduction in egg per gram of faeces.

Comparative anthelmintic efficacy of Arundo donax, Areca catechu, and Ferula assa-foetida against Haemonchus contortus / Eficácia anthelmíntica comparativa de Arundo donax, Areca catechu e Ferula assa-foetida contra Haemonchus contortus

Abstract In the present study, anthelmintic activities of Arundo (A.) donax L., Areca (Ar.) catechu L., and Ferula (F.) assa-foetida L. were determined. Leaves of A. donax L., latex of F. assa-foetida L. and seeds of Ar. catechu L. in different solvent fractions were subjected to in vitro (egg hatch assay; EHA, and adult motility assay; AMA) and in vivo (faecal egg count reduction test; FECRT) tests of anthelmintic activity using Haemonchus contortus model. In the AMA, crude aqueous methanol extracts (CAME) and ethyl acetate fractions of F. assa-foetida at 10 hr post-treatment showed maximum mortality of H. contortus at 12.5-50 mg mL-1. In the EHA, CAME of F. assa-foetida was identified as a potent ovicide based on its low LC50 (16.9 µg mL-1), followed in order by Ar. catechu and A. donax. Results from the FECRT also showed the extract of F. assa-foetida L. to be more effective than those of Ar. catechu L. and A. donax L., against the gastrointestinal parasitic nematodes. Chloroform and ethyl acetate fractions showed better anthelmintic activities against the adult worms in vitro, while CAME of these plants were better than their crude powders in vivo. It is recommended to document and investigate indigenous knowledge of possible medicinal plants to plan scientific trials that may justify their endorsement.

Resumo No presente estudo, as atividades anti-helmínticas de Arundo (A.) donax L., Areca (Ar.) Catechu L. e Ferula (F.) assa-foetida L. foram determinadas. Folhas de A. donax L., látex de F. assa-foetida L. e sementes de Ar. catechu L. em diferentes frações de solvente foram submetidos a testes in vitro (teste de eclosão de ovos, EHA e ensaio de motilidade em adultos, AMA); e in vivo (teste de redução da contagem de ovos fecais, FECRT) de atividade anti-helmíntica, usando-se Haemonchus contortus. Na AMA, extratos aquosos brutos de metanol (CAME) e frações de acetato de etila de F. assa-foetida. Dez horas pós-tratamento, apresentaram mortalidade máxima de H. contortus em 12,5-50 mg mL-1. No EHA, CAME de F. assa-foetida foi identificado como um ovicida potente baseado em seu baixo LC50 (16,9 µg mL-1), seguido em ordem por Ar. catechu e A. donax. Os resultados do FECRT também mostraram que o extrato de F. assa-foetida L. é mais eficaz do que o de Ar. catechu L. e A. donax L., contra nematoides parasitas gastrointestinais. As frações clorofórmio e acetato de etila mostraram melhores atividades anti-helmínticas contra vermes adultos in vitro, enquanto o CAME dessas plantas foi melhor do que o pó bruto in vivo. Recomenda-se documentar e investigar o conhecimento indígena de possíveis plantas medicinais para planejar ensaios científicos que possam justificar seu endosso.

Anthelmintic activity of Calotropis procera (Ait.) Ait. F. flowers in sheep.

The anthelmintic activity of Calotropis (C.) procera flowers in comparison with levamisole was evaluated through in vitro and in vivo studies. In vitro studies revealed anthelmintic effects (P<0.05) of crude aqueous (CAE) and crude methanolic extracts (CME) of Calotropis procera flowers on live Haemonchus (H.) contortus as evident from their mortality or temporary paralysis. For in vivo studies, Calotropis procera flowers were administered as crude powder (CP), CAE and CME to sheep naturally infected with mixed species of gastrointestinal nematodes. Egg count percent reduction (ECR) was recorded as 88.4 and 77.8% in sheep treated with CAE and CP at 3gkg(-1) body weight on day 7 and 10 post-treatment (PT), respectively. CME was least effective resulting in 20.9% reduction in ECR on day 7 PT. It was found that Calotropis procera flowers possess good anthelmintic activity against nematodes, yet it was lower than that exhibited by levamisole (97.8-100%). It is suggested that further research on large scale be carried out involving a large number of animals, doses higher than those used in the current study, identification of active principles, and standardization of dose and toxicity studies for drug development.

Prevalence of tick infestation (Rhipicephalus sanguineus and Hyalomma anatolicum anatolicum) in dogs in Punjab, Pakistan.

This study was conducted to identify ticks that infest dogs in the Rawalpindi region of Punjab, Pakistan. A total of 525 dogs were examined between June and September 2010. Of these, 60 (11.42%) were infested with ticks. The morphological features of the ticks were identified by using a stereomicroscope to confirm their identity using morphological keys. Two species were observed, namely: Rhipicephalus sanguineus and Hyalomma anatolicum anatolicum. The prevalence rate of these species was 98.33% and 1.66%, respectively. Of the 508 specimens collected, 265 male, 224 female ticks and 19 nymphs were identified. No larvae were collected from the infested dogs during the study period. There was no significant difference observed during the study months.