Background: This was a preliminary study to test for the presence of multiple antibiotic-resistant extended spectrum β-lactamase (ESBL) producing bacteria in Malaysian urban surface waters. Although the literature review revealed several published papers on clinical ESBL isolates in Malaysia, none were found on ESBL isolates obtained from local surface waters. Methods: Isolated bacterial species were tested for resistance to cefotaxime, amoxicillin/clavulanate and aztreonam, and susceptibility to imipenem and meropenem using antibiotic susceptibility testing (AST) by disc diffusion. This served as a screening step to detect bacteria that could be potential ESBL species. 16S ribose ribonucleic acid (rRNA) polymerase chain reaction (PCR) testing with two clusters of bla (β-lactamase) gene primers was used to test for the bla genes CTX-M (Groups 1, 2, 9), OXA-1, SHV and TEM. Results: A total of 19 isolates were found, possessing at least one of the bla genes tested for. There was a relatively high occurrence of CTX-M genes (84.2%) among these, followed by TEM genes (47.4%). The isolates were identified as Enterobacteriaceae (89.5%), predominantly Escherichia coli and Klebsiella pneumoniae. Conclusion: There appears to be a high occurrence of ESBL-bacteria in local surface waters, among these being opportunistic pathogens. The persistence and spread of these species in the environment poses a threat to exposed human populations.
Hazardous Substances ; Bacteria ; Water

Aims: The aims of this study were to isolate and characterize antimicrobial producing bacteria from tropical peat swamp forest soils. Methodology and results: Bacteria isolated from peat soil were screened for antimicrobial properties via agar overlay assay. Broth microdilution was performed using crude-cell free supernatant (CCFS) to determine the minimum inhibitory concentration (MIC). One isolate was selected due to its broad spectrum activity and identified as Burkholderia spp. with a maximum identity of 99% via 16s rRNA gene PCR. This isolate was able to produce antimicrobials that were active against several Gram positive bacteria, Gram negative bacteria and also yeast. The antimicrobial activity of the CCFS was stable at a pH range of 1 to 11, temperatures of −20 °C to 80 °C, and after treatment with several proteolytic enzymes: α-chymotrypsin, proteinase K and trypsin, indicating that the antimicrobial produced might not be proteinaceous in nature. It is possible that the isolate can produce polyketides, a type of antimicrobial compound produced by Burkholderia known to be resistant to proteolytic enzymes. However, further work needs to be done to confirm this. Conclusion, significance and impact of study: The presence of antimicrobial producing bacteria signified that tropical peat swamps are indeed a potential source for antimicrobials to combat infections.
Anti-Infective Agents

Aims: Cellulases are enzymes that convert cellulose into glucose molecules, and are produced by various microorganisms in the environment. Due to their importance to the biofuel industry, there is a need to screen for more efficient varieties of cellulases. In this study, leachate samples from a landfill site were screened for cellulolytic bacteria. Methodology and results: Leachate samples obtained from a landfill collection pond were cultured in an enriched cellulose medium. Two cellulolytic isolates, designated MAEPY1 and MAEPY2, were isolated and further characterized. Phenotypic profiles and phylogenetic analyses using sequences of 16S rRNA, gyrB and whole genome suggested that these isolates are new strains of the Paenibacillus genera. The crude enzyme extracts from both isolates have cellulose degradation activity at approximately 0.1-0.2 IU/mg under working conditions of pH 6 and 55 °C. Assays using other lignocellulosic substrates showed that the crude enzyme extracts also have high xylan degradation activity. Conclusion, significance and impact of study: Paenibacillus sp. are known to produce multiple enzymes for lignocellulolytic degradation and the present results suggest that isolates described in this study, MAEPY1 and MAEPY2, are excellent candidates deserving further study as potential producers of efficient cellulases for use in industries associated with cellulosic biomass.
Cellulases

Aims: Bacteria on chicken egg surfaces can be potential sources of food borne diseases. The aim of this study was to determine the prevalence of E. coli, Salmonella and enterococci on the surface of conventional broiler eggs, “Kampung” chicken eggs and carrying trays and to determine the antimicrobial resistant profile of these isolates. Methodology and results: Conventional broiler eggs, “Kampung” chicken eggs and carrying trays were sampled randomly from nine wet markets in Selangor, Malaysia. The surface of the eggs and carrying trays were swabbed and E. coli, Salmonella and enterococci were isolated using selective agars. Antimicrobial susceptibility testing (AST) was performed on the isolates against different antimicrobials via disk diffusion test. A large proportion of E. coli isolates (>50% of isolates from conventional broiler eggs and “Kampung” chicken eggs) was resistant to chloramphenicol and tetracycline whereas enterococci (>60% of isolates from conventional broiler eggs and “Kampung” chicken eggs) isolates were resistant to tetracycline and erythromycin. Salmonella isolates were found to be susceptible to all of the antimicrobials tested except for tetracycline. There was also presence of isolates showing multiple resistances in this study. E. coli isolates (8.8%) from the surface of “Kampung” chicken eggs were resistant against 10 different antimicrobials whereas 17.8% of the enterococci isolates from the surface of “Kampung” chicken eggs were resistant to 11 different antimicrobials. Conclusion, significance and impact of study: The presence of multiple-antimicrobial resistant bacteria especially on the surface of “Kampung” chicken eggs that are ready to be sold to consumers is a serious concern. However, further study has to be conducted to determine the ultimate source of the bacterial contamination before specific food safety measures can be introduced.
Drug Resistance, Microbial ; Salmonella