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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Characterization of genomic and plasmid DNA of Chlamydia psittaci isolates
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2001) Sreeja, R Nair; KAU; Mini, M
Four isolates of Chlamydia psittaci (M-28, M-430, M-121, P-156) obtained from ruminant abortion were subjected to restriction enzyme analysis and plasmid profiling to assess molecular level differences/homogenity among the isolates. The changes produced in developing chick embryo and the cytopathic effect in Me Coy cell line by the isolates were also investigated in this study. The C. psittaci isolates preserved in the Department of Microbiology were revived by passaging in embryonated chicken eggs through yolk sac route. Six to seven day-old developing chick embryo were used for propagation of the isolates. The isolates, M-28 and M-430 were found to produce death of the embryos within 10 days. The isolates M-121 and P-156 were not able to cause death within 10 days. The embryos appeared stunted with patchy haemorrhages all over the body and hyperemic legs. The yolk sac was thin walled, with deeply injected blood vessels and the yolk was thin and watery. M-28 and M-430 produced more severe lesions of the inoculated embryos compared with the other isolates. The isolates were grown in Me Coy cell line. All isolates revealed marked CPE, characteristic of C. psittaci, with rounding and clumping of cells to form syncytia at 48 h PI and formation of intra cytoplasmic inclusion bodies by 96 h PI. The isolates M-121 and P-156 took more time for the production of CPE compared with that of M-28 and M-430. The isolates were confirmed as C. psittaci by fluorescence antibody technique which revealed, apple green fluorescing chlamydial inclusions in the infected cell line by 96 h PI. The CE as well as cell line propagated isolates were used as a source of elementary bodies for DNA extraction. The purified EBs were prepared by urografin density gradient centrifugation. After obtaining sufficient amount of EBs, it was subjected to DNA extraction. The isolates M-28, M-430, M-121 and P-156 had DNA concentrations of 1710 ug/ml, 1130 Jlgl Iml, 1545 ug/ml and 1475 ug/ml respectively. Restriction enzyme digestion of all the isolates were done separately with Eeo RI, Hae III and Barn HI. Eeo RI cleaved DNA of the isolates M-430 and P-156 into rune fragments, M-28 into eight and M-121 into ten fragments respectively. The molecular size of bands for all isolates had variation in heavier fragments. Common bands were observed at five regions for all isolates. Hae III cleaved the caprine isolates into eight fragments and the bovine isolate M-121 into seven and P-156 into nine fragments. Variation could be observed on the molecular size of the fragments. Bam HI digested all isolates except M-121 into eight fragments. For M-121, seven fragments were obtained. Common bands were noticed at 0.2 and 0.7 kbp region. All enzymes were found to be useful in the differentiation of C. psittaci isolates as these REs produced variation as well as similarity in the restriction fragment sizes. Plasmid profiling revealed the absence of plasmids ill all the four isolates screened. Thus, a combination of phenotypic and genotypic methods could be used as epidemiological tools in the differentiation of C. psittaci strains of mammalian origin.
Open Access
Seroprevalence and Restriction enzyme analysis of Egg Drop Syndrome virus
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2000) Priya, P M; KAU; Krishnan Nair, G
In the present study, seroprevalence of EDS-76 was conducted in five districts of Kerala in duck and' chicken flocks using HI and ELISA. Out of 322 chicken and 281 duck sera samples screened, an overall incidence of 14.91 per cent and 26.69 per cent respectively were recorded. The high proportion of birds showing antibodies to EDS-76 reveals that the infection is widespread in Kerala and may be the major etiological factor associated with drop in egg production in poultry. Among the two serological tests namely, HI and ELISA employed for the detection of EDS-76 viral antibody, HI was found to be simple, sensitive and reliable. It is concluded that HI test could be used for the detection of EDS-76 infection in poultry flocks. Restriction DNA fingerprinting of the three indigenous strains were carried out in conjunction with the reference strain to check for any genetic variation between the strains. Comparison of the DNA fingerprint of all the four strains digested with restriction endonucleases BamHI, EcoRI and HindIII revealed identical banding pattern thereby conforming the genetic similarity of the strains.
Open Access
Nucleic acid and protein profile of Pasteurella multocida of avian origin
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2001) Rajalakshmi Radabai, S; KAU; Krishnan Nair, G
Four avian field isolates (one from quail and three from ducks) suggestive of Pasteurella sp. were compared with the avian Pasteurella multocida reference strain (LKO) for their biochemical properties, drug resistance patterns, whole cell and outer membrane protein profiles and restriction enzyme digestion pattern of chromosomal DNA. Biochemical characterisation revealed two biotypes among the four field isolates; namely P. multocida subsp. septica (quail isolate) and P. multocida subsp. multocida (duck isolates). The study of antibiogram using 14 chemotherapeutic agents revealed the presence of multiple drug resistance in all the four field isolates and reference strain. Whole cell protein profiles of the four field isolates and reference strain revealed 7 to 11 bands of molecular weights ranging from 197.4 kDa to 2.5 kDa. Polypeptide bands in the molecular weight range of 36 to 38 kDa (which were attributable to the protein-H), 29 to 32 kDa and 10 to 11 kDa were common to all the isolates of P. multocida. The major outer membrane proteins (OMPs) in the molecular weight range of 32 to 33 kDa and 27 kDa region were common to all the four field isolates and reference strain. II Unique protein bands in each of the isolate were indicative of variant forms of the same organism. Restriction enzyme analysis of chromosomal DNA yielded different restriction fragments in each of the isolates which were not easily distinguishable. Hence, of the tests used, phenotypic characters, biotype, antibiogram and protein profile were found to be more effective in differentiating the isolates rather than restriction enzyme digestion patterns of genomic DNA.
Open Access
Isolation and characterization of Chlamydia psittaci with emphasis on protein profile
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2001) Binu, K Mani; KAU; Mini, M
For isolation of Chlamydia psittaci 46 clinical cases associated with abortions in livestock from livestock farms and Veterinary hospitals in and around Thrissur were screened. Impression smear staining of the clinical materials USin9 Giemsa, Modified Ziehl Neelsen and Gimenez revealed two positive cases. Both were from aborted foetuses, one from bovine (M-121) and the other from caprine (M-430). Chick embryo inoculation by YS route was used for isolation of the organism. The organisms could be isolated only from the two cases positive by preliminary screening, as confirmed by staining of YS impression smears. The mortality rate in CE was more in case of M-430 compared to the other two isolates (M-121 and reference isolate, P-156). All the three isolates produced patchy haemorrhagic lesions in YS and skin of embryoS. An overall isolation percentage of 4.3 was obtained in this study. All the three isolates were resistant to sodium sulphadiazine. Pathogenicity studies indicated that, all the three isolates were of moderate virulence for mice and guinea pigs. Based on AGPT, the local isolates were confirmed as Chlamydia psittaci. All the three isolates caused rounding and swelling of fibroblast cells and syncytia formation at 24h PI in Mc Coy cell line. By 48 to 72 h PI, these effects became more prominent with detachment of cells from adjacent cells and from the glass surface. Purified EBs were prepared from infected Mc Coy cell line by Urografin gradient centrifugation. After confirming the presence of sufficient amount of protein by Biuret method, they were used for SDS-PAGE. A total of 12 polypeptide bands were obtained for both the bovine isolates while the caprine isolate gave 10 bands. Molecular weights of 148 kDa and 135 kDa were present only in P-156 and 152 kDa and 137 kDa were unique for M-121. Bands at 152 kDa, 148 kDa, 137 kDa, 135 kDa, 32 kDa, 18 kDa, 15 kDa and 7 kDa were lacking in M-430. This isolate was having unique bands of 155 kDa, 19 kDa, 12.2 kDa and 6.4 kDa.
Open Access
Characterisation of virus isolates from lesser whistling teals and channa species of fish
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2000) Pradeep, V; KAU; Krishnan Nair, G
Characterisation of virus isolates (T18 and T22) from lesser whistling teals (Dendrocygna javanica) and Channa species of fish (Fe and F12) was carried out to determine the similarities if any between the isolates and to identify the role of waterfowls in dissemination of these viruses. The virus isolates. preserved in the Department of Microbiology were revived by passaging through embryonated chicken! duck eggs through allantoic route. After the third passage, all the isolates were found to produce death of the embryos and the allantoic fluid collected agglutinated one percent chicken RBC. The isolates T18, Fs and F12 produced congestion of the embryo and CAM and the embryos showed sub-occipital and interdigital haemorrhages. Isolate T 22 also produced congestion of the embryo and CAM and the embryos were stunted. Liver of the embryos had yellowish brown patches. The EID50 of isolates were 3.2x10s, 5.6 x 105 , 1.65x 107 and 3.16 x 105 respectively for the isolates T18, T22. Fs and F12. The infectivity and haemagglutinating activity of all the isolates were retained at pH 7.2, but were completely lost at pH 3.2 and 9.0 and also by treatment at 56°C for 30 min. All the isolates were sensitive to chloroform indicating their enveloped nature. Pretreatment of chicken embryo fibroblast cultures with 1 OO~g! ml of luDR did not inhibit the multiplication of any of the isolates indicating they all had RNA genomes. All the isolates were resistant to treatment with brine indicating that they were capable of survlvinp at high salt concentration. The isolates T18 and Fa produced marked CPE in chicken embryo fibroblast culture with rounding and clumping of cells and syncytia formation. Marked cytoplasmic vacuolation was also observed. Inclusion bodies could not be detected either in nucleus or cytoplasm. For isolates T22 and F12, CPE developed later only and was not as prominent as for T18 and Fa. Rounding of cells and their fusion forming syncytia was noticed by 72 h. Cytoplasmic vacuolation though present was much less marked. Inclusion bodies were absent. Large polykaryocytes were produced by the isolates T18, T 22 and F12 in BHK-21 cell line within 24h after inoculation. Between 48-72h large syncytia were formed. Intracytoplasmic inclusions could be observed by 24h after infection, which were quite prominent by 96h. The isolate Fa failed to produce any CPE in BHK-21 cell line. Pathogenecity tests in day old and six-week-old chicken and ducklings showed that all the four isolates were non-pathogenic when given by the oral Isubcutaneous route or by both. Neither clinical signs or mortality could be observed in the birds. Virus isolation was possible from the cloacal swabs of six-week-old chicken for the isolates T18 and T22 up to the 14th and th day respectively. Antigenic relationship between the isolates was tested by gel diffusion and haemagglutination inhibition tests, which showed that the isolate T18 did not have any similarity with any of the other three isolates. The isolate T 22 showed antigenic similarity by both the tests. Fa showed similarity to T18 by HI test but not by immunodiffusion test. Isolate F12 was found to be distinct from the other three by HI test, but showed some similarity with them by immunodiffusion test. By sodium dodecyl sulphate polyacrylamide gel electrophoresis on 10 percent gels, 7-11 bands could be resolved for the different isolates. Three of the bands were common for all the four isolates and were having molecular weights similar to the three major proteins HN, NP and MP of avian paramyxoviruses, suggesting that the isolates belonged to the paramyxovirus group. Monoclonal antibody typing of the isolates T18 and T 22 at the Central Veterinary Laboratory, Surrey, England confirmed that both belonged to the paramyxovirus group with T18 belonging to group C (velogenic) and T 22 to group E (B1 and LaSota) viruses. The isolates Fe and F12 need to be further typed. It was concluded from the study that all the isolates were enveloped RNA viruses with T18 and T 22 being paramyxoviruses belonging to Group I. The properties of the isolates Fs and F12 resembled the paramyxoviruses and from the similarly in protein profile with the other two viruses can also be concluded to be paramyxoviruses.
Open Access
Molecular characterisation of salmonellae isolated from poultry
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2000) Tressa Mary, G; KAU; Punnose, K T
Five serotypes of salmonellae from avian sources were examined for biochemical properties, serology, drug resistance, plasmids, restriction enzyme pattern of plasmid as well as genomic DNA and pathogenicity. The biochemical characters are in confirmity with the characters described for the serotypes by the earlier workers. The study of antibiogram with 20 antibiotics/chemotherapeutic agents revealed the presence of multiple drug resistance in all the five serotypes. In the plasmid analysis, S enteritidis and S. branderup were found to be plasmid free. The number of plasmids in other serotypes ranged from two to three and the size ranged from 1.75 kb to 48.32 kb. Identical low molecular weight plasmids were present in both S. gallinarum. The presence of large plasmid in one of the S. gallinarum did not confer any additional detectable resistance character. S. typhimurium contained two plasmids of sizes 13.62 kb and 4.2 kb. Restriction enzyme analysis of plasmid DNA from three salmonellae with EcoRl and Hindlll yielded three different restriction pattern with each enzyme. Ethidium bromide and exposure to elevated temperature cured the plasmids present in the salmonellae within two days and 14 days respectively. Acridine orange and sodium dodecyl sulphate were found to be ineffective in curing the plasmid DNA. The elimination of plasmids resulted in the loss of resistance to antibiotics was demonstrated in S. typhimurium. In tests to assess the differences in pathogenicity between wild and cured isolates of S. typhimurium in day old chicks, only intraperitoneal route was found to be effective when compared to oral route. A relation of plasmids to virulence was noted only in S. typhimurium. Day old chicks were refractory to infection to S. gallinarum by both the routes. Plasmids encoding both resistance and virulence were observed in S typhimurium. Plasmid negative serotypes of S. enteritidis and S. branderup were found to be equally virulent as wild strains of S. typhimurium. So a definite correlation between virulence and plasmids could not be made. Restriction enzyme analysis of chromosomal DNA yielded bands which were indistinct and so uncomparable. Hence of the tests based on the analysis of genetic content plasmid profile was found ,to be efficient in typing the isolate rather than restriction enzyme analysis of genomic DNA.
Open Access
Homology between corynebacterium psedotuberculosis isolates from goats and standard reference strain
(Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2000) Mohan, P; KAU; Jayaprakasan, V
Corynebacterium pseudotuberculosis was isolated in pure culture on blood agar from pus collected from lymph nodes of goats, which suffered from caseous lymphadenitis. Three field isolates of C pseudotuberculosis were characterized and compared with standard reference" strain based on cultural, biochemical, toxigenicity, protein profile and restriction endonuclease digest analysis of chromosomal DNA. The morphological and cultural characters of all the field isolates and reference strain were similar and characteristics to the species. Biochemical reactions employed were also similar for all the strains except isolate F3, which fermented lactose. The biochemical characters were in confirmity with the characters described by the earlier workers. Three different types of toxin viz., culture filtrate (CF), whole cell lysate (WCL) and sodium chloride extract (SCE) were tested for dermonecro toxicity in rabbit skin. Among the preparations, the whole cell lysate produced a definite necrosis at the point of intradermal injection followed by culture filtrate and sodium chloride extract. The toxin preparations of the isolates invariably produced inflammatory and necrotic changes, the degree and severity of the reactions varied between samples. When the three toxin preparations of the isolates and reference strain were tested for synergistic haemolytic activity with Rhodococcus equi, the whole cell lysate produced maximum zone of haemolysis followed by culture filtrate and sodium chloride extract. The proteins presented in the different preparations were analyzed by SDS-P AGE. The protein profiles discerned by whole cell lysate, culture filtrate and sodium chloride extract were 25,9 and 9 protein bands with ranged on masses approximately 8-200KDa, 20-200KDa and 19-191KDa respectively. The 31.6KDa and 68 KDa proteins were found to be consistent in three toxin preparations. The DNA extracted from the isolates and reference strain were subjected to restriction endonuclease digestion employing Eco RI, Barn HI, BgI II, Eco RV and Pst I. The enzyme digest of DNA varied between the enzymes employed. There are no observable differences between the field isolates and reference strain, when the DNA digested with these five restriction enzymes. The enzymes Eco RI, Barn HI and BgI I presented similar restriction pattern whereas the DNA fragments generated by Eco RV and Pst I were different from the other three enzymes.

Kerala Agricultural University, Thrissur

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