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ThesisItem Open Access CERTAIN ASPECTS OF SPIROCERCOSIS WITH REFERENCE TO ITS PREVALENCE AND IMMUNODIAGNOSIS(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2006-12) JYOTHISREE, CHITITHOTI; HAFEEZ, Md.(Major); UDAY KUMAR, M; ALAHA SINGARI, NABSTRACT : The research work on “Certain aspects of Spirocercosis with reference to its prevalence and immunodiagnosis” was under taken to record the prevalence of spirocercosis in different regions of Andhra Pradesh besides the study of the tissue reactions, haematological and biochemical changes of the disease and the efficacy of certain chemotherapeutic drugs against the spirocercosis and also to prepare an antigen of Spirocerca lupi to detect spirocercosis by immuno diagnostic techniques. A total of 562 dogs were examined in five (5) different places of Adhra Pradesh. Based on faecal sample examination, the over all per cent prevalence of spirocercosis in dogs was 3.02. Among the different (5) places of Adhra Pradesh under this study the prevalence was more in Hyderabad (5.45%) followed by Tirupati(3.6%), Ongole(2%), Kurnool (1.75%) and Guntur (0%). The prevalence of spirocercosis based on post mortem examination was 10 per cent at Tirupati. The prevalence of spirocercosis was more in 1-5 years age group (3.83%) followed by above 5 years age group (2.64%). The prevalence was more in males (3.30%) than females (2.70%). The prevalence was more in non discripts (5.92%) followed by German shepherd (4.09%) and in Pomeranians (1.57%). The prevalence was more in Winter season (5.91%) followed by Rainy season (2.12%) and Summer (1.06%). Radiographic examination showed 47.05 per cent confirmity in suspected cases. The gross pathological lesions like pale lungs, putrid material in the thoracic cavity, exostosis in 7th, 8th and 9th thoracic vertebrae, 5-6 thick hard nodular masses in thoracic portion of aorta and hen egg size nodule in the terminal portion of oesophagus and proximal portion of stomach were generally observed in natural cases of spirocercosis. Histopathological lesions in oesophagus were cut section of parasite and parasitic ova surrounded by inflammatory cells and moderate edema in the serosal layer along with giant cells. Aorta showed infiltration of inflammatory cells in tunica intima, cut section of parasite along with parasitic ova. Severe infiltration of inflammatory cells neutrophils and plasma cells along with fibrous tissue in tunica media. Lung showed focal interstitial pneumonia. Liver showed inflammatory cells between the hepatic cards. Haemorrhages between the cardiac muscle fibres were also observed in the heart. Haematological and biochemical changes in blood and sera samples of naturally infected spirocercosis were low haemoglobin concentration, leukocytosis and Neutrophilia. Serum samples showed low level of Total protein, Albumin and Glucose and increased concentration of Alkaline phosphotase, Amylase and Creatinine. The efficacy of the drugs Ivermectin, Ivermectin plus Prednisalone and Doramectin was tested against naturally infected Canine spirocercosis. The therapeutic efficacy of the above drugs was found to be 100 per cent. Crude antigen of Spirocerca lupi revealed protein concentration as 11.38 mg/ml. Attempts were made to diagnose S.lupi infection in naturally infected dogs by conducting four serodiagnostic tests viz., Agar-Gel immuno diffusion test (AGID), Immuno Electrophoresis (IEP), Counter Immuno Electrophoresis (CIEP) and Passive Haemagglutination Test (PHA). 17 sera samples collected from dogs naturally infected with S.lupi infection were tested against crude S.lupi antigen using AGID, IEP, CIEP and PHA tests. These tests showed a sensitivity of 88.2, 82.3, 88.2 and 76.4 percent in detecting S.lupi infection in naturally infected dogs, respectively The whole homogenate of S.lupi antigen was subjected to 10 per cent SDS-PAGE for the analysis of polypeptide profile. SDS-PAGE revealed a total number of 7, 8 polypeptides, respectively. The apparent molecular weight of polypeptides of these ranged from < 25 KDa to 151 KDa.ThesisItem Open Access STUDIES ON CERTAIN ASPECTS OF BOLIARY AMPHISTOMOSIS IN BUFFALOES (Babalus bubalis)(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2006-02) MADHAVI, B; HAFEEZ, Md.(Major); RAMAKRISHNA REDDY, P; ALAHA SINGARI, NABSTRACT: We research work on "Studies on certain aspects of biliary amphistornosis in buffaloes" was under taken to study the prevalence of gigantocotylosis in naturally infected buffaloes, based on dung sample and slaughter house examination. Age, sex, breed, season wise prevalence at certain places of Chittoor and Nellore districts of Andhra Pradesh were surveyed-$ total of 221 buffaloes were examined in the present study. The present of prevalence in buffaloes was 38.91 per cent. Among the 12 places choosen the prevalence was more in Tirupati (41.67%) least in Chittoor (12.5%) in Chittoor District, whereas in Nellore District more prevalence was recorded in Nellore (48.15%) and least in Podalakur (35.71 %).The prevalence of biliary amphistomosis was more in above 4 years agc group (45%) and in females (40%). The prevalence was more in non-descriptive animals (40.68%) and also in rainy season (46.15%). Attempts were made to observe certain biochemical changes in - serum of naturally infected buffaloes. Serum total proteins (6.792 g/100 ml), albumins (3.256 g/ dl) and A/G ratio (0.921 g/dl) were decreased where as globulins (3.536 g/dl), conjugated bilirubin (0.093 g/dl) and total bilirubin (0.805 ddl) wcre increased.Attempts wcre madc to diagnose gigantocotylosis in naturally infected buffaloes by carrying out three sero diagnostic tests viz., counter current immunoelectrophoresis, ELISA and Dot - ELISA. The whole worm extract of antigen was prepared and the protein content of this antigen was prepared and the protein content of this antigen was estimated to be 11.2 mg/ml. Hyperimmune serum was raised against whole worm extract of Gigantocotyle explanaturn antigen, and serum samples selected randomly from naturally infected buffaloes with Gigantocotyle explanaturn were tested against whole worm extract of Gigantocotyle explanaturn. The CIEP, Dot - ELISA and ELISA tests showed a sensitivity of 85.8, 92 and 98 per cent respectively in detecting biliary amphistornosis in naturally infected buffaloes.ThesisItem Open Access Studies On Certain Apects Of Canine Ehrlichiosis(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2001-09) Lejeune Virapin Manigandan; Hafeez, Md.(Major); Chengalvarayulu, V; Nisar Ahmed, MThesisItem Open Access Studies On Myiasis Of Certain Domestic Animals(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2001-02) Jalajakshi, K; Hafeez, Md.(Major); Ramkrishna Reddy, P; Prabhu, M.R.LThesisItem Open Access Studies On Certain Aspects Of Tick Infestation In Crossbred Cattle In And Around Tirupati (Andhra Pradesh)(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2000-12) Rajendran, C; Hafeez, Md.(Major); Ramakrishna Reddy, P; Sreenivasulu, D; Kameswara Rao Panth, MThesisItem Open Access STUDIES ON BOVINE MICROFILARIOSIS WITH SPECIAL EMPHASIS ON PREVALENCE AND IMMUNODIAGNOSIS(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2009-10) LALITHA, Ch; Md. HAFEEZ, (Major); CHENGALVA RAYULU, V; SRILATHA, ChABSTRACT : The research work on “Studies on bovine microfilariosis with special emphasis on prevalence and immunodiagnosis” was undertaken to evaluate certain immunodiagnostic tests for large scale screening and field applicability in bovines. Attempts were made to study the prevalence of bovine microfilariosis in certain coastal districts of Andhra Pradesh. Blood and sera samples of 1628 bovines were collected from Vishakapatnam (575), Vizianagaram (362), Srikakulam (222), Prakasam (130) and West Godavari (339) districts of coastal Andhra Pradesh and screened by conventional techniques viz., wet blood film examination and modified Knott’s technique and immunological techniques viz., indirect ELISA and dot-ELISA. With modified Knott’s method microfilariae could be detected in 81 blood samples out of 1628 samples examined with the overall prevalence rate of 4.97 percent. The prevalence of microfilariosis in cattle and buffaloes was recorded as 4.4 percent and 5.1 percent, respectively. The higher rate of infection was recorded in West Godavari (6.4 %), followed by 4.97, 4.69, 4.6 and 3.6 percent in Vizianagaram, Vishakapatnam, Prakasam and Srikakulam districts, respectively. Highest prevalence observed in female bovines as 5.1 percent while none of the male animal was found positive for microfilariosis. Higher prevalence was noticed in the bovines with age group of above 9 years (9.09 %), followed by age groups of 6 to 9 years (5.7 %) and 3 to 6 years (2.6 %). Prevalence of bovine microfilariosis was recorded as 6.5, 3.6 and 3.1 percent in lactating animals, heifers and non-lactating animals, respectively. Cross bred cattle and Murrah buffaloes have an infection rate of 4.8 and 5.9 percent, respectively while the infection rates of indigenous cattle and non-descript buffaloes were 3.9 and 2.9 percent, respectively. Dot-ELISA was standardized to detect the circulating antibodies of bovine microfilariosis. The overall prevalence of bovine microfilariosis recorded was 11.3 and 11.1 percent by indirect ELISA and dot-ELISA, respectively. Indirect ELISA and dot-ELISA detected antibodies of microfilariosis in 80 and 79 sera samples out of 81 known positive samples for microfilariosis by modified Knott’s test. Out of 1547 samples found negative by modified Knott’s technique, indirect ELISA and dot-ELISA detected antibodies for microfilariosis in 104 (6.7 %) and 103 (6.6 %) samples, respectively. The results obtained by indirect ELISA and dot-ELISA were compared with the conventional diagnosis of microfilariosis. The indirect ELISA and dot-ELISA were found more sensitive assays for diagnosis of bovine microfilariosis than wet blood film and modified Knott’s methods. The present study also revealed the advantages of dot-ELISA for diagnosis of bovine microfilariosis.ThesisItem Open Access PREVALENCE OF BOVINE MICROFILARIOSIS IN RAYALASEEMA REGION OF ANDHRA PRADESH WITH SPECIAL REFERENCE TO IMMUNODIAGNOSIS(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2008-10) JAYALAKSHMI, J; Md. HAFEEZABSTRACT : “ Prevalence of bovine microfilariosis in Rayalaseema region of Andhra Pradesh with special reference to immunodiagnosis” was undertaken to study the prevalence of microfilariosis in bovine blood samples (1286) collected from Chittoor (360), Kadapa (298), Anantapur (352) and Kurnool (276) districts of Rayalaseema region of Andhra Pradesh, India by using conventional viz., wet mount/ blood smear examination Giemsa’s, leishman’s, JSB staining techniques and knott’s technique and also by Immunological techniques viz., CIEP and ELISA. Based on blood examinations 35 out of 1286 samples were found positive showing the prevalence as 2.72 percent. The incidence in cattle and buffaloes was recorded as 2.45 percent (15/614) and 2.97 percent (20/672), respectively. With respect to region, higher rate of infection was recorded in Kurnool (3.25%) followed by 3.12, 2.5 and 2.01 percent in Anantpaur, Chittoor & Kadapa districts, respectively. Higher prevalence was noticed in the age group of above 9 years (4.72 %) followed by age groups of 6 to 9years (2.98%), 3 to 6 years (2.05%) and below 3 years (1.04%). Further the incidence was higher in rainy season (3.89%) than summer (2.87%) and winter (1.25%) seasons. The infection was higher in females animals (35/1254) compared to male animals (0/32) showing the incidence as 2.79 percent and 0 percent respectively. Cross bred cattle and Murrah buffaloes showed an infection rate of 3.04 percent (14/460) and 3.67 percent (15/408), respectively, while non descriptive animals showed an infection rate of 0.64 percent (1/154) and 1.89 percent (5/264), respectively in both the breeds. An incidence of 3.47 percent (27/777), 2.65 percent (5/264) and 0.47 percent (1/230) was recorded in lactating, non lactating animals and heifers, respectively. Morphological characterization of microfilaria by Giemsa’s and JSB staining techniques revealed long, slender microfilaria with a blunt anterior and tapering posterior ends. Antigen used for immunodiagnosis was prepared from Setaria cervi adult worm with a protein content of 5.6mg/ml and excretory-secretory antigen with 2.2 mg/ml. SDS – PAGE profile of polypeptides revealed 15 polypeptides ranging from 7.5 to 205 kDa. The somatic antigen of Seteria cervi used in CIEP and ELISA gave positive results with knott’s technique positive sera in 32 and 34 out of 35 bovines showing a sensitivity of 91.4 and 97 percent, respectively. Similarly 8 and 10 out of 25 knott’s technique negative bovine sera were found positive in CIEP and ELISA showing a sensitivity of 32 and 40 percent, respectively. The results of CIEP and ELISA with sera from bovines found positive in Knott’s method and negative in Knott’s method in detecting microfilariosis confirmed that a certain percentage of bovines found negative in Knott’s method were found positive by both the serodiagnositc tests indicating that they have been exposed to infection but could not be detected in routine Knott’s technique (conventional) technique. It is therefore suggested that ELISA was sensitive in the immunodiagnosis of microfiariosisThesisItem Open Access HISTOCHEMICAL AND BIOCHEMICAL STUDIES OF BILIARY AMPHISTOMOSIS IN BUFFALOES (Bubalus bubalis) WITH SPECIAL REFERENCE TO CHARACTERIZATION OF Gigantocotyle explanatum ANTIGEN(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2005) USHA RANI, AMMINA; Md. HAFEEZ; RAMAKRISHNA REDDY, P; ALAHA SINGARI, NABSTRACT : The research work on “Histochemical and Biochemical studies of Biliary Amphistomosis in buffaloes (Bubalus bubalis) with special reference to characterization of Gigantocotyle explanatum antigen” was undertaken to carryout the histochemical and biochemical studies of biliary amphistomosis with an attempt to characterize Gigantocotyle explanatum antigen, besides recording the prevalence of biliary amphistomosis in certain places of Andhra Pradesh. A total of 238 buffaloes were examined in eight (8) places of Andhra Pradesh. Based on slaughter house examination, the per cent prevalence of biliary amphistomosis in buffaloes was 35.71. Among the eight (8) places under this study the prevalence was more in Renigunta (50.00%) followed by 42.10, 37.93, 37.03, 34.61, 32.00, 31.25 per cent at Guntur, Kadapa, Vizag, Vijayawada, Rajahmundry, Hyderabad and was least in Vijayanagaram (27.77%), respectively. The prevalence of biliary amphistomosis with regard to age, sex, breed and season were also studied. Among 238 buffaloes examined for biliary amphistomosis, 79 were below 6 months age group, out of which 18 were found positive with 22.78 per cent prevalence. In the age group of 6 months to 5 years, out of 60 buffaloes, 24 were found positive with 40 per cent prevalence and 99 of above 5 years age group, 43 were found positive with the prevalence rate of 43.43 per cent. Among the total (238) number of buffaloes examined for biliary amphistomosis, 116 were males in which 50 were found positive with a prevalence rate of 43.10 per cent and 122 were females in which 37 were found positive with a prevalence rate of 30.32 per cent. Among 238 buffaloes examined for biliary amphistomosis, 113 were cross-bred animals, out of which 42 were found positive with a prevalence rate of 37.16 per cent and 120 were non-descriptive, in which 47 were positive (39.16%). Among 238 buffaloes examined for biliary amphistomosis 109, 59 and 70 animals were examined in rainy, winter and summer seasons, out of which 45, 15 and 25 were found positive with a per cent prevalence of 41.28, 25.42 and 35.71, respectively. Histopathological examination of liver tissue sections in buffaloes naturally infected with biliary amphistomosis, bile ducts revealed thickening and desquammation of the bile ductular epithelium with periductular infiltration of cells. Periductular fibrosis was evident with mononuclear cell infiltration. Mild proliferation of bile duct was also observed. Sinusoidal haemorrhages with dilatation of sinusoids were noticed. Hepatic cells were swollen with granularity in the cytoplasm (cloudy swelling). Thickening of glissions capsule was found. Histochemical observation of frozen liver tissue sections in buffaloes naturally infected with biliary amphistomosis – alkaline phosphatase and acid phosphatase enzyme activity by Gomori’s method was seen in sinusoidal spaces and hepatic cells. Alkaline phosphatase activity was noticed as black colour where as acid phosphatase activity as brown colour in the stained infected liver tissue frozen sections and correlation with the histological findings of liver necrosis. In an attempt to estimate the biochemical constituents (total protein, albumin, globulin, A/G ratio, cholesterol, alkaline phosphatase) in the serum samples collected from naturally infected buffaloes, there was a decrease in the total protein, albumin, A/G ratio but slight increase in globulin, cholesterol and alkaline phosphatase levels. Crude antigen of Giganotocotyle explanatum having protein concentration of 10.47 mg/ml was purified by column chromatography with sephadex G -200 using 0.15 M PBS (PH 7.4) buffer. A total of 50 fractions were collected and were monitored for protein concentration using spectrophotometer at 280 nm. The purified fractions showing the maximum peak value F1, F2 and F3 were injected in rabbits to raise the hyperimmunesera and was confirmed by double immuno diffusion test. The whole homogenate and fractions of Gigantocotyle explanatum were subjected to 5 per cent SDS-PAGE for the analysis of polypeptide profile. SDS-PAGE revealed a total number of 17,16 and 23 polypeptides, respectively. The apparent molecular weight of the polypeptides of these ranged from <14 to 165 KDa.ThesisItem Open Access ON TOXOPLASMOSIS WITH SPECIAL REFERENCE TO SERODIAGNOSIS(Sri Venkateswara Veterinary University, TIRUPATI – 517 502,A.P, 2003-09) THIMMA REDDY, P M; RAMAKRISHNA REDDY, P (Major); ALHA SINGARI, N; NARASIMHA REDDY, YABSTRACT : The research work on "Studies on Toxoplasmosis with special reference to serodiagnosis" was under taken to evaluate certain serodiagnostic tests for large scale screening in sheep and pigs in selected places in Andhra Pradesh and Karnataka. In the present study, the oocysts typical of T. gondii by morphologic criteria were isolated from cats. Attempts were made to diagnose Toxoplasmosis in sheep and pigs by using modified agglutination test and Dot-Enzyme linked immunosorbent assay. Two diagnostic antigens were prepared from mice peritoneal fluid infected with Toxoplasma gondii tachyzoites of RH strain. The protein content of the whole formalin fixed tachyzoite antigen was 1.44 μg/μl. A total of 257 sera samples were examined by MAT. The percentage of seropositivity was 21.01. Among the sheep from organised farms (18.37%) and slaughter houses (21.83%) and both male (17.65%) and female (23.91%), there was no significant difference in seroprevalence (P > 0.05). The highest antibody titre of 1 in 8000 was observed in 2 sheep. MAT detected toxoplasma antibodies in 103 pigs out of 195 examined with a overall percentage of 52.82. The prevalence of toxoplasmosis was comparatively high in males (43.33%) than females (35.29%). The antibody titre of as high as 1:1600 was observed in 12 pigs. Dot-ELISA was standardized to detect toxoplasmosis in sheep and pigs for the first time. A total of 207 sheep sera samples were examined. The percentage of seropositivity was 23.67. Among the sheep from organised farms (36.60%) and slaughter houses (21.12%) and both male (23.71%) and female (26.64%), there was no significant difference in seroprevalence (P > 0.05). Dot-ELISA detected toxoplasma antibodies in 80 pigs out of 195 examined with a overall percentage of 52.82. The prevalence of toxoplasmosis was comparatively high in males (33.33%) than females (29.41%). The prevalence of toxoplasmosis in human beings was studied by using commercially available ELISA kit (sigma) in Bidar District of Karnataka. A total of 78 individuals were screened. The per cent prevalence in human beings was 33.33. Among the 6 categories of individuals suffering from various ailments, the prevalence was more in individuals suspected for acquired immuno deficiency syndrome (AIDS) and also in AIDS confirmed patients. Serodiagnostic tests such as MAT and Dot-ELISA detected anti-toxoplasma antibodies in sheep and pigs. A systematic study on toxoplasmosis was conducted in parts of Andhra Pradesh and Karnataka to document the prevalence of disease in sheep and pigs in view of the high zoonotic potential of these two important food animals in spreading the disease to human beings. The MAT and Dot-ELISA were found to be rapid, easy to perform, visually read, does not require costly instruments and equipments. Therefore these tests can be routinely employed in any National Survey Programme in detection of toxoplasma antibodies in livestock population.