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SUSTAIN drilling at Surtsey volcano, Iceland, tracks hydrothermal and microbiological interactions in basalt 50 years after eruption

The 2017 Surtsey Underwater volcanic System for Thermophiles, Alteration processes and INnovative concretes (SUSTAIN) drilling project at Surtsey volcano, sponsored in part by the International Continental Scientific Drilling Program (ICDP), provides precise observations of the hydrothermal, geochemical, geomagnetic, and microbiological changes that have occurred in basaltic tephra and minor intrusions since explosive and effusive eruptions produced the oceanic island in 1963–1967. Two vertically cored boreholes, to 152 and 192 m below the surface, were drilled using filtered, UV-sterilized seawater circulating fluid to minimize microbial contamination. These cores parallel a 181 m core drilled in 1979. Introductory investigations indicate changes in material properties and whole-rock compositions over the past 38 years. A Surtsey subsurface observatory installed to 181 m in one vertical borehole holds incubation experiments that monitor in situ mineralogical and microbial alteration processes at 25–124 ∘C. A third cored borehole, inclined 55∘ in a 264∘ azimuthal direction to 354 m measured depth, provides further insights into eruption processes, including the presence of a diatreme that extends at least 100 m into the seafloor beneath the Surtur crater. The SUSTAIN project provides the first time-lapse drilling record into a very young oceanic basaltic volcano over a range of temperatures, 25–141 ∘C from 1979 to 2017, and subaerial and submarine hydrothermal fluid compositions. Rigorous procedures undertaken during the drilling operation protected the sensitive environment of the Surtsey Natural Preserve.

Scientific Drilling

Freshwater bacteria are stoichiometrically flexible with a nutrient composition similar to seston

Although aquatic bacteria are assumed to be nutrient-rich, they out-compete other foodweb osmotrophs for nitrogen (N) and phosphorus (P) an apparent contradiction to resource ratio theory. This paradox could be resolved if aquatic bacteria were demonstrated to be nutrient-poor relative other portions of the planktonic food web. In a survey of >120 lakes in the upper Midwest of the USA, the nutrient content of bacteria was lower than previously reported and very similar to the Redfield ratio, with a mean biomass composition of 102:12:1 (C:N:P). Individual freshwater bacterial isolates grown under P-limiting and P-replete conditions had even higher C:P and N:P ratios with a mean community biomass composition ratio of 875C:179N:1P suggesting that individual strains can be extremely nutrient-poor, especially with respect to P. Cell-specific measurements of individual cells from one lake confirmed that low P content could be observed at the community level in natural systems with a mean biomass composition of 259C:69N:1P. Variability in bacterial stoichiometry is typically not recognized in the literature as most studies assume constant and nutrient-rich bacterial biomass composition. We present evidence that bacteria can be extremely P-poor in individual systems and in culture, suggesting that bacteria in freshwater ecosystems can either play a role as regenerators or consumers of inorganic nutrients and that this role could switch depending on the relationship between bacterial biomass stoichiometry and resource stoichiometry. This ability to switch roles between nutrient retention and regeneration likely facilitates processing of terrestrial organic matter in lakes and rivers and has important implications for a wide range of bacterially mediated biogeochemical processes.

Frontiers in Microbiology

Microbially induced anaerobic oxidation of magnetite to maghemite in a hydrocarbon-contaminated aquifer

Iron mineral transformations occurring in hydrocarbon-contaminated sites are linked to the biodegradation of the hydrocarbons. At a hydrocarbon-contaminated site near Bemidji, Minnesota, USA, measurements of magnetic susceptibility (MS) are useful for monitoring the natural attenuation of hydrocarbons related to iron cycling. However, a transient MS, previously observed at the site, remains poorly understood and the iron mineral phases acting as reactants and products associated with this MS perturbation remain largely unknown. To address these unknowns, we acquired mineral magnetism measurements, including hysteresis loops, backfield curves, and isothermal remanent magnetizations on sediment core samples retrieved from the site and magnetite-filled mineral packets installed within the aquifer. Our data show that the core samples and magnetite packs display decreasing magnetization with time and that this loss in magnetization is accompanied by increasing bulk coercivity consistent with decreased average grain size and/or partial oxidation. Low-temperature magnetometry on all samples displayed behavior consistent with magnetite, but samples within the plume also show evidence of maghemitization. This interpretation is supported by the occurrence of shrinkage cracks on the surface of the grains imaged via scanning electron microscopy. Magnetite transformation to maghemite typically occurs under oxic conditions, here, we propose that maghemitization occurs within the anoxic portions of the plume via microbially mediated anaerobic oxidation. Mineral dissolution also occurs within the plume. Microorganisms capable of such anaerobic oxidation have been identified within other areas at the Bemidji site, but additional microbiological studies are needed to link specific anaerobic iron oxidizers with this loss of magnetization.

Minnesota

Concentration and retention of Toxoplasma gondii oocysts by marine snails demonstrate a novel mechanism for transmission of terrestrial zoonotic pathogens in coastal ecosystems

The parasite T oxoplasma gondii is an environmentally persistent pathogen that can cause fatal disease in humans, terrestrial warm-blooded animals and aquatic mammals. Although an association between T . gondii exposure and prey specialization on marine snails was identified in threatened California sea otters, the ability of kelp-dwelling snails to transmit terrestrially derived pathogens has not been previously investigated. The objective of this study was to measure concentration and retention of T . gondii by marine snails in laboratory aquaria, and to test for natural T . gondii contamination in field-collected snails. Following exposure to T . gondii -containing seawater, oocysts were detected by microscopy in snail faeces and tissues for 10 and 3 days respectively. Nested polymerase chain reaction was also applied as a method for confirming putative T . gondii oocysts detected in snail faeces and tissues by microscopy. T oxoplasma gondii was not detected in field-collected snails. Results suggest that turban snails are competent transport hosts for T . gondii . By concentrating oocysts in faecal pellets, snails may facilitate entry of T . gondii into the nearshore marine food web. This novel mechanism also represents a general pathway by which marine transmission of terrestrially derived microorganisms can be mediated via pathogen concentration and retention by benthic invertebrates.

Environmental Microbiology

Ground-water quality in the carbonate-rock aquifer of the Great Basin, Nevada and Utah, 2003

The carbonate-rock aquifer of the Great Basin is named for the thick sequence of Paleozoic limestone and dolomite with lesser amounts of shale, sandstone, and quartzite. It lies primarily in the eastern half of the Great Basin and includes areas of eastern Nevada and western Utah as well as the Death Valley area of California and small parts of Arizona and Idaho. The carbonate-rock aquifer is contained within the Basin and Range Principal Aquifer, one of 16 principal aquifers selected for study by the U.S. Geological Survey’s National Water- Quality Assessment Program. Water samples from 30 ground-water sites (20 in Nevada and 10 in Utah) were collected in the summer of 2003 and analyzed for major anions and cations, nutrients, trace elements, dissolved organic carbon, volatile organic compounds (VOCs), pesticides, radon, and microbiology. Water samples from selected sites also were analyzed for the isotopes oxygen-18, deuterium, and tritium to determine recharge sources and the occurrence of water recharged since the early 1950s. Primary drinking-water standards were exceeded for several inorganic constituents in 30 water samples from the carbonate-rock aquifer. The maximum contaminant level was exceeded for concentrations of dissolved antimony (6 μg/L) in one sample, arsenic (10 μg/L) in eleven samples, and thallium (2 μg/L) in one sample. Secondary drinking-water regulations were exceeded for several inorganic constituents in water samples: chloride (250 mg/L) in five samples, fluoride (2 mg/L) in two samples, iron (0.3 mg/L) in four samples, manganese (0.05 mg/L) in one sample, sulfate (250 mg/L) in three samples, and total dissolved solids (500 mg/L) in seven samples. Six different pesticides or metabolites were detected at very low concentrations in the 30 water samples. The lack of VOC detections in water sampled from most of the sites is evidence thatVOCs are not common in the carbonate-rock aquifer. Arsenic values for water range from 0.7 to 45.7 μg/L, with a median value of 9.6 μg/L. Factors affecting arsenic concentration in the carbonate-rock aquifer in addition to geothermal heating are its natural occurrence in the aquifer material and time of travel along the flow path. Most of the chemical analyses, especially for VOCs and nutrients, indicate little, if any, effect of overlying land-use patterns on ground-water quality. The water quality in recharge areas for the aquifer where human activities are more intense may be affected by urban and/or agricultural land uses as evidenced by pesticide detections. The proximity of the carbonate-rock aquifer at these sites to the land surface and the potential for local recharge to occur through the fractured rock likely results in the occurrence of these and other land-surface related contaminants in the ground water. Water from sites sampled near outcrops of carbonate-rock aquifer likely has a much shorter residence time resulting in a potential for detection of anthropogenic or land-surface related compounds. Sites located in discharge areas of the flow systems or wells that are completed at a great depth below the land surface generally show no effects of land-use activities on water quality. Flow times within the carbonate-rock aquifer, away from recharge areas, are on the order of thousands of years, so any contaminants introduced at the land surface that will not degrade along the flow path have not reached the sampled sites in these areas.

Nevada, Utah

Spatially integrating microbiology and geochemistry to reveal complex environmental health issues: Anthrax in the contiguous United States

Maxent models were run using the B. anthracis presence data and/or the animal outbreak presence data. Models run using the animal outbreak data alone utilized two scales: the Outbreak State scale which included only states reporting animal anthrax outbreaks from 2001 to 2013 and the National scale which included all states in the contiguous United States. Three iterations of the environmental data were used and included the Sample Location dataset which utilized the environmental variable data with assigned latitude and longitude locations from the USGS NASGLP project; the Normalized dataset which scaled the environmental variables so that the values fell between 0 and 1; and the Interpolated dataset which provided an interpolation of the environmental variables averaged for each county and assigned to a point for that county at the centroid (rather than using the NASGLP latitude and longitude location). Two metrics were used to measure model performance including the widely used area under the curve (AUC) and an alternative method, the True Skill Statistic (TSS). The AUC gives the probability that a randomly chosen presence location has been correctly ranked higher than the absence/background site. AUC values at 0.5 or lower mean the ranking is no better than random, while the AUC values nearer to 1 mean the model is a better predictor. The TSS provides a comparison of how well the background predictions made by the model match the model results at the test dataset (presence) locations. TSS values near +1 means the model approaches perfect agreement, while values near −1 indicate the model is no better than random. Maxent models to determine the influence of environmental factors on the B. anthracis distribution using the PCR data yielded a low TSS, which suggested the model might be underfitting the data. This was not surprising due to the difficulty in recovering B. anthracis in soil samples as well as the samples themselves being discrete in nature and only capturing a snapshot in time. Therefore, the distribution of B. anthracis and its niche in the contiguous United States could not be determined in this study. However, efforts to investigate environmental factors that would have a higher potential of supporting an anthrax outbreak in wildlife and livestock yielded better results. Results showed that most of the Maxent models in this study performed best when using the Outbreak State scale. When the models were scaled up to the National scale, model performance declined, except for the Normalized variable dataset. At the Outbreak State scale, a large proportion of the area was predicted to be of higher probability for wildlife/livestock anthrax outbreaks, and the statistical measures assumed the model was underfitting the data. The model with the highest AUC and TSS scores for this study was the Outbreak State scale using Sample Location dataset (AUC = 0.918 and TSS = 0.82). Some of the variables found to be closely related to the occurrence of B. anthracis in this study included pH, drainage potential, and concentration of elements including Na, Ca, Sr, and Mg, which have also been found to be related to animal outbreaks or to the occurrence of B. anthracis in previous studies. The models in the current study indicated possible regions that have not had recent wildlife/livestock anthrax outbreaks but contained environmental conditions that could potentially support an outbreak if one were to occur (Michigan and Maine). This work provides an extension to the use of ecological niche modeling to outbreak potential in livestock/wildlife in the United States because it utilizes additional soil geochemistry data and has shown that further validation techniques, such as the TSS, should be considered in addition to AUC. Results from this study could be used by animal and public health officials to identify areas with a higher potential for anthrax outbreak in wildlife and livestock due to naturally occurring soil and environmental conditions.

Book chapter

Comparison of filters for concentrating microbial indicators and pathogens in lake-water samples

Bacterial indicators are used to indicate increased health risk from pathogens and to make beach closure and advisory decisions; however, beaches are seldom monitored for the pathogens themselves. Studies of sources and types of pathogens at beaches are needed to improve estimates of swimming-associated health risks. It would be advantageous and cost-effective, especially for studies conducted on a regional scale, to use a method that can simultaneously filter and concentrate all classes of pathogens from the large volumes of water needed to detect pathogens. In seven recovery experiments, stock cultures of viruses and protozoa were seeded into 10-liter lake water samples, and concentrations of naturally occurring bacterial indicators were used to determine recoveries. For the five filtration methods tested, the highest median recoveries were as follows: glass wool for adenovirus (4.7%); NanoCeram for enterovirus (14.5%) and MS2 coliphage (84%); continuous-flow centrifugation (CFC) plus Virocap (CFC+ViroCap) for Escherichia coli (68.3%) and Cryptosporidium (54%); automatic ultrafiltration (UF) for norovirus GII (2.4%); and dead-end UF for Enterococcus faecalis (80.5%), avian influenza virus (0.02%), and Giardia (57%). In evaluating filter performance in terms of both recovery and variability, the automatic UF resulted in the highest recovery while maintaining low variability for all nine microorganisms. The automatic UF was used to demonstrate that filtration can be scaled up to field deployment and the collection of 200-liter lake water samples.

Michigan, New York, Ohio, Wisconsin

Natural inactivation of Escherichia coli in anoxic and reduced groundwater

Aims Inactivation rates of E. coli in groundwater have most often been determined in aerobic and oxidized systems. This study examined E. coli inactivation rates in anaerobic and extremely reduced groundwater systems that have been identified as recharge zones. Methods and Results Groundwater from six artesian wells was diverted to above ground, flow through mesocosms that contained laboratory grown E. coli in diffusion chambers. All groundwater was anaerobic and extremely reduced (ORP < - 300 mV). Cells were plated onto mTEC agar during 21 day incubation periods. All data fit a bi-phasic inactivation model, with > 95% of the E. coli population being inactivated < 11.0 hrs (mean k = 0.488&plusmn; 0.188 h &minus;1 ). Conclusions The groundwater geochemical conditions enhanced the inactivation of E. coli to rates approximately 21-fold greater than previously published inactivation rate in groundwater (mean k = 0.023 &plusmn; 0.030 h &minus;1 ). Also, mTEC agar inhibits E. coli growth following exposure to anaerobic and reduced groundwater. Significance and Impact of the Study Aquifer recharge zones with geochemical characteristics observed in this study complement above ground engineered processes (e.g., filtration, disinfection), while increasing the overall indicator microorganism log-reduction rate of a facility.

Journal of Applied Microbiology

A method adapting microarray technology for signature tagged mutagenesis of Dusulfovibrio dusulfuricans G20 and Shewanella oneidensis MR-1 in anaerobic sediment survival experiments

Signature-tagged mutagenesis (STM) is a powerful technique that can be used to identify genes expressed by bacteria during exposure to conditions in their natural environments. To date, there have been no reports of studies in which this approach was used to study organisms of environmental, rather than pathogenic, significance. We used a mini-Tn 10 transposon-bearing plasmid, pBSL180, that efficiently and randomly mutagenized Desulfovibrio desulfuricans G20 in addition to Shewanella oneidensis MR-1. Using these organisms as model sediment-dwelling anaerobic bacteria, we developed a new screening system, modified from former STM procedures, to identify genes that are critical for sediment survival. The screening system uses microarray technology to visualize tags from input and output pools, allowing us to identify those lost during sediment incubations. While the majority of data on survival genes identified will be presented in future papers, we report here on chemotaxis-related genes identified by our STM method in both bacteria in order to validate our method. This system may be applicable to the study of numerous environmental bacteria, allowing us to identify functions and roles of survival genes in various habitats.

Applied and Environmental Microbiology

The effect of natural organic matter on mercury methylation by Desulfobulbus propionicus 1pr3

Methylation of tracer and ambient mercury ( 200 Hg and 202 Hg, respectively) equilibrated with four different natural organic matter (NOM) isolates was investigated in vivo using the Hg-methylating sulfate-reducing bacterium Desulfobulbus propionicus 1pr3. Desulfobulbus cultures grown fermentatively with environmentally representative concentrations of dissolved NOM isolates, Hg[II], and HS − were assayed for absolute methylmercury (MeHg) concentration and conversion of Hg(II) to MeHg relative to total unfiltered Hg(II). Results showed the 200 Hg tracer was methylated more efficiently in the presence of hydrophobic NOM isolates than in the presence of transphilic NOM, or in the absence of NOM. Different NOM isolates were associated with variable methylation efficiencies for either the 202 Hg tracer or ambient 200 Hg. One hydrophobic NOM, F1 HpoA derived from dissolved organic matter from the Florida Everglades, was equilibrated for different times with Hg tracer, which resulted in different methylation rates. A 5 day equilibration with F1 HpoA resulted in more MeHg production than either the 4 h or 30 day equilibration periods, suggesting a time dependence for NOM-enhanced Hg bioavailability for methylation.

Frontiers in Microbiology

The relative contribution of methanotrophs to microbial communities and carbon cycling in soil overlying a coal-bed methane seep

Seepage of coal-bed methane (CBM) through soils is a potential source of atmospheric CH 4 and also a likely source of ancient (i.e. 14 C-dead) carbon to soil microbial communities. Natural abundance 13 C and 14 C compositions of bacterial membrane phospholipid fatty acids (PLFAs) and soil gas CO 2 and CH 4 were used to assess the incorporation of CBM-derived carbon into methanotrophs and other members of the soil microbial community. Concentrations of type I and type II methanotroph PLFA biomarkers (16:1ω8c and 18:1ω8c, respectively) were elevated in CBM-impacted soils compared with a control site. Comparison of PLFA and 16s rDNA data suggested type I and II methanotroph populations were well estimated and overestimated by their PLFA biomarkers, respectively. The δ 13 C values of PLFAs common in type I and II methanotrophs were as negative as −67‰ and consistent with the assimilation of CBM. PLFAs more indicative of nonmethanotrophic bacteria had δ 13 C values that were intermediate indicating assimilation of both plant- and CBM-derived carbon. Δ 14 C values of select PLFAs (−351 to −936‰) indicated similar patterns of CBM assimilation by methanotrophs and nonmethanotrophs and were used to estimate that 35–91% of carbon assimilated by nonmethanotrophs was derived from CBM depending on time of sampling and soil depth.

FEMS Microbiology Ecology

Resistance, resilience, and recovery of dryland soil bacterial communities across multiple disturbances

Dryland ecosystems are sensitive to perturbations and generally slow to recover post disturbance. The microorganisms residing in dryland soils are especially important as they contribute to soil structure and nutrient cycling. Disturbance can have particularly strong effects on dryland soil structure and function, yet the natural resistance and recovery of the microbial components of dryland soils has not been well documented. In this study, the recovery of surface soil bacterial communities from multiple physical and environmental disturbances is assessed. Samples were collected from three field sites in the vicinity of Moab, UT, United States, 6 to 7 years after physical and climate disturbance manipulations had been terminated, allowing for the assessment of community recovery. Additionally, samples were collected in a transect that included three habitat patches: the canopy zone soils under the dominant shrubs, the interspace soils that are colonized by biological soil crusts, and edge soils at the plot borders. Field site and habitat patch were significant factors structuring the bacterial communities, illustrating that sites and habitats harbored unique soil microbiomes. Across the different sites and disturbance treatments, there was evidence of significant bacterial community recovery, as bacterial biomass and diversity were not significantly different than control plots. There was, however, a small number of 16S rRNA gene amplicon sequence variants that distinguished particular treatments, suggesting that legacy effects of the disturbances still remained. Taken together, these data suggest that dryland bacterial communities may possess a previously unappreciated potential to recover within years of the original disturbance.

Frontiers in Microbiology

Anaerobic oxalate degradation: Widespread natural occurrence in aquatic sediments

Significant concentrations of oxalate (dissolved plus particulate) were present in sediments taken from a diversity of aquatic environments, ranging from 0.1 to 0.7 mmol/liter of sediment. These included pelagic and littoral sediments from two freshwater lakes (Searsville Lake, Calif., and Lake Tahoe, Calif.), a hypersaline, meromictic, alkaline lake (Big Soda Lake, Nev.), and a South San Francisco Bay mud flat and salt marsh. The oxalate concentration of several plant species which are potential detrital inputs to these aquatic sediments ranged from 0.1 to 5.0% (wt/wt). In experiments with litter bags, the oxalate content of Myriophyllum sp. samples buried in freshwater littoral sediments decreased to 7% of the original value in 175 days. This suggests that plant detritus is a potential source of the oxalate within these sediments. [ 14 C]oxalic acid was anaerobically degraded to 14 CO 2 in all sediment types tested, with higher rates evident in littoral sediments than in the pelagic sediments of the lakes studied. The turnover time of the added [ 14 C]oxalate was less than 1 day in Searsville Lake littoral sediments. The total sediment oxalate concentration did not vary significantly between littoral and pelagic sediments and therefore did not appear to be controlling the rate of oxalate degradation. However, depth profiles of [ 14 C]oxalate mineralization and dissolved oxalate concentration were closely correlated in freshwater littoral sediments; both were greatest in the surface sediments (0 to 5 cm) and decreased with depth. The dissolved oxalate concentration (9.1 μmol/liter of sediment) was only 3% of the total extractable oxalate (277 μmol/liter of sediment) at the sediment surface. These results suggest that anaerobic oxalate degradation is a widespread phenomenon in aquatic sediments and may be limited by the dissolved oxalate concentration within these sediments.

California, Nevada

Exposure to synthetic hydraulic fracturing waste influences the mucosal bacterial community structure of the brook trout (Salvelinus fontinalis) epidermis

Production of natural gas using unconventional technologies has risen as demand for alternative fuels has increased. Impacts on the environment from waste generated from these processes are largely unexplored. In particular, the outcomes of organismal exposure to hydraulic fracturing waste have not been rigorously evaluated. We evaluated the effects of exposure to surrogate hydraulic fracturing waste (HF waste) on mucosal bacterial community structure of the brook trout ( Salvelinus fontinalis ) epidermis. Brook trout are fish native to streams at risk to HF waste exposure. Here, fish were exposed to four treatments (control, 0.00%; low, 0.01%; medium, 0.10%; and high, 1.0% concentrations) of surrogate HF waste synthesized to mimic concentrations documented in the field. Epidermal mucus samples were collected and assessed 15 days post-exposure to determine if the associated bacterial community varied among treatments. We observed differences in epidermal mucosal bacterial community composition at multiple taxonomic scales among treatments. These community changes reflected compositional differences in taxa dominance and community similarity rather than losses or gains in taxonomic richness. The dominant bacterial genus that explained the greatest variation in community structure between exposed and unexposed fish was Flavobacterium . Two genera associated with salmonid diseases, Flavobacterium and Pseudomonas , were statistically more abundant in high treatments than controls. These results suggest that exposure to low levels of HF waste influences bacterial colonization and may lead to a disruption that favors bacterial populations associated with fish disease.

Pennsylvania

Characterization of pH dependent Mn(II) oxidation strategies and formation of a bixbyite-like phase by Mesorhizobium australicum T-G1

Despite the ubiquity of Mn oxides in natural environments, there are only a few observations of biological Mn(II) oxidation at pH < 6. The lack of low pH Mn-oxidizing bacteria (MOB) isolates limits our understanding of how pH influences biological Mn(II) oxidation in extreme environments. Here, we report that a novel MOB isolate, Mesorhizobium australicum strain T-G1, isolated from an acidic and metalliferous uranium mining area, can oxidize Mn(II) at both acidic and neutral pH using different enzymatic pathways. X-ray diffraction, Raman spectroscopy, and scanning electron microscopy with energy dispersive X-ray spectroscopy revealed that T-G1 initiated bixbyite-like Mn oxide formation at pH 5.5 which coincided with multi-copper oxidase expression from early exponential phase to late stationary phase. In contrast, reactive oxygen species (ROS), particularly superoxide, appeared to be more important for T-G1 mediated Mn(II) oxidation at neutral pH. ROS was produced in parallel with the occurrence of Mn(II) oxidation at pH 7.2 from early stationary phase. Solid phase Mn oxides did not precipitate, which is consistent with the presence of a high amount of H 2 O 2 and lower activity of catalase in the liquid culture at pH 7.2. Our results show that M. australicum T-G1, an acid tolerant MOB, can initiate Mn(II) oxidation by varying its oxidation mechanisms depending on the pH and may play an important role in low pH manganese biogeochemical cycling.

Frontiers in Microbiology

Mixotrophic iron-oxidizing Thiomonas isolates from an acid mine drainage-affected creek

Natural attenuation of heavy metals occurs via coupled microbial iron cycling and metal precipitation in creeks impacted by acid mine drainage (AMD). Here, we describe the isolation, characterization, and genomic sequencing of two iron-oxidizing bacteria (FeOB) species: Thiomonas ferrovorans FB-6 and Thiomonas metallidurans FB-Cd, isolated from slightly acidic (pH 6.3), Fe-rich, AMD-impacted creek sediments. These strains precipitated amorphous iron oxides, lepidocrocite, goethite, and magnetite or maghemite and grew at a pH optimum of 5.5. While Thiomonas spp. are known as mixotrophic sulfur oxidizers and As oxidizers, the FB strains oxidized Fe, which suggests they can efficiently remove Fe and other metals via coprecipitation. Previous evidence for Thiomonas sp. Fe oxidation is largely ambiguous, possibly because of difficulty demonstrating Fe oxidation in heterotrophic/mixotrophic organisms. Therefore, we also conducted a genomic analysis to identify genetic mechanisms of Fe oxidation, other metal transformations, and additional adaptations, comparing the two FB strain genomes with 12 other Thiomonas genomes. The FB strains fall within a relatively novel group of Thiomonas strains that includes another strain (b6) with solid evidence of Fe oxidation. Most Thiomonas isolates, including the FB strains, have the putative iron oxidation gene cyc2 , but only the two FB strains possess the putative Fe oxidase genes mtoAB . The two FB strain genomes contain the highest numbers of strain-specific gene clusters, greatly increasing the known Thiomonas genetic potential. Our results revealed that the FB strains are two distinct novel species of Thiomonas with the genetic potential for bioremediation of AMD via iron oxidation.

Thuringia

Bacterial community diversity and potential eco-physiological roles in toxigenic blooms composed of Microcystis, Aphanizomenon or Planktothrix

Cyanobacterial toxicity, cyanotoxins, and their impact on aquatic ecosystems and human health are well documented. In comparison, less is known about bloom-associated bacterial communities. Co-occurring bacteria can influence bloom development, physiology and collapse, and may also provide a niche for pathogenic bacteria. Existing research focuses on the cyanosphere of Microcystis -dominated blooms, despite the increasing prevalence of filamentous genera ( Aphanizomenon and Planktothrix ). This pilot study aimed to broaden our understanding of the bacterial consortia attached to morphologically distinct cyanobacteria (coccoid and filamentous) dominating phytoplankton communities and to explore their potential roles in amplifying the impacts of cyanobacterial blooms. We investigated four shallow freshwater bodies across three continents and two climate zones: an urban pond in the USA, a dammed reservoir and a natural lake in Poland, and an urban water body in Singapore. Amplicon sequencing (16S rRNA gene) was used to characterize bacterial communities, while shotgun metagenomics identified nitrogen- and phosphorus-cycling genes to infer potential eco-physiological functions. Cyanobacteria dominated bacterioplankton assemblages at all sites (>35.6%), with bloom composition influencing toxigenic profiles. A mixed bloom of Microcystis , Snowella , and Aphanizomenon had the broadest range of cyanotoxin synthetase genes ( mcy E, cyr J, ana F and sxt A). Microcystis blooms correlated with increased Roseomonas , while Planktothrix co-occurred with Flavobacterium – both bacteria likely contribute to nutrient-cycling within blooms and represent potential opportunistic pathogens for aquatic organisms and humans. The Microcystis cyanosphere exhibited the highest number of significant positive correlations with bacteria (19 relations), compared to Planktothrix and Aphanizomenon (11 and 2 relations, respectively). Non-diazotrophic blooms of Microcystis and Planktothrix showed greater abundances of nitrogen – ( ure B, gln A, nar B, and nar HZ) and phosphorus-cycling genes ( pho BHPR and ppk 1), indicating a strong dependence on associated bacteria for nutrient acquisition compared to diazotrophic Aphanizomenon . These findings suggest that Aphanizomenon -dominated blooms may be sustained by simpler microbiomes. Our results provide preliminary evidence of cyanosphere heterogeneity potentially shaped by the dominance or coexistence of three morphologically and eco-physiologically distinct genera of cyanobacteria. A comprehensive knowledge of the taxonomy and functional roles of bloom-associated microbiomes is therefore essential to understand bloom activity, evaluate the environmental threat, and develop effective strategies for prevention and mitigation.

Frontiers in Microbiology

Transport behavior of groundwater protozoa and protozoan-sized microspheres in sandy aquifer sediments

Transport behaviors of unidentified flagellated protozoa (flagellates) and flagellate-sized carboxylated microspheres in sandy, organically contaminated aquifer sediments were investigated in a small-scale (1 to 4-m travel distance) natural-gradient tracer test on Cape Cod and in flow-through columns packed with sieved (0.5-to 1.0-mm grain size) aquifer sediments. The minute (average in situ cell size, 2 to 3 μ m) flagellates, which are relatively abundant in the Cape Cod aquifer, were isolated from core samples, grown in a grass extract medium, labeled with hydroethidine (a vital eukaryotic stain), and coinjected into aquifer sediments along with bromide, a conservative tracer. The 2- μ m flagellates appeared to be near the optimal size for transport, judging from flowthrough column experiments involving a polydispersed (0.7 to 6.2 μ m in diameter) suspension of carboxylated microspheres. However, immobilization within the aquifer sediments accounted for a log unit reduction over the first meter of travel compared with a log unit reduction over the first 10 m of travel for indigenous, free-living groundwater bacteria in earlier tests. High rates of flagellate immobilization in the presence of aquifer sediments also was observed in the laboratory. However, immobilization rates for the laboratory-grown flagellates (initially 4 to 5 μ m) injected into the aquifer were not constant and decreased noticeably with increasing time and distance of travel. The decrease in propensity for grain surfaces was accompanied by a decrease in cell size, as the flagellates presumably readapted to aquifer conditions. Retardation and apparent dispersion were generally at least twofold greater than those observed earlier for indigenous groundwater bacteria but were much closer to those observed for highly surface active carboxylated latex microspheres. Field and laboratory results suggest that 2- μ m carboxylated microspheres may be useful as analogs in investigating several abiotic aspects of flagellate transport behavior in groundwater.

Massachusetts