Isolation and identification of Streptomyces sp. Act4Zk, a good producer of Staurosporine and some derivatives.

In this study, strain Streptomyces sp. Act4Zk was isolated based on a method developed for the isolation of myxobacteria. Due to the low efficiency of the majority of conventional DNA extraction techniques, for molecular identification of the strain Streptomyces sp. Act4Zk, a new technique for DNA extraction of Actinobacteria was developed. In order to explore potential bioactivities of the strain, extracts of the fermented broth culture were prepared by an organic solvent (i.e. ethyl acetate) extraction method using. These ethyl acetate extracts were subjected to HPLC fractionation against standard micro-organisms, followed by LC/MS analysis. Based on morphological, physiological, biochemical and 16S rRNA gene sequence data, strain Streptomyces sp. Act4Zk is likely to be a new species of Streptomyces, close to Streptomyces genecies and Streptomyces roseolilacinus. Antimicrobial assay indicated high antifungal activity as well as antibacterial activity against Mycobacterium smegmatis and Gram-positive bacteria for the new strain. HPLC and LC/MS analyses of the extracts led to the identification of three different compounds and confirmed our hypothesis that the interesting species of the genus Streptomyces being a good producer of staurosporine and some derivatives.

Evaluation of potential probiotic characteristics and antibacterial effects of strains of Pediococcus species isolated from broiler chickens.

Pediococcus spp. were isolated from poultry rectum, faeces and food as good probiotic candidates in order to select strains to be used as probiotic in poultry feed. A total of 168 lactic acid bacteria were isolated and 51 isolates including 31 Lactobacillus spp. and 20 Pediococcus spp. were able to survive in low pH and bile salt concentration. The Pediococcus spp. were identified and their ability to form biofilm, adhesion to Caco-2 cells and antimicrobial activities against enteric pathogenic bacteria were determined. The results showed the presence of two strains, Pediococcus acidilactici P17 and P19 in rectal swab samples from 21-d old chickens with significant antibacterial activities against Salmonella enteritidis and Escherichia coli. The results suggest that only a few isolates of Pediococcus with potential probiotic activities are present in the poultry industry.

Molecular identification and detection of virulence genes among Pseudomonas aeruginosa isolated from different infectious origins.

BACKGROUND AND OBJECTIVES: Pseudomonas aeruginosa possesses a variety of virulence factors that may contribute to its pathogenicity. The aim of this study was to evaluate oprI, oprL and toxA genes for PCR identification of clinical P. aeruginosa. In order to find out any relation between special virulence factors and special manifestation of P. aeruginosa infections, we detected virulence factors among these isolates by PCR. Ribotyping was used to evaluate the clonal relationship between strains with the same genetic patterns of the genes studied. MATERIALS AND METHODS: In this study, 268 isolates of P. aeruginosa were recovered from burn, wound and pulmonary tract infections. The prevalence of oprI, oprL, toxA, lasB, exoS and nan1 genes was determined by PCR. One hundred and four isolates were selected randomly to investigate clonal diversity of the isolates with ribotyping using SmaI. RESULTS AND CONCLUSIONS: All P. aeruginosa isolates in this study carried oprI, oprL and lasB genes. Difference between exoS prevalence in isolates from pulmonary tract and burn isolates was statistically significant. Prevalence of nan1 and toxA gene was significantly higher in pulmonary tract and burn isolates, respectively. Ribotyping showed that most of the isolates (87%) belonged to clone A and B. Detection of oprI, oprL and toxA genes by PCR is recommended for molecular identification of P. aeruginosa. Determination of different virulence genes of P. aeruginosa isolates suggests that they are associated with different levels of intrinsic virulence and pathogenicity. Ribotyping showed that strains with the same genetic patterns of the genes do not necessarily have similar ribotype patterns.

The homogeneity of vanB gene cluster among enterococcal isolates in Iran.

AIMS: This study aimed to analyse the diversity of the vanB gene cluster in enterococcal species isolated from sewage treatment plants (STP) in Tehran, Iran. METHODS AND RESULTS: The enterococcal isolates were collected from three sewage treatment plants in Tehran, Iran, during 2005. A total of 203 enterococcal isolates, collected over six rounds of sampling from three STPs were tested for the presence of vanB gene. Long-PCR showed that amongst the isolates, three Enterococcus faecium, one Enterococcus gallinarum and one Enterococcus casseliflavus harboured the complete vanB gene cluster. Restriction fragment length polymorphism (RFLP) of the vanB1 gene cluster (5900 bp) from the isolates showed an identical pattern to a standard strain of Enterococcus faecalis (V583). None of the isolates were able to transfer the vanB gene in conjugation experiments. Different pulsed-field gel electrophoresis patterns were obtained for the three E. faecium isolates with vanB gene clusters. CONCLUSIONS: Our results indicated that the dissemination of vanB is not widespread in Tehran. Although only a few vanB positive isolates were detected, vanB was found in several enterococcal species. SIGNIFICANCE AND IMPACT OF THE STUDY: In view of the lack of information on vanB resistance genes and their diversity in Iran, knowledge of the global dissemination of vanB genes in Enterococcus spp. is noteworthy.