Browsing by Author "Joshi, Dev Raj"
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Publication Antibacterial Compounds of Actinomycetes Isolated From Altitude Soils(Nepal Health Research Council, 2024) Aryal, Sagar; Adhikari, Rameshwar; Regmi, Balmukunda; Joshi, Dev RajBackground: Bacterial infection is a global threat due to antibiotic resistance. This demands the urgent need for novel antibiotics, and soil actinomycetes could be the potential candidate. The key objective of our study was to detect antibacterial compounds from selected actinomycetes species isolated from high-altitude soil samples. Methods: Three strains Streptomyces sp. 12923, Streptomyces sp. 13102, and Nocardia sp. 13105 were subjected to fermentation using International Streptomyces Project (ISP) 2 medium. Crude extracts of each isolate were recovered by Rotary evaporator. Crude extracts were fractionated in HPLC and fractions were collected in a 96-well plate to evaluate the antibacterial activity of each of the 19 fractions against a test organism E. coli BW25113. Crude extracts of three strains were analyzed using UPLC-MS/MS for antibacterial compounds. The LC-MS/MS data were processed using Metabo Scape software and features were annotated using different libraries in the software. Results: For Streptomyces sp. 12923, fractions of the crude extract with the lowest OD600 0.472, 0.484 showed higher antibacterial activity against E. coli BW25113T, while the inhibitory action against same test organism was shown better by fractions OD600 0.250, 0.329, and OD600 0.273, 0.326 for Streptomyces sp. 13102 and Nocardia sp. 13105, respectively. The antibacterial compounds detected included Mayamycin and Mayamycin B from Streptomyces sp. 12923, Nocardamine and Streptazone D from Streptomyces sp. 13102, and Nocardimicin B, 4-O-methylmelleolide, Spathullin B and Nannozinone B from Nocardia sp. 13105. Conclusions: The identification of these compounds from high-altitude actinomycetes further strengthens the claim that actinomycetes are rich sources of bioactive compounds. Keywords: Actinomycetes; antibacterial; metabolites; Nocardia; Streptomyces.Publication Distribution of MecA and Erm Genes among Methicillin-resistant Staphylococcus Aureus with Inducible Resistance to Clindamycin(Nepal Health Research Council, 2023) Dudha Raj Malla Thakuri,; Anil Pokhrel,; Ritu Amatya,; Bashyal, Narayan Sharma; Neupane, Mary; KC, Sudeep; Joshi, Dev Raj; Khanal, SantoshAbstract Background: The emergence of Methicillin-resistant Staphylococcus aureus and its ability to confer cross-resistance to macrolide-lincosamide-streptogramin B has complicated the treatment against it. Gene-based studies among phenotypic methicillin-resistant isolates with inducible resistance to clindamycin are less available in Nepal. This work was undertaken to detect the mecA and erm genes among such phenotypes isolated from clinical samples. Methods: S. aureus isolated from different clinical samples was identified by standard microbiological procedures (Gram-staining, colony morphology, and different biochemical tests). Methicillin-resistant and inducible resistant to clindamycin phenotypes were detected by using cefoxitin disc (30 µg) and a double disk diffusion test according to the Clinical and Laboratory Standards Institute guidelines and mecA and erm genes were detected by polymerase chain reaction. Results: Among 120 S. aureus isolates, 51.67% (n=62) were MRSA, and the prevalence of inducibly-resistant, constitutively-resistant and Macrolide-Streptogramin phenotypes were 15.83% (n=19), 28.33% (n=34) and 15.83% (n=19) respectively. While 35.84% (n=43) of isolates showed sensitivity to both antibiotics, erythromycin and clindamycin. Out of 14 inducibly-resistant phenotypes, 57.14% (n=8) were found carrying ermC and 28.57% (n=4) phenotypes contained both ermA and ermC. All phenotypes were positive for the mecA gene. Conclusions: Macrolides-Lincosamide-Streptogramin B resistance was predominant among methicillin-resistant S. aureus. While all isolates with inducible clindamycin resistance harbored mecA gene, most of them also harbored ermC gene. The higher prevalence of inducible-resistant to clindamycin indicated the need for rational use of antimicrobial agents. Keywords: Erm gene; iMLSB resistance; mecA; methicillin-resistance; MRSA