Dynamics of Marine Bacterial Community Diversity of the Coastal Waters of the Reefs, Inlets, and Wastewater Outfalls of Southeast Florida

Coastal waters adjacent to populated southeast Florida possess different habitats (reefs, oceanic inlets, sewage outfalls) that may affect the composition of their inherent microbiomes. To determine variation according to site, season, and depth, over the course of 1 year, we characterized the bacterioplankton communities within 38 nearshore seawater samples derived from the Florida Area Coastal Environment (FACE) water quality survey. Six distinct coastal locales were profiled – the Port Everglades and Hillsboro Inlets, Hollywood and Broward wastewater outfalls, and associated reef sites using culture-independent, high-throughput pyrosequencing of the 16S rRNA V4 region. More than 227,000 sequences helped describe longitudinal taxonomic profiles of marine bacteria and archaea. There were 4447 unique operational taxonomic units (OTUs) identified with a mean OTU count of 5986 OTUs across all sites. Bacterial taxa varied significantly by season and by site using weighted and unweighted Unifrac, but depth was only supported by weighted Unifrac, suggesting a change due to presence/absence of certain OTUs. Abundant microbial taxa across all samples included Synechococcus, Pelagibacteraceae, Bacteroidetes, and various Proteobacteria. Unifrac analysis confirmed significant differences at inlet sites relative to reef and outfalls. Inlet-based bacterioplankton significantly differed in greater abundances of Rhodobacteraceae and Cryomorphaceae, and depletion of SAR406 sequences. This study also found higher counts of Firmicutes, Chloroflexi, and wastewater associated SBR1093 bacteria at the outfall and reef sites compared to inlet sites. This study profiles local bacterioplankton populations in a much broader context, beyond culturing and quantitative PCR, and expands upon the work completed by the National Oceanic and Atmospheric Administration FACE program

Boynton-Delray Coastal Water Quality Monitoring Program

This report discusses a sequence of six cruises in the vicinity of the Boynton-Delray (South Central) treated-wastewater plant outfall plume (26°27\u2743 N, 80°2\u2732 W), the Boynton Inlet (26°32\u2743 N, 80°2\u2730 W), and the Lake Worth Lagoon, Palm Beach County, Florida. The sampling cruises took place on June 5-6, 2007; August 28-29, 2007; October 18-19, 2007; February 14 and 18, 2008; May 19-20, 2008; and July 11-13, 2008. Water was sampled at 18 locations at the surface, middle, and near the seafloor (where there was sufficient depth) for a total of 45 samples; these samples were analyzed for a variety of nutrients and related parameters. The water sampling unit contained a conductivity-temperature-depth (CTD) instrument from which data were obtained at each sampling site. Synchronal ocean current data were measured by a nearby acoustic Doppler current profiler (ADCP) instrument. The inlet measurements were consistently lower in salinity and more acidic (lower in pH) than the coastal ocean and were warmer during the May and, especially, during the February cruises. For most analytes, viz., nitrite+nitrate (N+N), total suspended solids (TSS), chlorophyll-a, silica (Si), and total dissolved nitrogen (TDN), the lagoon concentrations were significantly higher than the coastal ocean; the inlet concentrations appeared to be consistent with lagoon water with partial mixing with the coastal ocean, as expected. Estimates of the nutrient flux to the coastal ocean were computed: approximately 1,500 kg of dissolved nitrogen (N), 2,350 kg of silicate (Si), 33 kg of orthophosphate (P), and 59 kg of ammonium (NH4) per day were delivered to the coastal ocean through the inlet. The outfall boil at South Central outfall (the smallest in volume of the six outfalls in southeast Florida) is only visible under ideal conditions. In the six cruises described in this document, the outfall boil could be found in only one cruise (August 28-29, 2007). Elevated concentrations of nutrients (N+N, P, Si, and P) at the outfall vicinity were measured, and these concentrations decreased rapidly away from the outfall for most analytes, to become undistinguished from the background within 3 km or less. Not finding the boil, however, in five of six cruises meant that the waters with the highest concentrations were probably missed. When the boil was sampled in August 2007, N+N, P, and total dissolved phosphorus (TDP) concentrations at the boil were roughly the same as from the inlet. For other analytes (chlorophyll-a, TSS, Si, and dissolved organic carbon [DOC]), the concentrations at or near the outfall were significantly less than those from the lagoon and inlet on most of the cruises. The coastal ocean appeared to be significantly impacted by the Boynton Inlet and less so from the inlet. A suggestion of a source to the south was seen in some analytes. Measurements from the Gulf Stream Reef area were the lowest in the study, and may provide “background” concentrations for this region. As expected, the coastal ocean was warmer and more stratified in the summer compared to the winter, e.g., whereas no thermocline was noted in the CTD data from February 2007, a strong thermocline was observed in most casts during July 2008. In certain cases (e.g., N+N in June 2007, pH in July 2008), an increase in the concentration (decrease for pH) from north to south implied a source from the south, e.g., the Boca Raton Inlet or Boca Raton outfall

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

Beaches and Coastal Environments

This chapter summarizes the rationale for using microbial source tracking (MST) methods at beach sites and coastal water bodies (Sect. 20.1), as MST methods are especially useful for evaluating waters impacted by nonpoint sources of pollution. This chapter also describes the most common traditional and alternative MST markers used at beach sites (Sect. 20.2). Two case studies are presented (Sect. 20.3) that describe the use of both biological/chemical MST methods and physical MST methods for identifying sources of microbes at two marine beach sites in USA, one located on the west coast (California) and the other located on the east coast (Florida). The chapter closes with discussion and recommendations concerning the utility and application of MST tools at beach sites impacted by nonpoint-source pollution (Sect. 20.4). Although this chapter focuses on marine beaches, an incredible wealth of MST data has been gathered at freshwater beaches (Byappanahalli et al. 2006; Harwood et al. 2005; Jenkins et al. 2005; Scott et al. 2002; Stapleton et al. 2009; Whitman and Nevers 2003; Whitman et al. 2004), and a comprehensive review of beach studies merits the inclusion of MST work within freshwater systems. The use of MST in freshwater systems is further discussed in Chaps. 18, 19, and 21

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