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Anand Agricultural University, Anand
Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur
AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.
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ThesisItem Open Access ISOLATION AND CHARACTERIZATION OF AEROMONAS SPECIES FROM POULTRY MEAT(AAU, Anand, 2009) SMITA; Brahmbhatt, M. N.The aim of the present study was to isolate, identify and characterize Aeromonas species from poultry meat. A total of 120 samples were processed for estimating prevalence oi Aeromonas spp. Out of those 120 samples, 66 isolates oi Aeromonas were identified. On the basis of biochemical characterization 47 isolates of A. sobria, 11 isolates of A. hydrophila and 8 isolates of A. caviae were detected. When the source wise study of Aeromonas was conducted it was found that maximum number of Aeromonas isolates were recovered from heart (87.5 per cent) followed by liver (66.66 per cent), thigh muscle and chest muscle (40 per cent each). When different selective culture media were evaluated for isolation of Aeromonas spp. from poultry meat it was found that percent recovery of Aeromonas isolates were more from Ampicillin Dextrin Agar (89.39 per cent), followed by Aeromonas Starch DNAse agar (68.18 per cent) and Aeromonas isolation media (18.18 per cent). Specificity of PCR assay for detection of A. hydrophila and A. sobria was performed by testing against different gram positive and gram negative organisms.Primers were found to be specific for A. hydrophila and A. sobria. All the isolates of A. sobria (47) and A. hydrophila (11) which were identified on the basis of biochemical characterization were subjected to PCR and confirmed. All 66 Aeromonas isolates were tested for presence of aerolysin, haemolysin and enterotoxin gene. None of the isolate showed presence of aerolysin and enterotoxin while overall prevalence of haemolysin gene was 78.78 per cent. In A. hydrophila, 54.54 per cent; A. caviae, 37.5 per cent and in A. sobria, 91.48 per cent isolates were found to possess haemolysin gene. All the isolates of Aeromonas were subjected to antimicrobial drug sensitivity test against gentamicin, chloramphenicol, ciprofloxacin, kanamycin, bacitracin, rifampicin, tetracycline and erythromycin. Maximum sensitivity pattern was recorded with chloramphenicol (86.36per cent), gentamicin (81.82 per cent), ciprofloxacin (60.61 per cent), kanamycin (34.85 per cent), bacitracin (25.76 per cent), tetracycline (16.67 per cent), rifampicin (15.15 per cent) and erythromycin (9.09 per cent). The resistance pattern of Aeromonas isolated from chicken meat to various antibiotics was observed as bacitracin (74.24 per cent) followed by rifampicin (71.21 per cent), kanamycin (65.15 per cent), erythromycin (53.03 per cent), tetracycline (33.33 per cent), ciprofloxacin (22.73 per cent) and gentamicin (18.18 per cent). It was concluded that overall prevalence of Aeromonas spp. in poultry meat was 55 per cent which is a matter of concern from public health point of view and needs proper attention. The overall prevalence of 78.78 per cent of Aeromonas isolates showing presence of haemolysin gene (Virulence gene) in poultry meat is also a matter of concern and this study reveals that poultry meat could be a potential threat to public health. Hence, more attention for implementation of HACCP concept from food safety point of view is required. Antibiotic sensitivity pattern of Aeromonas isolates revealed that chloramphenicol was found to be most effective drug (86.36per cent) followed by gentamicin (81.82 per cent), ciprofloxacin (60.61 per cent), kanamycin (34.85 per cent), bacitracin (25.76 per cent), tetracycline (16.67 per cent), rifampicin (15.15 per cent) and erythromycin (9.09 per cent).ThesisItem Open Access MEAT SPECIATION BY MOLECULAR AND SEROLOGICAL TECHNIQUES(AAU, Anand, 2002) Thumbar, J. M.; BRAHMBHATT, M. N.Among several methods of meat speciation, method based on DNA especially Polymerase Chain Reaction (PCR) provide potentially more information than others. The present study was under taken for meat speciation by molecular and serological techniques. Meat samples from meat species (cattle, buffalo, sheep, goat, pig and chicken) were utilized for molecular analysis. Genomic DNA was isolated from eight meat samples of each species as per method described by Sambrook et al. (1989) with some modifications. Actin multigene family was amplified by PCR using a pair of family specific primers (forward primer-5' CCT ACA ACA GCA TCA TGA AGT G 3'and reverse primer-5' GCT GAT CCA CAT CTG CTG GAA G 3'). PCR cycling protocol included to initial denaturation at 95 °C for 5 minutes then followed by 30 cycles of 95°C for 1 minute, 48°C for 1 minute, 72°C for 1 minutes and final extension at 72°C for 1 minute, 48°C for 1 minute, 72°C for 1 minutes and final extension at 72°C for 5 minutes. PCR products were resolved by agarose gel electrophoresis it produced a characteristic band pattern for each species. PCR band pattern generated for cattle included one major band of 0.328 kb. The PCR band pattern for buffalo included two bands of 0.328 and 0.242 kb. Sheep and goat produced identical band patterns having two bands of 0.328 and 0.242 kb and one faint band of 0.765 kb. Pig also revealed pattern identical to buffalo. A band of approximately 0.330 kb size was common in all livestock species. Chicken exhibited a characteristic pattern with six bands, three bands of high intensity (approximately 0.365, 0.334 and 0.242 kb) and three faint bands (approximately 0.547, 0.480 and 0.113 kb). Out of six species explored, four were found to have characteristic PCR band pattern. Thus, PCR was considered to be a potential technique for meat species identification and speciation. Actin gene band patterns were identical in both the sexes for all the species studies. PCR was also found to be consistent and effective tool as it remained same for heat treated and putrified (unpreserved) of meat. An agar gel precipitation technique was successfully used for identification of meat species, cross-reaction study and detection of adulteration level in meat. Hyper immune sera were raised in rabbits by intramuscular injection of meat extract. Anti buffalo meat sera were cross-reacted to cattle, sheep and goat meat extract. At same time anti cattle meat sera were also found to be crossreacted with buffalo, sheep and goat meat extract. Species specific sera were made by absorption technique. These sera were used to detect level of adulteration. By AGPT, up to 10 % level of adulteration between buffalo or cattle meat with sheep and goat meat was successfully detected. Counter immunoelectrophoresis technique was also successfully used to detect the meat species. This method was found to be rapid as compare to the AGPT.ThesisItem Open Access HUMAN AND ANIMAL INFECTIONS CAUSED BY CANDIDA ALBICANS(AAU, Anand, 2001) Jadhav, Vijay Jayawant; Pal, MahendraThe aim of the present study was to elucidate the etiologic significance of C. albicans in various clinical disorders of man as well as animals. Mycological examination of 293 clinical samples (136 from man, 97 from animals and 60 visceral organs of chicks) from suspected fungal infections was carried out for the presence of C. albicans. The various disorders encountered during the study period in man were otomycosis (84), cutaneous infection (20), oral infection (12), onychomycosis (10) and vulvovaginitis (10), while in animals were mastitis (49), stomatitis (30), dermatitis (11), otitis (7), The visceral organs included in the study were, gizzard (13), proventriculus (13), liver (12), lung (13), and crop (9). The isolates of C. albicans were subjected to in-vitro antifungal drug susceptibility test agaiiis: clotrimazole, fluconazole, amphotericin-B and nystatin. The results of this investigation indicated that frequency of occurrence of organism was higher in man (55.55 per cent) followed by dogs (22.22 per cent), chickens (14.81 per cent) and cows and buflaloes (3.71 per cent each) and most of the C. albicans isolates were contributed by oral infection. The study of in-vitro drug sensitivity of the clinical isolates of C. albicans showed that there was a wide variation in the resistant pattern ranging from 0.00 per cent to 51.85 per cent and clotrimazole was found to be the most effective drug (62.96 per cent). The observations of this preliminary study on in-vitro efficacy of six nutrient media suggested that Sabouraud dextrose agar is a cheap and good selective fungal medium. It was concluded that, the overall prevalence of C. albicans infection was higher in man as compared to animals. The good management practices of dairy cattle farm along with hygienic methods of milking followed at the Livestock Research Station at Anand, prevented the incidence of subclinical mastitis caused by C. albicans. Of all the drugs tested in-vitro by disc diffusion technique, clotrimazole was found to be the most effective chemotherapeutic agent, which may help in the management of the candidiasis. Being a cheap and selective fungal medium, Sabouraud dextrose agar can be recommended for the routine mycological work in the microbiology and public health laboratories. This appears to be the first record from India, which delineates the occurrence and etiologic significance of C. albicans in stomatitis of dogs and buffaloes.ThesisItem Open Access THE SANITARY QUALITY OF COMMUNITY DRINKING WATER SUPPLIES AT GUJARAT AGRICULTURAL UNIVERSITY, ANAND CAMPUS, ANAND(AAU, Anand, 1990) CHAURASIA, A. K.; Anjaria, J. M.The sanitary quality of community drinking water supplies of Gujarat Agricultural University, Anand Campus, Anand, was determined by bacteriological examination both quantitatively and qualitatively. The levels of index organisms such as coliforms, faecal streptococci and E.coli type I were extimated. Parasitic assessment of drinking water supplies was also made. An attempt was made to find out suitable indicator of faecal pollution as well as to draw correlation, if any, between pollution index and number of diarrhoeal cases occurring in human population at studied campus. At Gujarat Agricultural University, Anand Campus, there were two different sources of drinking water supplies, one in each of the two residential campuses i.e. Veterinary and Khetiwari; and study was carried out separately for both the campuses in two runs i.e. in rainy season (July 1989 to October 1989) and winter season (November 1989 to February 1990). Standard techniques were used for processing all the 130 tap water and four samples from source of water supplies. The quality of drinking water supplies at both the campuses during whole course of study, was not up to the standard as recommended by WHO.ThesisItem Open Access STUDIES ON THE OCCURRENCE OF AEROBIC MICRO-ORGANISMS IN MEAT ASSOCIATED WITH FOOD INFECTIONS AND INTOXICATIONS(AAU, Anand, 1990) Brahmbhatt, M. N.; Pal, MahendraThe present study was aimed to asses the bacterial load and types of bacterial organisms present in the fresh market meat samples of cattle, buffalo, sheep and goat. The study also includes the coliform plate count, fungal Isolation and antibiotics sensitivity pattern of the Isolates in vitro. Two hundred sixteen samples of fresh market meat were examined which included 54 each from cattle , buffalo, sheep and goat. The samples were collected randomly from 10 different butcher's shop in Anand city of Gujarat State.ThesisItem Open Access STUDIES ON THE INCIDENCE OF ASPERGILLOSIS IN POULTRY AND POULTRY HANDLERS(AAU, Anand, 1990) Savalia, C. V.; Pal, MahendraThe present stady was coiadacted to record the incidence of palmonary aspergillosis in the White Iieghom chicken, and in the poaltry handlers. In addition, environmental distribution of aspergilli was studied in feed, litter , soil and air of different pens on the All India Co-ordinated Research Project (AICRP) Poultry Farm under Gujarat Agricultural University, Anand Campus, Anand and Patel Poultry Parm, Navli, District Kheda, Gujarat. Pour hundred fifty specimens of the trachea, lungs and abdominal or thoracic air sacs of 150 White leghorn chicken: were investigated for the gross lesions, histopathologtoal examination and cultural isolation . None of the birds showed gross pathological lesions of the disease on autopsy. The histopathologioal examination also could not reveal the presence of the fungi in the tissues . However, 23.98 per cent (108 out of 450) specimens of the trachea, lungs and air sacs yielded aspergilli on the cultural isolation. This included 9.33 per cent of A.niger 7.55 per cent of A.fumigatus, 5.11 per cent of A.flavus, 1.33 per cent of A.terreus and 0.22 per cent each of A.vyolaceous. A.stelletus and A.chevalieri. Growths were identified on the basis of ths colonial and microscopic morphology.ThesisItem Open Access ROLE OF ZOOPHILIC DERMATOPHYTES IN THE ETIOLOGY OF HUMAN RINGWORM(AAU, Anand, 2001) Ahmed, Kakoli; PAL, MAHENDRAA clinico-epidemiological study of 124 clinically diagnosed cases of dermatophytosis was conducted at the Department of Veterinary Public Health, College of Veterinary Science and Animal Husbandry, Gujarat Agricultural University, Anand Campus, Anand, to ascertain the prevalence of zoophilic dermatophytes in the suspected patients with tinea infections, attending the Skin O. P. D. at the Shree Krishna Hospital, Pramukhswami Medical College, Karamsad, Anand and also other Skin Clinics in and around Anand. The. various types of tinea infections diagnosed clinically based on the typical lesions presented by 124 patients included tinea corporis (36), tinea unguium (25), tinea pedis (19), tinea cruris (15), tinea manuum (10), tinea capitis (8), tinea barbae (5), tinea faciei (4) along with mixed tinea infection (2). Detailed mycological investigation was performed on 124 clinical specimens (99 skin scrapings including hairs and 25 nail clippings) to elucidate the presence of zoophilic dermatophytes in the suspected patients. The overall prevalence rate of dermatophytosis was determined to be 19.35 per cent. Tinea corporis (37.5 per cent) was the most predominant clinical type among the patients followed by tinea manuum (16.67per cent), tinea capitis (12.5 per cent), tinea cruris,tinea faciei and tinea pedis (8.33 per cent); tinea barbae and tinea unguium (4.17 per cent each). The zoophilic dermatophytes identified as the causative agents, in order of their frequencies were T. mentagrophytes (41.67 per cent), T. verrucosum (29.16 per cent), M. canis (25.0 per cent) and T. simii (4.17 per cent). Maximum number of dermatophytic patients were between 31 to 40 years of age (41.67). The males (75.0 per cent) were affected by the zoophilic dermatophytes more than the females (25.0 per cent). Epidemiological investigations indicated that the infected patients were mainly from the low socio-economic background (33.33 per cent). Moreover, T. mentagrophytes (33.33 per cent) and T. verrucosum (25.0 per cent) played the major role in causing dermatophytosis in persons, residing in the villages whereas M. canis (16.67 per cent) was identified as the chief etiologic agent in the city dwellers. Further studies signified that patients from rural communities (66.67 per cent) with history of contact with animals were the frequent victims of zoophilic dermatophytes. Altogether the overall prevalence of zoophilic dermatophjrtes was higher in rural areas (66.67 per cent) than the urban areas (33.33 per cent). Among various occupational groups, with greater opportunity of exposure to animals, the most vulnerable group to be infected with the zoophilic species comprised of the dairy farmers working in the villages (37.5 per cent). Likewise in the urban localities, the animal handlers (16.67) were identified as the most susceptible group. However, even animal owners belonging to the urban areas were considered as a risk group. Through retrospective epidemiology the source of infection was established in 14 out of 24 positive patients, with a history of contact with animals. Isolation of similar zoophilic dermatophytes (14) from the human patients as well as their respective in-contact animals confirmed the zoonotic significance of the animal dermatophyties. It can be concluded that ringworm was prevalent (19.35 per cent) among the suspected patients, to a considerable extent. This potential mycotic dermatological hazard, affected the males more frequently than the females. Most of the dermatophytic cases were observed in adults of 31 to 40 years of age. The frequency of T. mentagrophytes in causing tinea infections in man was higher than the other zoophilic species namely T. verrucosum, M. canis and T. simii. Among all the clinical types of tinea, tinea corporis was recorded as the most common form in the positive patients. Maximum number of dermatophytic patients with history of close association with animals, were exclusively from the rural localities owing to their low socioeconomic status and poor personal hygiene. Moreover, T. mentagrophytes and T. verrucosum were detected primarily in persons residing in the villages while M. canis was recovered from the city dwellers. This was attributed to the close contact of the rural people to farm animals, which were principal sources of these dermatophytes. Increased contacts with pets led to the spread of ringworm to the urban people. Dermatophytosis due to zoophilic species was recognized as an occupational hazard, predominantly of the dairy farmers followed by animal handlers, owing to their increased exposure to animals due to occupational obligations. However, animal owners were also at greater risk. Animals of different species namely buffalo, cattle, camel, dog, goat, hen and horse were established as the source of infection in 14 positive patients, thereby establishing the role of zoophilic dermatophytes in the etiology of human ringworm. This heralded the need of thorough medical investigation of patients with cutaneous lesions having history of exposure to animals. As far as it could be ascertained, the isolation of T. mentagrophytes as the etiologic agent of ringworm in camel, dog and horse and T. simii in a hen constitutes the first report of its kind from Gujarat. Moreover, zoonotic. potential of these dermatophytes have also been established for the first time in Gujarat.ThesisItem Open Access PREVALENCE OF CRYPTOCOCCUS NEOFORMANS IN MAN, ANIMAL AND ENVIRONMENT(AAU, Anand, 2004) THOMES, LINI; Pal, MahendraThe aim of the present investigation was to study the prevalence of C. neoformans in man, animal and environment. Mycological examination of 217 clinical (120 of man, 97 of animals) and 127 environmental samples (96 of avian droppings and 31 of wood scrapings) was conducted to know the occurrence of C. neoformans. In addition, Petri plates of sunflower seed medium and Sabouraud dextrose agar were exposed to inside and outside environment of the zoo aviaries which inhabitated several species of birds. The result of this investigation indicated that frequency of occurrence of organism was higher in avian droppings (41.8 per cent) followed by man (25 per cent), cows (16.6 per cent), dogs (8.3 per cent) and air (8.3 per cent). No isolation could be made from the buffaloes, horses, goats and wood scrapings. Attempt was also made to isolate C. neoformans from 51 zoo workers and 47 bird fanciers who are exposed to avian excreta during the cleaning of cages. But failed to yield any isolation of C. neoformans. In vitro colonisation of fruits and vegetables was done to know whether these natural substrates can support the growth of the pathogen in the laboratory. In vitro colonization of papaya, water melon, grape fruit, carrot, tomato, banana, apple, brinjal and potato by known C. neoformans strains (human and animal origin) under laboratory conditions showed good and luxurient growth of the yeast. All the isolates of C. neoformans were subjected to in vitro antifungal drug susceptibility test against amphotericin B, clotrimazole, fluconazole and nystatin. There was a wide variation in the resistant pattern ranging from 0.00 per cent to 16.67 per cent and clotrimazole and fluconazole was found to be the most effective drug (75.0 per cent). Of all the drugs tested in vitro by disc diffusion techniques, clotrimazole and fluconazole was found to be most effective chemotherapeutic agent, which may help in the management of the cryptococcosis. The observations of this study suggested that sunflower seed agar is a more economical and good selective fungal medium for the study of C. neoformans. Sunflower seed agar can be recommended for the routine mycological work in the microbiology and public health laboratories. It was concluded that the overall prevalence of C. neoformans was higher in avian droppings as compared to man and animals. The good management practices of zoological garden with proper disposal of avian droppings can help to prevent the infection caused by C. neoformans.ThesisItem Open Access SEROPREVALENCE OF BRUCELLOSIS IN CATTLE, BUFFALO AND HUMAN BEING IN CENTRAL GUJARAT(AAU, Anand, 2003) VARASADA, RUPESHKUMAR NATVARLAL; Brahmbhatt, M. N.A study was carried out at the Department of Veterinary Public Health, Veterinary College, Anand to assess the seroprevalence of brucellosis in cattle, buffalo and human in four district of Central Gujarat viz. Ahemdabad, Anand, Kaira, and Vadodara during the period from January 2002 to December 2002. The study included screening of 595 serum samples of cattle and buffalo by i-ELISA, RBPT and STAT from above four districts where as milk base i-ELISA was used for screening of 650 village milk co-opcralivc socictiCvS for brucellosis. To detect prevalence of brucellosis in human PCR, RBPT and STAT were used. Characterizations of Brucella spp. involved in human infection were done by rep-PCR (Repetitive Element- PCR) and PCR-SSCP (Single Stranded Conformational Polymorphism) technique. Out of 595 (Anand (152), Vadodara (99), Ahemdabad (209) and Kaira (135)] serum samples tested 25.66, 19.19, 22.48, and 19.26 per cent reacted positive by i-ELlSA in respective district giving over all prevalence 22.01 per cent. Seroprevalence of brucellosis among cattle in the districts of Anand, Vadodara, Ahemdabad, and Kaira was 31.08, 23.61, 21.95 and 21.33 per cent respectively while among buffaloes prevalence were observed 20.51, 7.40, 23.26 and 16.67 per cent respectively by i-ELISA. RBPT gave 19.08, 16.16, 12.92 and 15.55 per cent prevalence among bovine in Anand, Vadodara, Ahemdabad, and Kaira districts giving over all prevalence of 16.80 per cent. Seroprevalance of brucellosis among cattle in the districts of Anand, Vadodara, Ahemdabad, and Kaira was as 24.32, 19.44, 17.07 and 20.00 per cent respectively while among buffaloes the prevalence was observed as 14.10, 7.40, 15.11 and 10.00 per cent respectively by RBPT. STAT gave 15.18, 14.14, 14.83 and 12.59 per cent prevalence among bovine in Anand, Vadodara, Ahemdabad, and Kaira districts giving over all prevalence of 14.03 per cent. Seroprevalance of brucellosis among cattle in the districts of Anand, Vadodara, Ahemdabad, and Kaira was 18.91, 16.67, 15.44 and 16.00 per cent respectively while among buffaloes the prevalence was 11.53, 7.40, 13.95 and 8.34 per cent recorded by STAT. Out of 344 serum samples of cattle (336 cows and 8 bulls) tested, 24.10%, 19.94% and 16.67% were found positive by i-ELISA, RBPT, and STAT tests respectively in cows, while 25.00.12.5, and 12.5 per cent were found positive in bulls by respective test. Out of 251 sera samples tested from buffaloes (230 female and 21 male), 18.70, 12.61 and 10.87 per cent were found positive by i-ELISA, RBPT, and STAT tests respectively in cows while 23.81, 14.29, and 14.29 per cent were found positive in bulls by respective test giving overall prevalence of 22.30, 17.27, and 14.57 per cent in female and 24.13, 13.79 and 13.79 per cent prevalence in male by the respective tests. Out of 87 serum samples tested from human beings, 60 were from occupationally exposed group [Veterinarian (25), Farmer (10), Animal handler (15), Slaughterhouse worker (10)] and 27 from patients with pyrexia of unknown origin. Among veterinarians, 1(4.0%) sample was positive by RBPT and STAT. One slaughterhouse worker (10.0%) has reacted positive by both RBPT and STAT. From among animal handler, 1(6.67%) serum sample was found positive by RBPT and none by STAT, and none of the farmers were positive, giving 5.00% and 3.34% overall prevalence in occupationally exposed groups by RBPT and STAT, respectively. From 27 patients with pyrexia of unknown origin group, 4(14.81%)) were positive by RBPT while 2(7.40%)) were positive by STAT. Overall prevalence of brucellosis in human being was 8.04 per cent by RBPT and 4.60 per cent by STAT. Out of 77 human blood samples tested PCR detects 25.97 per cent prevalence in human being. In professionally exposed group PCR detected 12 per cent prevalence in veterinarians, 20 per cent prevalence in farmers, 13 per cent prevalence in animal handler hence the overall prevalence in professional group was 14 per cent. While in pyrexia of unknown origin group PCR gave 48 per cent prevalence. Overall prevalence in human group was observed 25.97 per cent by PCR. In Bovine the relative sensitivity and specificity of the RBPT and STAT tests vis-a-vis i-ELISA was assessed and it was observed that the relative sensitivity o[ RBPT was 68.70 per cent and that o[ STAT was 63.36 per cent. In cattle the relative sensitivity of RBPT and STAT was 79.51 and 67.47 per cent respectively while in buffalo the relative sensitivity of RBPT was 64.58 per cent, and 56.25 per cent of STAT. The relative specificity was observed more than 99.00 per cent in all of the above cases. In case of human being, in comparison to RBPT, STAT showed 57.14 per cent sensitivity while in comparison to PCR sensitivity of RBPT and STAT for detection of brucellosis was observed 30 per cent and 15 per cent, respectively .The specificity was observed 100 per cent for all tests. The concordance among all tests in animal was observe more than 90 per cent while between PCR and RBPT was 81.81 per cent, between PCR and STAT was 77.92 per cent and between RBPT and STAT was 96.55. Fingerprinting of Brucella isolates with rep PCR (BOX PCR , ERIC PCR and REP PCR) generated distinct amplification bands ranging from 1-6, 1-6 and 2-10, respectively. With frequency ranging from 0.10-0.85, 0.05-0.80, and 0.05-60 with 85.78, 83.34 and 100 per cent polymorphic bands respectively. Dendogram observed by popgene analysis of results of BOX PCR, REP PCR and ERIC PCR showed that all isolates were divided into four major clusters by BOX PCR and REP PCR while in five major clusters by ERIC PCR according to simple similarity coefficient. Over all analysis divided all isolates into four major clusters. Network analysis of BOX PCR, ERIC PCR and REP PCR divides all isolates into 5, 8 and 8 clusters, respectively according to the mutation present in between various groups. Network graph of pulled analysis showed that only Brucella-15 86 Brucella-27, Brucella-59 85 Brucella-63, Brucella-69 8B Brucella-? 1 were 100 per cent similar to each other while rest of isolates appeared different from each other with minimum one to maximum 16 mutation in between. By analyzing rep-PCR fingerprinting, no group specific pattern was observed according to professional group or source of infection or exposure. Only single SSCP band pattern was observed by PCR-SSCP analysis indicating lack of variation in amplified product. In a survey of Vadodara district by milk base i-ELISA 4.15 per cent prevalence was observed in cow herds where as in buffalo herds it was 0.46 per cent and the prevalence among villages was 3.53 per cent.