Detection and modeling of Staphylococcus aureus and fecal bacteria in Hawaiian coastal waters and sands.

Microbial pollution of recreational waters leads to millions of skin, respiratory, and gastrointestinal illnesses globally. Fecal indicator bacteria (FIB) are monitored to assess recreational waters but may not reflect the presence of Staphylococcus aureus, a global leader in bacterial fatalities. Since many community-acquired S. aureus skin infections are associated with high recreational water usage, this study measured and modeled S. aureus, methicillin-resistant S. aureus (MRSA), and FIB (Enterococcus spp., Clostridium perfringens) concentrations in seawater and sand at six beaches in Hilo, Hawai'i, USA, over 37 sample dates from July 2016 to February 2019 using culturing techniques. Generalized linear models predicted bacterial concentrations with physicochemical and environmental data. Beach visitors were also surveyed on their preferred activities. S. aureus and FIB concentrations were roughly 6-78 times higher at beaches with freshwater discharge than at those without. Seawater concentrations of Enterococcus spp. were positively associated with MRSA but not S. aureus. Elevated S. aureus was associated with lower tidal heights, higher freshwater discharge, onsite sewage disposal system density, and turbidity. Regular monitoring of beaches with freshwater input, utilizing real-time water quality measurements with robust modeling techniques, and raising awareness among recreational water users may mitigate exposure to S. aureus, MRSA, and FIB. PRACTITIONER POINTS: Staphylococcus aureus and fecal bacteria concentrations were higher in seawater and sand at beaches with freshwater discharge. In seawater, Enterococcus spp. positively correlated with MRSA, but not S. aureus. Freshwater discharge, OSDS density, water turbidity, and tides significantly predicted bacterial concentrations in seawater and sand. Predictive bacterial models based upon physicochemical and environmental data developed in this study are readily available for user-friendly application.

Correction: Spatial variation and antecedent sea surface temperature conditions influence Hawaiian intertidal community structure.

[This corrects the article DOI: 10.1371/journal.pone.0286136.].

Detection and impact of sewage pollution on South Kohala's coral reefs, Hawai'i.

Sewage pollution from on-site sewage disposal systems and injection wells is impacting coral reefs worldwide. Our study documented the presence and impact of sewage on South Kohala's coral reefs, on Hawai'i Island, through benthic water quality and macroalgal sampling (fecal indicator bacteria, nutrients, δ15N macroalgal tissue), NO3- stable isotope mixing models, water motion measurements, and coral reef surveys. Sewage pollution was moderate on the offshore reef from benthic seeps, and water motion mixed and diluted it across the benthos. These conditions likely contribute to the dominance of turf algae cover, and the severity and prevalence of growth anomalies and algal overgrowth on corals. Use of multiple indicators and studying water motion was necessary to assess sewage pollution and identify environmental drivers associated with impaired coral health conditions. Methods used in this study can be utilized by natural resource managers to identify and reduce anthropogenic stressors to coral reefs.

A comparison of soil Staphylococcus aureus and fecal indicator bacteria concentrations across land uses in a Hawaiian watershed.

Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and fecal indicator bacteria (FIB; Enterococcus spp., Clostridium perfringens) concentrations increase in Hawaiian streams and estuaries following storms and pose a health threat to recreational water users. To reduce this risk, watershed bacteria sources need to be identified for management actions. This study's goals were to identify soil bacteria sources among different land uses and to determine if their concentrations were associated with different soil properties. Soil samples were collected three times on 24 d between October 2017 and November 2018 at urban, agriculture, and native-forest land uses in the Hilo Bay watershed, Hawai'i Island, Hawai'i. Soil bacteria concentrations were quantified using culturing techniques with selective media. Staphylococcus aureus, MRSA, and FIB were present in soil from all land uses. Bacteria concentrations were highest in urban soils and lowest in native-forest soils, with up to three orders of magnitude differences among land uses. Staphylococcus aureus, MRSA, and FIB soil concentrations were positively correlated with each other and with soil temperature and pH, but inversely correlated with soil moisture and organic matter content. Our results demonstrate that soils are a watershed bacteria source and that some soil properties affect their concentrations. Identifying these sources is critical for implementing management actions to reduce pathogen loads to estuaries and transmission to recreational water users.

Spatial distribution and sources of nutrients at two coastal developments in South Kohala, Hawai'i.

Nutrient sources to coastal waters with coral reefs are not well-characterized. This study documented spatial distributions of nutrients within coastal waters along two developments with coral reefs, and identified nutrient sources through nutrient mixing plots, δ15N measurements in macroalgal tissue, and NO3- stable isotope mixing models. Nutrients decreased from fresh groundwaters to offshore waters, with some surface waters higher in concentrations than benthic ones. Conservative and non-conservative mixing between fresh and ocean waters occurred, the latter suggestive of local nutrient sources and biological removal. δ15N in macroalgal tissue and NO3- concurred that fresh groundwater, ocean water, and fertilizers were dominant nutrient sources. Benthic salinity and NO3- + NO2- concentrations illustrated that submarine groundwater discharge delivered nutrients to reefs in pulses ranging from minutes to days. Information generated from this study is imperative for developing management actions to improve water quality and make coral reefs more resilient to stressors.

Environmental Surveillance and Characterization of Antibiotic Resistant Staphylococcus aureus at Coastal Beaches and Rivers on the Island of Hawai'i.

Staphylococcus aureus are human facultative pathogenic bacteria and can be found as contaminants in the environment. The aim of our study was to determine whether methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) isolated from coastal beach and river waters, anchialine pools, sand, and wastewater on the island of Hawai'i, Hawai'i, are a potential health risk. Samples were collected from three regions on Hawai'i Island from July to December 2020 during the COVID-19 pandemic and were characterized using whole-genome sequencing (WGS). From WGS data, multilocus sequence typing (MLST), SCCmec type, antimicrobial resistance genes, virulence factors, and plasmids were identified. Of the 361 samples, 98.1% were positive for Staphylococcus spp. and 7.2% were S. aureus positive (n = 26); nine MRSA and 27 MSSA strains were characterized; multiple isolates were chosen from the same sample in two sand and seven coastal beach water samples. The nine MRSA isolates were multi-drug resistant (6-9 genes) sequence type (ST) 8, clonal complex (CC) 8, SCCmec type IVa (USA300 clone), and were clonally related (0-16 SNP differences), and carried 16-19 virulence factors. The 27 MSSA isolates were grouped into eight CCs and 12 STs. Seventy-eight percent of the MSSA isolates carried 1-5 different antibiotic resistance genes and carried 5-19 virulence factors. We found S. aureus in coastal beach and river waters, anchialine pools, and sand at locations with limited human activity on the island of Hawai'i. This may be a public health hazard.

Spatial distribution of sewage pollution on a Hawaiian coral reef.

While sewage pollution is contributing to the global decline of coral reefs, its offshore extent and direct reef impacts from water column mixing and benthic seeps are poorly documented. We addressed this knowledge gap on a Hawaiian coral reef using sewage indicator and benthic cover measurements, macroalgal bioassays, and a pollution scoring tool. Fecal indicator bacteria (FIB) and nutrient concentrations were spatially variable in surface and benthic waters, with shoreline values being highest. Shoreline macroalgae δ15N and %N indicated high nitrogen loads containing sewage, while offshore surface and benthic values suggested lower nitrogen loads from environmental sources. Coral cover was negatively correlated with FIB, macroalgal δ15N, and nutrient concentrations. Benthic salinity and temperature measurements detected daily tidal groundwater pulses which may explain these associations. While pollution scores revealed that sewage was largely concentrated along the shoreline, results showed some reached the reef and may be contributing to its declining condition.

A multi-indicator approach for identifying shoreline sewage pollution hotspots adjacent to coral reefs.

Sewage pollution is contributing to the global decline of coral reefs. Identifying locations where it is entering waters near reefs is therefore a management priority. Our study documented shoreline sewage pollution hotspots in a coastal community with a fringing coral reef (Puako, Hawai'i) using dye tracer studies, sewage indicator measurements, and a pollution scoring tool. Sewage reached shoreline waters within 9 h to 3 d. Fecal indicator bacteria concentrations were high and variable, and δ15N macroalgal values were indicative of sewage at many stations. Shoreline nutrient concentrations were two times higher than those in upland groundwater. Pollution hotspots were identified with a scoring tool using three sewage indicators. It confirmed known locations of sewage pollution from dye tracer studies. Our study highlights the need for a multi-indicator approach and scoring tool to identify sewage pollution hotspots. This approach will be useful for other coastal communities grappling with sewage pollution.

Identifying nitrogen sources to thermal tide pools in Kapoho, Hawai'i, U.S.A, using a multi-stable isotope approach.

Nitrogen (N) enrichment often results in coastal eutrophication, even in remote areas like Hawai'i. Therefore, determining N sources to coastal waters is important for their management. This study identified N sources to tide pools in Kapoho, Hawai'i, and determined their relative importance using three stable isotopes (δ(15)N, δ(18)O, δ(11)B). Surface waters and macroalgal tissues were collected along 100-m onshore-offshore transects in areas of high groundwater input for three months at low tide. Water samples from possible N sources were also collected. Mixing model output, along with macroalgal δ(15)N values, indicated that agriculture soil (34%) was the largest anthropogenic N source followed by sewage (27%). These findings suggest that more effective fertilizer application techniques and upgrading sewage treatment systems can minimize N leaching into groundwater. Overall, our multi-stable isotope approach for identifying N sources was successful and may be useful in other coastal waters.

Differences in ecological structure, function, and native species abundance between native and invaded Hawaiian streams.

Poeciliids, one of the most invasive species worldwide, are found on almost every continent and have been identified as an "invasive species of concern" in the United States, New Zealand, and Australia. Despite their global prevalence, few studies have quantified their impacts on tropical stream ecosystem structure, function, and biodiversity. Utilizing Hawaiian streams as model ecosystems, we documented how ecological structure, function, and native species abundance differed between poeciliid-free and poeciliid-invaded tropical streams. Stream nutrient yields, benthic biofilm biomass, densities of macroinvertebrates and fish, and community structures of benthic algae, macroinvertebrates, and fish were compared between streams with and without established poeciliid populations on the island of Hawai'i, Hawaii, USA. Sum nitrate (sigmaNO3(-) = NO3(-) + NO2(-)), total nitrogen, and total organic carbon yields were eight times, six times, and five times higher, respectively, in poeciliid streams than in poeciliid-free streams. Benthic biofilm ash-free dry mass was 1.5x higher in poeciliid streams than in poeciliid-free streams. Percentage contributions of chironomids and hydroptilid caddisflies to macroinvertebrate densities were lower in poeciliid streams compared to poeciliid-free streams, while percentage contributions of Cheumatopsyche analis caddisflies, Dugesia sp. flatworms, and oligochaetes were higher. Additionally, mean densities of native gobies were two times lower in poeciliid streams than in poeciliid-free ones, with poeciliid densities being approximately eight times higher than native fish densities. Our results, coupled with the wide distribution of invasive poeciliids across Hawaii and elsewhere in the tropics, suggest that poeciliids may negatively impact the ecosystem structure, function, and native species abundance of tropical streams they invade. This underscores the need for increased public awareness to prevent future introductions and for developing and implementing effective eradication and restoration strategies.