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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 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 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 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.ThesisItem Open Access SEROEPIDEMIOLOGY AND ELECTROPHEROGRAM STUDIES OF BLUETONGUE VIRUS(AAU, Anand, 2000) Bhalodiya, M. B.; Jhala, M. K.Bluetongue (BT) is an insect transmitted viral disease of several species of domestic and wild ruminants. The disease is a cause for serious concern to livestock industry due to staggering direct and indirect economic losses. In many countries like India having considerable sheep population, the disease has become endemic. Severity of the infection depends upon the species and breed of animals, serotypes/strains of virus and prevalent ecosystem. The present study was aimed to find out prevalence of Bluetongue virus (BTV) antibodies in cattle and sheep of different regions of the Gujarat State. Avidin-Biotin enzyme-linked immunosorbent assay (AB-ELISA) and indirect enzyme-linked immunosorbent assay (I-ELISA) employed for antibody detection were also compared in terms of their sensitivity and specificity. An attempt was also made to standardize polyacrylamide gel electrophoresis (PAGE) for detecting BTV. Out of 527 cattle and sheep sera tested, 343 (65.08%) and 396 (75.14%) were found to be positive for BTV antibodies by AB-ELISA and IELISA respectively. Specieswise, 69.04 and 77.57 percent of cattle and 55.55 and 63.33 percent of sheep revealed antibodies to BTV by AB-ELISA and l-ELISA respectively. The highest prevalence rate was found in South Gujarat region (67.94 and 78.63 percent) by AB-ELISA and l-ELISA respectively and female animals (65.53 and 77.84 percent) showed more prevalence than male animals (64.35 and 70.79 percent). Districtwise seroprevalence study revealed highest incidence in Valasad district (68.51%), followed by Navsari (67.70%), Kutch (60.74%), Junagadh (57.69%) and Rajkot (51.72%) district by AB-ELISA, whereas IELISA yielded highest prevalence rate in Valasad (80.55%) followed by Navsari (77.82%), Rajkot (69.96%), Junagadh (69.23%)) and Kutch (66.96%>) district. In cattle, highest incidence was observed in Kankrej (75.52%)) by ABELISA, while HF cross-yielded highest positive samples (84.72%) by l-ELISA. Lowest incidence was seen in Gir breed by both the tests (51.51 and 66.66 percent), while in sheep, higher prevalence rate in native Patanwadi breed (64.28 and 73.21 percent) was observed than the crossbred (41.17 and 47.05 percent) by AB-ELISA and I-ELISA respectively. Comparison of l-ELISA and AB-ELISA for the detection of BTV antibodies, revealed that the former test was better in terms of sensitivity and specificity. Out of a total 527 serum samples tested 343 (65.08%) and (75.14%) reacted positively in AB-ELISA and l-ELISA respectively. l-ELISA detected BTV antibodies in 54 (10.25%) samples, which were negative by AB-ELISA. Relative sensitivity and specificity of AB-ELISA to l-ELISA were 86.63 and 99.23% respectively and overall agreement between both the tests was 89.56%. RNA-PAGE was employed for detecting BTV using a cell culture propagated BTV serotype 1. This was attempted essentially to standardize this highly sensitive technique, so as to use if routinely in future for the field samples. The study revealed 10 RNA segments with a migration pattern resembling to serotype 1 of BTV.ThesisItem Open Access CHARACTERIZATION OF DETERMINANT FOR TRANSFERABLE ANTIMICROBIAL DRUG RESISTANCE AND VIRULENCE ASSOCIATED FACTORS OF S. GALLINARUM(AAU, Anand, 1999) Patel, Ashvinkumar Ramabhai; Roy, A.In the present study characterization of antimicrobial drug resistance and its transferable nature in S. gallinarum isolates was carried out. Further curing of R-determinants and the effect of curing on virulence on virulence associated factros viz., colicinogeny and serum resistance was ascertained. Virulence of isolates were assayed by lethality and invasiveness assay and attempts were also made to isolate plasmid from S. gallinarum isolates. In vitro antimicrobial drug resistance against 10 commonly used antibiotics were detected. All the S. gallinarum isolates showed resistance against one or more drug tested. All the isolates were resistant to sulfamethoxazole while higher number of isolates showed resistance to nalidixic acid, gentamicin and enrofloxacin. Moderate number of isolates showed resistance against furazolidone, streptomycin, tetracycline and cotrimoxazole while resistance against pefloxacin was shown by lesser number of isolates. Least resistance isolates were observed for chloramphenicol. In vitro transfer of drug resistance was also studied. Out of 12 S. gallinarum isolates tested all (100 per cent) were found to transfer resistance against tetracycline, nine (75 per cent) isolates transferred en bloc resistance against tetracycline, gentamicin and sulfamethoxazole while three (25 per cent) isolates transferred resistance against only tetracyc1ine. In vivo transfer of drug resistance was carried out from two donor strains of S. gallinarum to recipient E. coli strain. Out of two S. gallinarum strains tested both revealed transfer of drug resistance from donor to recipient. Total 20 (57.1 per cent) isolates out of 35 isolates tested were found to produce bacteriocin. Twelve isolates were tested for transfer of colicinogeny, out of which six (56 per cent) were able to transfer of colicinogeny to recipient E. coli strain along with multiple drug resistance. Elimination of drug resistance (curing) markers in six strains of S. gallinarum were studied using different chemical and physical method. The chemical agents used for curing were ethidium bromide (EtBr) and novobiocin (Novo) ethidium bromide and novobiocin combination and sodium dodecyl sulfate (SDS). In present study the elimination of drug resistance by chemical agents observed frequently. Use of EtBr and SDS resulted in curing of drug resistance in all six strains while five and four strains were cured by EtBr and Novo combination and novobiocin alone respectively. In physical method of curing, 3 out of six S. gallinarum strains were eliminated drug resistance on 1 day of incubation at 45°C and higher number of curing frequencies, was observed on subsequent days. Thus resistance markers were more readily eliminated by incubation at 45°C for prolonged periods of time rather than by chemical agents. Resistance of six strains of S. gallinarum wild and their cured derivatives to bactericidal effect of chicken serum was studied. The study revealed that all the wild and cured strains tested were resistant to chicken serum. To know the role of virulence associated factors like colicinogeny and serum resistance, virulence assay of six selected wild and their cured derivatives was carried out by determining their lethality as well as invasiveness ability in day old chicks. In lethality assay wild strains (S-21, S-4 and S-27) caused mortality 66.66, 60.00 and 46.66 per cent respectively, while their cured derivatives caused mortality of 46.66, 20.00 and 20.00 per cent respectively. At the 15th day post infection (PI) surviving birds revealed infection as per the CFU count method. In invasiveness assay there was no significant difference in the CFU count among S. gallinarum wild. isolates and their cured derivatives was observed upto 12 day CFU count, but by the 16th day none of the chicks survived in wild strains. Thus it indicates that wild strains were more invasive and lethal as compared to their cured derivatives. All the surviving birds in cured group revealed infection at 16th PI. There was no apparent difference found between lesions of wild and the cured derivatives during virulence assay. Isolation of plasmids from 10 wild and their eight cured derivatives of S. gallinarum were carried out. Majority of the strains showed a plasmid of molecular weight 56 Md. In addition other small plasmids found were of 2.8 Md and 1.79 Md molecular weight.ThesisItem Open Access SEROPREVALENCE AND DIAGNOSIS OF BLUETONGUE VIRUS BY REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION(AAU, Anand, 1998) Hinsu, T. V.; Kher, H. N.Bluetongue (BT) is an insect transmitted viral disease of several species of domestic and wild ruminants. The disease is a cause for serious concern to livestock industry due to staggering direct and indirect economic losses. In many countries like India having considerable sheep population, the disease has become endemic. Severity of the infection depends upon the species and breed of animals, serotypes/strains of the virus and prevalent ecosystem. The present study was aimed to find out prevalence of Bluetongue virus (BTV) antibodies in domestic ruminants and of Epizootic haemorrhagic disease virus (EHDV) antibodies in cattle of Kutch district of Gujarat state. Agar gel immunodiffusion (AGIO) and competitive enzyme-linked immunosorbent assay (c-ELISA) employed for antibody detection were also compared in terms of their sensitivity and specificity. An attempt was also made to standardize Reverse transcriptase-polymerase chain reaction (RT-PCR) for detecting BTV nucleic acid. Out of 162 sera tested, 87 (53.70%) and 126 (77.78%) were found to be positive for BTV antibodies by AGID and c-ELISA respectively. Specieswise, 24.56 and 63.16% of sheep, 80.00 and 96.36% of goats and 58.00 and 74.00% of cattle revealed antibodies to BTV by AGIO and c-ELISA respectively. The highest prevalence rate was found in Northern-east region (70.69 and 86.21%), followed by Central (48.72 and 75.64%) and Southern-west region (30.77 and 65.38%) of the Kutch district, respectively by AGID and c-ELISA. Female animals (63.03 and 86.81%) showed more prevalence than male animals (36.63 and 66.20%), as determined by AGID and c-ELISA respectively. In sheep, higher prevalence rate in native Patanwadi breed (47.06 and 86.16%) was observed than the crossbreds (11.76 and 43.48%) by AGID and c-ELISA respectively. A total of 50 cattle sera tested for BTV antibodies were also tested for EHDV antibodies by EHDV-AGID test. Of these, 9 (18.00%) were found positive for EHDV antibodies. Amongst these positive sera, five reacted specifically to EHDV antigen, without crossreacting to BTV antigen. Comparison of c-ELISA and AGID tests for the detection of BTV antibodies, revealed that the former test was better in terms of sensitivity and specificity. Of the total 162 serum samples, 87 and 126 samples reacted positively in AGIO and c-ELISA respectively. Eighty six samples were found positive and 35 negative by both the tests; while 40 samples detected positive by c-ELISA were negative by AGIO. Only one sample reacted positively to AGIO but negative in c-ELISA. This sample turned out to be positive for EHDV antibodies. Relative sensitivity and specificity of AGIO to c-ELISA were 68.25 and 97.22% respectively and overall agreement between both the tests was 74.69%. RT-PCR was employed for detecting BTV nucleic acid using BTV groupspecific segment 7 prime and BHK-21 cells adopted BTV serotype 1. This was attempted essentially to standardize this highly sensitive technique, so as to use it routinely in future for the field samples. The study revealed an amplified product of 500 bp specific to the primer used in the study.ThesisItem Open Access ANTIGENIC CHARACTERIZATION OF EGG DROP SYNDROME-1976 (EDS-76) VIRUS(AAU, Anand, 1994) Bhalja, S. R.; Anjaria, J. M.Egg drop syndrome-1976 (EDS-76) virus has gained importance as one of the major causes of drop in egg production. EDS-78 virus has representative strains like 127 and BC14 which are identical serologically. The possibility of appearance of new isolates can not be ruled out by investigating the variations in immunogenic fractions and their interrelationship say provide indications to select a particular strain for vaccine production and to correlate specific antigenic fractions to immunogenicity of the virus isolate. The present work was on establishing antigenic relationship between strain 127 and two local isolates Sb and Kr of BDS-76 virus employing Haemagglutination inhibition (HI), Agar gel inmunodiffusion (AGID), Counter immunoelectrophoresis (CIEP) and Cross immunoeleotrophoresis (CrIEP). No much significant variation could be observed in the HI titres of sera against strain 127 and isolates Sb and Kr. Thus isolates of EDS-76 were indistinguishable based on HI test. Two common sharing lines were observed in strain 127 and Kr isolate, while one common sharing line was observed in Sb isolate with homologous as well as heterologous system in AGID. More number of precipitation lines were detected in ClEP than that in AGID in homologous and heterologous systens. Closed antigenic relationship was observed among strain 127 and isolates Sb and Kr. In standardized CrIEP using homologous and heterologous system of strain 127 and isolates Sb and Kr, four immuno precipitates were observed in each case. No apparent differences could be established among three isolates (strain 127 and isolates Sb and Kr of EDS-76 virus) by HI test, AGID, CIEP and CrIEP.