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20186
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Subject: Veterinary Microbiology × End date: 2018 × Start date: 2018 ×
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    ThesisItemOpen Access
    A COMPREHENSIVE APPROACH FOR DIAGNOSIS OF LEPTOSPIROSIS IN DOMESTIC PIGS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2018-09-30) P. S. RESHMA; M. Mini
    Leptospirosis is a worldwide zoonotic disease that in pigs primarily causes reproductive disturbances. Samples collected from Centre for Pig Production and Research (CPPR), Mannuthy and selected private farms in Thrissur were used for the present study. Serum samples (n=103) were tested using microscopic agglutination test (MAT) and an overall seropositivity of 35.92 per cent could be detected. The serogroup Pomona (45.95 per cent) was the most prevalent among the 37 positive samples followed by serogroups Grippotyphosa (24.32 per cent), Canicola (13.51 per cent), Icterohaemorrhagiae (10.81 per cent) and Tarassovi (5.41 per cent). Samples of serum (n=56), whole blood (n=52) and aborted foetus (n=7) when tested using PCR for the presence of lipl32 gene of Leptospira, DNA of three whole blood samples and one aborted foetus amplified the gene producing an expected 767 bp amplicon. Latex agglutination test (LAT) and indirect fluorescent antibody test (FAT) was standardised using seven-day-old reference cultures of Leptospira interrogans serovar Pomona as positive controls at 10-fold dilutions. However, none of the abortion samples tested using LAT and FAT were positive. Attempts for isolation from positive samples were also made, but Leptospira could not be isolated from any of the foetal membrane, liver, kidney samples from the PCR positive animals. Hence in the present study, a seroprevalence of 35.92 per cent for leptospirosis among pigs in Thrissur district was detected using MAT, and PCR was found to be the most sensitive method for directly detecting the presence of the organism in clinical samples, compared to LAT, FAT and culture.
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    ThesisItemOpen Access
    MOLECULAR DIAGNOSIS OF LEPTOSPIROSIS AND DETECTION OF LEPTOSPIRA SEROVARS PREVALENT IN DOMESTIC ANIMALS IN WAYANAD DISTRICT
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2018-12-10) SHIJU SHAJI; John, Koshy
    Leptospirosis has re-emerged as a prominent zoonotic disease of global distribution especially in developing countries causing severe complications in human beings and animals. Accurate identification of infecting serovars is necessary for adopting effective vaccines, as the immunity is serovar specific. Southern states like Kerala and Tamil Nadu is found to be endemic for leptospirosis. In districts like Wayanad of Kerala, conditions are congenial for occurrence of leptospires in environment as well as natural carriers. Chances of endemicity also are greater due to several factors including heavy rainfall, water holding soil, sharing of natural water resources by man and animals, and large number of rodents and stray dogs. Though many studies have identified the prevalent Leptospira serovars in many districts in Kerala, no study has been carried out in diagnosis and detection of prevalent serovars in Wayanad. . This scenario demanded for identification of prevalent serovars and detection of presence of infection in domestic animals in Wayanad. Hence, the present study was undertaken with the objective of detecting infection by PCR and identification of prevalent serovars by MAT. A total of 71 samples were collected from dogs, cattle and rats from Wayanad districts. Out of 71 samples, 25 samples were collected from animals suspecting leptospirosis brought to Teaching Veterinary Clinical Complex, Pookode. Six post mortem samples of dogs suspecting leptospirosis were collected from Department of Veterinary Pathology, College of Veterinary and Animal Sciences, Pookode. Four rat tissue samples were collected from rats captured from campus of College of Veterinary and Animal Sciences, Pookode. Thirty six serum samples were collected from different regions of Wayanad. All the samples collected were subjected to total DNA extraction and PCR was carried out using G1/G2 and lipL32 based primers. The test could detect amplicons in positive control but no positive results were yielded by clinical samples. For detection of prevalent serovars, MAT was conducted for 61 samples with 12 reference serovars. Twenty serum samples were found to be positive at 1: 200 serum dilution. Predominant serovar showed agglutination was Pyrogenes (7 samples), followed by Grippotyphosa (6 samples), and then Bataviae (3 samples), Icterohaemorrhageae (3 samples), Javanica (3 samples), Pomona (3 samples), Australis (2 samples), Canicola (2 samples). Out of 20 positive samples, nine samples have shown 54 agglutination with more than one serovar. In such cases, serovar with highest MAT titre was considered as infecting serovar. Mixed infections included Pyrogenes and Bataviae (3 samples), Pyrogenes and Javanica (2 samples), Grippotyphosa and Canicola (2 samples), Pyrogenes and Australis (1 sample), Grippotyphosa and Javanica (1 sample). The MAT titre of agglutinating serovars varied from 1:3200 to 1: 200. Highest MAT titre was 1:3200 against serovar Pyrogenes. Previously reported prevalent serovars in other parts of Kerala included Australis, Autumnalis, Pomona, Canicola etc. Though these serovars were reported in present study also, the predominant serovar revealed was Pyrogenes. Similarly, serovars present in mixed infections were also slightly different from those present in other parts of Kerala. As the serological data acquired from this study is novel and distinct, more detailed studies including more number of samples is required for confirmation of prevalent serovars in Wayanad district.
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    ThesisItemOpen Access
    A COMPREHENSIVE APPROACH FOR DIAGNOSIS OF LEPTOSPIROSIS IN DOMESTIC PIGS
    (College of Veterinary and animal Science,Mannuthy, 2018) P. S. RESHMA; M. Mini
    Leptospirosis is a worldwide zoonotic disease that in pigs primarily causes reproductive disturbances. Samples collected from Centre for Pig Production and Research (CPPR), Mannuthy and selected private farms in Thrissur were used for the present study. Serum samples (n=103) were tested using microscopic agglutination test (MAT) and an overall seropositivity of 35.92 per cent could be detected. The serogroup Pomona (45.95 per cent) was the most prevalent among the 37 positive samples followed by serogroups Grippotyphosa (24.32 per cent), Canicola (13.51 per cent), Icterohaemorrhagiae (10.81 per cent) and Tarassovi (5.41 per cent). Samples of serum (n=56), whole blood (n=52) and aborted foetus (n=7) when tested using PCR for the presence of lipl32 gene of Leptospira, DNA of three whole blood samples and one aborted foetus amplified the gene producing an expected 767 bp amplicon. Latex agglutination test (LAT) and indirect fluorescent antibody test (FAT) was standardised using seven-day-old reference cultures of Leptospira interrogans serovar Pomona as positive controls at 10-fold dilutions. However, none of the abortion samples tested using LAT and FAT were positive. Attempts for isolation from positive samples were also made, but Leptospira could not be isolated from any of the foetal membrane, liver, kidney samples from the PCR positive animals. Hence in the present study, a seroprevalence of 35.92 per cent for leptospirosis among pigs in Thrissur district was detected using MAT, and PCR was found to be the most sensitive method for directly detecting the presence of the organism in clinical samples, compared to LAT, FAT and culture.
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    ThesisItemOpen Access
    MOLECULAR DIAGNOSIS OF LEPTOSPIROSIS AND DETECTION OF LEPTOSPIRA SEROVARS PREVALENT IN DOMESTIC ANIMALS IN WAYANAD DISTRICT
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES POOKODE, WAYANAD, 2018) SHIJU SHAJI; Koshy John
    Leptospirosis has been identified as re-emerged potentially zoonotic disease with wide distribution in developing countries like India. In regions like Wayanad, leptospirosis is underdiagnosed and prevalent serovars are unknown due to lack of previous studies and due to lack of facilities for proper diagnosis. In this scenario, a comprehensive study was conducted for identification of prevalent serovars and detection of infection in domestic animals of Wayanad district by molecular methods. Total 71 samples including blood, serum, urine and tissue samples were collected from clinically suspected cases of leptospirosis and from different regions of Wayanad district. All the samples collected were subjected to total DNA extraction and PCR was carried out using G1/G2 and lipL32 based primers. The test could detect amplicons in positive control but no positive results were yielded by clinical samples.Out of total 62 serum samples tested by MAT, the standard serological test for leptospirosis, 20 samples have given positive result. The most predominant serovar identified was Pyrogenes which was followed by Grippotyphosa, Bataviae, Icterohaemorrhageae, Javanica, Pomona, Australis and Canicola. Mixed infections were also observed which included Pyrogenes and Bataviae, Pyrogenes and Javanica, Grippotyphosa and Canicola, Pyrogenes and Australis, Grippotyphosa and Javanica. The MAT titre of agglutinating serovars varied from 1:3200 to 1: 200. Highest MAT titre observed was 1:3200 which was against serovar Pyrogenes.
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    ThesisItemOpen Access
    DETECTION OF BOVINE CORONAVIRUS IN DIARRHOEIC CALVES
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2018) THANKSY S. AKKARA; M. Mini
    The present research was undertaken to detect the presence of coronavirus in faecal samples of calves with diarrhoea by RT-PCR and confirmation by TEM. One hundred-and-four faecal samples of diarrhoeic calves below three months were collected from farms and hospitals of Kerala Veterinary and Animal Sciences University as well as from selected private farms located in Thrissur and Palakkad districts. All the samples were screened for detecting bovine coronavirus (BCoV) by RT-PCR with focus on the conserved nucleocapsid (N) gene of BCoV. Among 104 samples collected, two (1.92 per cent) were found to be positive by RT-PCR and both were from calves in the age group 0-14 days. The PCR product was sequenced and analysed using BLAST. The results showed one hundred per cent homology to BCoV isolates from Croatia, Korea and France. The two samples positive in RT-PCR when further examined by TEM, revealed typical morphology of BCoV, thus confirming the presence of the virus. The phylogenetic analysis of the sequence of the BCoV was carried out using MEGA 7 software. The evolutionary history inferred by maximum likelihood method revealed that the sequence showed similarity to the sequences of Croatia, Korea, France and several other BCoV isolates and divergence to avian infectious bronchitis virus.
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    ThesisItemOpen Access
    ISOLATION AND MOLECULAR DETECTION OF Mycoplasma capricolum subsp. capripneumoniae ASSOCIATED WITH RESPIRATORY TRACT INFECTION IN GOATS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2018) ROSHIN M. REJI; M. Mini
    The present research was undertaken to detect the presence of mycoplasmas associated with respiratory infection in goats with a preference to Mccp, the exact cause of CCPP. Ninety-six nasal swabs were collected from goats with respiratory infections from selected veterinary hospitals and farms in Ernakulam, Thrissur, Palakkad, Malappuram and Wayanad districts. Postmortem samples including lungs, mediastinal lymphnodes and pleural fluid were collected from 10 animals with pneumonic lesions brought for postmortem examination in the Department of Veterinary Pathology. Nucleic acid based techniques like PCR and conventional methods like isolation were employed for the detection of mycoplasmas. Out of the 106 samples tested, 52 (49 per cent) were found to be positive by 16S rRNA genus specific PCR. Among the Mycoplasma genus positive samples, one was found to be positive for Mycoides cluster, Mycoides group and Mmc specific PCR and was confirmed as Mycoplasma mycoides subsp. capri by nucleotide sequencing. None of the samples were found to be positive in Mccp specific PCR. A representative sample from each of the 16 flocks was subjected to nucleotide sequencing and the results revealed that animals in 10 flocks were infected with M. ovipneumoniae, four with M. conjuctivae, one with M. capri and one with M. agalactiae. On inoculating samples on to PPLO medium (modified Hayflick’s medium), two samples produced typical fried egg like colonies on agar plate and was confirmed as Mycoplasma by 16S rRNA genus specific PCR. Later, it was confirmed as M. agalactiae by nucleotide sequencing. The isolation rate was found to be 3.84 per cent.