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Kerala Agricultural University, Thrissur
The history of agricultural education in Kerala can be traced back to the year 1896 when a scheme was evolved in the erstwhile Travancore State to train a few young men in scientific agriculture at the Demonstration Farm, Karamana, Thiruvananthapuram, presently, the Cropping Systems Research Centre under Kerala Agricultural University. Agriculture was introduced as an optional subject in the middle school classes in the State in 1922 when an Agricultural Middle School was started at Aluva, Ernakulam District. The popularity and usefulness of this school led to the starting of similar institutions at Kottarakkara and Konni in 1928 and 1931 respectively.
Agriculture was later introduced as an optional subject for Intermediate Course in 1953. In 1955, the erstwhile Government of Travancore-Cochin started the Agricultural College and Research Institute at Vellayani, Thiruvananthapuram and the College of Veterinary and Animal Sciences at Mannuthy, Thrissur for imparting higher education in agricultural and veterinary sciences, respectively. These institutions were brought under the direct administrative control of the Department of Agriculture and the Department of Animal Husbandry, respectively. With the formation of Kerala State in 1956, these two colleges were affiliated to the University of Kerala. The post-graduate programmes leading to M.Sc. (Ag), M.V.Sc. and Ph.D. degrees were started in 1961, 1962 and 1965 respectively.
On the recommendation of the Second National Education Commission (1964-66) headed by Dr. D.S. Kothari, the then Chairman of the University Grants Commission, one Agricultural University in each State was established. The State Agricultural Universities (SAUs) were established in India as an integral part of the National Agricultural Research System to give the much needed impetus to Agriculture Education and Research in the Country. As a result the Kerala Agricultural University (KAU) was established on 24th February 1971 by virtue of the Act 33 of 1971 and started functioning on 1st February 1972. The Kerala Agricultural University is the 15th in the series of the SAUs.
In accordance with the provisions of KAU Act of 1971, the Agricultural College and Research Institute at Vellayani, and the College of Veterinary and Animal Sciences, Mannuthy, were brought under the Kerala Agricultural University. In addition, twenty one agricultural and animal husbandry research stations were also transferred to the KAU for taking up research and extension programmes on various crops, animals, birds, etc.
During 2011, Kerala Agricultural University was trifurcated into Kerala Veterinary and Animal Sciences University (KVASU), Kerala University of Fisheries and Ocean Studies (KUFOS) and Kerala Agricultural University (KAU).
Now the University has seven colleges (four Agriculture, one Agricultural Engineering, one Forestry, one Co-operation Banking & Management), six RARSs, seven KVKs, 15 Research Stations and 16 Research and Extension Units under the faculties of Agriculture, Agricultural Engineering and Forestry. In addition, one Academy on Climate Change Adaptation and one Institute of Agricultural Technology offering M.Sc. (Integrated) Climate Change Adaptation and Diploma in Agricultural Sciences respectively are also functioning in Kerala Agricultural University.
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ThesisItem Open Access Bacteria associated with respiratory infections in poultry(Department of Veterinery Microbiology, College of Veterinary and Animal Science, Mannuthy, 2007) Jesto, George; KAU; Krishnan Nair, GThis study was undertaken to isolate and identify of bacteria from respiratory tract of poultry and to study the antibiogram of the isolates. Birds showing respiratory signs were sacrificed, postmortem examination was conducted and samples such as nasal, tracheal and air sac swabs and lungs were collected after taking all sterile precautions. A total of 105 samples were collected by sacrificing birds showing clinical signs. Isolation of causative bacteria was made by culturing on brain heart infusion agar, Mac Conkey agar and blood agar. For identification of isolates all the procedures were followed as described by Barrow and Feltham (1993). A total of 31 bacterial isolates were obtained from samples. A total of 12 Escherichia coli isolates were isolated and identified, 4 Pasteurella multocida isolates and 15 Staphylococcus sp. Isolates were isolated and identified biochemically. Out of 15 Staphylococcus sp. isolated and identified 11 isolates (73.33 per cent) were coagulase negative This result indicate that CoNS were more frequently isolated from staphylococcal infections although they do not possess the virulent coagulase activity. So importance must be given to CoNS also, as given to coagulase positive staphylococci and much study need to be diverted to find the virulence factors and role of them in producing bacterial infections in poultry. Multi drug resistance (resistance to at least three antimicrobials) was found among all E. coli isolates obtained in the study. Hence it may be concluded that the high level of resistance observed among poultry E coli isolates obtained in the study may be due to incorporation of antibiotics in feed as growth promoters. As 100 per cent sensitivity is shown to enrofloxacin and chloramphenicol by P. multocida isolates, these two drugs may be used for treating pasteurellois. Amoxycillin clavulanic acid (Ac) and cephalexin (Cp) was found to be the most effective antibiotic against Staphylococcus sp. in the study. The plasmid DNA content of the seven isolates of E. coli was analysed on agarose gel electrophoresis but correlation between the number of plasmids and antibiotic resistance could not be ascertained in this study. In conclusion, the results of this study provide evidence for significant antimicrobial resistance among bacterial isolates from birds. Long term prospective studies involving isolation, identification and antibiogram from more samples are required to identify novel pathogens causing respiratory disease in birds. Such studies provide data on temporal and spatial difference in antibiotic resistance patterns, which in turn helps the scientific community to design better disease control strategies.ThesisItem Open Access Development and evaluation of outer membrane protein vaccine against duck pasteurellosis(Department of Veterinary Microbiology, College of Veterinary and Animal Science, Mannuthy, 2007) Ranjini, A R; KAU; Krishnan, Nair GA research work was undertaken to prepare effective vaccines against P. multocida grown under different conditions and their immunopotency was assessed in one month old ducklings. The purity of the P. multocida A:l strain (DP1) was confirmed as per standard procedure. Pathogenicity of the isolate was assessed in six to eight weeks old mice. The isolate killed the mice with in 8 h intra peritoneally and within 24 h when injected subcutaneously. Whole cell protein of P. multocida (DP1) was extracted and they were subjected to discontinuous system of SDS- PAGE which revealed 20 to 26 visible protein bands of molecular weight ranging from 102 to 19 kDa. After growing P .multocida in iron sufficient and iron restricted medium its OMPs were extracted and they were analysed by SDS PAGE. In iron sufficient medium, 10 protein bands of MW which ranging from 91.84 to 19.02 kDa were revealed. In addition to the above a protein band with MW of 97.8 kDa were detected in iron restricted medium.The protein concentration was estimated by Modified Lowry method,and it was found to be 4mg/ml. The Median Lethal Dose (LDso) of P. multocida when determined in one, month old ducklings was found to be 10 -7.13 which contained 13 cells. Oil adjuvanted formalin inactivated vaccines were prepared from DPl grown under three different conditions viz., TSB, BHIB and BHIB supplemented with 100 u M 2,2' dipyridyl. Sterility, safety and potency test of the vaccines were done as per standard procedures. A total of 120 one month old ducklings were divided in to four groups with 30 birds in each group. The first three groups were vaccinated with ordinary bacterin, OMP vaccine prepared under iron sufficient condition and OMP vaccine prepared under iron restricted condition respectively and the fourth group served as control.The birds were vaccinated with 0.5millilitre of vaccine intramuscularly. The blood was collected from all the ducks on day 0,7, 14,21,28,45 and 60 day PV. Passive haemagglutination was done and the increase in antibody titre was observed from day 7 PV onwards for groups I,.Il and In. The highest antibody titre was obtained at 14th day PV and 21 st day PV for aMP vaccine prepared under iron restricted condition. All the vaccine groups had shown a significant difference from the control group at all stages of study. On homologous challenging OMP vaccine under iron restricted media on 42nd day PV afforded 60 per cent protection. On 60th day PV afforded 50 per cent protection. Though the aMP vaccine prepared under iron restricted condition was found to provide high antibody titre, the protection percentage afforded by it in comparison with ordinary bacterin and aMP vaccine prepared under iron sufficient media was low. Hence, the aMP vaccine prepared under iron restricted condition need to be subjected to further investigation.ThesisItem Open Access Evaluation of pathogenicity and antigenic relationship of salmonella isolates from poultry(Department of Veterinery microbiology, College of Veterinary and Animal Science, Mannuthy, 2007) Sunil, G; KAU; Koshy, John (Guide)In the present study the pathogenicity and the antigenic relationship of three local isolates of S. gallinarum was compared with that of a reference/known strain. The antigenic homogeneity/heterogeneity of these isolates was studied using SDS PAGE and AGID. All the four isolates when subjected to antibiogram revealed similar pattern. The only difference was with isolate X 4 which was susceptible to co-trimoxazole while all the other three were resistant. Except for the reference/known strain BGL, all the other three isolates were found pathogenic to mice. All the four isolates were pathogenic to day-old chicks. Isolates QS 1 and X 4 were most pathogenic. Major clinical signs were somnolence, weakness, inappetance and whitish diarrhea and were more prominent in IM inoculated group. All chicks that died during the experimental period gave positive Salmonella isolation from liver, spleen, heart blood, lungs, yolk and ceca. At the end of experimental period survived birds were sacrificed and they gave positive isolation only from ceca. All the four isolates were pathogenic to layers. The clinical signs observed were listlessness, inappetance, ruffled feathers, shrunken comb, greenish-yellow diarrhea with gradual weight loss and was more prominent in IV challenged group, followed by those inoculated by oral and IC routes. The most pathogenic strain was QS 1. Cloacal swabs examined for Salmonella revealed intermittent shedding in all groups. The highest re-isolation was obtained from liver. The egg production was affected in all test groups and was most severely affected in IV group. No significant difference was noticed between different routes of inoculation and among different isolates in re-isolation from egg. The re-isolates were confirmed as S. gallinarum by standard biochemical reactions. The gross and histopathological changes observed in experimental infection with all the isolates were the same. There was enlargement and necrosis of liver and spleen, with congestion of heart and lungs. Day-old chicks had omphalitis. Histopathologically there was congestion, necrosis and infiltration of inflammatory cells into the internal organs, including submucosa of intestine. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis analysis of the OMPs of all four isolates revealed 13 different polypeptide bands. These bands were of similar molecular weight in all the four isolates, indicating antigenic homogeneity. Agar gel immunodiffusion carried out using hyper immune serum raised against isolates BGL and QS 1 revealed lines of identity between the four isolates, indicating antigenic homogeneity by serology.ThesisItem Open Access Detection of rotavirus in the faeces of diarrhoeic calves by reverse transcriptase- polymerase chain reaction and silver staining(Department of Veterinary and Microbiology, College of Veterinary and Animal Science, Mannuthy, 2007) Ambily, R; KAU; Koshy JohnA study was undertaken to detect the presence of rotavirus in the faeces of diarrhoeic calves by RT-PCR and RNA-PAGE. Agar Gel Immune Diffusion was performed to detect the presence of rotaviral antigens in faecal samples using hyperimmune serum raised in rabbits. The protein profile of BRV was analyzed using SDS-PAGE. Attempts were made to isolate BRV from faecal samples in MDBK cell line. One hundred and twenty four faecal samples of diarrhoeic calves were collected from University Livestock Farm, Mannuthy, Veterinary Hospitals of Kerala Agricultural University, some dairy farms in Thrissur district and also from individual farmers in and around Thrissur. Twenty samples each were collected from adult cattle above one year of age with diarrhoea and normal healthy calves. All these samples were screened for the presence of BRV by RNA-PAGE, RT-PCR and AGID. Among 124 faecal samples collected 29 (23.39 per cent) samples were detected as positive by RNA-PAGE. The clustered arrangement of the 11 segments of the genome showed a 4:2:3:2 migration pattern, typical of group A bovine rotavirus. Reverse Transcriptase – Polymerase Chain Reaction could detect BRV in 35 (28.23 per cent) samples. By using AGID, only 16 (12.90 per cent) samples were found positive. Among the various tests employed, RT-PCR was found to be more sensitive in the diagnosis of BRV infections. All the 20 faecal samples from adult cattle with diarrhoea were tested negative by the three methods. Rotavirus could not be detected in the faeces of healthy calves by any of the tests employed. The protein profile of BRV revealed nine polypeptides having molecular weight in the range of 16.5 to 131 kDa. The age-wise distribution of BRV infection in calves was studied. It was found that the occurrence of infection was most common in zero to two weeks of age (39.39 per cent) followed by two to four weeks of age (37.04 per cent). The faecal samples which were found positive by RNA-PAGE was inoculated into MDBK cell line. But the attempts to isolate BRV were found unsuccessful.ThesisItem Open Access Molecular methods based detection of pathogenic mycoplasmas of chicken(Department of Veterinery Microbiology, College of Veterinary and Animal Sciences, 2006) Dipu, M K; KAU; Jayaprakasan, VA study was undertaken for the detection of Mycoplasma DNA from the specimens by Polymerase Chain Reaction (PCR), differentiate the three significantly pathogenic mycoplasmas of chicken namely M. gallisepticum, M. synoviae and M. iowae from the other less pathogenic ones based on the result of polymerase chain reaction. Attempts were also carried out to isolate Mycoplasma from clinical samples testing positive by polymerase chain reaction and from a few randomly selected negative samples and to comprehend the different strains of M. gallisepticum, if any, among chicken of various age groups. Out of a total of 225 birds subjected for the study 25 were found positive for the presence of avian Mycoplasma by genus-specific PCR. Thirty samples from these twenty-five birds were positive. Tracheal swabs form all these birds were positive. Fifteen isolates were obtained from these twenty five birds when the tracheal swabs were directly streaked onto BHI agar whereas when these tracheal swabs were collected in BHI broth initially and later subcultured onto BHI agar following a positive result in PCR / colour change of the broth, only 13 of them yielded colonies. Five of the isolates were found to be M. gallisepticum by MG-PCR and these were obtained upon direct inoculation of BHI agar with tracheal swabs from birds. Those samples collected in BHI broth yielded MG colonies only in two cases. Non-MG colonies were found interspersed with MG colonies in agar plates and thus the selective isolation of MG colonies must be performed prior to subculture. The utility of PBS and buffered peptone water supplemented with glycerol as transit media for clinical samples intended for PCR was evidenced. The isolates obtained were successfully lyophilized and stored at –70°C throughout the span of the study. The PCR-RFLP pattern of the obtained MG isolates revealed uniformity among the isolates. None of the samples yielded positive result in MS-PCR and MI-PCR. The sensitivity and usefulness of molecular methods based detection of pathogenic mycoplasmas of chicken over isolation techniques could be appreciated.ThesisItem Open Access Polymerase chain reaction based screening of bovine mastitis milk to detect leptospira and mycoplasma(Department of Veterinery Microbiology, College of Veterinary and Animal Science, Mannuthy, 2007) Jibi, M G; KAU; Krishnan Nair, GIn the present study an attempt has been made to detect the Leptospira and Mycoplasma from bovine mastitis milk by PCR. Simultaneously trials were also made to isolate these fastidious organisms from these milk samples. Direct screening of milk sample by PCR could not successfully detect the DNA of either of these organisms. All the previous studies had revealed that inhibitory factors present in the milk might interfere with the amplification of target DNA present in the milk. To overcome this difficulty initially all the milk samples were preenriched in the broth media, subsequently the target DNA was extracted. The genus specific primer A and B were used to detect the leptospires in milk samples. Out of fifty haemagalactic milk samples screened by PCR only one sample was positive for Leptospira. Fletcher’s semisolid media that could support the growth of reference strain of Leptospira pomona was used for the isolation study. But all the attempts to recover the organism from milk sample were unsuccessful. For the detection of Mycoplasma, genus specific primers MGSO and GPO3 were used. Out of the fifty samples screened only one sample was positive for Mycoplasma. But the M.bovis specific primers PpMB 920-1 and PpMB 920-2 failed to amplify the DNA that was found positive in the genus specific PCR. Since several species of Mycoplasma was found to be associated with the mastitis, organisms detected in this study might not be M.bovis. The media used for the isolation of Mycoplasma were BHI broth and BHI agar. Media were standardized with the reference strain and were found to be capable of supporting the growth of organisms. But all the attempts to isolate the Mycoplasma from milk samples were unsuccessful. It was concluded that PCR is a rapid and sensitive technique to detect the fastidious organisms that are difficult to isolate in the ordinary laboratory conditions. But one difficulty encountered with the PCR from milk samples was the extraction of good quality template DNA that contained fewer inhibitors. Since the rapidity and ease of PCR is largely dependent upon the extraction procedure adopted in each bio materials, the protocol followed in this study needs some modifications.ThesisItem Open Access Detection of infectious bovine rhinotracheitis virus by immunofluorescence and polymerase chain reaction(Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2008) Anupama, K; KAU; Koshy, JohnA study was undertaken to detect the presence of IBR virus in clinical samples by PCR and immunofluorescence and to compare the efficacy of these tests. Attempts were also made to isolate the virus from clinical samples in MDBK cell line. A total of 60 samples from suspected animals and 5 samples from healthy animals were collected from various sources like, University Livestock Farms, Veterinary Hospital, Mannuthy and some private dairy farms in Thrissur district. The samples included nasal and vaginal swabs, aborted materials and semen. All these samples were screened for the presence of IBR virus by PCR and immunofluorescence. The TK gene based primers were used for PCR and the positive control generated an amplicon of size 298bp. No sample was found positive by PCR. Immunofluorescence was performed on smears prepared from clinical samples using FITC conjugated polyclonal antibovine IgG. This test detected viral antigen in one of the samples, indicated by focal, bright yellowish green fluorescence associated with the cytoplasm of epithelial cells. The inability of PCR for detection of viral DNA in sample tested positive by immunofluorescence may be due to the presence of non-specific inhibitors in clinical sample. These results suggest that even though PCR is highly sensitive compared to other diagnostic tests, false negative results cannot be excluded. Immunofluorescence on smears prepared from clinical samples can be considered as a rapid diagnostic method for IBR. Attempts to isolate the virus from clinical samples in MDBK cell line were found unsuccessful.ThesisItem Open Access Development and evaluation of whole cell and membrane protein vaccines against mycoplasma gallisepticum(Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2009) Surya, Sankar; KAU; Krishnan Nair, GA study was carried out to develop and evaluate the efficacy of vaccines against M. gallisepticum infections in Gallus domesticus using whole cell and membrane proteins produced under iron sufficient and restricted conditions, incorporating the adjuvants aluminium hydroxide and saponin either alone or in combination. A total of 50 samples were collected in broth and in addition 25 tracheal swabs were streaked onto PPLO agar plates also, from birds showing respiratory ailments. Mycoplasma genus specific PCR revealed twelve positive samples and on subjecting these samples to species specific PCR, a total of four samples showed positive results. From four species specific PCR samples, two isolates could be obtained, named as MG UPF-1 and MG UPF-2, which were sub cultured and subjected to morphological, biochemical and serological characterization. The SDS-PAGE profile of whole cell proteins of the two isolates revealed similar profile, indicating their homogeneity. Whole cell and membrane proteins were isolated from cultures grown under iron sufficient and iron deficient conditions. Both the whole cell and membrane proteins revealed bands ranging from 24 kDa to 200 kDa on SDS-PAGE. The whole cell proteins had two prominent bands at 75 kDa and 35 kDa and eight faint bands, which the membrane proteins lacked. The whole cell and membrane proteins produced under iron restricted conditions had an additional band of 52 kDa, which was not seen in the respective protein groups isolated under iron sufficient condition. Most of the proteins resolved on SDS-PAGE in the four sets of proteins were found to be antigenic as found out by western blot analysis. The optimum concentration of the proteins used for vaccine preparation was 84.2 µg per dose of the vaccine. Formalin inactivated vaccines were formulated with saponin and aluminium hydroxide alone and in combination at a final concentration of 100 µg/dose and 25 per cent v/v respectively, thereby producing eight different sets of vaccines. The sterility and safety testing of the vaccines were carried before vaccination trials. The infective dose of M. gallisepticum cells for challenge studies was found to be 1.1 x 105. For vaccination studies, 240 chicks of five weeks age were divided into 10 groups and were inoculated with eight different sets of vaccines. One group was inoculated with commercial vaccine and other was kept as control. Potency testing for humoral response was conducted with HI test and CMI response with LMIT and Blastogenic calorimetry. The mean HI titre of groups vaccinated with iron sufficient and restricted saponin adjuvanated vaccines showed an increase in their titre at first week post booster vaccination followed by decline of the titre on subsequent collections on third and fifth week post booster vaccination. The mean HI titres of groups vaccinated with iron sufficient and restricted saponin-aluminium hydroxide adjuvanated vaccines maintained significantly high HI titres during the entire study period. Commercial vaccine group also showed a similar response. The control group showed significantly less titre from other vaccinated groups during the entire time period. Leukocyte migration inhibition test and Blastogenic calorimetry were carried out to assess the CMI response evoked by different sets of vaccines. A significant CMI response was shown by all vaccinated groups except commercial vaccine and control group. To assess the protective response evoked by different vaccines, two challenge studies were conducted two weeks apart. All the vaccinated birds were having significantly lower lesion score when compared to the control group. Mycoplasma gallisepticum was re-isolated from all the control birds with lesions during the first challenge study and from 70 per cent of the birds in the second challenge study. A latex agglutination test was developed using iron restricted M. gallisepticum whole cell proteins. When compared with HI test, LAT had a sensitivity of 95 per cent and specificity of 93 per cent. The test was statistically significant as indicated by a kappa value greater than 0.81. The saponin-aluminium hydroxide adjuvanated whole cell and membrane protein vaccines had elicited similar humoral immune response than vaccines adjuvanated with saponin alone. With regard to CMI response, the experimental vaccines produced a significant CMI response whereas, commercial vaccines failed to evoke a protective CMI response. In brief, saponin-aluminium hydroxide adjuvanated whole cell and membrane protein vaccines, especially those produced under iron restricted conditions, could be an effective tool to afford protection against M. gallisepticum infection, as it would stimulate both humoral and cell mediated immune responses in host Latex agglutination test was found to be an effective tool for flock screening of birds for M. gallisepticum antibodies and the test is simple, rapid, and easy to conduct, with applicability under field conditions.ThesisItem Open Access Comparative efficacy of certain diagnostic tests on detection of paratuberculosis in cattle(Department of Veterinary Microbiology,College of Veterinary and Animal Sciences, Mannuthy, 2010) Remya, Raveendran; KAU; Priya, P MA study was conducted to compare the efficacy of ZN acid fast staining, and IS900 PCR to detect bovine paratuberculosis. Indirect ELISA was also performed to assess the seroconversion. A total of 58 faecal samples and sera samples were collected from University Livestock Farm, Mannuthy; Cattle Breeding Farm, Thumburmuzhy; Livestock Research Station, Thiruvazhamkunnu and one private farm in Thrissur. The faecal samples were used to conduct the ZN acid fast staining and IS900 PCR. Indirect ELISA was performed using the sera samples. Among 58 samples collected, 10 samples (17.24 per cent) were positive in IS900 PCR and 6 samples (10.34 per cent) were positive in ZN acid fast staining. Two samples (3.45 per cent) were positive in indirect ELISA. The two ELISA positive samples were positive in IS900 PCR and one was positive in ZN acid fast staining. Among the six acid fast positive samples, five samples were positive in IS900 PCR but one sample was negative. Although IS900 PCR detected maximum number of positive samples than acid fast staining, statistically there was no significant difference. Since the seroconverted animals are very less at the initial stage of infection, indirect ELISA cannot be used for serodiagnosis for subclinical bovine paratuberculosis. ZN acid fast staining is a rapid, cheap, easy and field oriented diagnostic technique for detecting subclinical paratuberculosis. At the same time, IS900 PCR is a rapid and sensitive molecular based method for detecting subclinical paratuberculosis. Hence it was concluded that a combination of ZN acid fast staining and IS900 PCR was found to be very useful in diagnosing subclinical cases of bovine paratuberculosis.