Deep-Sea Archaea Fix and Share Nitrogen in Methane-Consuming Microbial Consortia

Nitrogen-fixing (diazotrophic) microorganisms regulate productivity in diverse ecosystems; however, the identities of diazotrophs are unknown in many oceanic environments. Using single-cell–resolution nanometer secondary ion mass spectrometry images of ^(15)N incorporation, we showed that deep-sea anaerobic methane-oxidizing archaea fix N_2, as well as structurally similar CN^–, and share the products with sulfate-reducing bacterial symbionts. These archaeal/bacterial consortia are already recognized as the major sink of methane in benthic ecosystems, and we now identify them as a source of bioavailable nitrogen as well. The archaea maintain their methane oxidation rates while fixing N_2 but reduce their growth, probably in compensation for the energetic burden of diazotrophy. This finding extends the demonstrated lower limits of respiratory energy capable of fueling N_2 fixation and reveals a link between the global carbon, nitrogen, and sulfur cycles

Analyzing Gene Expression from Marine Microbial Communities using Environmental Transcriptomics

Analogous to metagenomics, environmental transcriptomics (metatranscriptomics) retrieves and sequences environmental mRNAs from a microbial assemblage without prior knowledge of what genes the community might be expressing. Thus it provides the most unbiased perspective on community gene expression in situ. Environmental transcriptomics protocols are technically difficult since prokaryotic mRNAs generally lack the poly(A) tails that make isolation of eukaryotic messages relatively straightforward 1 and because of the relatively short half lives of mRNAs 2. In addition, mRNAs are much less abundant than rRNAs in total RNA extracts, thus an rRNA background often overwhelms mRNA signals. However, techniques for overcoming some of these difficulties have recently been developed. A procedure for analyzing environmental transcriptomes by creating clone libraries using random primers to reverse-transcribe and amplify environmental mRNAs was recently described was successful in two different natural environments, but results were biased by selection of the random primers used to initiate cDNA synthesis 3. Advances in linear amplification of mRNA obviate the need for random primers in the amplification step and make it possible to use less starting material decreasing the collection and processing time of samples and thereby minimizing RNA degradation 4. In vitro transcription methods for amplifying mRNA involve polyadenylating the mRNA and incorporating a T7 promoter onto the 3 end of the transcript. Amplified RNA (aRNA) can then be converted to double stranded cDNA using random hexamers and directly sequenced by pyrosequencing 5. A first use of this method at Station ALOHA demonstrated its utility for characterizing microbial community gene expression 6

Algae and Clay Water Additives Differentially Impact Survival and Microbial Community Structure in Sablefish (Anoplopoma fimbria) Rearing Tanks

Algae, or “greenwater,” is a traditional water additive used in finfish aquaculture but it is associated with high costs and potentially harmful bacterial growth. “Claywater,” a mix of clay and seawater, has been explored as a replacement for greenwater. In some fish species, however, claywater reduces survival rates, but the mechanisms are not understood. A link between water additive and microbial community composition may exist. In this study, the effects of different water additives on the microbial communities of larval sablefish were studied. Three treatments were evaluated: a traditional greenwater additive, a claywater additive, and a greenwater additive switched to claywater after 1 week. Microbial communities were characterized using 16S rRNA gene sequencing, and sablefish survival and growth were recorded. Tank seawater microbial communities were significantly influenced by water additive (treatment). Sablefish microbiomes were significantly but weakly influenced by treatment, and there were time-specific differences within the claywater treatment. Sablefish, from the treatment that was switched after 1 week, maintained microbiomes that were more similar to the initial greenwater treatment. In general, sablefish were dominated by Vibrionaceae operational taxonomic units (OTUs). Variability in the sablefish microbiomes between tanks from the same treatment was high, especially in the claywater treatment, which may have confounded treatment effects. Larvae in the claywater treatment had significantly lower survival rates compared to greenwater and greenwater-claywater treatments, but no treatment effect was observed on sablefish growth (length). Overall, results suggest that claywater does not negatively impact survival or the microbial community of sablefish when preceded by 1 week of greenwater

Soil bacterial community and functional shifts in response to altered snowpack in moist acidic tundra of northern Alaska

Abstract. Soil microbial communities play a central role in the cycling of carbon (C) in Arctic tundra ecosystems, which contain a large portion of the global C pool. Climate change predictions for Arctic regions include increased temperature and precipitation (i.e. more snow), resulting in increased winter soil insulation, increased soil temperature and moisture, and shifting plant community composition. We utilized an 18-year snow fence study site designed to examine the effects of increased winter precipitation on Arctic tundra soil bacterial communities within the context of expected ecosystem response to climate change. Soil was collected from three pre-established treatment zones representing varying degrees of snow accumulation, where deep snow  ∼ 100 % and intermediate snow  ∼ 50 % increased snowpack relative to the control, and low snow ∼ 25 % decreased snowpack relative to the control. Soil physical properties (temperature, moisture, active layer thaw depth) were measured, and samples were analysed for C concentration, nitrogen (N) concentration, and pH. Soil microbial community DNA was extracted and the 16S rRNA gene was sequenced to reveal phylogenetic community differences between samples and determine how soil bacterial communities might respond (structurally and functionally) to changes in winter precipitation and soil chemistry. We analysed relative abundance changes of the six most abundant phyla (ranging from 82 to 96 % of total detected phyla per sample) and found four (Acidobacteria, Actinobacteria, Verrucomicrobia, and Chloroflexi) responded to deepened snow. All six phyla correlated with at least one of the soil chemical properties (% C, % N, C : N, pH); however, a single predictor was not identified, suggesting that each bacterial phylum responds differently to soil characteristics. Overall, bacterial community structure (beta diversity) was found to be associated with snow accumulation treatment and all soil chemical properties. Bacterial functional potential was inferred using ancestral state reconstruction to approximate functional gene abundance, revealing a decreased abundance of genes required for soil organic matter (SOM) decomposition in the organic layers of the deep snow accumulation zones. These results suggest that predicted climate change scenarios may result in altered soil bacterial community structure and function, and indicate a reduction in decomposition potential, alleviated temperature limitations on extracellular enzymatic efficiency, or both. The fate of stored C in Arctic soils ultimately depends on the balance between these mechanisms. </jats:p

Ultraviolet disinfection impacts the microbial community composition and function of treated wastewater effluent and the receiving urban river

BackgroundIn the United States, an estimated 14,748 wastewater treatment plants (WWTPs) provide wastewater collection, treatment, and disposal service to more than 230 million people. The quality of treated wastewater is often assessed by the presence or absence of fecal indicator bacteria. UV disinfection of wastewater is a common final treatment step used by many wastewater treatment plants in order to reduce fecal coliform bacteria and other pathogens; however, its potential impacts on the total effluent bacterial community are seemingly varied. This is especially important given that urban WWTPs typically return treated effluent to coastal and riverine environments and thus are a major source of microorganisms, genes, and chemical compounds to these systems. Following rainfall, stormflow conditions can result in substantial increases to effluent flow into combined systems.MethodsHere, we conducted a lab-scale UV disinfection on WWTP effluent using UV dosage of 100 mJ/cm2and monitored the active microbiome in UV-treated effluent and untreated effluent over the course of 48 h post-exposure using 16S rRNA sequencing. In addition, we simulated stormflow conditions with effluent UV-treated and untreated effluent additions to river water and compared the microbial communities to those in baseflow river water. We also tracked the functional profiles of genes involved in tetracycline resistance (tetW)and nitrification (amoA) in these microcosms using RT-qPCR.ResultsWe showed that while some organisms, such as members of the Bacteroidetes, are inhibited by UV disinfection and overall diversity of the microbial community decreases following treatment, many organisms not only survive, but remain active. These include common WWTP-derived organisms such asComamonadaceaeandPseudomonas.When combined with river water to mimic stormflow conditions, these organisms can persist in the environment and potentially enhance microbial functions such as nitrification and antibiotic resistance.</jats:sec

Ultraviolet disinfection impacts the microbial community composition and function of treated wastewater effluent and the receiving urban river

Background In the United States, an estimated 14,748 wastewater treatment plants (WWTPs) provide wastewater collection, treatment, and disposal service to more than 230 million people. The quality of treated wastewater is often assessed by the presence or absence of fecal indicator bacteria. UV disinfection of wastewater is a common final treatment step used by many wastewater treatment plants in order to reduce fecal coliform bacteria and other pathogens; however, its potential impacts on the total effluent bacterial community are seemingly varied. This is especially important given that urban WWTPs typically return treated effluent to coastal and riverine environments and thus are a major source of microorganisms, genes, and chemical compounds to these systems. Following rainfall, stormflow conditions can result in substantial increases to effluent flow into combined systems. Methods Here, we conducted a lab-scale UV disinfection on WWTP effluent using UV dosage of 100 mJ/cm2 and monitored the active microbiome in UV-treated effluent and untreated effluent over the course of 48 h post-exposure using 16S rRNA sequencing. In addition, we simulated stormflow conditions with effluent UV-treated and untreated effluent additions to river water and compared the microbial communities to those in baseflow river water. We also tracked the functional profiles of genes involved in tetracycline resistance (tetW) and nitrification (amoA) in these microcosms using RT-qPCR. Results We showed that while some organisms, such as members of the Bacteroidetes, are inhibited by UV disinfection and overall diversity of the microbial community decreases following treatment, many organisms not only survive, but remain active. These include common WWTP-derived organisms such as Comamonadaceae and Pseudomonas. When combined with river water to mimic stormflow conditions, these organisms can persist in the environment and potentially enhance microbial functions such as nitrification and antibiotic resistance

Ultraviolet disinfection impacts the microbial community composition and function of treated wastewater effluent and the receiving urban river

Background. In the United States, an estimated 14,748 wastewater treatment plants (WWTPs) provide wastewater collection, treatment, and disposal service to more than 230 million people. The quality of treated wastewater is often assessed by the presence or absence of fecal indicator bacteria. UV disinfection of wastewater is a common final treatment step used by many wastewater treatment plants in order to reduce fecal coliform bacteria and other pathogens; however, its potential impacts on the total effluent bacterial community are seemingly varied. This is especially important given that urban wastewater treatment plants (WWTPs) typically return treated effluent to coastal and riverine environments and thus are a major source of microorganisms, genes, and chemical compounds to these systems. Following rainfall, stormflow conditions can result in substantial increases to effluent flow into these systems. Methods. Here, we conducted a lab-scale UV disinfection on WWTP effluent using UV dosage of 100 mJ/cm2 and monitored the active microbiome in UV-treated effluent and untreated effluent over the course of 48h post-exposure using 16S rRNA sequencing. In addition, we simulated stormflow conditions with effluent UV-treated and untreated effluent additions to river water and compared the microbial communities to those in baseflow river water. We also tracked the functional profiles of genes involved in tetracycline resistance (tetW) and nitrification (amoA) in these microcosms using qPCR. Results. We showed that while some organisms, such as members of the Bacteroidetes, are inhibited by UV disinfection and overall diversity of the microbial community decreases following treatment, many organisms not only survive, but remain active. These include common WWTP-derived organisms such as Comamonadaceae and Pseudomonas. When combined with river water to mimic stormflow conditions, these organisms can persist in the environment and potentially enhance microbial functions such as nitrification and antibiotic resistance.</jats:p

Effects of greenwater and claywater regimes on early exogenous feeding in larval sablefish (Anoplopoma fimbria)

Marine fish larvae often survive better with algae in their rearing water, but algae is expensive and cheaper alternatives should be explored. This study tested the effects of algae and clay, a less-expensive water additive, on feeding, growth and survival during the first week of exogenous feeding in sablefish (Anoplopoma fimbria). After three days with algae or clay, larvae with algae fed better than those with clay. On the fourth day, half of the algae tanks were transitioned to clay, and half of the clay tanks were transitioned to algae. When the transition was 90 % complete, feeding was better if algae was used for the first three days, regardless of whether the larvae were transitioning to clay on day-4 or kept on algae, so the previous water additive (algae or clay) predicted feeding rates better than the additive at the time of the feeding trial. Benefits of the first three days with algae did not last indefinitely, however, as larvae that transitioned from algae to clay on the fourth day stopped showing feeding benefits by the fifth and sixth days. In a separate experiment, different algae-clay mixtures were compared: i) 100 % clay, ii) 25 % algae / 75 % clay, iii) 50 % algae / 50 % clay, and iv) 100 % algae. The third and fourth treatments fed and survived better relative to the first treatment, and did not differ from each other. In another experiment, larval feeding was higher in 100 % algae than in 50 % algae / 0 % clay, highlighting the importance of the clay component of the 50 % algae / 50 % clay mixture. Thus, algae use during the first week of exogenous feeding was halved by using an algae-clay mixture, without impacting larval feeding, growth, or survival. This study provides methods to reduce the reliance on expensive algae during the larval period