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The polymerase chain reaction technique is used to identify the amoA gene encoding at the active site of ammonium monooxygenase from pure cultures of chemolithotrophic ammonium-oxidizing bacteria and the natural bacterial population of an unfiltered seawater sample. By partial sequencing the amplified DNA from Nitrosomonas europeae, Nitrosomonas crytolerans and Nitrosomonas oceanus, the amoA genes are detected.

Abstract Background Staphylococcus aureus including methicillin resistant S. aureus, MRSA, are human colonizing bacteria that commonly cause opportunistic infections primarily involving the skin in otherwise healthy individuals. These infections have been linked to close contact and sharing of common facilities such as locker rooms, schools and prisons Waterborne exposure and transmission routes have not been traditionally associated with S. aureus infections. Coastal marine waters and beaches used for recreation are potential locations for the combination of high numbers of people with close contact and therefore could contribute to the exposure to and infection by these organisms. The primary aim of this study was to evaluate the amount and characteristics of the shedding of methicillin sensitive S. aureus, MSSA and MRSA by human bathers in marine waters. Results Nasal cultures were collected from bathers, and water samples were collected from two sets of pools designed to isolate and quantify MSSA and MRSA shed by adults and toddlers during exposure to marine water. A combination of selective growth media and biochemical and polymerase chain reaction analysis was used to identify and perform limited characterization of the S. aureus isolated from the water and the participants. Twelve of 15 MRSA isolates collected from the water had identical genetic characteristics as the organisms isolated from the participants exposed to that water while the remaining 3 MRSA were without matching nasal isolates from participants. The amount of S. aureus shed per person corresponded to 105 to 106 CFU per person per 15-minute bathing period, with 15 to 20% of this quantity testing positive for MRSA. Conclusions This is the first report of a comparison of human colonizing organisms with bacteria from human exposed marine water attempting to confirm that participants shed their own colonizing MSSA and MRSA into their bathing milieu. These findings clearly demonstrate that adults and toddlers shed their colonizing organisms into marine waters and therefore can be sources of potentially pathogenic S. aureus and MRSA in recreational marine waters. Additional research is needed to evaluate recreational beaches and marine waters as potential exposure and transmission pathways for MRSA.

Data suggesting that fecal indicating bacteria may persist and/or regrow in sand has raised concerns that fecal indicators may become uncoupled from sources of human fecal pollution. To investigate this possibility, wet and dry beach sand, beach water, riverine water, canal water, and raw sewage samples were screened by PCR for certain pathogenic microbes and molecular markers of human fecal pollution. The targets included in this study were human specific Bacteroides (HF8 marker), human-specific enterococci (esp gene), Staphylococcus aureus, Escherichia coli 0157:H7, Campylobacter jejuni, and adenovirus. Sewage samples were also tested for Salmonella species. The results were compared to concentrations of enterococci, Escherichia coli, and Bacteroides species, as determined by membrane filtration methods. Molecular analysis yielded positive results for human specific Bacteroides, and S. aureus, in samples of raw sewage. Two of the environmental samples were positive for human specific Bacteroides and one was positive for S. aureus. The PCR screen was negative for other samples and targets, despite exceedance of EPA single sample guidelines for recreational waters on several of the sample dates (5/11 dates). However, estimates of the number of cells delivered to the PCR reaction suggested that few of the samples met the detection limit of the PCR reaction due to a variety of factors. The analysis indicated a need to improve nucleic acid processing in order to enable better delivery of DNA to downstream molecular methods. [ABSTRACT FROM AUTHOR]