Molecular characterization of environmental non-Tuberculous mycobacteria using PCR- RFLP analysis of 441 Bp heat shock protein 65 fragments

Non- Tuberculous Mycobacteria are environmental opportunistic pathogens that can be found in various terrestrial and aquatic habitats. There are an epidemiological links between species isolated in tap water and those isolated from patients. hsp65 gene has more variability in its sequences, compared to the some more conserved genes in NTM, for identification of mycobacteria to species level. In this study, the prevalence of NTM in Isfahan City water samples was determined using culture, biochemical tests and PCR-RFLP analyses of hsp65 gene. Eighty-five water samples were collected and cultured. The mycobacterial isolates were identified by conventional biochemical tests. A 441 bp fragment of hsp65 genes was amplified and digested by two restriction enzymes, BstEII and HaeII. Digested products were analyzed using polyacrilamid gel electrophoresis [PAGE]. 25.9% of the water samples contained different species of NTM. Dominant isolates were M. fortuitum [26.7%], M. chelonae like organism [13.3%] and M. mucogenicum [13.3%]. Nineteen isolates of Mycobacteria were differentiated using hsp65 genes PCR-RFLP. Three isolates could not be identified at the species level because their RFLP patterns were different from other known PCR-RFLP profiles. There were different hsp65 gene PCR-RFLP profiles produced by digestion with BstEII and HaeIII. This study showed that PCR-RFLP of hsp65 gene in mycobacteria is more reliable method for identification of NTM at the specie level than conventional phenotypic methods [P<0.05]. In comparing of RFLP patterns of this study to other investigation, some minor differences were negligible

[ study of antibacterial effect of humulus lupulus on some of gram positive and gram negative bacteria]

In recent years, a new view has been stablished toward pharmaceutical plants and applicational use of them all over the world and researchers, in various medical courses are investigating about this issue. On this basis, antibacterial effect of Humulus lupulus, the pharmaceutical plant which has an important place in traditional medicine of Iran, was evaluated on some of bacteria. Based on scientific methods after being dried, prepared extract by using macerasion method. Antibacterial effects of ethanol extract were studied on Staphylococcus aureus [1112PTCC], Escherichia coli [1276PTCC], Bacillus subtilis [1023PTCC], Psedomonas aeroginosa [1430PTCC], by using spreading method, well plate method and MIC. The ethanol extract of Humulus lupulus showed antimicrobial activity on S. aureus and B. subtilis. The extract exhibited antimicrobial activity at a range of 125 mg/ml to 62/5 mg/ml against S. aureus and B. subtilis respectively. With the increase of worldwide antibiotic resistance of many strains of disease producing bacteria, there is a need to access new and complementary approaches to antibiotic therapy. Extract of Humulus lupulus has antibacterial effect. The antimicrobial activities of Humulus lupulus against a variety of gram positive bacteria confirm its use for treatment of infectious