Loading...
Thesis
Browse
Search Results
ThesisItem Open Access IN VITRO EVALUATION OF ANTHELMINTIC PROPERTIES OF THE METHANOLIC AND HYDRO ALCOHOLIC EXTRACTS OF ANNONA MURICATA ON HAEMONCHUS CONTORTUS.(SRI VENKATESWARA VETERINARY UNIVERSITY, TIRUPATI - 517 502. (A.P.) INDIA, 2023-06) SRI MOUNIKA .G; SRINIVASA RAO .K (MAJOR); SREEDEVI .c; Ravi Kumar .PHaemonchus contortus is the most pathogenic nematode in sheep and has developed resistance to all commercial anthelmintics throughout the world. In this context, the use of medicinal plants represented as an alternative to chemical anthelmintics. The present study was conducted to evaluate the in vitro anthelmintic effect of crude hydroalcoholic (HE) and methanol extracts (ME) of Annona muricata fruit peel, seed and bark against eggs, infective larvae, and adults of H. contortus by egg hatch assay (EHA), the larval paralysis assay (LPA) and worm motility inhibition assay (WMIA) at different concentrations (0.3 to 81 mg/mL) using albendazole as a reference drug. All extracts exhibited significant (P<0.001) dose and or time dependant anthelmintic effects by inhibiting egg hatching and causing larval paralysis, and mortality of worms compared to negative control, and the effect was comparable with albendazole. At higher (81 mg/mL) concentration, A. muricata extracts showed 96 to 100% efficacy in EHA and 100% efficacy in LPA. In WMIA, worms were completely immobilized in extracts within 1 h post-exposure. Overall performance of the hydroalcoholic extracts of the plant on egg, larva, and adult H. contortus was better than the methanol extract of the same plant except for the seed. At lower (9 and 3 mg/mL) concentrations extracts were effective on egg and larval stage of H. contortus, but did not inhibit the motility of worms. The respective effective dose 50% (ED50) with 95% confidence intervals were estimated. The hydroalcoholic extracts of A. muricata fruit peel (ED50=1.28 mg/mL) and seed (ED50=1.29 mg/mL) significantly inhibited egg hatching, whereas fruit peel extracts significantly inhibited larval motility (ED50=2.71 mg/mL) and adult worm motility (ED50=12.63 mg/mL), indicating their anthelmintic activity. Quantitative and qualitative phytochemical screening of extracts revealed the presence of alkaloids, flavonoids, tannins, amino acids, terpenoids and saponins, which could be accountable for the anthelmintic activity observed. Alkaloids were exclusively present in fruit peel HE. The results confirm the traditional use of A. muricata as a natural anthelmintic, however, further in vivo studies are warranted to validate these effectsThesisItem Open Access MORPHOLOGICAL AND MOLECULAR IDENTIFICATION OF VISCERAL SCHISTOSOMES IN BUFFALOES OF ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2023-04) VEERENDRA .U; JYOTHISREE .CH (MAJOR); SREEDEVI .C; SAMATHA .V; VINOO .RThe present study was undertaken to determine the prevalence of visceral schistosomosis caused by Schistosoma species in Andhra Pradesh. For this, a total of 467 slaughtered buffaloes mesenteries and livers collected from abattoirs of Krishna district were sampled for a period of one year. On examination, 95 buffaloes were found to be infected with Schistosoma species giving an overall prevalence of 20.34%. The prevalence of schistosomosis was significantly (P<0.001) higher in adults (>2.5yr; 31.95%) than in young (<2.5yr; 13.75%) animals and non-significantly high in females (26.02%) than in males (19.28%). The prevalence was significantly (P<0.001) high during rainy season (28.47%) followed by winter (21.73%) and summer (10.96%) seasons. Month wise prevalence of schistosomosis in the study area revealed highest prevalence of schistosomosis in July (34.21%) and lowest prevalence during May (5.88%). Most of the infections were mild (48.42%) followed by moderate (36.84%) and severe (14.73%) infection. Mild to moderate infections were noticed in young animals and severe infection was noticed in adults. The worms were milky white in colour. Male worms were stout and carried the females in a groove like a gutter i.e, gynaecophoric canal. Based on three to six testicles, a smooth cuticle, a conical projection at the back of the male, and spindle-shaped eggs in the female's uterus the worms were identified as Schistosoma spindale. Grossly infected livers showed petechiae to patchy haemorrhages, necrotic foci, and whitish worms entrenched in portal veins. The serosal surface of the small intestines had white grey nodules and moderate haemorrhages with necrotic foci on the mucosa. The liver sections revealed moderate to severe congestion of blood vessels and mild degenerative changes in hepatocytes along with adult flukes in the central veins and capillaries of sinusoidal spaces. Infected intestines displayed moderate to severe congestion of blood vessels, haemorrhages in mucosa and the cut section of the parasite in the muscularis mucosa. Few sections of liver and intestine were evident with fibrous tissue proliferation surrounding the parasitic ova and development of microgranulomas. Trabecular thickening, and haemosiderin engulfed macrophages were present in mesenteric lymph nodes along with degenerated parasitic ova. The morphologically identified adult fluke were confirmed through amplification of 28S rRNA gene that yielded 330bp product specific for S. spindale. The sequencing and blast analysis of two S. spindale AP isolates revealed 100% homology with Asian populations of S. spindale. Results confirm that schistosomosis is endemic in buffaloes in Andhra PradeshThesisItem Open Access EVALUATION OF NATURAL RESISTANCE STATUS OF SHEEP AGAINST HAEMONCHUS CONTORTUS INFECTION(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2023-02) RAJESHWARI; SREEDEVI, C(MAJOR); JYOTHI SREE, CH; RAVI KUMAR, P; SUDHAKAR, KAnthelmintic resistance is a major concern throughout the world, especially for the control of Haemonchus contortus. Selection of genetically resistant animals is one alternative to control infection that minimize herd use of anthelmintics. The aim of the current study was to explore associations between phenotypic traits for H. contortus and polymorphisms in the DRB1 region of major histocompatibility complex and GALNT6 gene involved in gastrointestinal mucus production in a Nellore (n=95), Deccani (n=28) and Kenguri (n=34) sheep naturally infected with H. contortus. Blood and faecal samples were collected to evaluate faecal worm egg counts (FEC), packed cell volume (PCV), haemoglobin (Hb), eosinophilia and for DNA isolation. Animals were grouped into high FEC (susceptible) and low FEC (resistant) group based on EPG counts. FEC and circulating eosinophilia were higher in susceptible group. Log FEC was negatively correlated (P < 0.01) with PCV, and Hb estimates. The second exon of DRB1 and intron variant of GALNT6 genes were amplified by polymerase chain reaction from DNA samples of resistant and susceptible sheep. PCR products were characterized by the restriction fragment length polymorphism technique using restriction enzymes. Two genotypes (‘bb’ and ‘ab’) were observed following digestion of Ovar-DRB1 with PstI. Genotype frequencies were 0.78 for ‘bb’ and 0.22 for ‘ab’. The frequency of ‘a’ and ‘b’ alleles were 0.11 and 0.89, respectively. The genotype frequencies differed significantly between both the groups (P<0.05). A generalized linear model was used to test effects of genotype on FEC, Hb and PCV. The ‘bb’ genotypes had higher (P<0.05) log FEC value than ‘ab’ genotypes and ‘b’ allele was linked with susceptibility to haemonchosis in sheep. The mean FEC of Nellore sheep was high (P<0.01) indicating susceptibility of the breed and also in which the frequency of ‘b’ allele was more compared to other two breeds. Ovar-DRB1 genotypes associated to FEC did not affect PCV and Hb. Digestion of GALNT6 amplicons with AluI revealed undigested single fragment referred to ‘aa’ genotypes. Since monomorphic no further association studies could be conducted for this gene. In conclusion, Ovar-DRB1 could be a reliable marker for selecting sheep resistant against H. contortus.ThesisItem Open Access IN VITRO EVALUATION OF ANTICOCCIDIAL PROPERTIES OF SELECTED EDIBLE NUT BY-PRODUCTS ON EIMERIA OOCYSTS OF POULTRY(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-06) SUDHA RANI, MOGILI; Jyothi Sree, Ch.(MAJOR); Sreedevi, C; Bharavi, KThe present study was conducted to evaluate the effect of methanolic extracts from cashew (Anacardium occidentale) nut peel, green coconut (Cocos nucifera) endocarp, almond (Terminalia catappa) hulls and partially developed groundnut (Arachis hypogea) kernel on sporulation and morphology of oocysts of Eimeria species of poultry. Sporulation inhibition bioassay was used to evaluate the activity of four extracts on the sporulation of coccidian oocysts. In this assay, unsporulated oocysts were exposed to five (300, 100, 30, 10, and 3 mg/ml) concentrations of each extract in distilled water in Petri dishes and cell-culture plates while salinomycin, ethopabate, formalin and potassium dichromate solution served as control groups. The Petri dishes and cell-culture plates were partially covered to allow the passage of oxygen and incubated at 25-29°C for 48 h, providing 80% humidity. The sporulation of the oocyst was confirmed by examining sporocysts under microscope at 40x at the end of incubation. The methalolic extracts of A. occidentale peel and T. catappa hulls showed significant (P<0.01) inhibitory effect (88.58±0.39 and 79.24±0.83%, respectively) at highest concentration of 300 mg/ml on sporulation of Eimeria oocysts in comparison with negative control, with reference drugs i.e., salinomycin, ethopabate and formalin (10%) (92.91±1.80, 84.64±2.73 and 98.03±1.04%, respectively) being more effective than the plant extracts. The inhibition of sporulation was not dose dependent for A. occidentale nut peel and T. catappa hulls extracts. At highest concentration (300 mg/ml) the rate of sporulation inhibition by C. nucifera endocarp and partially developed A. hypogea kernels was 8.07±2.34 and 1.43±0.75 per cent respectively, which was imperceptible in comparison with the reference drugs. Extracts of A. occidentale peel induced 50% inhibition at lower concentration (1.099 μg/ml) compared to T. catappa extract (7.627 μg/ml). All four extracts also damaged the normal morphology and shape of the Eimeria oocysts. Thin Layer Chromatography analysis and qualitative phytochemical screening of extracts revealed presence of flavonoids, tannins, saponins, carbohydrates, triterpenoids and alkaloids that might be responsible for the anticoccidial effects noticed. In conclusion, this in vitro study suggests that A. occidentale peel and T. catappa hulls have promising anticoccidial effects.ThesisItem Open Access CROSS SECTIONAL STUDY TO DETERMINE APPARENT OCCURRENCE OF FAMILY Anaplasmataceae IN CANINES IN AND AROUND KRISHNA DISTRICT, ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-05) SREE DIVYA DURGA, G; JEYABAL, L(MAJOR); SREEDEVI, C; SUDHAKAR, KVector-borne diseases (VBDs) are indeed a major health concern for dogs around the world, with the potential to spread zoonoses. The VBDs represent a varied group of diseases, including anaplasmosis and ehrlichiosis, which can cause serious, even life-threatening clinical conditions in dogs. In the present study, prevalence of blood parasites of canines belonging to the family Anaplasmataceae was carried out by blood smear examination in correlation with suitable haemogram and genomic DNA amplification by PCR. A total of 54 blood samples were collected at Teaching Veterinary Clinical Complex, NTR College of Veterinary Science, Gannavaram and nearby private veterinary clinics over a period of nine months (March to November, 2021) from dogs presented with tick infestation and clinical symptoms suggestive of diseases caused by Anaplasmataceae pathogens. The blood samples were initially screened for the presence of Anaplasmataceae species by microscopic examination, which revealed 14.81 (8/54) and 1.85 (1/54) per cent prevalence of Anaplamataceae species (Ehrlichia canis) by Leishman and acridine orange staining technique. None of the samples were positive for E. ewingii, Anaplasma platys, and A. phagocytophilum. DNA from whole blood was isolated to amplify E. canis virB9, E. ewingii p28, A. phagocytophilum 16SrRNA and A. platys 16SrRNA genes. Out of fifty four, 46.29 per cent of dogs were found positive for at least one pathogen of Anaplamataceae by PCR with E. canis (40.74%) being the most prevalent species followed by A. platys (5.56%). Sequences of E. canis and A. platys obtained from dogs demonstrated 95.75% and 99.86% homology with the published sequences of E. canis and A. platys, respectively. There was close similarity between clinical and haematological signs of E. canis and A. platys infections. Pyrexia and lymphadenopathy were frequent signs noticed in infected dogs. Based on clinical signs and haematological findings, dogs found positive for ehrlichiosis were graded as acute, chronic and sub clinical cases. In acute case, fever, anorexia, reduction in all the erythrogram values along with relative monocytosis, neutropenia, lymphocytosis, leucopenia and mild thrombocytopenia was observed. In chronic cases, the erythrogram values were found in the normal range with marginal leucopenia, relative neutrophilia, mild monocytosis and thrombocytopenia. In subclinical cases, the leucogram was found in normal range with severe reduction of erythrogram values. Because of low prevalence of A. platys we could not reveal any correlation between infected dogs and hematological changes but platelets count in infected dogs was found normal. Ehrlichia canis infection was more prevalent (P>0.05) in females and German shepherds dogs. The study reported the prevalence of A. platys for first time in Andhra Pradesh. Further study involving larger number of dogs from other districts of Andhra Pradesh may be beneficial for the effective characterization of Ehrlichia and Anaplasma species in Andhra Pradesh.ThesisItem Open Access MOLECULAR CHARACTERIZATION OF SARCOCYSTIS spp. ISOLATED FROM PIGS IN COASTAL DISTRICTS OF ANDHRA PRADESH, INDIA(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-01) MADHURI, GATTA; Jyothi Sree, Ch (MAJOR); Sreedevi, C; Sudhakar, KThe present study was carried out to determine the prevalence, morphological and molecular characteristics of Sarcocystis species in pigs in coastal Andhra Pradesh by microscopy and polymerase chain reaction using tissue samples collected from 163 slaughtered pigs for a period of one year from January 2021 to December 2021. Data regarding age, sex, breed and area of the animals were noted. Microscopic examination of tissue samples collected from different predilection sites such as heart, tongue, oesophagus, intercostals and diaphragmatic pillar muscles from each of 163 slaughtered pigs by intact cyst isolation and pepsin digestion method revealed sarcocysts in 6 (3.68%) and 20 (12.26%) pigs samples, respectively. Tissue samples positive for sarcocysts were fixed in 10% formalin for histopathological examination. Sarcocysts were microscopic with a smooth and thin cyst wall and were filled with banana-shaped bradyzoites. No thick walled cysts were identified. The morphological features of the cystic stage were suggestive of S. suihominis. Histopathological examination of affected muscle tissues revealed mild leucocytic infiltration, myositis and hyaline degeneration along with fatty degeneration. DNA was isolated from each pig tissue sample separately and was separately used to amplify partial segment of 18S rRNA gene of Sarcocystis spp. with the estimated amplicon size of 915 bp. Out of 163 samples, 34 (20.85 %) samples were found to be positive for Sarcocystis species. Subsequent digestion of PCR amplicons with SspI, two different patterns were visualized on electrophoresis, one with 915 bp (S. suihominis) and other with 650 bp and 265 bp (S. meischeriana). Of the 34 positive pigs, 31 (91.2%) were found to be positive for S. suihominis infection, one (2.9%) was found to be positive for S. meischeriana and two pigs (5.9%) were positive for mixed species infection with S. suihominis and S. meischeriana. Sequences of S. suihominis obtained from pigs demonstrated 90-100% identity with the Indian isolates of S. suihominis as single monophyletic group along with Italian isolate of S. suihominis. The prevalence was high in Krishna (36.36%) followed by Guntur (22.22%), Visakhapatnam (20.0%), East Godavari (18.18%), Srikakulam (15.83%), Nellore (14.28%), West Godavari, (8.33%), Prakasam (7.69%) and Vijayanagaram (7.14%) districts of coastal Andhra Pradesh. Age and breed influenced the prevalence of Sarcocystis infection being more prevalent in Non descriptive pigs (P<0.01) and adults (P<0.05). No significant difference was observed between the prevalence of male and female pigs. Overall, the prevalence of S. suihominis (20.24%; 33/163) was more compared to that of S. miescheriana (1.84%; 3/163) in the study area possessing public health risk.ThesisItem Open Access OCCURENCE OF OVINE INTRACELLULAR HAEMOPROTOZOAN PARASITES IN ANDHRA PRADESH AND THEIR ASSOCIATION WITH DRB1 GENE POLYMORPHISM(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-03) SAI SUMANTH, P; SREEDEVI, C (MAJOR); JYOTHI SREE, CH; SUDHAKAR, KThe present study was carried out to detect Babesia spp. and Theileria spp. in sheep from Andhra Pradesh and to determine the relationship between ovine MHC-DRB1 gene polymorphism and genetic resistance/susceptibility to babesiosis and theileriosis. A total of 249 blood samples including healthy (n=224) and those showing clinical signs (n=25) suggestive of haemoprotozoan infection were collected from sheep from different zones in Andhra Pradesh. The blood samples were initially processed to detect Babesia and Theileria spp. by microscopic examination. Microscopic examination of Giemsa-stained thin blood smears revealed 12.9 (32/249) per cent prevalence of Theileia spp. DNA was isolated from each sample by modified high salt method and amplified by polymerase chain reaction for simultaneous detection of 18S rRNA gene of Babesia and Theileria spp. The Babesia/Theileria genus specific PCR amplified only 430 bp 18S rRNA specific to Theileria spp. in 34.9 (87/249) per cent of sheep samples. None of the samples yielded amplification specific for Babesia spp. or mixed infections. Samples positive for Theileria spp. were further subjected to semi-nested PCR for species specific differentiation. Out of 87 theileriosis sheep, 76 (87.6%) were found to be positive for T. luwenshuni infection and eleven sheep (12.4%) were positive for mixed infection with T. luwenshuni and T.ovis. Overall, the prevalence of T. luwenshuni and T. ovis in sheep was 34.9 (87/249) and 4.4 (11/249) per cent, respectively. Age was found to be significantly (P=0.05) associated with positivity for Theileria infection in sheep being high in adult animals. The prevalence of theileriosis in sheep was not significant with respect to gender, breed and region. The Ovar-DRB1 exon 2 of 249 sheep were amplified by nested PCR from DNA samples. The 296 bp PCR products were digested by REnase BstNI, which generated three genotypes ‘aa’ (86 bp, 87 bp and 123 bp), ‘ab’ (86 bp, 87 bp, 123 bp, and 210 bp) and ‘bb’ (86 bp and 210 bp) with frequencies of 72.0, 22.0 and 6.0 per cent, respectively and two alleles ‘a’ and ‘b’ with frequencies of 0.82 and 0.18, respectively. The χ2 test of independence revealed a significant association (P<0.05) between genotypes and infection status in sheep. Though the frequency of ‘aa’ genotype was more in both the healthy (66.7) and infected (81.6) group, a significantly high frequency of ‘bb’ (6.8) and ‘ab’ (26.5) genotypes was observed compared to the infected group. A proportion of 39.7 per cent of ‘aa’, 23.2 per cent of ‘ab’ and 21.4 per cent of ‘bb’ genotypes were positive for theileriosis among the respective genotype sheep. The frequencies of ‘bb’ genotypes were significantly (P<0.05) higher in the healthy group compared with the theileriosis sheep, which implied that there was an association between these genotypes and theileriosis resistance or susceptibility. Among theileriosis positive sheep the genotype frequencies of ‘aa’, ‘ab’ and ‘bb’ pattern were 81.6 (71), 14.9 (13) and 3.4 (3) per cent, respectively. The frequency of ‘a’ and ‘b’ allele among infected sheep was 0.89 and 0.11 and is not consistent with Hardy-Weinberg equilibrium (P<0.01). The corresponding allele frequencies in the healthy sheep were 0.80 and 0.20.ThesisItem Open Access STATUS OF RESISTANCE TO CYPERMETHRIN AND IVERMECTIN IN THE BROWN DOG TICK (Rhipicephalus sanguineus sensu lato) IN ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2021-03) PRATHYUSHA, S; SREEDEVI, C (MAJOR); JYOTHISREE, Ch; SUDHAKAR, KThe brown dog tick, Rhipicephalus sanguineus sensu lato, is an important disease vector of veterinary and public health importance that is distributed worldwide. The intensive use of acaricides for tick control exerts a strong selective pressure for brown dog ticks to become resistant to them. Information on acaricide resistance in R. sanguineus s.l. populations from India is meager and no such data from Andhra Pradesh though veterinarians frequently report treatment failures. The present study was aimed to characterize the level of resistance in R. sanguineus s.l. against most commonly used drugs, alpha cypermethrin and ivermectin in Andhra Pradesh and identify the mutation in sodium channel gene that corresponds with resistance of R. sanguineus s.l. to alpha cypermethrin. Fourteen tick populations were collected from naturally infested dogs at veterinary clinics, residence and stray dog in nine state municipal corporations/municipalities. Interviews were conducted with dog owners to gather information on the history of dog treatment with acaricides. The larval packet test (LPT) and larval immersion test (LIT) was used on the progeny of adult female ticks infesting dogs to test for resistance to cypermethrin and ivermectin respectively. Mortality data was analyzed by probit analysis to determine the median lethal concentrations (LC50) for each tick population and resistance was characterised based on relative susceptibility of the different tick populations against each acaricide. Compared with the most susceptible population, thirteen (92.6%) and six (42.9%) tick populations were classified as resistant to cypermethrin and ivermectin respectively. An inter-population variation in the phenotypic level of cypermethrin and ivermectin resistance was evident (resistance ratio at LC50% ranged from 1.55 to 13.29 and from 1.16 to 4.79, respectively). Tick populations classified as resistant were collected from dogs known to have been treated with cypermethrin and ivermectin. A molecular assay targeting a point mutation in the sodium channel was optimized to identify ticks expressing cypermethrin resistance. Then, all field collected phenotypically cypermethrin-resistant populations were evaluated employing this molecular assay to determine genotypes. As confirmed by DNA sequencing, a single nucleotide polymorphism of thymine to cytosine on domain III segment VI of the sodium channel was present at a high rate in phenotypically resistant tick populations that was not present in the susceptible population. The results of this study further emphasize the necessity to maintain acaricide efficacy through rotation of active ingredients used to control ticks.ThesisItem Open Access DETECTION AND MOLECULAR CHARACTERIZATION OF SOIL TRANSMITTED HELMINTHS OF ZOONOTIC SIGNIFICANCE IN ANDHRA PRADESH(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA, 2019-12) GNANI CHARITHA, V; CHENGALVA RAYULU, V (MAJOR); MALA KONDAIAH, P; ASWANI KUMAR, K; JAGADEESH BABU, AThe present study was carried out to determine the prevalence of soil transmitted helminths of zoonotic significance among canine faecal and soil samples of Andhra Pradesh (A.P) along with molecular characterization of Toxocara and Ancylostoma spp. The overall prevalence of gastrointestinal (GI) parasites was 83.5 percent (n=745) out of 892 canine faecal samples screened by conventional microscopy. Significantly (P ≤ 0.01) higher prevalence of GI parasites was recorded in Costal Andhra (87.1%) than Rayalseema (75.2%) region of A.P. Eleven parasitic species including five nematodes, four cestodes and two protozoans were isolated from the faecal samples. The most ubiquitous parasites were Ancylostoma spp. (33.5%) and Toxocara canis (20.7%) followed by Strogyloides stercoralis (5.04%), Cystoisospora spp. (3.69%), Toxascaris leonina (3.2%), Trichuris vulpis (2.9%), Taenia spp. (2.5%), Dipylidium caninum (2.1%), Entamoeba spp. (1.23%), Diphyllobothrium latum (0.45%) and Spirometra spp. (0.34%). Concurrent mixed infection was recorded in 7.7% of dogs. Dog associated risk factors such as sex, age, breed and domestication along with effect of urbanization and seasonal influences on the prevalence of parasites were analyzed. The overall prevalence of soil transmitted parasites was 43.5 percent out of 390 soil samples screened. High contamination index of Toxocara spp. (18.5%) followed by hookworms (12.3%) was recorded. Rural soils (48.7%) were found more contaminated than urban areas (44.0%). More number of soil samples were positive for different parasitic stages collected from the garden soils (75.5%) followed by play/school grounds (65.2%), animal dwelling areas (37.3%), parks (34.0%) and veterinary dispensaries (24.0%). The prevalence of Ancylostoma spp. and Isospora spp. were positively correlated (P <0.05) with rainfall while Toxocara and Toxascaris spp. were independent of temperature influence. Responses retrieved from the questionnaire survey revealed that more than half of the pet owners (63.7%) were unaware of zoonotic soil transmitted helminths. Genomic DNA (gDNA) of 151 faecal samples were screened for Toxocara spp. using PCR-RFLP targeting ITS-2 gene. Upon genotyping of PCR amplicons (n=50) with RsaI enzyme, all the canine faecal isolates were identified as T. Canis (287 bp and 244bp). Amplification of gDNA from soil samples (n=72) yielded different product sizes viz., ~540bp (ITS-2) and ~500bp (ITS-2). Genotyping of randomly selected ~540bp product (n=32) from soil isolates with RsaI enzyme confirmed T. cati (37.5%) with three fragments (286bp, 150bp and 103bp) and T. canis (62.5%) with two fragments (287bp and 244bp). Sequencing and phylogentic analysis of ITS-2 products (~540bp) revealed 100% homozygous within the species and with the other geographical isolates of T. canis and T. cati. However, the other amplified product of ~500bp (n=21) with RsaI enzyme yielded two fragments (263bp and 219bp) and the sequencing result revealed 81.8 percent identity with T. cati. The phylogenetic analysis of ITS-2 sequences of 500bp products of A.P constituted a different sub branch and diverging from T. cati, T. canis, T. malayseinsis and T. vitulorum thus categorizing as a separate group or variant. Randomly selected 169 faecal and 48 soil samples from A.P were screened for detection of Ancylostoma spp. using a two step semi-nested PCR targeting ITS-1, ITS-2 and 5.8S gene. On gel electrophoresis, 132 (78.10%) samples showed primary PCR band at ~450bp size while secondary amplicons were observed at ~410bp size. Twenty one out of 48 soil PCR products were found positive in first round PCR and as well as in semi-nested PCR. Genotyping of semi-nested PCR products with BfaI and AhdI restriction enzymes revealed A. caninum (43/50) and A. ceylanicum (7/50) from faecal and only A. caninum (21/21) from soil isolates. The phylogeny of A.P isolates of A. caninum and A. ceylanicum revealed 100% homology within the species and with the other geographical isolates. Taken together, present data suggest the potential role of pet/stary dogs as being the main sources of contamination and signifies the need of integrated approaches to minimize the risk at different settings.
