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Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar
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ThesisItem Restricted Detection of anthelmintic resistance against gastrointestinal nematodes in sheep and goats(Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar, 2024-04) Santosh; Santosh; Vohra, Sukhdeep; Vohra, SukhdeepThe study was conducted to evaluate the status of anthelmintic resistance in gastro intestinal nematodes (GINs) in sheep and goats at two organised farms (CSBF and UCBF) located in Hisar, Haryana. Sheep and goats reared at these farms exhibited reduced efficacy for multiple anthelmintics following treatments with Fenbendazole (FBZ), Closantel (CLS), Morantel (MOR), and Ivermectin (IVM) in the Faecal Egg Count Reduction Test (FECRT). The results suggested that the overall efficacy was highest for CLS at 89.41% and 77.23% in sheep and goats, respectively, on the 14th day post-treatment, and least for FBZ at 58.73% and 64.55% in sheep and goats, respectively. The pre-treatment faecal culture revealed Haemonchus contortus, Trichostrongylus spp. Strongyloides spp., and Oesophagostomum spp.; however, in post-treatment samples, H. contortus was predominantly observed. ED50 values of EHA demonstrated the presence of thiabendazole resistance in one sheep farm (UCBF) and two goat farms (CSBF and UCBF). Further, the 180 infective larvae of H. contortus from both sheep and goat farms were subjected to allele-specific PCR (As-PCR) for accurate diagnosis of BZ resistance. The As-PCR revealed 40% homozygous resistant (rr), 46.11% heterozygous (rS), and 13.88% homozygous susceptible (SS), with the overall frequency of the resistant (r) allele being 63.5% and for the susceptible allele (s) being 36.5%. The survey indicated that the GINs of sheep and goats on the farms have developed multiple anthelmintic resistance to FBZ, CLS, MOR, and IVM, and the condition is alarming on the farm.ThesisItem Open Access Molecular characterization of eimeria species of buffalo-calves in Haryana(LUVAS, 2017) Nain, Navjot; Gupta, S.K.To study the prevalence of Eimeria species in Haryana, a total of 427 faecal samples were collected from buffalo calves of four districts namely, Fatehabad, Hisar, Bhiwani and Sirsa. Parasitological examination was carried out using saturated salt (NaCl) solution and sporulation was done by using 2.5% potassium dichromate solution to identify the species. Over-all prevalence was 57.87%. Based on morphological characteristics and sporulation time, eleven Eimeria species were identified, these include: E. bareillyi (38.46%), E. bovis (34.81%), E. zuernii (31.98%), E. subspherica (29.29%), E. canadensis (19.43%), E. alabamensis (12.9%), E. ellipsoidalis (9.31%), E. cylindrica (8.09%), E. auburnensis (7.29%), E. wyomingensis (6.07%) and E. pellita (4.45%). Prevalence among male calves (69.39%) was significantly higher (p< 0.01) as compared to females (48.05%). Calves of 0-3 months of age showed significantly higher (p< 0.01) prevalence (63.14%) than 4-6 months old calves (43.47%). Similarly, calves of organized farms showed significantly higher (p< 0.01) rate of prevalence (70.86%) than calves of unorganised sector (52.33%). Out of 247 positive samples, 118 (47.77%) had monoinfection of Eimeria, 40 (16.19%) had two species, 65 (26.31%) had three species and 24 (9.72%) had multiple species. Eimeria pellita has been recorded for the first time in Haryana. Based on amplification of ITS-1 region of rRNA. five Eimeria species were confirmed to occur in Haryana. Four Eimeria species subjected to phylogenetic analysis revealed that Eimeria bovis and Eimeria zuernii Indian isolates are closely related to Turkey and Japan isolates. Eimeria cylindica of Indian origin formed a separate clade with Japanese isolate, whereas, E. alabamensis Indian isolate shared the same clade with Japanese isolate.ThesisItem Open Access Detection, Morphological characterization and epidemiological investigations on parasitic ciliates in buffaloes(LUVAS, 2017) Kalkal, Hardeep; Sangwan, A.KA survey was conducted to study the epidemiology of parasitic ciliates in buffaloes from Hisar and Karnal districts of Haryana. Morphological study was also done to differentiate parasitic ciliates found in pigs and buffaloes. A total of 800 samples were collected (400 from each district) and were collected randomly. A total of 50 fecal samples were collected from pigs for comparison with buffalo parasitic ciliates. The samples were processed by direct smear, sedimentation and floatation techniques. There was significant difference between size of parasitic ciliates in pigs and buffaloes and main morphological character of buffalo parasitic ciliate was presence of ridge which runs from one end to other end and location of Cytostome (mouth) which was opposite to progression direction and in pig parasitic ciliates the Cytostome (mouth) was located at tapering anterior end and with the direction of progression it is concluded the parasitic ciliates in buffalo as B. sulcata. The overall prevalence of B. sulcata was 59.12% whereas; it was 54.5% in Hisar and 63.7% in Karnal. Age and sex had no effect on prevalence of B. sulcata. The risk of prevalence of B. sulcata was lower (OD=0.522) in buffaloes raised on Pakka floor as compared to Katcha floor. The animals with diarrhoea were at greater risk (OD=6.30) of having B. sulcata infection as compared to animals without diarrhoea. There was significant association between degree of intensity of infection with diarrhoea i.e. as degree of intensity increased, the per cent of diarrhoea positive cases also increased. This study provides evidence that B. sulcata can be one of the factors of diarrhoea.ThesisItem Open Access Studies on epidemiology and anthelmintic resistance in gastrointestinal nematodes of cattle under different management systems in Haryana(LUVAS, 2017) Sukhjeet Singh; Satyavir SinghThe epidemiology of gastro intestinal nematodes of cattle was studied for one year starting from June, 2016 to May, 2017 to know the prevalence of nematodes in cattle (>3 years of age) in relation to temperature, rainfall and relative humidity under different management systems i.e. small holder dairy unit, commercial dairy farm, large organised farm and Gaushala in Haryana. The eggs of strongyle spp. and Strongyloides sp. were found in faeces of naturally infected cattle throughout the study period under different cattle management systems. The eggs of strongyle spp. were found higher as compared to Strongyloides sp. throughout the year. However, the egg per gram of these animals was found very low under all management systems throughout the year indicating very low grade of infection. Out of four systems, incidence and severity of gastro intestinal nematodes was found statistically higher (P< 0.05) in small holder dairy production system as compared to other three systems. The infection of gastro intestinal nematodes was higher in small holder dairy production system in the rainy season, while in Gaushala and large organized dairy farm it was found to be higher in summer season. In commercial dairy farm it was almost equal in summer and winter season.For anthelmintic resistance studies, twenty four cattle under small holder dairy units with egg per gram (EPG) of more than or equal to 150 were divided into 4 groups of 6 animals each. Groups I, II and III were administered fenbendazole (5mg/ kg body weight, orally), morantal (10mg/ kg body weight, orally) and ivermectin (0.2mg/ kg body weight, subcutaneously), respectively. Group IV served as the untreated control. Faecal samples were collected on 14th day after treatment from animals of all groups and individual faecal egg counts were determined by modified McMaster technique. Faecal egg count reduction percentage in groups I, II and III were 87.32, 100 and 100%, respectively. Identification of infective larvae from pre treatment faecal cultures revealed the predominance of Haemonchus spp. Larvae identified in post treatment culture of fenbendazole treated group were only Haemonchus spp. No larvae were recovered from group II and III.This indicates presence of slight fenbendazole resistance to Haemonchus spp. and no resistance to morantel and ivermectin in cattle. This is the first report of fenbendazole resistance to Haemonchus spp. in cattle from India.ThesisItem Restricted Studies on the efficacy of diclazuril alone and in combination with turmeric powder and vitamin K on caecal coccidiosis in broiler chicks(LUVAS, 2008) Vijay Kumar; Rajinder KumarThe studies on the efficacy of diclazuril alone and in combination with turmeric powder and vitamin K on caecal coccidiosis in broiler chicks was viii conducted in the department of veterinary parasitology, CCS HAU, Hisar. A total of 90, day old, broiler chicks were divided randomly into nine groups of 10 birds each. These birds were provided with diclazuril @ 1.0 ppm in feed, turmeric powder @ 7.0 g/ kg in feed and vitamin K @ 1g/ 150 lit of drinking water for 14 days, starting from day 15 of age till the age of day 28. These medicaments were provided alone as well as in combination. Afterwards, the birds were infected artificially with Eimeria tenella infection. The effect was monitored in terms of mean weight gain, oocyst index, hematology, serum protein profiling, mortality, global index and histopathological studies. The birds medicated with diclazuril performed better in terms of mean weight gain, oocyst output, oocyst index and lesion scores whereas the results were not encouraging when turmeric or vitamin K were provided alone. The hematological finding were recorded to be improved for turmeric and vitamin K groups, though no such obvious effects were observed for serum protein profiling and histopathological examination. On the basis of global index percentage, diclazuril was found to be an effective anticoccidial. However even more exaggerated anticoccidial efficacy of diclazuril was observed when conjuncted with turmeric and vitamin K. There was tremendous reduction in the mortality of all the medicated birds during the experiment.ThesisItem Restricted Efficacy of probiotics and medicinal plants on Eimeria tenella infection of chickens and their effect on development of immunity(LUVAS, 2007) Hadimani, Santosh; Gupta, S.K.Anticoccidial efficacy of probiotics (Lactobacillus acidophilus-1 g/kg, Saccharomyces cerevisiae-1 g/kg) alone and in combination with salinomycin (60 ppm), Neem (Azadirachta indica) and Papaya (Carica papaya) against Eimeria tenella, using criteria mentioned in WAAVP Guidelines along with the effect of these agents on host immunity during the infection were studied. A total of 108 cage reared birds randomly allocated to nine groups were used, out of which all groups except the negative control received an infection of 50,000 sporulated oocysts of Eimeria tenella per bird on 21 days of age. Lesion scoring, analysis of serum protein profile, total lipids, carotenoids, haematocrit was done on day 7 post inoculation (p.i.). On day 8 and 9 p.i., OPG counts were estimated. Serum Ig Y levels were determined using 11th day p.i. sera. DTH test was done on 14th day p.i. All probiotic fed groups produced a significant fall in oocyst output with reduction being 79, 84, 82, and 79 % of positive control respectively for Lactobacillus acidophilus, Lactobacillus acidophilus + salinomycin, Saccharomyces cerevisiae and Saccharomyces cerevisiae + salinomycin groups. The least fall in PCV was experienced by Lactobacillus acidophilus group and the rest of the groups except yeast alone group recorded significantly lower falls compared to the positive control. Only Lactobacillus acidophilus group fared poorly in MWG and FCR whereas all other were far better than positive control and yeast groups proved even better than negative control. Only Lactobacillus acidophilus + salinomycin group recorded significantly elevated serum Ig Y levels while other groups showed insignificantly elevated levels. All probiotic fed groups showed significantly enhanced CMI responses, Saccharomyces cerevisiae + salinomycin group being the highest. Among the medicinal plant groups Papaya group resisted the infection well with its oocyst output being the lowest (88 % reduction), significantly less fall in PCV, good weight gain and FCR. Neem group performed well with respect to parasitological parameters but failed miserably in performance parameters which may be ascribed to systemic toxicity. The combination of these two powders did not go down well with this infection as the group performed no better than the positive control. Papaya groups recorded significantly elevated serum Ig Y levels whereas Neem group had the highest CMI response. Probiotics, Papaya and Neem certainly seemed to have anticoccidial effects with apparent immuno-modulatory mechanisms involved.ThesisItem Restricted Quantitative xenodiagnosis of theileria annulata under different cattle management systems(LUVAS, 2008) P.Prasanna Kumar; Sangwan, Arun KumarBlood smears and ticks were collected from three large organized dairy farms, six commercial dairy farms, nine small holder dairy production units and twelve Gaushalas located at Hisar and surrounding areas in Haryana, India. These ticks were examined for the presence of Theileria annulata by using methyl-green pyronin staining method and blood smears were examined for the presence of T. annulata piroplasms and any other haemoprotozoan parasite using Giemsa staining method. Epidemiological variables related to host, vector and environment were also observed under different cattle management systems. A total of 3816 ticks were collected and identified from the four different cattle management systems. Out of that 1280 ticks were Hyalomma anatolicum anatolicum, 2455 were Boophilus microplus and 81 were Ornithodoros savigniyi. A total of 1200 H. a. anatolicum ticks (211 males and 989 females) were screened for the presence of Theileria. In large organized dairy farms 11.9 %, in commercial dairy farms 7.14 %, in small holder dairy production units 5.32 % and in Gaushalas 5.38 % ticks were positive for Theileria. Frequency distribution of Theileria positive acini per positive tick showed that the higher frequency was much more in small holder dairy production units when compared to other systems as more than 25 positive acini per positive tick were seen only in small holder dairy production units. Theileria prevalence in Hyalomma ticks was highest in large organized dairy farms and lowest in small holder dairy units and Gaushalas while Theileria intensity was lowest in large organized farms and highest in small holder dairy units. The lesser intensity and higher prevalence of T. annulata in Hyalomma ticks is epidemiologically a good situation as it leads to premunity in the population whereas higher intensity and lower prevalence leads to more clinical cases of theileriosis. Thus, small holder dairy units are at a greater risk of theileriosis than organized farms. The overall prevalence of Theileria organisms in cattle was estimated to be 48.89 % by blood smear examination that included large organized dairy farms 50.76 %, commercial dairy farms 48.03 %, small holder dairy production units 59 % and Gaushalas 39.16 %. Per cent parasitaemia was highest in small holder dairy unit 0.08% followed by commercial farms 0.03% and lowest in large organized dairy farms 0.02% and Gaushalas 0.02%. The lower parasitaemia of T. annulata in cattle is epidemiologically a good situation as it shows premune status of the animals. All the four different cattle management systems were mainly infested with Boophilus microplus and Hyalomma anatolicum anatolicum of which B. microplus was more abundant. Prevalence and abundance of Theileria in ticks and prevalence of Theileria in cattle was significantly higher in mixed (cross bred and indigenous) cattle populations maintained on the same premises than the indigenous cattle population alone or cross bred cattle population alone. Intensity and abundance of Theileria in ticks and prevalence of Theileria in cattle was significantly higher in cattle of mixed feeding (stall feeding and grazing) than that of stall feeding alone. Intensity and abundance of T. annulata in ticks was significantly lower in the dairies where acaricide use was regular.ThesisItem Restricted Comparison of monoclonal antibody based latex agglutination test with PCR for diagnosis of Trypanosoma evansi in domestic animals(LUVAS, 2009) Shyma K.P.; Gupta, S. K.Trypanosoma evansi, a blood protozoan parasite causes a serious disease known as ‘surra’ in domestic and wild animals. It is an arthropod borne disease and Tabanus spp. has been implicated as the main vector. It is the most widely geographically distributed pathogenic trypanosome in Africa, South and Central America and Asia. In India, T. evansi infection is widely prevalent in different parts and is of significant economic importance in livestock production. Though trypanosomosis has been studied since many years, its definite diagnosis still suffers from low sensitivity and specificity. Therefore, the present investigation has been carried out with the aim of detecting T. evansi in cattle, buffaloes and equines in the state of Haryana by parasitological (WBF), antigen-detecting (MAb-LAT) and DNA-detecting (TE-PCR) tests and to compare these tests for their relative sensitivity and specificity. A field isolate of T. evansi collected from an infected cattle was propagated in rats and PCR was standardized using DNA isolated from infected rat blood. The assay employed synthetic oligonulceotide primers (21 mer sense and 22 mer antisense) targeted to a repetitive nuclear DNA sequence of T. evansi. TE-PCR positive signal (227bp) was obtained with template DNA content of 12 trypanosomes extracted from whole blood sample. Field serum samples were screened using monoclonal antibody based latex agglutination test (MAb-LAT) and blood samples by WBF and TE-PCR. Out of 205 blood and sera samples from cattle (n=88), buffaloes (n=46) and equines (n=71) examined, 1.95% were found positive for T. evansi infection by WBF while 59.51 and 60.49 per cent found positive by MAb-LAT and TE-PCR, respectively. D-SN and D-SP values of MAb-LAT using TE-PCR as reference test were 88.43% and 81.48% respectively. D-SN and D-SP values of TE-PCR were 87.70% and 82.50% respectively using MAb-LAT as reference test. Prevalence of T. evansi was 3.41%, 60.23% and 65.91% in cattle; 2.17%, 78.26%, and 76.09% in buffaloes and nil, 46.48% and 43.66% in equines, by WBF, MAb-LAT and TE-PCR, respectively. A good correlation was found between MAb-LAT and TE-PCR. It was inferred that MAb-LAT, being simple to perform, rapid, convenient, cost-effective could be quite suitable for field-level diagnosis and screening of trypanosomosis. However, PCR on the other hand has its role in monitoring the efficacy of trypanocidal treatment.ThesisItem Restricted Assessment and biochemical characterization of acaricide resistance in livestock ticks of Haryana(LUVAS, 2014) Gaur, Ruchi Singh; Sangwan, Arun KumarObjectives of the study were three fold: (i) Assessment of acaricide resistance in Rhipicephalus (Boophilus) microplus and Hyalomma anatolicum against deltamethrin and diazinon , (ii) Esterase profile pattern in susceptible and resistant ticks and (iii) antioxidant status/activity in ticks as influenced by acaricides exposure. The ticks were collected from various areas of Hisar and adjoining districts. Sufficient number of engorged females of H. anatolicum for acaricide resistance studies could only be collected from the Churu district of Rajasthan, adjoining Hisar. Rhipicephalus. (Boophilus). microplus ticks of Tohana (Hisar) were found resistant to both the acaricides while the ticks of Agroha (Hisar) were found susceptible using adult immersion test (AIT). The same results was obtained by using standard method of larval packet test (LPT). The H. anatolicum ticks of Tara Nagar (Churu) were found susceptible while of Churu city were resistant by using adult immersion test (AIT). The same results were obtained by larval packet test except that of ticks of Churu city were also found susceptible. A comparative analysis of the esterase patterns in susceptible and resistant ticks using non-denaturing polyacrylamide gel electrophoresis revealed six types of esterases (dubbed as EST-1b, EST-2b, EST-3b, EST-4b, EST-5b and EST-6b) in R.(B).microplus and four types of esterases (EST-1h, EST-2h, EST-3h, EST-4h) in H.anatolicum using α-Naphthyl acetate substrate. Qualitative expression of EST-4b while quantitative expression of EST-5b and EST-6b might be responsible for deltamethrin and diazinon resistance in R.(B). microplus while qualitative expression EST-4h was probably responsible for diazinon resistance in H. anatolicum. The data suggest that these AChEs may represent an important detoxification strategy developed to overcome the effects of acaricides. In acaricide exposed ticks increases in superoxide dismutase (SOD) activity as the concentration of exposure to acaricides increases suggests better combating of the harmful effects of free radical generation. The decrease in reduced glutathione (GSH) activity as the concentration of exposure to acaricides increases suggests utilization of glutathione for combating the oxidative stress caused by acaricides.
