Molecular characterization of environmental mycobacterial species from leprosy endemic tribal regions of North Purulia District, West Bengal.
Background: The aim of the present study was to isolate and characterize nontuberculous mycobacteria (NTM) on Lowenstein-Jensen media supplemented with glycerol or pyruvate on two different temperatures from soil samples from leprosy endemic tribal areas of Purulia.
Methods: Mycobacterium leprae DNA was isolated from these samples followed by polymerase chain reaction (PCR) amplification using RLEP gene target specific to M. leprae. DNA was extracted from NTM cultures by lysis method. The presence of Mycobacterial DNA was confirmed by PCR using universal mycobacterial primer as 16S rRNA. NCBI nBlast was used for the authentication of NTMs, and phylogenetic tree was constructed using M. leprae and NTM species.
Statistical Analysis Used: The percentile method and phylogenetic tree were used as stastical tool in this research article.
Results: The rapid-growing mycobacteria (RGM) species, 4 (80%) was obtained more than that of slow growing mycobacteria (SGM) 1 (20%) supplemented on glycerol at 30°C followed by SGM species 8 (62%) were recovered more than RGM at 37°C. Similarly, SGM species 2 (100%) were recovered on supplemented with pyruvate at 30°C and no RGM growth when supplemented with pyruvate. Further, the recovery of RGM species 3 (60%) was better on supplemented with pyruvate than SGM species at 37°C. Mycobacterium timonense was first time isolated from Indian soil samples. Highest numbers of NTM were isolated from bathing place than washing and sitting places along with M. leprae PCR positivity. Phylogenetic tree showed a close genetic evolutionary association between Mycobacterium simiae and M. leprae in the leprosy endemic environment.
Conclusion: Several NTM was isolated from soil of leprosy endemic area which might have role in susceptibility of leprosy. Phylogenetic tree revealed a closed association of M. simiae with M. leprae in the environment and might be maintaining the leprosy endemicity in north block of Purulia.