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Open Access
PREVALENCE AND RISK FACTORS ASSOCIATED WITH BOVINE TICK INFESTATION IN TWO DIFFERENT AGRO-CLIMATIC ZONES IN ANDHRA PRADESH
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA, 2019-12) SUNEETHA, KANDI; SREEDEVI, C (MAJOR); JYOTHISREE, Ch.; Muralidhar, M
The present study was carried out to identify the tick species and risk factors associated with tick prevalence in bovines in Andhra Pradesh, India. A total of 3720 animals, including 1942 cattle and 1778 buffaloes in 344 livestock farms covering tropical wet and dry zone (North coastal zone) and tropical semi-arid zone (scarce rainfall zone of Rayalaseema) were examined for the presence of ticks. Overall, 10,371 ticks representing three genera were collected. Analysis on morphology and DNA sequence of 16S ribosomal RNA and cytochrome C oxidase subunit I (COI) four species: Haemaphysalis bispinosa (n = 5704), Rhipicephalus microplus (n = 2533), Hyalomma anatolicum (n = 1272) and H. isaaci (n = 862) were identified with single infestations being the rule. Results revealed that the molecular and morphological data were consistent in the identification of four tick species. The constructed COI phylogenetic tree further identified R. microplus tick samples as R. microplus clade C. Haemaphysalis bispinosa was the predominant species in two zones and the occurrence of H. anatolicum was restricted to semi-arid zone. The overall percentage of tick infested bovines was 25.4 per cent and was 25.8 and 25.0 per cent in cattle and buffaloes respectively. Out of 947 infested bovines, 397 were infested with H. bispinosa, 253 with R. microplus, 175 with H. anatolicum and 122 with H. isaaci. The variation in the prevalence of tick infestation was not significant (P>0.05) in two zones but the prevalence was significantly different among districts of semi-arid zone (P<0.05) and wet and dry zone (P<0.05). The median tick burden per animal significantly (P<0.01) differed between two zones, however, the difference was not significant (P>0.05) among cattle and buffaloes. Age, gender and breed significantly (P<0.01) influenced the prevalence of tick infestation. The prevalence was higher in young and male animals and was lower in indigenous animals. Analysis of questionnaire data by binary logistic regression revealed that the loose housing system, absence of rural poultry, soft floor, presence of boundary wall and rearing multiple ruminant species were potential risk factors associated with higher tick prevalence in livestock farms. Data from the results of this study provide a guideline in the planning of effective control strategies for ticks and tick-borne diseases in bovine population of the studied area.
Open Access
PREVALENCE AND RISK FACTORS ASSOCIATED WITH GASTRO-INTESTINAL PARASITIC INFECTION IN DOGS FROM CENTRAL COASTAL DISTRICTS, ANDHRA PRADESH WITH SPECIAL REFERENCE TO ANTHELMINTIC RESISTANCE
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA, 2019-02) ROJA, KURAMDAS; SREEDEVI, C(MAJOR); MALAKONDAIAH, P; SUDHAKAR, K
Coprological examination was used to estimate the prevalence of gastrointestinal parasites in dogs from central coastal districts of Andhra Pradesh, India for a period of one year. On examination of faecal samples of 1907 dogs, an overall prevalence of 52.2 per cent was observed. Dogs harbouring one parasite was more common (49.7%) than those harbouring two (2.5%). The most prevalent species was Ancylostoma caninum followed by Toxocara canis, Cystoisospora ohioensis, Dipylidium caninum, Toxascaris leonina, Capillaria aerophila, Diphyllobothrium latum, C. canis, Entamoeba spp., Trichuris vulpis and Cryptosporidium spp. The overall prevalence was high (P<0.001) in young dogs (71.1%) than that in adult dogs (28.0%). The infection with helminth parasites (P<0.001) and co-infections (P<0.01) were more frequently found in young dogs. Ancylostoma caninum, T. canis, T. leonina, D. caninum, and Cystoisopora spp. were significantly more prevalent in young dogs. However, the prevalence of all the parasite species showed a significant decreasing trend with increasing age. Statistically there was no significant (P>0.05) difference between prevalence of parasites and sex, breed and medium of dogs. The prevalence of GI parasitic infection was significantly (P<0.001) high in stray dogs (65.9%) owing to higher prevalence of T. canis, D. latum, C. aerophila and C. canis. The prevalence of A. caninum (31.6%) was highly significant (P<0.001) in pet dogs. Statistically significant (P<0.01) relationship between the season and prevalence of parasites was observed in the study area, the infection being more prevalent during rainy season (62.4%) than that in winter (54.4%) and summer (39.9%). The prevalence of T. canis was significantly (P<0.001) high during rainy season whereas A. caninum, D. caninum and C. canis showed statistically significant higher prevalence during winter season. Zoonotic risk factors for canine GI parasitic infection were assessed by obtaining information viz. origin/owner’s address, age, sex, breed, living condition of the dog, history of anthelmintic usage and owner’s awareness on zoonoses from the pet and kennel owners in the form of questionnaire. The analysis revealed that the owners from study region are unaware about parasite, its mode of transmission, prophylactic medication and control. The living condition of the dogs, age, season and lack of zoonotic awareness of owners were identified as risk factors for different parasitic infection using binary logistic regression. The higher prevalence of zoonotic species in dogs in the studied region, associated with unawareness of the owners regarding parasitic zoonoses indicated that the risk of zoonotic infection by canine intestinal parasite may be high. Further egg hatch assay was used to evaluate the efficacy of thiabendazole in the treatment of canine ancylostomosis using A. caninum eggs isolated from naturally infected dogs. The results of the EHA revealed highly significant (P<0.01) inhibitory effect on egg hatching rates with thiabendazole (LD50 = 0.0049 μg/mL) reflecting benzimidazoles high effectiveness.
Open Access
EPIDEMIOLOGICAL SURVEILLANCE OF ECTOPARASITIC INFECTIONS IN DOGS IN KRISHNA - GODAVARI ZONE OF ANDHRA PRADESH
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA, 2019-01) SOWJANYA, MANTRI; MALAKONDAIAH, P(MAJOR); SREEDEVI, C; SATHEESH, K
The present study was investigated to determine the prevalence of ectoparasites in dogs of Krishna – Godavari zone in Andhra Pradesh and to study the alterations in haematology, mineral status and enzyme profile in canine demodicosis. Examination of 2197 dogs in different Veterinary Hospitals and Veterinary Poly Clinics in East Godavari, West Godavari, Krishna, Guntur and Prakasam districts revealed 56.9 per cent prevalence. The infection was highly prevalent in Guntur (60.2%) followed by west Godavari (57.5%), Krishna (57.3%), Prakasam (55.2%) and East Godavari (52.1%) districts. The species of ectoparasites identified were Ctenocephalides canis (6.4%) Ctenocephalides felis (3.9%), Heterodoxus spiniger (1.9%), Rhipicephalus sanguineus (26.8%), Sarcoptes scabei var canis (8.7%), Demodex canis (6.1%) and Chrysomyia bezziana (1.7%). Statistically significant (P<0.05) difference between the prevalence of infected dogs and species of ectoparasites was observed. The overall prevalence of ectoparasites was 56.7 and 57.1 per cent in juvenile and adult dogs respectively. There was no significant (P>0.05) association between the prevalence of infection and age groups of dogs though the infection was more in adult dogs compared to juveniles. In both the age groups, arachnids were more prevalent than insects. There was no significant (P>0.05) association between prevalence of ectoparasites and sex of dogs. The prevalence of ectoparasites among males and females was 57.7 and 56.0 per cent respectively. The overall prevalence was higher in males compared to females whereas ticks showed highest prevalence in females (28.3%) than in males (25.7%). The prevalence of ectoparasites in pure breed and cross breed was 56.0 and 58.5 per cent respectively. There was no significant (P>0.05) association between infection of parasites and breeds of dogs. The prevalence of Ctenocephalides canis, Ctenocephalides felis, Rhipicephalus sanguineus, Sarcoptes scabei var canis were higher in cross breed dogs whereas Heterodoxus spiniger, Demodex canis, Chrysomyia bezziana were higher in pure breed dogs. The prevalence of ectoparasites in winter, summer and rainy was 55.9, 46.6 and 70.4 per cent respectively. Significant (P<0.05) relationship between the seasonality and breed of dogs was observed in the present study. The prevalence was highest in rainy season followed by winter and summer. The ticks, mites and maggots were more prevalent in rainy whereas insects were more prevalent in winter season. Haemato-biochemical, enzyme and mineral profile alterations were found in demodicosis dogs compared to that of healthy dogs. There was significant (P<0.05) decrease in Hb, PCV and TEC whereas significant increase in TLC of demodicosis dogs. There was non-significant (P>0.05) elevation in ALT and AST of demodicosis infected dogs. The level of calcium and zinc showed significant (P<0.05) decrease in demodicosis infected dogs whereas copper showed significant (P<0.05) elevation. No significant (P>0.05) alterations in the level of phosphorous of demodicosis infected dogs was observed.
Open Access
IN VITRO EVALUATION OF ANTHELMINTIC PROPERTIES OF THE HYDRO-ALCOHOLIC EXTRACTS OF SELECTED MEDICINAL PLANTS ON Haemonchus contortus
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517502. (A.P.) INDIA, 2018-12) TULASI, DAVULURI; SREEDEVI, C(MAJOR); MALAKONDAIAH, P; SRINIVASA RAO, G
In recent years, there has been growing interest in alternative therapies and the therapeutic use of natural products especially of medicinal plants for control of parasites. In the present study in vitro assays such as egg hatch assay (EHA), larval paralysis assay (LPA) and adult worm motility inhibition assay (WMIA) were conducted to determine the anthelmintic efficiency of hydro-alcoholic extracts of Anacardium occidentale shell, Illicium verum fruit and Artocarpus heterophyllus seed on eggs, infective larvae (L3) and adult worms of Haemonchu contortus, in comparison to albendazole. Among three extracts, A. occidentale shell showed significant (P<0.01) inhibitory effect on egg hatching and larval motility followed by A. heterophyllus seed and I. verum fruit extracts. Extracts of A. occidentale shell induced 50% inhibition at lower concentration (0.0255 mg/mL) compared to I. verum fruit extract (0.0612 mg/mL) and A. heterophyllus seed (0.0372 mg/mL) extracts. The LD50 value of reference drug albendazole (positive control) was 0.237 µg/mL. Extracts of A. occidentale shell required maximum of 0.5 mg/mL, whereas extracts of I. verum fruit and A. heterophyllus seed required maximum concentration of 4 and 2 mg/mL respectively, to induce 100 per cent egg hatch inhibition. Similarly A. occidentale shell showed maximum activity on motility of L3 larvae (LD50 = 0.196 mg/mL) with 100 per cent paralysis while A. heterophyllus seed (LD50 = 0.666 mg/mL) and I. verum fruit (LD50 = 1.418 mg/mL) exhibited 84.67±1.76 and 72.66±1.76 per cent paralysis respectively, at higher tested concentration of 6 mg/mL. In WMIA, three extracts induced significant (P<0.001) mortality of adult worms; however the activity of A. occidentale shell was higher (100%) than I. verum fruit (36.6±3.3%) and A. heterophyllus seed (70.00±5.7%) at a concentration of 6 mg/mL within 1 h post exposure. Anacardium occidentale extract revealed better LD50 (1.0365 mg/mL) values in comparison with I. verum fruit (LD50 = 3.848 mg/mL) and A. heterophyllus seed (LD50 = 2.398 mg/mL) in the WMIA. Three plant extracts exhibited significant (P<0.001) dose dependant anthelmintic responses by inhibiting egg hatching and causing paralysis of larvae and mortality of worms. In vitro effect of these extracts on lactate dehydrogenase activity of H. contortus was also studied. All extracts significantly (P<0.01) inhibited the activity of LDH catalysing the oxidation of lactate. Maximum level of inhibition of LDH activity was noticed in A. occidentale shell extract treated worms. Thin Layer Chromatography analysis and qualitative phytochemical screening of three extracts revealed presence of alkaloids, flavonoids, tannins, saponins, carbohydrates and proteins which might be responsible for the anthelmintic effects noticed. Overall, this in vitro study suggests that these three plants have promising anthelmintic effects.
Open Access
ASSESMENT OF ACARICIDAL ACTIVITY OF NANOSCALE ZnO ENCAPSULATED PIPERINE FORMULATION AGAINST RHIPICEPHALUS MICROPLUS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-03) SNIGDHA, KANCHARANA; CHENGALVA RAYULU, V(MAJOR); SRINIVASA RAO, K; SREEDEVI, B
ABSTRACT: The present study was undertaken with an aim to synthesize and evaluate the acaricidal activity of nanoscale zinc oxide piperine formulation against Rhipicephalus microplus ticks. Nanoscale zinc oxide piperine formulation (NZPF) was prepared by using 0.1% zinc oxide nanoparticles (ZnONPs) solution and 20% piperine solution employing encapsulation technique. The synthesised NZPF was characterized by employing UV–Vis spectroscopy, Fourier Transformed Infrared (FT-IR) analysis, X-ray Diffraction (XRD), Dynamic Light Scattering (DLS), Scanning Electron Microscopy (SEM) and Energy Dispersion Spectroscopy (EDS) analysis. The maximum absorbance peak of the Localized Surface Plasma Resonance (LSPR) of ZnONPs and NZPF was observed at 240 nm and 345 nm, respectively by using UV-Vis Spectrophotometer. The FT-IR spectra peaks of ZnONPs and NZPF were found to be 3844, 3739, 3619, 2351, 1694 and 1526 cm-1 and1633, 1583, 1490, 1448 and1436 cm-1, respectively. The XRD analysis showed strong peaks of 31.854°, 34.497°, 36.327°, 47.64°, 56.70°, 63.060°and 68.10° corresponds to Bragg’s reflections at (1 0 0), (0 0 2), (1 0 1), (1 0 2), (1 1 0) and (1 0 3) planes conforming the wurtzite crystalline structure of NZPF. Hydrodynamic diameter and zeta potential of the hydrosol of ZnONPs and NZPF were found to be 38.6nm and - 28.8 mV and 75nm and -36.4 mV, respectively with DLS technique. The EDS micrograph of ZnONPs showed the peaks of zinc and oxygen elements confirming the chemical composition and presence of chemical constituents (Zn77.51 % and O 22.49%).The SEM images revealed that the synthesized ZnONPs were spherical in shape with mean size of 30nm. Whereas synthesized NZPF were in rectangular rod shape with occasional agglomeration, highly poly dispersed and measured 1-2 μm. Acaricidal activity of deltamethrin, piperine, ZnONPs and NZPF on Rhipicephalus microplus was evaluated by two bioassays viz., Larval Packet Test (LPT) and Adult Immersion Test (AIT). LPT with a discriminating dose of deltamethrin (75 ppm) showed 59% mortality of R. microplus larvae. Total mortality (100%) was seen against R. microplus larvae at concentrations of 9ppm, 8ppm and 7 ppm with piperine, ZnONPs and NZPF, respectively. AIT with a discriminating dose of deltamethrin (75ppm) against adult R. microplus showed a mortality of 40%, oviposition inhibition of 78.309% and the lowest egg mass weight with17.8±1.31 mg. Mortality rate and oviposition inhibition of R. microplus were 100% whereas egg mass and reproductive index were completely nil with both piperine and ZnONPs at a concentration of 20ppm and NZPF at a concentration of 15 ppm. The egg mass values and reproductive indices were inversely proportional to the piperine, ZnONPs and NZPF concentrations whereas oviposition inhibition percent was directly proportional to the piperine, ZnONPs and NZPF concentrations. LC50 values of LPT and AIT were at the lowest concentration of 1.312 ppm and 3.505 ppm for NZPF whereas LC99 values were seen at the lowest concentration of 12.690 ppm and 33.741 ppm for ZnONPs. NZPF showed a potent ovulation inhibitory activity with significantly (P<0.05) lower IC50 and IC99 values compared to ZnONPs and piperine. Both LPT and AIT results clearly indicate the development of resistance in R. microplus ticks against deltamethrin. Synthesised NZPF, ZnONPs and piperine were found to have significantly (P<0.05) higher acaricidal activity. However, NZPF had high acaricidal efficacy at lower concentrations than pure phytochemical piperine, ZnONPs and deltamethrin. NZPF could be potential alternative to routine chemical acaricides for control of tick infestation of cattle in the wake of development of acaricidal resistance, residual effect and environmental pollution. Synthesis of NZPF and evaluation for its acaricidal activity in vitro is probably the first kind of report and further detailed study must be carried out before their application in vivo.
Open Access
SYNTHESIS OF MYCOGENIC SILVER NANOPARTICLES AND EVALUATION OF THEIR ACARICIDAL ACTIVITY AGAINST RHIPICEPHALUS MICROPLUS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-01) PRIYADARSHINI, A.T; SRINIVASA RAO, K(MAJOR); CHENGALVA RAYULU, V; PRASAD, T.N.V.K.V
Open Access
EVALUATION OF IN VITRO ANTHELMINTIC ACTIVITY OF CHITOSAN ENCAPSULATED ALBENDAZOLE AGAINST GASTROINTESTINAL NEMATODES OF SHEEP
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2017-02) RAMYA, V; CHENGALVA RAYULU, V(MAJOR); VENU, R; ALPHA RAJ, M
ABSTRACT: Present investigation was carried out to evaluate the anthelmintic activity of chitosan encapsulated albendazole with phytochemical combinations against GI nematodes of sheep. Various microspheres including chitosan alginate (CS-ALG), chitosan encapsulated albendazole (CS-ABZ), chitosan encapsulated albendazole in combination with five phytochemicals viz., caffeic acid, cinnamic acid, curcumin, piperine and syringic acid were synthesized and characterized. Phytochemical loaded CS-ABZ microspheres were designated as chitosan encapsulated albendazole with caffeic acid (CS-ABZ-CAA), chitosan encapsulated albendazole with cinnamic acid (CS-ABZ-CIA), chitosan encapsulated albendazole with curcumin (CS-ABZ-CUR), chitosan encapsulated albendazole with piperine (CS-ABZ-PIP) and chitosan encapsulated albendazole with syringic acid (CS-ABZ-SYN). Synthesized microspheres were spherical with a diameter ranging from 0.470 to 0.634 mm. The mean diameter of CS-ALG and CS-ABZ were significantly (P<0.05) lower than CSABZ- CIA. Majority of unloaded CS-ALG microspheres had a diameter of 0.3 mm. Loading of CS-ALG with ABZ and phytochemicals resulted in microspheres with 0.3 to 0.8 mm diameter. Maximum concentration of ABZ was achieved in CS-ABZ microspheres (55.56mg/100mg). Addition of phytochemicals decreased the loading of ABZ in the microspheres. The highest concentration of ABZ was observed in CSABZ- CUR (19.53 mg/100mg) and the lowest in CS-ABZ-CIA (11.63 mg/100mg). Phytochemical concentration (mg/100mg) of microspheres ranged from 0.78 (CS-ABZ-CAA) to 1.18 (CS-ABZ-SYA). Loading efficacy of CS-ABZ, CS-ABZCAA, CS-ABZ-CIA, CS-ABZ-CUR, CS-ABZ-PIP and CS-ABZ-SYN estimated in this study was 69.45, 23.63, 14.54, 23.42, 23.23 and 19.28 percent, respectively. The highest albendazole loading efficacy (69.45%) achieved in CS-ABZ decreased with the addition of phytochemicals. The release of ABZ from the microspheres under simulated gastric conditions was minimum at pH 1.2 (1.93 to 6.65%) and maximum at pH 6.8 (10.13 to 51.73%). Adding phytochemicals to the microspheres improved the release of ABZ in basic pH with the highest release from PIP microspheres (51.73%) followed by CAA (39.04%) and SYA (38.10%) while CUR and CIA had no effect. Release of ABZ from all the microspheres followed Krosmeyer-Peppas model (Q=ktn) with R2 ranging from 0.845 to 0.915. The release from ABZ, CAA, CIA, PIP and SYA was predominantly through anomalous diffusion while release from CUR microspheres followed zero order. Examination of sheep faecal samples collected during the investigation revealed the presence of eggs of Haemonchus contortus, Strogyloides spp., Bunostomum spp. and Trichuris spp. Egg hatch test (EHT) with LC50 (μg mL-1) values of ABZ (0.51), CS-ABZ (0.40) and phytochemicals (0.57 to 1.30) indicated the development of anthelmintic resistance of GI nematodes. Whereas the LC50 (μg mL-1) of CS-ABZ-CUR (0.09) and CS-ABZ-PIP (0.08) were significantly (P<0.05) lower than ABZ and CS-ABZ. The LC50 (μg mL-1) values of CS-ABZ-CAA (0.27), CS-ABZ-CIA (0.24), CS-ABZ-CUR (0.25), CS-ABZ-PIP (0.17) and CSABZ- SYA (0.26) obtained with larval development test (LDT) were significantly (P<0.05) lower than CS-ABZ (0.41) and pure ABZ (0.46). There was no significant (P>0.05) difference between the LC50 values of various phytochemical loaded microspheres. Coproculture of positive faecal samples for GI nematodes revealed the presence of third stage larvae of H. contortus and Strongyloides spp. No third stage larvae were observed from the wells after LDT with CS-ABZ-CAA, CS-ABZ-CIA, CS-ABZ-CUR, CS-ABZ-PIP and CS-ABZ-SYA. However, live third stage larvae of H. contortus were recovered from the wells cultured with ABZ, CS-ABZ and pure phytochemicals. Pure ABZ, CS-ABZ and pure phytochemicals were found least effective against GI nematodes of sheep as evidenced by EHT and LDT. CS-ABZ-PIP and CSABZ- CUR microspheres synthesized in the present study were found to be better alternatives to routine chemical anthelmintics for treatment of GI nematodes. These microspheres can overcome ABZ anthelmintic resistance due to synergistic combination with phytochemicals. Chitosan encapsulated albendazole with phytochemical combination is an effective novel formulation with improved anthelmintic activity and release kinetics.
Open Access
PREVALENCE, HAEMATOLOGICAL, BIOCHEMICAL AND HISTOPATHOLOGICAL OBSERVATIONS OF PARAMPHISTOMOSIS OF DOMESTIC RUMINANTS IN TIRUPATI
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-12) PREETHI, M; VENU, R(MAJOR); SRINIVASA RAO, K; SRILATHA, Ch; VINOD KUMAR, N
ABSTRACT: The present study on ‘prevalence, haematological, biochemical and histopathological observations of paramphistomosis of domestic ruminants in Tirupati’ was conducted in cattle, sheep and goats. A total of 2133 samples (dung, blood, serum, tissues and amphistome specimens) from slaughtered domestic ruminants was examined. In direct faecal smear examination, an overall prevalence of 24.29 percent of paramphistomosis infection was recorded, whereas in faecal sedimentation method, 32.51 percent was observed. In cattle, the prevalence of infection by direct faecal smear and faecal sedimentation methods were 17.43 and 31.19 percent, respectively. In sheep, the prevalence rates were recorded higher than cattle. In goats, the prevalence of paramphistomosis by direct faecal smear examination was 20.66 percent, whereas by faecal sedimentation method, it was 30.52 percent. Out of 109 cattle carcasses, in 47 cases (43.12) amphistomes were found in rumen, reticulum and bileduct during slaughterhouse examination. In sheep and goat, the prevalence rates were 42.15 and 40.85 percent, respectively by slaughterhouse examination. Age-wise, higher prevalence was recorded in cattle of 2-4 years followed by older animals of above 4 years and young animals of <2 years. Slightly higher prevalence (26.79%) was noticed in >1- 2 years old sheep than <1year age group. In goats, the prevalence of infection was lower in the age group of <1year, when compared to their counterparts in sheep. Based on slaughterhouse study, the sheep of <1 year old were more infected (56.28%) than >1-2 years old sheep (36.85%). In contrast, the higher prevalence was noticed in >1-2 years (63.27%) than < 1 year old goats (21.74%). Sex-wise, the prevalence of infection in male cattle was slightly higher than females by direct faecal smear and slaughterhouse examinations, whereas in sedimentation method, female animals showed higher infection than male. The prevalence of infection in female sheep was higher than male sheep. Overall, statistically no significance difference was observed between male and females, in respect to their diagnostic method. Blood samples were screened for haematological observations such as PCV, Hb and RBC counts. Statistically, significant difference was noted between immature amphistomosis and uninfected groups of cattle, sheep and goats. No significant difference was noted between infected and uninfected animals of cattle and sheep in relation to PCV, Hb and RBC values, but in goats significant difference was observed. In DLC, statistically, no significant difference was observed in immature, infected and uninfected amphistomosis cattle. In immature and uninfected sheep, neutrophil and monocyte values showed significant alterations, but in goats it was in neutrophil and lymphocytes. Serum sample screening revealed, low levels of total proteins and albumins in immature amphistomosis infected cattle, sheep and goats. Gross and histopathological changes of amphistome infected rumen, reticulum and bile duct were observed. The adult amphistome parasites invading the rumen mucosa by sucker and plugging of mucosa of bile duct was noticed in cattle. In the bile duct of cattle, massive and extensive infiltration of plasma cells, lymphocytes and monocytes in between acinar structures was recorded. Hyperplasia of bile duct, degenerative changes and necrotic changes in acinar mucosal lining epithelial cells were appreciated prominently. In immature amphistomosis, duodenum of sheep revealed thickened and edematous; mucosal surface was severely congested, petechial haemorrhages and necrosis were also noticed. Histopathological studies of infected duodenum of sheep revealed, plugging of the mucosa with sucker and extensive infiltration of macrophages and plasma cells deep into the mucosa. Based on morphology, 5 amphistomes were recognized viz., Cotylophoron cotylophorum, Paramphistomum cervi, Gastrothylax crumenifer, Fischoederius elongatus and Gigantocotyle explanatum. Mixed infections were noticed higher at 50, 73.6 and 62 percent, in cattle, sheep and goat, respectively. In immature amphistomosis infected ruminants, haematological and biochemical parameters were estimated and showed a great variation and microscopic examination of duodenal wall scrapings revealed immature amphistomes.
Open Access
IDENTIFICATION OF SARCOCYSTIS SPECIES IN CATTLE (Bos taurus) BY PCR-RFLP
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2016-10) MOUNIKA, K; SREEDEVI, C(MAJOR); VENU, R; SRINIVASA RAO, T
ABSTRACT: The present study was carried out to determine the prevalence of bovine sarcocystosis and identify various species of Sarcocystis from cattle in different regions of Chittoor district, Andhra Pradesh by PCR-RFLP. Macroscopic examination and microscopic examination by pepsin-HCl digestion method of 150 slaughtered cattle from Tirupati, Chandragiri, Chittoor, Renigunta and Pakala regions in Chittoor district revealed an overall 91.33 per cent of prevalence of sarcocystosis. The infection was highly prevalent in Tirupati (97.14 %) compared to that of Chandragiri (92.5 %), Renigunta (90.0 %),Chittoor (80.0 %), and Pakala (70.0 %) regions of Chittoor district. The prevalence of macroscopic and microscopic sarcocysts was 6.57 and 93.43 per cent respectively. Macroscopic cysts were exclusively observed in oesophagus. The prevalence of infection increased with advance in age and there was a significant relationship between the prevalence of infection and age groups of cattle (P<0.001). There was no significant difference (P>0.001) between the prevalence of Sarcocystis infection in male (91.76 %) and female (90.76 %) cattle. Genomic DNA was extracted separately from bradyzoites of all 137 cattle that were positive for sarcocystosis and amplified 18S rRNA gene of Sarcocystis that yielded PCR product of 900 bp. Digestion of 18S rRNA gene PCR products was performed with restriction endonuclease BseLI to detect different species of Sarcocystis. On digestion with restriction endonuclease three different patterns were observed on agarose gel electrophoresis, one with 513 bp and 343 bp and other with 525 bp, 241 bp and 141 bp which were referred to Sarcocystis cruzi and S. hirsuta respectively. While another with 532 bp and 335 bp was referred to S. fusiformis. S. cruzi (93.43 %) was significantly more prevalent in Chittoor district in comparison with the S. hirsuta (4.38 %) and S. fusiformis (2.19 %). Infection of cattle with S. hominis was not observed in the study area. These findings provide evidence that Sarcocystis species of cattle and water buffaloes are not strictly intermediate host specific but might occasionally infect water buffaloes and cattle, respectively where both hosts occur and natural cross transmission through dogs are possible. S. fusiformis is able to use the cattle as an intermediate host and is not restricted to buffalo. PCR-RFLP was helpful in studying the molecular epidemiology of sarcocystosis in cattle and was a good method in discriminating between species of Sarcocystis.

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