Heterotrophic Microbes Research Articles

Carbohydrates and carbohydrate degradation gene abundance and transcription in Atlantic waters of the Arctic

Carbohydrates are chemically and structurally diverse, represent a substantial fraction of marine organic matter and are key substrates for heterotrophic microbes. Studies on carbohydrate utilisation by marine microbes have been centred on phytoplankton blooms in temperate regions, while far less is known from high-latitude waters and during later seasonal stages. Here, we combine glycan microarrays and analytical chromatography with metagenomics and metatranscriptomics to show the spatial heterogeneity in glycan distribution and potential carbohydrate utilisation by microbes in Atlantic waters of the Arctic. The composition and abundance of monomers and glycan structures in POM varied with location and depth. Complex fucose-containing sulfated polysaccharides, known to accumulate in the ocean, were consistently detected, while the more labile β-1,3-glucan exhibited a patchy distribution. Through ‘omics analysis, we identify variations in the abundance and transcription of carbohydrate degradation-related genes across samples at the community and population level. The populations contributing the most to transcription were taxonomically related to those known as primary responders and key carbohydrate degraders in temperate ecosystems, such as NS4 Marine Group and Formosa. The unique transcription profiles for these populations suggest distinct substrate utilisation potentials, with predicted glycan targets corresponding to those structurally identified in POM from the same sampling sites. By combining cutting-edge technologies and protocols, we provide insights into the carbohydrate component of the carbon cycle in the Arctic during late summer and present a high-quality dataset that will be of great value for future comparative analyses.

Epidermal microorganisms contributed to the toxic mechanism of nZVI and TCEP in earthworms by robbing metal elements and nutrients

Disrupting effects of pollutants on symbiotic microbiota have been regarded as an important mechanism of host toxicity, with most current research focusing on the intestinal microbiota. In fact, the epidermal microbiota, which participates in the nutrient exchange between hosts and environments, could play a crucial role in host toxicity via community changes. To compare the contributions of intestinal and epidermal symbiotic microorganisms to host toxicity, this study designed single and combined scenarios of soil contamination [nano zero-valent iron (nZVI) and tris (2-chloroethyl) phosphate (TCEP)], and revealed the coupling mechanisms between intestinal/epidermal symbiotic bacterial communities and earthworm toxicological endpoints. Microbiome analysis showed that 15% of intestinal microbes were highly correlated with host endpoints, compared to 45% of epidermal microbes showing a similar correlation. Functional comparisons revealed that key species on the epidermis were mainly heterotrophic microbes with genetic abilities to utilize metal elements and carbohydrate nutrients. Further verifications demonstrated that when facing the co-contamination of nZVI and TCEP, certain symbiotic microorganisms became dominant and consumed zinc, copper, and manganese along with saccharides and amino acids, which may be responsible for the nutritional deficiencies in the host earthworms. The findings can enrich the understanding of the coupling relationship between symbiotic microorganisms and host toxicity, highlighting the importance of epidermal microorganisms in host resistance to environmental pollution.

Sulfamethoxazole removal from wastewater via anoxic/oxic moving bed biofilm reactor: Degradation pathways and toxicity assessment

The effects of sulfamethoxazole (SMZ), an antibiotic commonly detected in the water environment, on the performance of a single staged anoxic/oxic moving bed biofilm reactor (A/O MBBR), was investigated. The anoxic zone played a key role in the removal of SMZ with a percentage of contribution accounting for around 85% in the overall removal. Denitrifying heterotrophic microbes present in the anoxic zone showed relatively more resistance to higher SMZ loads. It was found that in extracellular polymeric substances, protein content was increased consistently with the increase in SMZ concentration. Based on the detected biotransformation products, four degradation pathways were proposed and the toxicity was evaluated. Metagenomic analysis revealed that at higher SMZ load the activity of genera, such as Proteobacteria and Actinobacteria was significantly affected. In summary, proper design and operation of staged A/O MBBR can offer a resilient and robust treatment towards SMZ removal from wastewater.

Interplay between autotrophic and heterotrophic prokaryotic metabolism in the bathypelagic realm revealed by metatranscriptomic analyses

BackgroundHeterotrophic microbes inhabiting the dark ocean largely depend on the settling of organic matter from the sunlit ocean. However, this sinking of organic materials is insufficient to cover their demand for energy and alternative sources such as chemoautotrophy have been proposed. Reduced sulfur compounds, such as thiosulfate, are a potential energy source for both auto- and heterotrophic marine prokaryotes.MethodsSeawater samples were collected from Labrador Sea Water (LSW, ~ 2000 m depth) in the North Atlantic and incubated in the dark at in situ temperature unamended, amended with 1 µM thiosulfate, or with 1 µM thiosulfate plus 10 µM glucose and 10 µM acetate (thiosulfate plus dissolved organic matter, DOM). Inorganic carbon fixation was measured in the different treatments and samples for metatranscriptomic analyses were collected after 1 h and 72 h of incubation.ResultsAmendment of LSW with thiosulfate and thiosulfate plus DOM enhanced prokaryotic inorganic carbon fixation. The energy generated via chemoautotrophy and heterotrophy in the amended prokaryotic communities was used for the biosynthesis of glycogen and phospholipids as storage molecules. The addition of thiosulfate stimulated unclassified bacteria, sulfur-oxidizing Deltaproteobacteria (SAR324 cluster bacteria), Epsilonproteobacteria (Sulfurimonas sp.), and Gammaproteobacteria (SUP05 cluster bacteria), whereas, the amendment with thiosulfate plus DOM stimulated typically copiotrophic Gammaproteobacteria (closely related to Vibrio sp. and Pseudoalteromonas sp.).ConclusionsThe gene expression pattern of thiosulfate utilizing microbes specifically of genes involved in energy production via sulfur oxidation and coupled to CO2 fixation pathways coincided with the change in the transcriptional profile of the heterotrophic prokaryotic community (genes involved in promoting energy storage), suggesting a fine-tuned metabolic interplay between chemoautotrophic and heterotrophic microbes in the dark ocean.13R5usaxM1iwstSWbHDM6jVideo

Multidimensional optimization for accelerating light-powered biocatalysis in Rhodopseudomonas palustris

BackgroundWhole-cell biocatalysis has been exploited to convert a variety of substrates into high-value bulk or chiral fine chemicals. However, the traditional whole-cell biocatalysis typically utilizes the heterotrophic microbes as the biocatalyst, which requires carbohydrates to power the cofactor (ATP, NAD (P)H) regeneration.ResultsIn this study, we sought to harness purple non-sulfur photosynthetic bacterium (PNSB) as the biocatalyst to achieve light-driven cofactor regeneration for cascade biocatalysis. We substantially improved the performance of Rhodopseudomonas palustris-based biocatalysis using a highly active and conditional expression system, blocking the side-reactions, controlling the feeding strategy, and attenuating the light shading effect. Under light-anaerobic conditions, we found that 50 mM ferulic acid could be completely converted to vanillyl alcohol using the recombinant strain with 100% efficiency, and > 99.9% conversion of 50 mM p-coumaric acid to p-hydroxybenzyl alcohol was similarly achieved. Moreover, we examined the isoprenol utilization pathway for pinene synthesis and 92% conversion of 30 mM isoprenol to pinene was obtained.ConclusionsTaken together, these results suggested that R. palustris could be a promising host for light-powered biotransformation, which offers an efficient approach for synthesizing value-added chemicals in a green and sustainable manner.Graphical

Exploring Biomarker Signatures in Glaciovolcanic Environments: Implications for the Search for Life on Mars

Glaciovolcanic systems, where hydrothermal heat interacts with ice, offer favorable conditions for life by providing liquid water, nutrients, and physicochemical gradients (Cousins and Crawford 2011). Despite significant climate change, evidence for glaciovolcanism is widespread through Mars’ history. Such sites may have offered refugia for life after Mars lost much of its surface water, representing some of the most recent habitable areas and promising sites to recover biomarkers. We examined a terrestrial glaciovolcanic site to study the indigenous biological community structure, the supporting physicochemical parameters, and the distribution of biomarkers within the geologic context. The insights will help refine Mars exploration of analogous sites. Method: We studied a partially subglacial hydrothermal area at the summit of the active Kverkfjöll volcano, Iceland. The heated ground has created a large ice-damned meltwater lake with shoreline hot springs, thermal streams, and mud pots of variable activity. We collected water and sediment samples aseptically across the breadth of features, including the lake (surface & depth). Samples were kept at -4°C. Environmental parameters were measured at each sample site. Sediment samples were split for mineralogical and organic analysis. Mineralogy was measured by X-ray Diffraction. Organic samples were freeze-dried and extracted with a Bligh & Dyer method (Bligh and Dyer 1959). Extracts were divided into hydrocarbons, neutral lipids, glycolipids, and phospholipid fatty acids (PLFA) with hexane, dichloromethane, acetone, and methanol. Gas Chromatography/Mass Spectrometry was used to analyze hydrocarbons and PLFA as fatty acid methyl esters. Results: Environmental: Fluids ranged from acidic to alkaline (pH 3-9), low to high temperature (8-87°C), and severely dysoxic to oxic (0.5-5 mg/L dissolved O). Mineralogy comprised a dioctahedral swelling clay, heulandite, and minor quartz, anatase, and pyrite. The assemblage suggests argillic-grade alteration at 100-140°C (Fulignati 2020), confirming that the glaciovolcanic conditions were within theoretical boundaries for life (<150°C (Merino et al. 2019)), and formed minerals capable of protecting organic matter (e.g., swelling clay). Lipids: PLFA are essential components of cell membranes and degrade rapidly upon cell death. PLFA profiles thus provide insight into the composition and distribution of the viable community. Results revealed a diversity of PLFA with low molecular weights and several bacterial-diagnostic structures, indicating an active prokaryote-dominated biosphere. Molecular patterns correlated (p<0.05) with pH, temperature, and oxygen, suggesting homeoviscous adaptations or community composition variations. In either case, microbes demonstrate adaptability to extreme conditions in glaciovolcanic settings. Hydrocarbons are inert and used as molecular fossils. We detected multiple patterns attributable to microbial biosynthesis, including abundant C17 (straight and branched) diagnostic of cyanobacteria; high concentrations of short-chain n-alkane/alkene doublets (<C22) with an even carbon preference (CPI<1) indicating diverse microbial input; and a narrow unresolved complex mixture with elution parameters characteristic of microbial heterotrophy (Finkel et al. 2023). The pattern of even-numbered alkane/alkene pairs has been scarcely reported and offers insight into rare hydrocarbon distributions. Interestingly, the hydrocarbon and PLFA patterns do not align. Thus, PLFA are not a significant source of hydrocarbons here and we suggest direct microbial synthesis dominates. Implications of this hypothesis will be discussed in the context of palaeobiological reconstruction. Conclusion: Our site provides an array of physicochemical niches for diverse life modes and a mineral assemblage amenable to organic preservation. Analogous sites on Mars should be explored, but more work is needed to understand preservation biases of hydrocarbon sources and the implications for interpretation.

Microbial degradation and assimilation of veratric acid in oxic and anoxic groundwaters.

Microbial communities are key players in groundwater ecosystems. In this dark environment, heterotrophic microbes rely on biomass produced by the activity of lithoautotrophs or on the degradation of organic matter seeping from the surface. Most studies on bacterial diversity in groundwater habitats are based on 16S gene sequencing and full genome reconstructions showing potential metabolic pathways used in these habitats. However, molecular-based studies do not allow for the assessment of population dynamics over time or the assimilation of specific compounds and their biochemical transformation by microbial communities. Therefore, in this study, we combined DNA-, phospholipid fatty acid-, and metabolomic-stable isotope probing to target and identify heterotrophic bacteria in the groundwater setting of the Hainich Critical Zone Exploratory (CZE), focusing on 2 aquifers with different physico-chemical conditions (oxic and anoxic). We incubated groundwater from 4 different wells using either 13C-labeled veratric acid (a lignin-derived compound) (single labeling) or a combination of 13CO2 and D-labeled veratric acid (dual labeling). Our results show that heterotrophic activities dominate all groundwater sites. We identified bacteria with the potential to break down veratric acid (Sphingobium or Microbacterium). We observed differences in heterotrophic activities between the oxic and anoxic aquifers, indicating local adaptations of bacterial populations. The dual labeling experiments suggested that the serine pathway is an important carbon assimilation pathway and that organic matter was an important source of hydrogen in the newly produced lipids. These experiments also yielded different labeled taxa compared to the single labeling experiments, showing that there exists a complex interaction network in the groundwater habitats.

The role of clay minerals in the preservation of Precambrian organic-walled microfossils.

Precambrian organic-walled microfossils (OWMs) are primarily preserved in mudstones and shales that are low in total organic carbon (TOC). Recent work suggests that high TOC may hinder OWM preservation, perhaps because it interferes with chemical interactions involving certain clay minerals that inhibit the decay of microorganisms. To test if clay mineralogy controls OWM preservation, and if TOC moderates the effect of clay minerals, we compared OWM preservational quality (measured by pitting on fossil surfaces and the deterioration of wall margins) to TOC, total clay, and specific clay mineral concentrations in 78 shale samples from 11 lithologic units ranging in age from ca. 1650 to 650 million years ago. We found that the probability of finding well-preserved microfossils positively correlates with total clay concentrations and confirmed that it negatively correlates with TOC concentrations. However, we found no evidence that TOC influences the effect of clay mineral concentrations on OWM preservation, supporting an independent role of both factors on preservation. Within the total clay fraction, well-preserved microfossils are more likely to occur in shales with high illite concentrations and low berthierine/chamosite concentrations; however, the magnitude of their effect on preservation is small. Therefore, there is little evidence that bulk clay chemistry is important in OWM preservation. Instead, we propose that OWM preservation is largely regulated by physical properties that isolate organic remains from microbial degradation such as food scarcity (low TOC) and low sediment permeability (high total clay content): low TOC increases the diffusive distances between potential carbon sources and heterotrophic microbes (or their degradative enzymes), while high clay concentrations reduce sediment pore space, thereby limiting the diffusion of oxidants and degradative enzymes to the sites of decay.

The effect of polyvinyl chloride (PVC) color on biofilm development and biofilm-heavy metal chemodynamics in the aquatic environment

Plastic surfaces are colonized by microorganisms and biofilms are formed in the natural aquatic environment. As the biofilm develops, it changes the density and buoyancy of the plastic-biofilm complex, results in plastic sinking, and increases the heavy metals accumulated by biofilm's mobility and availability in aquatic ecosystems. In this experiment, biofilms were cultured on five colors of polyvinyl chloride (PVC; transparent, green, blue, red, black) in an aquatic environment to investigate the effects of plastic color on biofilm formation and development (Phase 1) and to study the effects of being sunk below the photic zone on biofilm (Phase 2). The PVC color significantly affected the biofilm formation rate but had no impact on the final biofilm biomass. After sinking the biofilm-PVC below the photic zone in Phase 2, the layer of diatoms on the biofilm surface began to disintegrate, and the biomass and Chlorophyll-a (Chla) content of the biofilm decreased, except on the red PVC. Below the photic zone, the microbial community of the biofilm changed from primarily autotrophic microbes to mostly heterotrophic microbes. Microbial diversity increased and extracellular polymeric substances (EPS) content decreased. The primary factor leading to microbial diversity and community structure changes was water depth rather than PVC color. The changes induced in the biofilm led to an increase in the concentration of all heavy metals in the biofilm, related to the increase in microbial diversity. This study provides new insights into the biofilm formation process and the effects on a biofilm when it sinks below the photic zone.

Ecology theory disentangles microbial dichotomies.

Microbes are often discussed in terms of dichotomies such as copiotrophic/oligotrophic and fast/slow-growing microbes, defined using the characterisation of microbial growth in isolated cultures. The dichotomies are usually qualitative and/or study-specific, sometimes precluding clear-cut results interpretation. We can unravel microbial dichotomies as life history strategies by combining ecology theory with Monod curves, a laboratory mathematical tool of bacterial physiology that relates the specific growth rate of a microbe with the concentration of a limiting nutrient. Fitting of Monod curves provides quantities that directly correspond to key parameters in ecological theories addressing species coexistence and diversity, such as r/K selection theory, resource competition and community structure theory and the CSR triangle of life strategies. The resulting model allows us to reconcile the copiotrophic/oligotrophic and fast/slow-growing dichotomies as different subsamples of a life history strategy triangle that also includes r/K strategists. We also used the number of known carbon sources together with community structure theory to partially explain the diversity of heterotrophic microbes observed in metagenomics experiments. In sum, we propose a theoretical framework for the study of natural microbial communities that unifies several existing proposals. Its application would require the integration of metagenomics, metametabolomics, Monod curves and carbon source data.

Can we manage microbial systems to enhance carbon storage?

Climate change is an urgent environmental issue with wide-ranging impacts on ecosystems and society. Microbes are instrumental in maintaining the balance between carbon (C) accumulation and loss in the biosphere, actively regulating greenhouse gas fluxes from vast reservoirs of organic C stored in soils, sediments and oceans. Heterotrophic microbes exhibit varying capacities to access, degrade and metabolise organic C-leading to variations in remineralisation and turnover rates. The present challenge lies in effectively translating this accumulated knowledge into strategies that effectively steer the fate of organic C towards prolonged sequestration. In this article, we discuss three ecological scenarios that offer potential avenues for shaping C turnover rates in the environment. Specifically, we explore the promotion of slow-cycling microbial byproducts, the facilitation of higher carbon use efficiency, and the influence of biotic interactions. The ability to harness and control these processes relies on the integration of ecological principles and management practices, combined with advances in economically viable technologies to effectively manage microbial systems in the environment.

Stoichiometry of carbon, nitrogen, and phosphorus released from the leaf litter of various temperate tree species.

Dissolved organic matter and inorganic nutrients released from forest leaf litter through leaching are the important energy and nutrient sources that support the production of aquatic food webs. Leaf litter-derived dissolved organic carbon (DOC) is a critical energy source for aquatic heterotrophic microbes, and inorganic nitrogen and phosphorus can enhance primary production. In this study, we experimentally measured the release efficiencies and amounts of DOC, total dissolved nitrogen (TDN), and total dissolved phosphorus (TDP) of the leaf litter from 11 temperate tree species by soaking the leaf litter in water for 28 days. We found that the maximal release efficiency (% of element released per estimated mass of the element) was the highest for P and lowest for N. These efficiencies were species-specific. Additionally, the DOC:TDP, DOC:TDN, and TDN:TDP ratios varied among the leachate of different leaf litter species. DOC:TDP increased with the C:P ratio in leaf litter biomass but is considerably lower; TDN:TDP was lower than the N:P ratio in leaf litter biomass as well; DOC:TDN ratio was higher than the C:N ratio in leaf litter biomass. These results suggest that the ratios of DOC to dissolved N and P nutrients released into water are related to, but not the same as, the stoichiometry of leaf litter biomass. Based on these findings, we concluded that changes in the vegetations with different leaf litter stoichiometry can alter the relative importance of detrital and grazing food chains in aquatic ecosystems.

Identification of microbial metabolic functional guilds from large genomic datasets.

Heterotrophic microbes play an important role in the Earth System as key drivers of major biogeochemical cycles. Specifically, the consumption rate of organic matter is set by the interaction between diverse microbial communities and the chemical and physical environment in which they reside. Modeling these dynamics requires reducing the complexity of microbial communities and linking directly with biogeochemical functions. Microbial metabolic functional guilds provide one approach for reducing microbial complexity and incorporating microbial biogeochemical functions into models. However, we lack a way to identify these guilds. In this study, we present a method for defining metabolic functional guilds from annotated genomes, which are derived from both uncultured and cultured organisms. This method utilizes an Aspect Bernoulli (AB) model and was tested on three large genomic datasets with 1,733-3,840 genomes each. Ecologically relevant microbial metabolic functional guilds were identified including guilds related to DMSP degradation, dissimilatory nitrate reduction to ammonia, and motile copiotrophy. This method presents a way to generate hypotheses about functions co-occurring within individual microbes without relying on cultured representatives. Applying the concept of metabolic functional guilds to environmental samples will provide new insight into the role that heterotrophic microbial communities play in setting rates of carbon cycling.

Environmental ammonia analysis based on exclusive nitrification by nitrifying biofilm screened from natural bioresource

Biofilm-based biological nitrification is widely used for ammonia removal, while hasn't been explored for ammonia analysis. The stumbling block is the coexist of nitrifying and heterotrophic microbes in real environment resulting in non-specific sensing. Herein, an exclusive ammonia sensing nitrifying biofilm was screened from natural bioresource, and a bioreaction-detection system for the on-line analysis of environmental ammonia based on biological nitrification was reported. The nitrifying microbes were aggregated into a nitrifying biofilm through a result-oriented bioresource enrichment strategy. The predominant nitrifying population and progressive surface reaction in the plug flow bioreactor led to the exclusive and exhaustive ammonia biodegradation for the establishment of a novel analytical method. The on-line ammonia monitoring prototype achieved complete biodegradation for determining ammonium nitrogen within 5 min and showed exceptional reliability in long-term real sample measurements without frequent calibration. This work offers a low-threshold natural screening paradigm for developing sustainable bioresource-based analytical technologies.

Land use differentially alters microbial interactions and detritivore feeding during leaf decomposition in headwater streams

Abstract Human‐driven land conversion from forest to agriculture introduces nutrients, like nitrogen (N) and phosphorus (P), which upon entering nearby streams can stimulate growth and metabolism of microbial autotrophs (algae) and heterotrophs (fungi and bacteria). Recent evidence indicates that algae also can stimulate microbial heterotrophs associated with decaying plant litter by exuding labile carbon (C), defined as the priming effect on decomposition. These autotroph–heterotroph interactions may further alter detritivore feeding ecology, suggesting many complex mechanisms of land‐use change on stream ecosystem functions. In order to investigate the effects of land use on autotroph–heterotroph interactions, we used a full‐factorial design manipulating labile C (glucose and acetate) and nutrients (N and P) to test their effects during red maple litter decomposition within four forested and four pasture agriculture streams in the Point Remove catchment, Arkansas. We deployed nutrient‐diffusing substrates (NDS) containing litter and either agar, agar + labile C, agar + NP, or agar + CNP for 20 days and measured microbial biomass and metabolism, and detrital mass loss. Additionally, we fed litter to Tipula sp. to assess shifts in consumption and egestion rates. Both microbial heterotrophs and autotrophs responded to NDS manipulations, but not land use. Heterotrophic microbes showed evidence of nutrient limitation and negative priming, while algal biomass was negatively associated with heterotrophs. Tipula consumption and egestion rates were both elevated when feeding on litter incubated in agriculture streams, but only egestion responded to NDS manipulations, suggesting a change in assimilation efficiencies. Cumulatively, we suggest that altered stream conditions, such as those resulting from land‐use change, can alter food webs and detritivore feeding more heavily than microbial interactions during litter decomposition in stream ecosystems. These results add to the literature demonstrating shifts in autotroph–heterotroph interactions with anthropogenic nutrient pollution, and the implications for detritivore feeding and overall C cycling in streams.

Linking DOM characteristics to microbial community: The potential role of DOM mineralization for arsenic release in shallow groundwater

Dissolved organic matter (DOM) play critical roles in arsenic (As) biotransformation in groundwater, but its compositional characteristics and interactions with indigenous microbial communities remain unclear. In this study, DOM signatures coupled with taxonomy and functions of microbial community were characterized in As-enriched groundwater by excitation-emission matrix, Fourier transform ion cyclotron resonance mass spectrometry and metagenomic sequencing. Results showed that As concentrations were significantly positively correlated with DOM humification (r = 0.707, p < 0.01) and the most dominant humic acid-like DOM components (r = 0.789, p < 0.01). Molecular characterization further demonstrated high DOM oxidation degree, with the prevalence of unsaturated oxygen-low aromatics, nitrogen (N1/N2)-containing compounds and unique CHO molecules in high As groundwater. These DOM properties were consistent with microbial composition and functional potentials. Both taxonomy and binning analyses demonstrated the dominance of Pseudomonas stutzeri, Microbacterium and Sphingobium xenophagum in As-enriched groundwater which possessed abundant As-reducing gene, with organic carbon degrading genes capable of labile to recalcitrant compounds degradation and high potentials of organic nitrogen mineralization to generate ammonium. Besides, most assembled bins in high As groundwater presented strong fermentation potentials which could facilitate carbon utilization by heterotrophic microbes. This study provides better insight into the potential role of DOM mineralization for As release in groundwater system.

Performance and microbial analysis of a fluidized bed membrane bioreactor operated in the partial nitrification and anammox (PN/A) mode for polishing anaerobically treated effluent to industrial discharge standard

This study evaluates the performance and microbial dynamics of an ORP-controlled fluidized bed membrane bioreactor impregnated with partial nitrification and anammox culture (FMBRPN/A) for polishing the low COD/NH4+ anaerobically treated effluent of the seafood processing industry. Results showed that the highest average removal of COD and total nitrogen were 68.6% and 75.0% at COD/NH4+ 0.44 and 0.33, respectively. Heterotrophic denitrification accounted for only 3.0–7.1% of the overall nitrogen removals at COD/NH4+ 0.33–0.55. A low nitrite oxidation rate observed indicated that almost all NO3- came from anammox reactions. Data on ammonia oxidation rate, nitrite oxidation rate, and anammox nitrogen removal rate pointed to the competitive advantage between heterotrophic and nitrifying/anammox consortia at higher and lower COD/NH4+ ratios. When salinity threat at 5 g/L NaCl in the influent was imposed to FMBRPN/A, nitrogen removal was almost completely eliminated but quickly recovered to 25.5% as organic removal was not affected. Activities of the nitrogenous autotrophic organisms and heterotrophic microbes were strongly dependent on the presence of organic and salinity in the wastewater. Microbial analysis of biofilm formed on the membrane surface showed a significant difference in the succession of dominant bacteria during the experiments. Next Generation Sequencing indicated the dynamic shifting proportions of ammonium oxidizing bacteria (AOB), nitrite oxidization bacteria (NOB), denitrifying bacteria (DNB), and anammox bacteria (AMX) ranging from 49.4% to 64.2% at various COD/NH4+ ratios, which suggested their syntrophic relationship. Superior membrane retention of FMBRPN/A for heterotrophic and slow-growing autotrophic cells facilitated system stability and efficiency to meet the industrial effluent standard.

Larvae of an invasive scarab increase greenhouse gas emissions from soils and recruit gut mycobiota involved in C and N transformations.

Soil-derived prokaryotic gut communities of the Japanese beetle Popillia japonica Newman (JB) larval gut include heterotrophic, ammonia-oxidizing, and methanogenic microbes potentially capable of promoting greenhouse gas (GHG) emissions. However, no research has directly explored GHG emissions or the eukaryotic microbiota associated with the larval gut of this invasive species. In particular, fungi are frequently associated with the insect gut where they produce digestive enzymes and aid in nutrient acquisition. Using a series of laboratory and field experiments, this study aimed to (1) assess the impact of JB larvae on soil GHG emissions; (2) characterize gut mycobiota associated with these larvae; and (3) examine how soil biological and physicochemical characteristics influence variation in both GHG emissions and the composition of larval gut mycobiota. Manipulative laboratory experiments consisted of microcosms containing increasing densities of JB larvae alone or in clean (uninfested) soil. Field experiments included 10 locations across Indiana and Wisconsin where gas samples from soils, as well as JB and their associated soil were collected to analyze soil GHG emissions, and mycobiota (ITS survey), respectively. In laboratory trials, emission rates of CO2, CH4, and N2O from infested soil were ≥ 6.3× higher per larva than emissions from JB larvae alone whereas CO2 emission rates from soils previously infested by JB larvae were 1.3× higher than emissions from JB larvae alone. In the field, JB larval density was a significant predictor of CO2 emissions from infested soils, and both CO2 and CH4 emissions were higher in previously infested soils. We found that geographic location had the greatest influence on variation in larval gut mycobiota, although the effects of compartment (i.e., soil, midgut and hindgut) were also significant. There was substantial overlap in the composition and prevalence of the core fungal mycobiota across compartments with prominent fungal taxa being associated with cellulose degradation and prokaryotic methane production/consumption. Soil physicochemical characteristics such as organic matter, cation exchange capacity, sand, and water holding capacity, were also correlated with both soil GHG emission, and fungal a-diversity within the JB larval gut. Conclusions: Results indicate JB larvae promote GHG emissions from the soil directly through metabolic activities, and indirectly by creating soil conditions that favor GHG-associated microbial activity. Fungal communities associated with the JB larval gut are primarily influenced by adaptation to local soils, with many prominent members of that consortium potentially contributing to C and N transformations capable of influencing GHG emissions from infested soil.

Effects of global change on the ability of stream biofilm to dissipate the herbicide glyphosate

The herbicide glyphosate is contaminating a large number of freshwater ecosystems worldwide and its fate and effects remains uncertain in light of the effects of global change. The present study examines how variations in water temperature and light availability relative to global change affect the ability of stream biofilms to degrade the herbicide glyphosate. Biofilms were exposed in microcosms to two levels of water temperature simulating global warming (Ambient = 19–22 °C and Warm = 21–24 °C) and three levels of light representative of riparian habitat destruction due to land use change (Dark = 0, Intermediate = 600, High = 1200 μmol photons m−2 s−1). Biofilms were acclimated to six different experimental treatments, namely i) ambient temperature without light (AMB_D), ii) ambient temperature and intermediate light (AMB_IL), iii) ambient temperature and high light (AMB_HL), iv) warm temperature without light (WARM_D), v) warm temperature and intermediate light (WARM_IL) and vi) warm temperature and high light (WARM_HL). The ability of biofilms to degrade 50 μg L−1 of glyphosate was tested. Results showed that water temperature increase, but not light availability increase, significantly increased aminomethyl phosphonic acid (AMPA) production by biofilms. However, the combined increase of temperature and light generated the shortest time to dissipate half of the glyphosate supplied and/or half of the maximum AMPA produced (6.4 and 5.4 days, respectively) by biofilms. Despite light had a major effect in modulating biofilm structural and functional descriptors, the response of certain descriptors (i. e. chlorophyll-a concentration, bacterial density and diversity, nutrient content and PHO activity) to light availability increase depended on water temperature. Specifically, the biofilms in the WARM_HL treatment displayed the highest Glucosidase: Peptidase and Glucosidase: Phosphatase enzyme activity ratios and the lowest biomass C: N molar ratios compared to the other treatments. According to these results, warmer temperatures and high light availability could have been exacerbating the decomposition of organic C compounds in biofilms, including the use of glyphosate as a C source for microbial heterotrophs. This study shows that ecoenzymatic stoichiometry and xenobiotic biodegradation approaches can be combined to better understand the functioning of biofilms in pesticide-polluted streams.

Resolving the fate of trace metals during microbial remineralization of phytoplankton biomass in precursor banded iron formation sediments

Banded Iron Formations (BIFs) have long been considered a sedimentary record of seawater trace metal composition during the Precambrian. However, recent work has suggested that much of the trace metal composition of BIFs could have been derived from phytoplankton biomass, not seawater. In this model, phytoplankton biomass settles from the photic zone to the seafloor sediments, where it is then oxidized by heterotrophic microbes, such as dissimilatory Fe(III) reducing (DIR) bacteria, for energy generation. Remineralization of this biomass released the trace metals associated with organic molecules from phytoplankton (i.e., in metalloproteins), allowing these metals to be captured by Fe oxyhydroxides and preserved in BIFs. While there is compelling evidence that the phytoplankton biomass served as a trace metal shuttle to precursor BIF sediments, it is unclear whether the degradation of biomass by DIR bacteria would liberate the biogenic trace metals as the model proposes. This work tests this hypothesis by using anoxic incubations of a model DIR bacterium (Shewanella oneidensis MR-1) with phytoplankton biomass as energy and carbon sources and ferrihydrite, a poorly crystalline Fe(III) oxyhydroxide (Fe(OH)3), as electron acceptor. Our results show that while S. oneidensis MR-1 can consume some of the carbon substrates found in phytoplankton biomass, there is no evidence that S. oneidensis MR-1 degraded metalloproteins which would have liberated trace metals. In the context of the Precambrian, these data imply that other heterotrophic bacteria, such as fermenters, may have had a larger role in the liberation of trace metals from dead biomass during early BIF development.