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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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, G
A 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.
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 M
A 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.
Open Access
Isolation and characterization of bacteria associated with gastroenteritis in weaned Piglets
(College of Veterinary and Animal Sciences, Mannuthy, 2009) Atulya, M; KAU; Koshy, John
Gastroenteritis is a leading cause of morbidity and mortality in piglets. A number of factors are involved in post weaning diarrhoea in piglets. A comprehensive study was performed to examine the incidence, bacterial etiology, drug sensitivity, plasmid profile and pathogenicity of the bacteria isolated from weaned piglets with gastroenteritis prevalent in and around Kerala Agricultural university. Samples were taken only from piglets with diarrhoea that had not been previously treated with antibiotics. Rectal swabs were collected from live diarrhoeic piglets and intestinal contents, pieces of jejunum, ileum, colon, mesenteric lymph nodes, liver, spleen and stomach were collected during post mortem examination after taking all sterile precautions. Isolation of causative bacteria was made by culturing on Brain Heart Infusion Agar, Mac Conkey agar, Mannitol Salt agar, Blood agar, Brucella agar and Cooked Meat medium. The identification of isolates was carried out as per standard protocols. All the procedures of biochemical testing were followed as described by Barrow and Feltham (1993). For classification of Staphylococcus isolates, a system suggested by Baird Parker (1965) was considered. A total of 53 bacterial isolates were identified to species level from 82 samples tested for pathogens. Six different microorganisms were encountered in this study, with Escherichia coli being dominant, followed by Pseudomonas aeruginosa, Staphylococcus subgroups BPIII and BPV, Serratia fonticola, Salmonella typhimurium and Aeromonas hydrophila. Thirteen different serotypes of E. coli were encountered, with O69 being dominant and the others were O38, O5, O84, O132, O80, O79, O56, O41, O25, O109 and O103 and two were untypable. Majority of the isolates exhibited multidrug resistance. Plasmid profile of the Gram negative isolates were determined and 80.43 per cent were found to bear plasmids. Pathogenicity of the isolates was determined by performing in vivo mice pathogenicity test.
Open Access
Evaluation of whole cell antigen and outer memberane protein based latex agglutination test for serodiagnosis of canine leptospirosis
(College of Horticulture, Vellanikkara, 2010) Sariprabha, P; KAU; Koshy, John
A study was undertaken to evaluate the efficacy of whole cell antigen and OMP based Latex Agglutination Test (LAT) for the serodiagnosis of canine leptospirosis. Serum samples were collected from University Veterinary Hospitals of Mannuthy and Kokkali. These samples were subjected to serologic testing by Microscopic Agglutination Test (MAT) and LAT and the results were compared. A total of 60 serum samples were screened in this study. MAT detected a prevalence rate of 76.67 per cent. The most predominant serovar was Australis (38.33 per cent) followed by Grippotyphosa (18.33 per cent), Pomona (18.33 per cent), Canicola (15.0 per cent), Icterohaemorrhagiae (13.33 per cent), Javanica (10.0 per cent), Patoc (8.33 per cent), Autumnalis (6.67 per cent) and Pyrogenes (6.67 per cent). The sensitivity and specificity of whole cell antigen were 97.83 per cent and 75.86 per cent respectively. On the other hand, the sensitivity and specificity of OMP based LAT were 95.65 per cent and 86.21 per cent respectively. As the kappa values of both LATs were greater than 0.81, the tests indicate perfect agreement with MAT. From this results, OMP–based LAT developed for the detection of leptospiral antibodies was proved to be a very useful rapid test for immunodiagnosis. OMP–based LAT is an extremely simple and inexpensive test that does not require expertise or sophisticated equipments and could also be used for the detection of leptospiral antibodies in place of MAT, which requires live leptospiral cultures, expertise, time.
Open Access
Isolation and characterization of Pasteurella Multocida from animals and birds
(College of Veterinary and Animal Sciences, Mannuthy, 2010) Ambili, K; KAU; Krishnan Nair, G
A total of 284 samples comprising of tracheal, nasal and pharyngeal swabs, heart blood and tissues like liver, spleen, heart and lungs were processed for isolation of P. multocida.Twenty isolates were obtained from different species of animals and birds, which were characterized as P. multocida by morphological, cultural and biochemical tests. Reference strains of P. multocida P52 and DP1were used for comparison. All the isolates were found to be pathogenic for mice.Based on the variation in fermentation patterns of arabinose, dulcitol, sorbitol, xylose and trehalose the 20 isolates could be grouped into eight biovars. Three biotypes P. multocida subsp gallicida, P. multocida subsp septica and P. multocida subsp. multocida were observed among the twenty isolates.All isolates were uniformly sensitive to norfloxacin, gentamicin, cefotaxime, nitrofurantoin, erythromycin and chloramphenicol. All were resistant to metronidazole and sulphadiazine.A species-specific PCR assay using primer pair KMT1SP6 and KMT1T7 was used to confirm the identity of the isolates. All the isolates obtained were confirmed as serogroup A P. multocida when subjected to multiplex PCR, using PM– specific, Cap A, Cap B, Cap D and Cap F primer pairs. Pasteurella multocida could be detected in only 6 out of 75 clinical samples tested by species specific PCR (PM-PCR). The entire samples tested positive by PM-PCR were confirmed as type-A P. multocida by multiplex PCR.The PCR product (460 bp) of 20 isolates and 5 clinical samples amplified by using primer pairs KMT1SP6 and KMT1T7 when used for reamplification with nested PCR primers, a product of 214 bp size was observed. A highproportion of the clinical samples previously found negative by PM-PCR gave positive results in nested PCR.All the 20 isolates of P. multocida, which were found to be positive by PM-PCR, were subjected to REP-PCR, using the primer pairs REP-1 and REP-2. Pasteurella multocida isolates demonstrated 3 distinct REP profiles, indicating heterogeneity among the isolates.The genomic DNA isolated from all the twenty isolates of P. multocida from different animals and birds were subjected to REA with restriction enzymes Hpa II and Hha I. Four different banding patterns were observed among the 20 isolates of P. multocida when they were subjected to REA with Hpa II, while five REA profiles were obtained with Hha I.In conclusion, PCR assays could be used for the rapid identification and serogrouping of P. multocida from both cultures and clinical samples. The different molecular techniques used in the present study showed genetic heterogeneity among the isolates of P.multocida serogroup A. The results suggested that there was no correlation between the results obtained in REP-PCR and REA. Also among the enzymes used in DNA fingerprinting of P. multocida isolates by REA, Hha I was found to have more discriminatory power. Since only lesser number of samples were used in the current study, a definite conclusion could not be drawn in this basis.
Open Access
Antioxidant potential of malphigia glabra(acerola) berries in rats
(College of Veterinary and Animal Sciences , Mannuthy, 2009) Arul Mary Luveena, A; KAU; Karthiayini, K
The present study was designed to assess the effect of aqueous extract of mature fruits of Malphigia glabra (Acerola) berries on paracetamol induced hepatotoxicity in rats. Forty-two adult male Wistar albino rats were used for the experiment. The rats were randomly divided into five groups with eight animals each in G1, G2 and G5 and nine animals in G3 and G4. Group G1 served as normal control rats. The G2 (untreated vehicle alone) rats were administered with 40 % sucrose syrup p.o. @ 10 ml/kg b.w. on day one and two and then at three days interval for 21 days. The G3 group of rats were administered with paracetamol @ 2 g/kg b.w. p.o., on day one and two and at every three days interval upto day 18. The G4 (curative group) rats were administered with paracetamol p.o. @ 2 g/kg b.w. on day one and two and at every three days interval upto day 18 and Acerola berry extract (20 ml/ kg b.w., p.o.) for 21 days. The G5 (protective group) rats were given Acerola berry extract p.o. @ 20 ml/ kg b.w. for 17 days and paracetamol p.o. @ 2 g/kg b.w. on day 18 and 19. Body weight was recorded at weekly intervals. Blood samples were collected from eight animals in each group on day zero, four, 10, and 21. In G5 group, an additional blood collection was made on 18th day of the experiment before administration of paracetamol. The haematological parameters such as total erythrocyte count (TEC), haemoglobin (Hb) concentration, total leukocyte count (TLC) and differential leukocyte count (DLC) and serum biochemical parameters such as alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), total cholesterol, total protein, albumin, globulin, albumin:globulin, blood urea nitrogen (BUN), total bilirubin, direct bilirubin, indirect bilirubin, reduced glutathione were analysed. One animal each from G3 and G4 was euthanized on day four and rest of the rats were euthanized on day 21. Levels of liver reduced glutathione, lipid peroxides and superoxide dismutase (SOD) on 21st day were estimated. Representative samples of liver and kidney tissues collected on day four and 21 were subjected to histopathological examination. Administration of paracetamol in G3 group caused a significant (P 0.05) increase in the levels of serum ALT, AST, ALP, total cholesterol, BUN and total, direct and indirect bilirubin while reduced glutathione content was significantly reduced. The activities of liver reduced glutathione and SOD were also decreased significantly whereas the liver lipid peroxide content was significantly increased. Haematological analysis showed significantly decreased TEC and Hb concentration and a significantly increased TLC with monocytosis. No significant (P> 0.05) variation was observed in body weight, and in the levels of serum total protein, albumin, globulin and albumin:globulin. Histopathology indicated necrosis of hepatic cells, diffused haemorrhage, central venous congestion and focal coagulation in liver; while tubular dilatation and congestion was observed in kidney. Administration of Acerola berry extract along with paracetamol in G4 (curative group) rats effectively reversed the levels of serum ALT, AST, ALP, total cholesterol, BUN, total bilirubin (direct and indirect), reduced glutathione (in liver and serum), liver SOD and lipid peroxides in liver to normalcy signifying the antioxidant and hepatocurative effect of the extract. However, the active antioxidant components of Malphigia glabra such as vitamin C and polyphenols has a short half life, so that the berry extract could not produce any prophylactic effect against paracetamol induced toxicity in G5 (protective group) rats, evidenced by toxic range of values.

Kerala Agricultural University, Thrissur

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