APPLICATION OF POLYMERASE CHAIN REACTION FOR DIAGNOSIS AND MOLECULAR EPIDEMIOLOGY OF ANTHRAX IN LIVESTOCK IN KARNATAKA SHASHIKALA, N.
Loading...
Date
2018-01
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR – 585 401
Abstract
The present study employed the microscopy and the PCR for the detection of B
.anthracis by targeting virulence genes for protective antigen (PA) and capsule (CAP)
located on two plasmids, pXO1 and pXO2 respectively. Further, Ba813 gene of B.
anthracis and nhe gene of B. cereus were targeted for species specific identification. In
all, 14 blood smears from 5 different locations (Bellary, Koppal, Davangere,
Doddaballapura and Chamarajnagar) were subjected to Gram’s and Polychrome
methylene blue (PMB) staining. Of these, 2 smears from Doddaballapura were negative
for anthrax bacilli by both the staining techniques. These blood smears and the blood
samples were also found negative by PCR. Further, out of 10 blood samples from
Bellary, Koppal, Doddaballapura and Chamrajnagar, 5 were PCR positive. Out of 14
blood smears from Davanagere, Doddaballapura and Chamarajanagar, only 4 smears
revealed PCR positivity. All the 5 ear peice samples collected from Bellary, Koppal and
Tumkur were found negative by PCR. None of the 5 soil samples from Bellary, Koppal
and Chamarajnagara were PCR positive. The B. cereus isolate showed amplification for
both PA as well as nhe genes. Sequencing and phylogenetic analysis of the PA gene
revealed that the PA gene sequences of the B. cereus were homologous to that of B.
anthracis. This indicated close genetic relatedness between B. anthracis and B. cereus.
The high genetic homology (98%) among B. anthracis isolates was revealed. The Ba813
and CAP gene based phylogenetic analysis indicated probable prevalence of two strains
of B. anthracis in the outbreak of anthrax in Bellary