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Sharma, R. K.
- Molecular Characterization of Rhodococcus equi Isolates in Equines
Authors
1 Department of Microbiology, R.S. Pura, Sher-E-Kashmir University of Agricultural Sciences and Technology, Jammu, Jammu and Kashmir, IN
Source
Veterinary World, Vol 10, No 1 (2017), Pagination: 6-10Abstract
Aim: The aim was to determine the occurrence of Rhodococcus equi in equines and their environment in Jammu (R.S. Pura, Katra), molecular characterization and to determine the antibiotic resistance pattern of R. equi.Materials and Methods: A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of R. equi isolates was done by 16S rRNA gene amplification followed by virulence associated protein A (Vap A) gene amplification. Antibiogram was performed against five antibiotics, viz., amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin.
Results: During the study, 9 R. equi isolates were identified on the basis of cultural and biochemical tests. In the polymerase chain reaction based detection, 3 among the 9 rhodococcal isolates were positive for species-specific 16S rRNA gene and revealed amplicon of 450 bp for confirmation of 16S rRNA gene. None of the sample was found positive for Vap A gene. In antibiogram, R. equi isolates were found sensitive for amoxicillin, while some isolates were also found resistant to the most conventional antibiotic penicillin G.
Conclusion: From this study, it was concluded that R. equi infection is prevalent in equines in Jammu region of India and the indiscriminate use of the antibiotics is leading toward the development of resistant strains of R. equi.
Keywords
16S rRNA, Polymerase Chain Reaction, Rhodococcus equi.References
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- Virulence Gene Profiling of Porcine Pasteurella multocida Isolates of Assam
Authors
1 KVK Churachandpur, ICAR Manipur Centre, Imphal, Manipur, IN
2 Department of Microbiology, College of Veterinary Science, AAU, Khanapara, Guwahati - 781 022, Assam, IN
3 Department of Veterinary Epidemiology and Preventive Medicine, CVSc, CAU, Aizawl, Mizoram, IN
4 Department of Veterinary Public Health, College of Veterinary Science, AAU, Khanapara, Guwahati - 781 022, Assam, IN
Source
Veterinary World, Vol 11, No 3 (2018), Pagination: 348-354Abstract
Aim: The present study was conducted to detect and identify the virulence genes in Pasteurella multocida isolates of porcine origin from Assam.
Materials and Methods: A total of 21 porcine P. multocida isolates were subjected to capsular typing and detection of virulence-associated genes (pfhA, tbpA, hgbB, toxA, oma87, ompH, and nanB) using various polymerase chain reaction (PCR) methods reported elsewhere. Further, pathogenicity of the porcine isolates of P. multocida was studied in mice. For each strain of P. multocida selected for pathogenicity trial, the group of mice was injected intraperitoneally (i/p) with 0.1 ml of the inoculum prepared from respective field isolates, containing 109 organisms per ml.
Results: Capsular typing of the isolates by multiplex PCR showed two capsular types, type A (66.66%) and type D (33.33%). All the isolates were positive for outer membrane protein genes, oma87 and ompH genes. Iron acquisition genes, tbpA and hgbB, were detected in 14.28% and 19.04% of the isolates. The dermonecrotoxin encoding gene, toxA, was present in 23.80% of the isolates. Filamentous hemagglutinin encoding gene, pfhA, was detected in 28.57%. The virulence gene distribution pattern of the isolates indicates the important role of the genes in disease pathogenesis.
Conclusion: From the present study, it can be concluded that toxA gene is an important marker gene for defining the pathogenic potential of P. multocida strains in swine.