Sediment metagenomics reveals the impacts of poultry industry wastewater on antibiotic resistance and nitrogen cycling genes in tidal creek ecosystems

The intensification of the poultry industry may lead to the increased spread of antibiotic resistance genes (ARGs) in the environment. However, the impacts of wastewater discharge from poultry processing plants on the sediment resistome are relatively unexplored. Furthermore, its relationships with important biogeochemical pathways, such as the N cycle, are virtually unknown. The overall objective of this study was to examine the abundance and diversity of antibiotic resistance and N cycling genes in sediment microbial communities impacted by poultry industry wastewater. We performed a metagenomic investigation of sediments in an impacted and a reference tidal creek. We also quantified the abundance of the clinical class 1 integron-integrase gene (intI1) through qPCR as a secondary marker of anthropogenic contamination. Abundance and diversity of ARGs were substantially higher in the impacted tidal creek, especially near the wastewater discharge. Abundances of ARGs conferring resistance to macrolides, tetracyclines, and streptogramins were also higher in the impacted creek than the reference creek. From the N cycling genes detected in the metagenomes, nrfA, the genetic marker for dissimilatory nitrate reduction to ammonia (DNRA), had the strongest positive relationship with the total abundance of ARGs, which may indicate an increased potential of eutrophication in ARG-impacted ecosystems due to nitrogen retention. This study demonstrates that wastewater discharge from a poultry processing plant can increase the spread of ARGs, which may result in negative impacts on ecosystem health

The role of oyster in nitrous oxide emissions from oyster reefs

Oyster microbiomes may play a significant role in the biogeochemical N cycle and N2O emissions in estuarine and coastal ecosystems. Rates of denitrification and N2O production were measured from live oysters and oyster shells collected at a coastal lagoon and a tidal estuary in the Chesapeake Bay. Live oysters had the highest rates of denitrification and N2O production in both study sites. This indicates that oysters are an import N remover and a N2O source in coastal ecosystems

Linking DNRA community structure and activity in a shallow lagoonal estuarine system

Dissimilatory nitrate reduction to ammonium (DNRA) and denitrification are two nitrate respiration pathways in the microbial nitrogen cycle. Diversity and abundance of denitrifying bacteria have been extensively examined in various ecosystems. However, studies on DNRA bacterial diversity are limited, and the linkage between the structure and activity of DNRA communities has yet to be discovered. We examined the composition, diversity, abundance, and activities of DNRA communities at five sites along a salinity gradient in the New River Estuary, North Carolina, USA, a shallow temporal/lagoonal estuarine system. Sediment slurry incubation experiments with N-15-nitrate were conducted to measure potential DNRA rates, while the abundance of DNRA communities was calculated using quantitative PCR of nrfA genes encoding cytochrome C nitrite reductase, commonly found in DNRA bacteria. A pyrosequencing method targeting nrfA genes was developed using an Ion Torrent sequencer to examine the diversity and composition of DNRA communities within the estuarine sediment community. We found higher levels of nrfA gene abundance and DNRA activities in sediments with higher percent organic content. Pyrosequencing analysis of nrfA genes revealed spatial variation of DNRA communities along the salinity gradient of the New River Estuary. Percent abundance of dominant populations was found to have significant influence on overall activities of DNRA communities. Abundance of dominant DNRA bacteria and organic carbon availability are important regulators of DNRA activities in the eutrophic New River Estuary

Reconstructing primary production in a changing estuary: A mass balance modeling approach

Estuarine primary production (PP) is a critical rate process for understanding ecosystem function and response to environmental change. PP is fundamentally linked to estuarine eutrophication, and as such should respond to ongoing efforts to reduce nutrient inputs to estuaries globally. However, concurrent changes including warming, altered hydrology, reduced input of sediments, and emergence of harmful algal blooms (HABs) could interact with nutrient management to produce unexpected changes in PP. Despite its fundamental importance, estuarine PP is rarely measured. We reconstructed PP in the York River Estuary with a novel mass balance model based on dissolved inorganic nitrogen (DIN) for the period 1994–2018. Modeled PP compared well to previous estimates and demonstrated a long-term increase and down-estuary shift over the study period. This increase occurred despite reductions in discharge, flushing time, DIN loading, and DIN standing stock over the same period. Increased PP corresponded to increased water temperature, decreased turbidity and light attenuation, and increased photic depth and assimilation ratio, suggesting that phytoplankton in the York River Estuary have become more efficient at converting nutrients into biomass primarily due to a release from light limitation. The increase in PP also coincided with the increasing occurrence of late summer HABs in the lower York River Estuary, including the emergence of a second bloom-forming dinoflagellate in 2007. Results demonstrate how changes concurrent with nutrient management could alter expected system responses and illustrate the utility of the mass balance approach for estimating critical rate processes like PP in the absence of observations

The feeding ecology and microbiome of the pteropod Limacina helicina antarctica

The pteropod (pelagic snail) Limacina helicina antarctica is a dominant grazer alongthe Western Antarctic Peninsula (WAP) and plays an important role in regional food web dynamics and biogeochemical cycling. For the first time, we examined the gut microbiome and feeding ecology of L. h. antarctica based on 16S and 18S rRNA gene sequences of gut contents in the WAP during austral summer. Eukaryotic gut contents of L. h. antarctica indicate that this species predominantly feeds on diatoms and dinoflagellates, supplementing its diet with ciliates and foraminifera. Mollicutes bacteria were a consistent component of the gut microbiome. Determining the gut microbiome and feeding ecology of L. h.antarctica aids in identifying the underlying mechanisms controlling pteropod abundance and distribution in a region of rapid environmental change

Contrasting controls on seasonal and spatial distribution of marine cable bacteria (Candidatus Electrothrix) and Beggiatoaceae in seasonally hypoxic Chesapeake Bay

Marine cable bacteria (Candidatus Electrothrix) and large colorless sulfur-oxidizing bacteria (e.g., Beggiatoaceae) are widespread thiotrophs in coastal environments but may exert different influences on biogeochemical cycling. Yet, the factors governing their niche partitioning remain poorly understood. To map their distribution and evaluate their growth constraints in a natural setting, we examined surface sediments across seasons at two sites with contrasting levels of seasonal oxygen depletion in Chesapeake Bay using microscopy coupled with 16S rRNA gene amplicon sequencing and biogeochemical characterization. We found that cable bacteria, dominated by a single phylotype closely affiliated to Candidatus Electrothrix communis, flourished during winter and spring at a central channel site which experiences summer anoxia. Here, cable bacteria density was positively correlated with surface sediment chlorophyll, a proxy of phytodetritus sedimentation. Cable bacteria were also present with a lower areal density at an adjacent shoal site which supports bioturbating macrofauna. Beggiatoaceae were more abundant at this site, where their biomass was positively correlated with sediment respiration, but additionally potentially inhibited by sulfide accumulation which was evident during one summer. A springtime phytodetritus sedimentation event was associated with a proliferation of Beggiatoaceae and multiple Candidatus Electrothrix phylotypes, with cable bacteria reaching 1000 m length cm−2. These observations indicate the potential impact of a spring bloom in driving a hot moment of cryptic sulfur cycling. Our results suggest complex interactions between benthic thiotroph populations, with bioturbation and seasonal oscillations in bottom water dissolved oxygen, sediment sulfide, and organic matter influx as important drivers of their distribution

Antibiotic Effects on Microbial Communities Responsible for Greenhouse Gas Emissions

Nitrous oxide (N2O) is a powerful greenhouse gas generated by nitrification and denitrification. The goal of this project is to examine the effects of antibiotics on microbial communities responsible for N2O emissions from terrestrial and aquatic ecosystems. We conducted laboratory and mesocosm experiments in soil samples. Higher N2O production was observed in soils exposed to tetracycline. This was associated with reduction of bacterial denitrifiers abundance and enhanced fungal abundance

Trace Metal Availability Affects Greenhouse Gas Emissions and Microbial Functional Group Abundance in Freshwater Wetland Sediments

We investigated the effects of trace metal additions on microbial nitrogen (N) and carbon (C) cycling using freshwater wetland sediment microcosms amended with micromolar concentrations of copper (Cu), molybdenum (Mo), iron (Fe), and all combinations thereof. In addition to monitoring inorganic N transformations (NO3–, NO2–, N2O, NH4+) and carbon mineralization (CO2, CH4), we tracked changes in functional gene abundance associated with denitrification (nirS, nirK, nosZ), dissimilatory nitrate reduction to ammonium (DNRA; nrfA), and methanogenesis (mcrA). With regards to N cycling, greater availability of Cu led to more complete denitrification (i.e., less N2O accumulation) and a higher abundance of the nirK and nosZ genes, which encode for Cu-dependent reductases. In contrast, we found sparse biochemical evidence of DNRA activity and no consistent effect of the trace metal additions on nrfA gene abundance. With regards to C mineralization, CO2 production was unaffected, but the amendments stimulated net CH4 production and Mo additions led to increased mcrA gene abundance. These findings demonstrate that trace metal effects on sediment microbial physiology can impact community-level function. We observed direct and indirect effects on both N and C biogeochemistry that resulted in increased production of greenhouse gasses, which may have been mediated through the documented changes in microbial community composition and shifts in functional group abundance. Overall, this work supports a more nuanced consideration of metal effects on environmental microbial communities that recognizes the key role that metal limitation plays in microbial physiology