Thumbnail Image

Theses

Browse

20103
Reset filters
Subject: Veterinary Microbiology × Author: KAU × End date: 2011 × Start date: 2010 × Type: Thesis ×
Reset filters

Search Results

Now showing 1 - 3 of 3
  • Thumbnail Image
    ThesisItemOpen 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.
  • Thumbnail Image
    ThesisItemOpen 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.
  • Thumbnail Image
    ThesisItemOpen 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.