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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 CHARACTERISTICS OF Staphylococcus aureus FROM BOVINE MILK(AAU, Anand, 1990) Purohit, Jayantilal Hargovind; JHALA, V. M.The present study was undertaken with a view to know the occurrence of Staphylococcus aureus In bovine milk In relation to species, managemental conditions, breed, method of milking, parity, stage of lactation and Involvement of the quarters as well as to observe the relationships among the certain characteristics, Including enterotoxigenicIty, of S.aureus. The isolates were also phage typed to know the possible origin. The milk samples were collected from the animals maintained at six different farms comprising of four GAU farms and two private farms. The cows were maintained at four farms whereas buffaloes were maintained at remaining two farms. A total of 925 milk samples (758 from cows and 167 from buffaloes) from the individual quarters of 234 animals comprising of 191 cows and 43 buffaloes were collected and processed for isolation and identification of S.aureus. Of these, 94 quarters (10.16 per cent) of 67 animals (28.63 per cent) revealed the presence of S.aureus. The incidence of S.aureus was more commonly encountered amongst the cows on animal basis (31.94 per cent) as well as on quarter basis (11.35 per cent) than those of buffaloes (13.95 and 4.79 per cent, respectively).ThesisItem Open Access STUDIES ON IMMUNOMODULATION BY LEVAMISOLE ALONG WITH VACCINATION IN CHICKS AGAINST RANIKHET DISEASE AND IN CALVES AGAINST HAEMORRHAGIC SEPTICAEMIA(AAU, Anand, 1985) VYAS, GIRISH P.; DHOLAKIA, P. M.The present study was aimed assessment of the immunomodulatory effect of levamisole along with vaccination in chicks against RanikhetDisease (R.D.) and in calves against Haemorrhagic Septicaemia (H.S.) in relation to serum antibody titres, effect of dose of levamisole, total immunity period and electrophoretic pattern of serum proteins after immunization and treatment with levamisole. In all 560 serum samples from 140 chicks belonging to Central Poultry Research Station and 100 serum samples from 20 calves belonging to Livestock Research Station of Gujarat Agricultural University, Anand Campus, Anand were subjected to Haemaggulutination Inhibition (H.I.) test, Passive Haemagglutination (PHA) test, Sodium sulfite precipitation test and Agar gel electrophoresis.ThesisItem Open Access STUDIES ON MYCOTIC MASTITIS IN EXOTIC AND CROSSBRED CATTLE WITH SPECIAL REFERENCE TO ITS INCIDENCE AND DIAGNOSIS(AAU, Anand, 1984) SIMARIA, MULJI B.; DHOLAKIA, P. M.The present study was taken up to assess the incidence of mycotic infections of under in apparently healthy quarters of lactating cows and diagnosis in milk samples from clinical cases, Simultaneously, the prevalence of subclinical mastitis (SCM), and pathogenicity of fungal strains were also studied. The milk samples from 150 lactating cows (53 Jersey, 30 Kankrej and 67 crossbred) were collected. In addition to this 82 milk samples from clinical cases of mastitis received in Department of Bacteriology were also included in the present study. Subclinical mastitis (SCM) was recorded as 15.33 and 5.27 per cent on animal and quarter basis with California Mastitis Test. Although, incidence of SCM was highest in Jersey herd (8.82% quarterwise), fungal isolation was as low as 2.45 per cent, Hence CMT did not prove efficient enough to detect fungal infection in apparently normal udders.ThesisItem Open Access STUDIES ON POSTVACCINAL IMMUNITY TO BRUCELLA ABORTUS IN VACCINATED EXOTIC CATTLE(AAU, Anand, 1987) RAJPUT, H. A.; DHOLAKIA, P. M.The disease brucellosis caused by the infection of Brucella abortus in cattle adversely affects the economy of livestock rearing due to losses of calves created by abortions in suffering animals along with loss in milk production. The disease is world wide in distribution and calves lost due to abortion creat adverse economic impact on the animal husbandry development. Being zoonotic in nature the disease also occupies a special importance. It is therefore essential to diagnose the disease in herd as early as possible for its effective control. It is now well-established that the brucella organisms being the facultative intracellular organism the cellular immunity also plays a major role in the host resistance in addition to humoral immunity.ThesisItem Open Access STUDIES ON ADVANCED METHODS OF ASCERTAINING BRUCELLOSIS IN CATTLE(AAU, Anand, 1985) PATEL, J. B.; JHALA, V. M.The disease Brucellosis caused by the infection of Brucella abortus in cattle adversely affects the economy of livestock rearing due to losses of calves created by abortions in suffering animals along with loss in milk production. The disease is worldwide in distribution and calves lost due to abortions create highly adverse economic impact on the animal husbandry development. The disease also occupies a special importance being zoonotic in nature. It is# therefore, essential to diagnose the disease in a herd as early as possible for its effective control. It is now well established that the Brucella organisms being the facultative Intracellular organisms the cellular immunity plays a major role in the host resistance. In view of the above facts, the present work was carried out in cows at Livestock Research Station, Gujarat Agricultural University, Sardar Krushinagar, employing following six parametars to study their efficacy. (1) Screening of individual cow by milk ring test using ABR i.e. Aboirtus Bang Ring Antigen. (2) Above cows and the cows which were not in milk but having the history of abortion, were subjected to serum plate agglutination test. (3) All above sera samples were further examined by Serum Tube Agglutination Test (STAT). (4) Fourteen sera samples from positive, doubtful and negative reactors to STAT were sent to International Brucella Laboratory, Indian Veterinary Research Institute, Izatnagar for confirmation by complement fixation test. (5) Determination of conglutinin (K) and immunoconglutinin (IK) level in sera of positive, doubtful and negative reactors to STAT . (6) Measurement of cell mediated immunity in vitro by leucocyte migration inhibition test. Prom the results obtained it could be concluded that:- i)The serological tests namely milk ring test plate agglutination test, tube agglutination test may serve as useful tool for diagnosis of brucellosis in a herd. ii) The level of K and IK in sera of positive, doubtful or negative reactors to STAT have no significant difference and there was no significant correlation with agglutination titre, hence the determination of K and IK may not be of use in diagnosis of Brucellosis. iii) The Brucella organisms are being the facultative intracellular micro organisms, the cell mediated immunity plays an important role in resistance. The measurement of cell mediated immunity in vitro by leucocyte migration inhibition test led to the conclusion that this test may be of very great value as a modern advanced method for the diagnosis of Brucellosis where cellular immunity is considered to play a significant role.ThesisItem Open Access STUDIES ON MICROFLORA OF BOVINE SEMEN AND THEIR EFFECT ON SEMEN QUALITY(AAU, Anand, 1983) KHER, HIRABHAI N.; DHOLAKIA, P. M.Tho present study ims alaed to assess the baoterial load and types of organisms presant In saaien and their relation to semen quality. The study also included antibiotie sensitivity pattern of the isolates in vitro. Totally 45 bulls belonging to (a) Regional A. I. Centre, Rajkot, (b) Central Semen Collection Station, Mehsana, and (c) A. I. Centre, Godhra were studied during the year 1982-83. Bacterial load in the range frota 630 to 14375 organisms per ml for neat semen and in the range from 12300 to 130000 per ml for preputial washings had been encountered. The bacterial load from neat semen was high in summer season followed by monsoon and winter season, while that of preputial washings was high in monsoon season followed by summer and winter season. The semen quality was found fairly good , irrespective of bacterial load of ejaculates. Within the centre, bacterial load of neat semen and preputial washings was higher in buffalo bulls than that of cow bulls and was not varying according to type of sheath.ThesisItem Open Access ON THE OCCURRENCE OF GRAM NEGATIVE BACTERIAL INFECTIONS IN HATCHERIES WITH AN ATTEMPT TO ISOLATE SALMONELLA ORGANISMS(AAU, Anand, 1988) HATHI, A. V.; Dholakia, P. M.Present study was aimed to isolate and identify the bacterial organisms with emphasis on gram negative bacteria present in cloacal swab of appeirently healthy layers, droppings and litter samples of layer house, poultry feeds, fluff in the hatohery, air of the hatchery and fresh egg shell + CAM. The study also included serotyping of E.coli cultures and antibiotic sensitivity pattern of Escherichia ooli isolates in vitro. Totally 47, 51 and 65 samples of various sources were collected from the (A) Intensive Poultry Development Block, Makarba (Ahmedabad), (B) I.P.D.B., Baroda and (C) I.P.D.B., Surat, respectively. Of 47 collected samples of I.P.D.B. Makarba (Ahmedabad), yielded 90 bacterial isolates while of 51 samples of I.P.D.B. Baroda, 37 samples were sterile for bacterial organisms. Rest of the samples yielded 28 bacterial isolates whereas 65 samples of I.P.D.B. Surat yielded 107 bacterial isolates.ThesisItem Open Access DETECTION AND DIFFERENTIATION OF MAREK'S DISEASE VIRUS SEROTYPES BY POLYMERASE CHAIN REACTION (PCR) AND CHARACTERIZATION BY DNA SEQUENCING OF THE PCR PRODUCT(AAU, Anand, 2006) KALYANI, IRSADULLAKHAN HABIBULLAKHAN; Purohut, J. H.Marek's disease (MD) is a lymphoproliferative disorder of chicken characterized by oncogenic transformation of T cells that infiltrate lymphoid tissues, peripheral nerves and visceral organs, resulting in a complex pathogenesis that usually leads to death of the affected chicken. The causative agent of MD, Marek's disease virus (MDV) a cell associated herpesvirus, is ubiquitous in almost all poultry population raised under commercial conditions. The present study was under taken to assess incidence of MD as per the farm records on the basis of clinical signs and post mortem lesions, detection of MDV from clinical cases exhibiting frank lesions of MD by PCR, comparing the efficacy of different primers in detecting the MDV as well as differentiating the serotypes in tissues and feather follicle and genetic characterization by sequencing of the PCR amplified fragment carrying 132 bp repeats. During June to November 2005 outbreaks of MD were suspected in flocks of egg type birds on farms situated in and around Anand district located of central Gujarat as well as from farms around Mahuva located in Saurastra region of Gujarat state. The strength of individual affected flocks varied from 4500 to 25000 birds with a total population of 4,26,991 birds. Of these, 14110 birds died due to MD (as per the farm records of history, symptoms and postmorterm lesions) with an over all mortality of 3.30 per cent during the months of June to November 2005. During the investigation, a total of thirty six commercial poultry farms and forty nine batches of layer birds suffered from mortality due to MD, in addition to Central Poultry Research Station (CPRS) Veterinary College Anand, where mortality occured in layer birds. Mortality in commercial poultry farms ranged between 0.6 to 7.3 per cent. A total of 34 clinical samples (17 feather follicles and 17 spleen ) from 27 birds suspected of MD were collected by making visits to the various commercial poultry farms and also from CPRS Veterinary College Anand. Suspected clinical samples and four MD vaccine strains, including three HVT (MDV-3) vaccines and one SB-I(MDV-2) vaccine were processed for DNA extraction. DNA was extracted from samples of feather follicle using proteinase K mixture, where as DNeasy Tissue Kit (QIAGEN Pvt. Ltd) was used for tissue samples. Sample showing acceptable purity and concentration were subjected to PCR amplification using six different primers. Total of six pairs of primers were used, which were selected to amplify different gene segments. The primer pair BamHl/BamH2 specifically amplifying a 434 bp segment from BamHI fragment (of MDV-1) containing 132 bp repeats located in TRL and IRL region of MDV genome, detected 32 samples positive indicating presence of double 132 bp repeats in the amplified segment. All the three HVT vaccines and one SB-1 vaccine failed to produce the amplification as the selected primer is specific for MDV-1 only. Primer pair AGAM1/AGAM2 and P1/P2, both targeting the antigen A gene (UL44) produced 314 bp and 199 bp products respectively in 30 out of 34 clinical samples tested, however, none of the HVT vaccine or SB-1 vaccine yielded positive result, suggesting the specificity of these primers for MDV-1. Primer set M1.1/M1.8 amplified a 318 bp product as against expected 247 bp product in 30 samples. To confirm the result, these primers were subjected to NCBI BLAST, and it was found that the primer specific segment of 318 bp does exist in pubhshed sequence of Md5, and Mdl IBAC. None of the HVT as well as SB-1 vaccine produce any amplification indicating the specificity of primer in detecting MDV-1. Only two field samples produced expected amplicon of 505 bp when tested with MDV-3 specific HVT1/HVT2 to detect the vaccine virus (HVT). All the three HVT vaccines produced the amplicon of 505 bp, whereas SB-1 vaccine was negative, proving specificity of this primer pair for HVT. Out of the six primer pairs, BamHl/ BamH2 and AGA1/AGA2 detected MDV in maximum number (32) of field samples. Primers AGAM1/AGAM2, P1/P2 and Ml.l/M.1.8 detected the same number (30) of samples positive. Primer pair HVTl/ HVT-2 detected MDV-3 in two feather follicle samples indicating possible excretion of the vaccine virus. PCR products (434 bp) of MDV genome amplified by BamHl/BamH2 primers from two field samples (M3, M4) having good quality DNA (i.e. 1.7-1.9 OD at 260/280 nm) as detennined by spectrophotometry were used for sequencing by ABI PRISM® 310 Genetic Analyzer (Applied Biosystems, USA), using BigDye® Terminator v3.1 Cycle sequencing kit (Applied Biosystems, USA). The sequence obtained revealed two 132 bp repeats in the 378 bp stretch indicating virulent nature of the field MDV. The alignment of 378 bp sequence of M3 and M4 with previously published sequences of the MDV isolates Md5, Mdll, HPRS-16 and CDs of tumourogenicity associated m-RNA and CDs published from BamH gene family revealed that the field isolates of present study shared complete homology (100%). Results of sequencing along with history of vaccination in the affected birds and involvement of different organs on postmortem examination indicated about the increase in virulence of MDV circulating in field causing recent outbreaks.ThesisItem Open Access ASSESSMENT OF NOVEL N-GENE TARGET FOR DETECTION OF PESTE DES PETITS RUMINANTS (PPR) VIRUS BY RT-PCR AND CHARACTERIZATION BY RFLP PROFILE(AAU, Anand, 2006) CHANDRA, VARTIKA; Jhala, M. K.Peste des Petits Ruminants (PPR) is an economically important viral disease of sheep and goats, characterized by high fever, ocular and nasal discharge, pneumonia, necrosis and ulceration of the mucous membrane and inflammation of gastrointestinal tract leading to severe diarrhea. The disease is caused by a Morbillivirus of the family Paramyxoviridae. The disease is highly contagious causing varying degree of morbidity and mortality in susceptible animals. In India, severity of the disease is more pronounced in goats than in sheep and with a combined susceptible population of about 200 million, it is one of the major threats to the small ruminant population of the country. The present study was aimed to study the incidence of Peste des petits ruminants virus (PPRV) in goats of selected areas of Gujarat by sandwich ELISA and to derive estimates of overall, locationwise, agewise and sexwise incidence and samplewise positivity rates. In addition, the study also involved standardization and application of novel Nucleocapsid (N) gene based RT-PCR, for diagnosis of PPR. Further, an attempt was also made to access possible genetic variation among the field and vaccine PPRVs by RFLP profile. A total of 46 clinical samples from 31 goats suspected of PPRV from selected areas of Gujarat were tested by sandwich-ELISA, of which 23 animals were found positive yielding an overall incidence rate of 74.19 percent. Out of 18 animals suspected of PPR at Clinical Service Complex, Veterinary College. Anand, 15 (83.33%) were found positive; whereas four (80%) out of five samples from Koth, two (66.67%)) out of three samples from Vasna and two (40%) out of five samples from Harsoli yielded positive results. Out of the 31 animals, 13, 11, 6 and one animals belonged to age groups of 0-12, 12-24, 24-48 and >48 months, respectively, showing the respective incidence of PPRV as 84.61, 63.63, 66.67 and 100 per cent. Incidence rate of PPR by sandwich-ELISA was 80%) and 71.43%) in males and females, respectively. The PPRV antigen could be detected slightly more in blood samples (73.33%) than in nasal swabs (66.67%o), in case where both the sample types were taken from the same animals. All the 46 clinical samples, a reference vaccine virus (Sungri isolate) and a blood sample from apparently healthy goat were processed for RNA extraction using TRI Reagent . RNA samples showing acceptable purity and concentration were reverse transcribed with random hexamers to generate cDNA templates, which were subjected to PCR amplification using in silico designed N-gene specific primer pair N1-N2. Reference vaccine virus as well as fifteen (32.61%) of the 46 clinical samples, including fourteen blood samples and one nasal swab, produced the desired amplion of approximately 463 bp with primer pair N1-N2. The amplicons from all the positive samples and the vaccine virus migrated similarly in the gel. Thirty one (67.39%)) samples including 12 blood samples, 17 nasal swabs and two tissue samples as well as the blood sample taken from apparently healthy goat failed to produce the targeted amplification with the primer pair N1-N2. Out of 46 clinical samples tested for PPRV, 31 and 15 samples were positive by sandwich-ELlSA and RT-PCR, respectively. "None of the samples negative in sandwich- ELISA yielded positive amplification in RT-PCR. Relative to sandwich-ELlSA, sensitivity and specificity of the RT-PCR was 48.39 and 100.00 per cent, respectively. Overall agreement between the two tests was 65.22 per cent. RT-PCR products (463 bp) amplified with primer pair N1-N2, were further digested with two restriction enzymes viz. Pst-I and Alu-I for RFLP analysis. RT-PCR products of the representative samples from each of the three locations and the reference vaccine virus produced similar RFLP patterns with each of the two REs, indicating absence of genetic variation at the corresponding restriction sites of these two REs. Digestion of RT-PCR products with Alu-l yielded a fragment of 395 bp, whereas the other fragment was not visible. Similarly, digestion with Pst-I yielded a single fragment of405bp.