Shedding of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus from adult and pediatric bathers in marine waters

<p>Abstract</p> <p>Background</p> <p><it>Staphylococcus aureus </it>including methicillin resistant <it>S. aureus</it>, 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 <it>S. aureus </it>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 <it>S. aureus</it>, MSSA and MRSA by human bathers in marine waters.</p> <p>Results</p> <p>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 <it>S. aureus </it>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 <it>S. aureus </it>shed per person corresponded to 10⁵to 10⁶CFU per person per 15-minute bathing period, with 15 to 20% of this quantity testing positive for MRSA.</p> <p>Conclusions</p> <p>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 <it>S. aureus </it>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.</p

The coastal environment and human health : microbial indicators, pathogens, sentinels and reservoirs

© 2008 Author et al. This is an open access article distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Environmental Health 7 (2008): S3, doi:10.1186/1476-069X-7-S2-S3.Innovative research relating oceans and human health is advancing our understanding of disease-causing organisms in coastal ecosystems. Novel techniques are elucidating the loading, transport and fate of pathogens in coastal ecosystems, and identifying sources of contamination. This research is facilitating improved risk assessments for seafood consumers and those who use the oceans for recreation. A number of challenges still remain and define future directions of research and public policy. Sample processing and molecular detection techniques need to be advanced to allow rapid and specific identification of microbes of public health concern from complex environmental samples. Water quality standards need to be updated to more accurately reflect health risks and to provide managers with improved tools for decision-making. Greater discrimination of virulent versus harmless microbes is needed to identify environmental reservoirs of pathogens and factors leading to human infections. Investigations must include examination of microbial community dynamics that may be important from a human health perspective. Further research is needed to evaluate the ecology of non-enteric water-transmitted diseases. Sentinels should also be established and monitored, providing early warning of dangers to ecosystem health. Taken together, this effort will provide more reliable information about public health risks associated with beaches and seafood consumption, and how human activities can affect their exposure to disease-causing organisms from the oceans.The Oceans and Human Health Initiative research described within this paper is supported by the National Science Foundation, The National Institute for Environmental Health Sciences and the National Oceanic and Atmospheric Administration. Grant numbers are: NIEHS P50 ES012742 and NSF OCE- 043072 (RJG, LAA-Z, MFP), NSF OCE04-32479 and NIEHS P50 ES012740 (RSF), NSF OCE-0432368 and NIEHS P50 ES12736 (HMS-G), NIEHS P50 ES012762 and NSF OCE-0434087 (JSM)

Detection and characterization of ammonia monooxygenase genes from nitrifying bacteria and natural waters

The chemolithotrophic ammonium-oxidizing bacteria are involved in the oxidation of ammonium to nitrite by the enzyme ammonia monooxygenase, which is a key step in the process of nitrification. The active site component of this membrane-associated multi-enzyme complex is encoded by the amoA gene. Since amoA encodes a critical function, it provides a useful target for tracking ammonium-oxidizing bacteria in the environment. The Polymerase Chain Reaction, DNA hybridization, Single Stranded Conformational Polymorphism Analysis, and DNA sequencing were used in combination to detect and characterize amoA sequences from pure cultures of ammonium-oxidizing bacteria and from native populations of ammonium-oxidizing bacteria in natural waters. It was found that some ammonium-oxidizer cultures contained several different amoA sequences. This methodology was sensitive and specific enough to detect a variety of different amoA sequences from natural waters at their in situ concentrations. It could detect variations in the composition of the amoA population from different water sample locations. A novel amoA sequence was also identified that appears to be widespread in marine waters. The particulate methane monooxygenase genes of methane-oxidizing bacteria were found to show significant similarity of structure, organization, and sequence to all the known ammonia monooxygenase genes of ammonium-oxidizing bacteria, including the ones identified in this study. This suggests an evolutionary relationship between these genes. The methodology used in this study has the potential for tracking the distribution, composition, and abundance of nitrifying bacteria in the environment, including novel species not yet identified, without the need for isolating or culturing these organisms

Integrating microbial source tracking with quantitative microbial risk assessment to evaluate site specific risk based thresholds at two South Florida beaches

Quantitative microbial risk assessment (QMRA) can be used to evaluate health risks associated with recreational beach use. This study developed a site-specific risk assessment using a novel approach that combined quantitative PCR-based measurement of microbial source tracking (MST) genetic markers (human, dog, and gull fecal bacteria) with a QMRA analysis of potential pathogen risk. Water samples ( n = 24) from two recreational beaches were collected and analyzed for MST markers as part of a broader Beach Exposure And Child Health Study that examined child behavior interactions with the beach environment. We report here the measurements of fecal bacteria MST markers in the environmental DNA extracts of those samples and a QMRA analysis of potential health risks utilizing the results from the MST measurements in the water samples. Human-specific Bacteroides was enumerated by the HF183 Taqman qPCR assay, gull-specific Catellicoccus was enumerated by the Gull2 qPCR assay, and dog-specific Bacteroides was enumerated by the DogBact qPCR assay. Derived reference pathogen doses, calculated from the MST marker concentrations detected in recreational waters, were used to estimate the risk of gastrointestinal illness for both children and adults. Dose–response equations were used to estimate the probability of the risk of infection (P inf ) per a swimming exposure event. Based on the QMRA simulations presented in this study, the GI risk from swimming or playing in water containing a mixture of human and non-human fecal sources appear to be primarily driven by the human fecal source. However, the estimated median GI health risk for both beaches never exceeded the U.S. EPA risk threshold of 32 illnesses per 1,000 recreation events. Our research suggests that utilizing QMRA together with MST can further extend our understanding of potential recreational bather risk by identifying the source contributing the greatest risk in a particular location, therefore informing beach management responses and decision-making

Fecal indicator bacteria levels at a marine beach before, during, and after the COVID-19 shutdown period and associations with decomposing seaweed and human presence

Studies are limited that evaluate seaweed as a source of bacteria to beach waters. The objective of the current study was to evaluate whether seaweed, along with humans and other animals, could be the cause of beach advisories due to elevated levels of enterococci. The monitoring period occurred a year prior to and through the COVID-19 beach shutdown period, which provided a unique opportunity to evaluate bacteria levels during prolonged periods without recreational activity. Samples of water, sediment, and seaweed were measured for enterococci by culture and qPCR, in addition to microbial source tracking by qPCR of fecal bacteria markers from humans, dogs, and birds. During periods of elevated enterococci levels in water, these analyses were supplemented by chemical source tracking of human-associated excretion markers (caffeine, sucralose, acetaminophen, ibuprofen, and naproxen). Results show that enterococci with elevated levels of human fecal markers persist in the seaweed and sediment and are the likely contributor to elevated levels of bacteria to the nearshore waters. During the shutdown period the elevated levels of enterococci in the sediment were isolated to the seaweed stranding areas. During periods when the beaches were open, enterococci were distributed more uniformly in sediment across the supratidal and intertidal zones. It is hypothesized from this study that human foot traffic may be responsible for the spread of enterococci throughout these areas. Overall, this study found high levels of enterococci in decomposing seaweed supporting the hypothesis that decomposing seaweed provides an additional substrate for enterococci to grow. [Display omitted] •Elevated levels of enterococci were associated with decomposition of seaweed.•Human-specific Bacteroides marker (HF183) was higher in water, sediment and seaweed.•During the shutdown, high enterococci in sediment was confined to integration zone•When beach was open, enterococci was distributed throughout sediment zones.•Spread of sediment enterococci may be due to human foot traffic through seaweed zone

Trends in regional enterococci levels at marine beaches and correlations with environmental, global oceanic changes, community populations, and wastewater infrastructure

An increase in the number of advisories issued for recreational beaches across south Florida (due to the fecal indicator bacteria, enterococci) has been observed in recent years. To evaluate the possible reasons for this increase, we reviewed weekly monitoring data for 18 beaches in Miami-Dade County, Florida, for the years 2000–2019. Our objective was to evaluate this dataset for trends in enterococci levels and correlations with various factors that might have influenced enterococci levels at these beaches. For statistical analyses, we divided the 20-year period of record into 5-year increments (2000–2004, 2005–2009, 2010–2014, and 2015–2019). The Wilcoxon rank sum test was used to identify statistically significant differences between the geometric mean of different periods. When all 18 beaches were collectively considered, a significant increase (p = 0.03) in enterococci was observed during 2015–2019, compared to the prior 15-year period of record. To better understand the potential causes for this increase, correlations were evaluated with environmental parameters (rainfall, air temperature, and water temperature), global oceanic changes (sea level and Sargassum), community populations (county population estimates and beach visitation numbers), and wastewater infrastructure (sewage effluent flow rates to ocean outfalls and deep well injection). In relation to the enterococci geometric mean, the correlation with Sargassum was statistically significant at a 95% confidence interval (p = 0.035). Population (p = 0.078), air temperature (p = 0.092), and sea level (p = 0.098) were statistically significant at 90% confidence intervals. Rainfall, water temperature, beach visitation numbers, and sewage effluent flow rates via deep well injection had positive correlations but were not significant factors. Sewage effluent flow rates to ocean outfalls had a negative correlation. [Display omitted] •Enterococci bacteria were evaluated for trends over 20 years.•Levels were observed to increase systematically across 18 beaches.•The increase was observed during the last five years.•The highest correlation was with Sargassum volumes.•Significant correlations were observed with sea level, population, and air temperature

Viable cell sorting of dinoflagellates by multiparametric flow cytometry

Sinigalliano C.D., Winshell J., Guerrero M.A., Scorzetti G., Fell J.W., Eaton R.W., Brand L. and Rein K.S. 2009. Viable cell sorting of dinoflagellates by multiparametric flow cytometry. Phycologia 48: 249-257. DOI: 10.2216/08-51.1. Electronic cell sorting for isolation and culture of dinoflagellates and other marine eukaryotic phytoplankton was compared to the traditional method of manually picking cells using a micropipette. Trauma to electronically sorted cells was not a limiting factor, as fragile dinoflagellates, such as Karenia brevis (Dinophyceae), survived electronic cell sorting to yield viable cells. The rate of successful isolation of large-scale (> 4 litres) cultures was higher for manual picking than for electronic cell sorting (2% vs 0.5%, respectively). However, manual picking of cells is more labor intensive and time consuming. Most manually isolated cells required repicking, as the cultures were determined not to be unialgal after a single round of isolation; whereas, no cultures obtained in this study from electronic single-cell sorting required resorting. A broad flow cytometric gating logic was employed to enhance species diversity. The percentages of unique genotypes produced by manual picking or electronic cell sorting were similar (57% vs 54%, respectively), and each approach produced a variety of dinoflagellate or raphidophyte genera. Alternatively, a highly restrictive gating logic was successfully used to target K. brevis from a natural bloom sample. Direct electronic single-cell sorting was more successful than utilizing a pre-enrichment sort followed by electronic single-cell sorting. The appropriate recovery medium may enhance the rate of successful isolations. Seventy percent of isolated cells were recovered in a new medium (RE) reported here, which was optimized for axenic dinoflagellate cultures. The greatest limiting factor to the throughput of electronic cell sorting is the need for manual postsort culture maintenance and assessment of the large number of isolated cells. However, when combined with newly developed automated methods for growth screening, electronic single-cell sorting has the potential to accelerate the discovery of new algal strains