Geographic variability in amoeboid protists and other microbial groups in the water column of the lower Hudson River Estuary.

In comparison to other groups of planktonic microorganisms, relatively little is known about the role of amoeboid protists (amebas) in planktonic ecosystems. This study describes the !rst geographic survey of the abundance and biomass of amebas in an estuarine water column. Samples collected in the lower Hudson River Estuary were used to investigate relationships between ameba abundance and biomass and hydrographic variables (temperature, salinity, and turbidity), water depth (surface and near bottom), distance from mid-channel to shore, phytoplankton biomass (chlorophyll fluorescence) and the occurrence of other heterotrophic microbial groups (heterotrophic bacteria, nano flagellates, and ciliates) in the plankton. Although salinity increased signi!cantly towards the mouth of the estuary, there were no signi!cant differences in the abundance or biomass of any microbial group in surface samples collected at three stations separated by 44 km along the estuary's mid-channel. Peak biomass values for all microbial groups were found at the station closest to shore, however, cross-channel trends in microbial abundance and biomass were not statistically significant. Although ameba abundance and biomass in most samples were low compared to other microbial groups, clear patterns in ameba distribution were nevertheless found. Unlike other microbial groups examined, ameba numbers and biomass greatly increased in near bottom water compared to surface samples. Ameba abundance and biomass (in surface samples) were also strongly related to increasing turbidity. The different relationships of ameba abundance and biomass with turbidity suggest a rising contribution of large amebas in microbial communities of the Hudson estuary when turbidity increases. These results, emphasizing the importance of particle concentration as attachment and feeding surfaces for amebas, will help identify the environmental conditions when amebas are most likely to contribute significantly to estuarine bacterivory and C-flux

Particle Association of Enterococcus and Total Bacteria in the Lower Hudson River Estuary, USA

Bacterial particle association has important consequences for water-quality monitoring and modeling. Parti-cle association can change vertical and horizontal transport of bacterial cells, as well as patterns of persis-tence and production. In this study, the abundance and particle association of total bacteria and the fe-cal-indicator, Enterococcus, were quantified between June and October 2008 in the lower Hudson River Es-tuary (HRE). Twelve sites were sampled, including mid-channel, near shore, and tributary habitats, plus a sewage outfall. Total bacterial cell counts averaged 9.2 × 109 ± 6.4 × 109 cell·l–1 (1 standard deviation), com-parable to previous sampling in the HRE. Unlike earlier studies, bacterial abundance did not change consis-tently along the north/south estuarine salinity gradient. Enterococcus concentrations were highly variable, but mid-channel stations had significantly lower values than other habitat categories. Counts of total bacteria and Enterococci were both correlated with turbidity, which was also significantly lower at mid-channel sta-tions. A larger fraction of Enterococci were associated with particles (52.9% ± 20.9%, 1 standard deviation) than in the pool of total bacteria (23.8% ± 15.0%). This high frequency of particle association, relative to total bacteria, could cause Enterococci to be preferentially retained near input sources because of enhanced deposition to bottom sediments, where they would be available for later resuspension. In turn, retention and resuspension in nearshore environments may explain the observed cross-channel variability of turbidity and Enterococci. Assessments and predictive models of estuarine water quality may be improved by incorporat-ing cross-channel variability and the effects of particle association on key indicators

Particle association of Enterococcus sp. increases growth rates and simulated persistence in water columns of varying light attenuation and turbulent diffusivity

Predicting water quality and the human health risks associated with sewage-derived microbes requires understanding the fate and transport of these contaminants. Sewage-derived pathogen risks are typically assessed and monitored by measuring concentrations of fecal indicating bacteria (FIB), like Enterococcus sp. Previous research demonstrated that a high fraction of FIB is particle-associated, which can alter FIB dynamics within secondary water bodies. In this study, we experimentally quantified the effect of par- ticle association on dark, temperature- and light-dependent growth and sinking rates of enterococci. Particle association significantly increased dark growth rates, light-dependent growth rates (i.e. decreased mortality), and sinking rates, relative to free-living enterococci. Simulations using a novel, 1- dimensional model parameterized by these rates indicate greater persistence (T90) for particle-associated enterococci in water bodies across a wide range of diffuse attenuation coefficients of light (Kd) and turbulent diffusivity (D) values. In addition, persistence of both fractions increased in simulated turbid and turbulent waters, compared to clear and/or quiescent conditions. Simulated persistence of both fractions also increased when enterococci discharges occurred later in a diel cycle (towards sunset, as opposed to sunrise), especially for the free-living population, because later discharges under our model conditions allowed both fractions to mix deeper before inactivation via sunlight. Model sensitivity testing revealed that T90 variability was greatest when dark growth rates were altered, suggesting that future empirical studies should focus on quantifying these rates for free-living and particle-associated sewage- derived microbes. Despite greater sensitivity of T90 to variability in dark growth rates, omitting light- dependent growth rates from simulations dramatically influenced T90 values. Our results demonstrate that particle association can increase enterococci persistence in receiving waters and highlight the importance of incorporating particle association in future water quality models

Coupling of fog and marine microbial content in the near-shore coastal environment

Microbes in the atmosphere (microbial aerosols) play an important role in climate and provide an ecological and biogeochemical connection between oceanic, atmospheric, and terrestrial environments. However, the sources and environmental factors controlling the concentration, diversity, transport, and viability of microbial aerosols are poorly understood. This study examined culturable microbial aerosols from a coastal environment in Maine (USA) and determined the effect of onshore wind speed and fog presence on deposition rate, source, and community composition. During fog events with low onshore winds (<2 m s−1) the near-shore deposition of microbial aerosols (microbial fallout) decreased with increasing wind speeds, whereas microbial fallout rates under clear conditions and comparable low wind speeds showed no wind speed dependence. Mean aerosol particle size also increased with onshore wind speed when fog was present, indicating increased shoreward transport of larger aerosol particles. 16S rRNA sequencing of culturable ocean surface bacteria and microbial aerosols deposited onshore resulted in the detection of 31 bacterial genera, with 5 dominant genera (Vibrio, Bacillus, Pseudoalteromonas, Psychrobacter, Salinibacterium) making up 66 % of all sequences. The sequence library from microbial aerosol isolates, as with libraries found in other coastal/marine aerosol studies, was dominated at the phylum level by Proteobacteria, with additional representation from Firmicutes, Actinobacteria and Bacteroidetes. Seventy-five percent of the culturable microbial aerosols falling out under foggy conditions were most similar to GenBank-published sequences detected in marine environments. Using a 97 % similarity cut-off, sequence libraries from ocean surface and fog isolates shared eight operational taxonomic units (OTU's) in total, three of which were the most dominant OTU's in the library, representing large fractions of the ocean (28 %) and fog (21 %) libraries. The fog and ocean surface libraries were significantly more similar in microbial community composition than clear (non-foggy) and ocean surface libraries, according to both Jaccard and Sorenson indices. These findings provide the first evidence of a difference in community composition and microbial culturability of aerosols associated with fog compared to clear conditions. The data support a dual role for fog in enhancing the fallout of viable microbial aerosols via increased gravitational settling rates and decreased aerosolization stress on the organisms, which may include relief from UV inactivation, desiccation, and oligotrophic microconditions. This study provides a strong case for ocean to terrestrial transport of microbes and a potential connection between water quality and air quality at coastal sites

Antibiotic resistant bacteria in the Hudson River Estuary linked to wet weather sewage contamination

Heterotrophic bacteria resistant to tetracycline and ampicillin were assessed in waterways of the New York City metropolitan area using culture-dependent approaches and 16S rRNA gene sequence analysis of resultant isolates. Resistant microbes were detected at all 10 sampling sites in monthly research cruises on the lower Hudson River Estuary (HRE), with highest concentrations detected at nearshore sites. Higher frequency sampling was conducted in Flushing Bay, to enumerate resistant microbes under both dry and wet weather conditions. Concentrations of ampicillin- and tetracycline- resistant bacteria, in paired samples, were positively correlated with one another and increased following precipitation. Counts of the fecal indicator, Enterococcus, were positively correlated with levels of resistant bacteria, suggesting a shared sewage-associated source. Analysis of 16S rRNA from isolates identified a phylogenetically diverse group of resistant bacteria, including genera containing opportunistic pathogens. The occurrence of Enterobacteriaceae, a family of enteric bacteria, was found to be significantly higher in resistant isolates compared to total heterotrophic bacteria and increased following precipitation. This study is the first to document the widespread distribution of antibiotic-resistant bacteria in the HRE and to demonstrate clearly a link between the abundance of antibiotic-resistant bacteria and levels of sewage-associated bacteria in an estuary

Diatom vertical migration within land-fast Arctic sea-ice.

Light levels inside first-year, landfast sea ice were experimentally altered by manipulating overlying snow depths. Irradiance available for ice algae growing near the ice-bottom, and under the ice, was highly dependent on snow depths ranging from 0 to >30 cm. Importantly, algal vertical distributions also changed under different irradiances. Under thick snow (low light), the majority of algae were found several cm above the ice–seawater interface, while progressively more were found nearer the interface at locations with thinner overlying snow (higher light). Short-term field experiments suggested that ice algae were able to reposition themselves within the ice column within 3 days after manipulating snow depths. Laboratory gliding rate measurements of a cultured ice diatom suggested that it is capable of daily cm-scale movement. Vertical migration may help ice diatoms balance opposing light and nutrient resource gradients, similar to strategies used by some benthic and pelagic algae. Moreover, when ice algae congregate near the ice–seawater interface, they may be especially susceptible to loss from the ice environment. Vertical repositioning in response to changing light dynamics may be a mechanism to optimize between vertically-opposing environmental factors and help explain the connection between melting snow cover and export of biomass from sea ice

Challenges to managing microbial fecal pollution in coastal environments: extra-enteric ecology and microbial exchange among water, sediment and air.

Human population growth, especially in coastal ur- ban cities, increases the potential for fecal pollution of adjacent waterways, requiring continued advances in pollution monitoring and management. Infections remain the largest health risk from contact with fecal- and sewage-polluted wa- ters, and a small number of fecal indicator bacteria (FIB) are used as primary pollution assessment tools. While FIB con- tinue to be useful tools, some of the assumptions about the behavior of FIB in the environment, and the associated path- ways for pathogen exposure, have come into question. Research into the extra-enteric ecology of these indicators has identified management-relevant complexities including particle association, prolonged environmental persistence, and multidirectional microbial exchange among water, sedi- ment, and air. These complexities provide opportunities for improving current monitoring and modeling strategies and to better understand exposure pathways for sewage-related infections

Onshore Wind Speed Modulates Microbial Aerosols along an Urban Waterfront

Wind blowing over aquatic and terrestrial surfaces produces aerosols, which include microbial aerosols. We studied the effect of onshore wind speeds on aerosol concentrations as well as total and culturable microbial aerosols (bacterial and viral) at an urban waterfront (New York, NY, USA). We used two distinct methods to characterize microbial aerosol responses to wind speed: A culture-based exposure-plate method measuring viable bacterial deposition near-shore (CFU accumulation rate); and a culture-independent aerosol sampler-based method measuring total bacterial and viral aerosols (cells m−3 air). While ambient coarse (\u3e2 µm) and fine (0.3–2 µm) aerosol particle number concentrations (regulated indicators of air quality) decreased with increasing onshore wind speeds, total and depositing culturable bacterial aerosols and total viral aerosols increased. Taxonomic identification of the 16S rDNA of bacterial aerosol isolates suggested both terrestrial and aquatic sources. Wind appears to increase microbial aerosol number concentrations in the near-shore environment by onshore transport at low wind speeds (s−1 ), and increased local production and transport of new microbial aerosols from adjacent water surfaces at higher wind speeds (\u3e4 m s−1 ). This study demonstrates a wind-modulated microbial connection between water and air in the coastal urban environment, with implications for public health management and urban microbial ecology

Sediment Resuspension and Associated Extracellular Enzyme Activities Measured ex situ: A Mechanism for Benthic-Pelagic Coupling in the Deep Gulf of Mexico.

Sediment resuspension caused by near-bed currents mediates exchange processes between the seafloor and the overlying water column, known as benthic-pelagic coupling. To investigate the effects of sediment resuspension on microbial enzyme activities in bottom waters (<500 m), we conducted onboard erosion experiments using sediment cores taken with a multi-corer from six deep-sea sites in the northern Gulf of Mexico. We then incubated the core-top water with resuspended sediments in roller tanks to simulate bottom water conditions following sediment resuspension. Bacterial cell abundance, particulate organic matter content, and potential rates of three hydrolytic enzymes (leucine aminopeptidases – PEP; β-glucosidases – GLU, lipases – LIP) were monitored during the experimentally-generated erosion events and subsequently in the roller tanks to examine whether resuspension of deep-sea sediments enhances activities of extracellular enzymes in overlying waters. Surficial sediments were resuspended at critical shear stress velocities between 1.4 and 1.7 cm s−1, which parallel bottom water currents of 28 and 34 cm s−1. Only one of our nine cores resisted experimentally generated bottom shear stresses and remained undisturbed, possibly as a result of oil residues from natural hydrocarbon seeps at the investigated site. The most notable enzymatic responses to sediment resuspension were found for LIP activities that increased in overlying waters of all eight of our resuspended cores and remained at high levels during the roller tank incubations. PEP and GLU showed orders of magnitude lower rates and more variable responses to experimentally resuspended sediments compared with LIP. We also found a disconnect between enzyme activities and bacterial cell numbers, indicating a major role of extracellular enzymes physically disconnected from microbial cells in our experiments. Our results demonstrate that sediment resuspension may promote organic matter breakdown in bottom waters by supplying extracellular enzymes without requiring a bacterial growth response. The marked increase in LIP activity suggests that resuspended enzymes may affect the degradation of petroleum hydrocarbons, including those from the natural seeps that are abundant in the investigation area

Quantifying Per-Cell Chlorophyll a in Natural Picophytoplankton Populations Using Fluorescence-Activated Cell Sorting.

Marine phytoplankton play a central role in global biogeochemical cycling, carbon export, and the overall functioning of marine ecosystems. While chlorophyll a (Chl a) is widely used as a proxy for phytoplankton biomass, identifying the proportion of Chl a attributable to different phytoplankton groups remains a major challenge in oceanography, especially for the picophytoplankton groups that often represent the majority of phytoplankton biomass in the open ocean. We describe a method for measuring picophytoplankton per-cell Chl a in field samples using fluorescence-activated cell sorting followed by solvent-based Chl a extraction and fluorescence quantification. Applying this method to surface samples from the Gulf of Mexico, we determined per-cell Chl a to be 0.24 ± 0.07, 0.6 ± 0.33, and 26.36 ± 20.9 fg Chl a cell⁻¹ for Prochlorococcus, Synechococcus, and PPE, respectively (mean ± SD). Measurements of per-cell Chl a using this method are precise to within 1.7, 2.1, and 3.1% for Prochlorococcus, Synechococcus, and PPE, respectively. We demonstrate that this approach can be used to obtain estimates of group-specific Chl a for Prochlorococcus, Synechococcus, and picophytoeukaryotes, the latter two of which cannot be captured by existing methods. We also demonstrate that measurements of per-cell Chl a made using this method in field samples are sufficiently precise to capture relationships between per-cell Chl a and cytometer red fluorescence, providing a bridge between biomass estimates from cell counts and bulk measurements of total Chl a