Microbial Mat Samples Research Articles

Assessing microbial diversity in Yellowstone National Park hot springs using a field deployable automated nucleic acid extraction system

Microbial diversity estimation involves extracting nucleic acids from intricate sample matrices. Preparing nucleic acid samples is time-consuming, necessitating effective cell lysis and obtaining pure, inhibitor-free nucleic acid purifications before further use. An automated system offers advantages for field deployment due to its ease of use and quick autonomous results. This is especially beneficial for rapid measurement of in situ microbial diversity in remote areas. Our study aimed to assess microbial diversity of Yellowstone hot springs using a field-deployable lab in a resource-limited remote setting and demonstrate on-site nucleic acid sample processing and sequencing. We collected microbial mat and sediment samples from several Yellowstone National Park hot springs, focusing on the Five Sister Springs (FSS), spring LNN010, and Octopus Spring (OS). The samples were processed for DNA extraction on-site and further sequenced in the lab for microbial diversity. In addition, DNA extracted from one sample was sequenced and analyzed on-site as proof-of-concept. Using either Illumina or Oxford Nanopore Technology sequencing, we found similar microbial diversities. Bacteria (over 90%) were predominant at the FSS and OS sites, with archaea accounting for less than 10%. Metagenomic results were taxonomically categorized based on the closest known organism with a sequenced genome. The dominant archaeal community member was Candidatus Caldiarchaeum subterraneum, and among bacteria, Roseiflexus sp. RS-1 was abundant in mat samples. Interestingly, Bacterium HR17 was also frequently found, suggesting the need for more research on this newly recognized bacterial community member. The presence of Bacterium HR17 in these hot springs suggests its potential role in nitrogen cycling, informing both ecological understanding and industrial potential. This pioneering study assessed the microbiome of Yellowstone hot springs in about 8-9 hours using an automated system for nucleic acid extraction. By its deployment, the system’s value in elucidating the microbial diversity of extreme environments without the need to bring samples to the lab for processing had been highlighted. Sample processing and sequencing had been included in the benefits of the field-deployable lab, and the Nanopore platform had been utilized.

Genome-Centric Metatranscriptomics Reveals Multiple Co-occurring Routes for Hydrocarbon Degradation in Chronically Contaminated Marine Microbial Mats.

Long-term hydrocarbon pollution is a devious threat to aquatic and marine ecosystems. However, microbial responses to chronic pollution remain poorly understood. Combining genome-centric metagenomic and metatranscriptomic analyses of microbial mat samples that experienced chronic hydrocarbon pollution for more than 80 years, we analyzed the transcriptomic activity of alkane and aromatic hydrocarbon degradation pathways at the population level. Consistent with the fluctuating and stratified redox conditions of the habitat, both aerobic and anaerobic hydrocarbon degradation pathways were expressed by taxonomically and metabolically contrasted lineages including members of Bacteroidiales, Desulfobacteraceae, Pseudomonadales; Alcanivoraceae and Halieaceae populations with (photo)-heterotrophic, sulfur- and organohalide-based metabolisms, providing evidence for the co-occurrence and activity of aerobic and anaerobic hydrocarbon degradation pathways in shallow marine microbial mats. In addition, our results suggest that aerobic alkane degradation in long-term pollution involved bacterial families that are naturally widely distributed in marine habitats, but hydrocarbon concentration and composition were found to be a strong structuring factor of their intrafamily diversity and transcriptomic activities.

Preliminary assessment of microbial mats in seawater metal remediation.

Microbial mats have a special role in the removal of metals in marine environment. This study aimed to experimentally determine the efficiency of Cr removal from seawater by microbial mats. The effect of Cr on the microphytobenthic community and the influence of an aerated condition on the removal of metal and microorganisms were also considered. Hence, microbial mat samples were subdivided in 4 groups: submerged in Cr (chromium 2mg/L without aeration), Cr + O2 (chromium 2mg/L with aeration), SW + O2 (filtered seawater with aeration), and a control: SW (filtered seawater with neither chromium nor aeration). Water and microbial mat subsamples were used to determine Cr concentrations, organic matter content, granulometry, physicochemical parameters, chlorophyll a, phaeopigments, and quantitative analysis of the microphytobenthic community. Cr removal efficiency from seawater was 95% for the Cr treatment and 99% for the Cr + O2 treatment. The abundance of cyanobacteria tended to decrease between the initial and final days of the assay, whereas the opposite trend was observed for diatoms. Two aspects are worth noting in the paper: microbial mats efficiently removed Cr from seawater at a concentration of 2mg Cr/L and the removal of Cr by microbial mats was more efficient with water aeration.

Authigenic clays as precursors to carbonate precipitation in saline lakes of Salar de Llamara, Northern Chile

The Salar de Llamara, Atacama Desert is host to saline lakes known as Puquios, which are possible analogs for lakes on early Earth or Mars. Here we examine associations between microbial communities, Mg-clay minerals, and carbonates in microbial mat samples from the Puquios using scanning electron microscopy, energy dispersive X-ray spectroscopy, focused ion beam nanotomography, and transmission electron microscopy. We observe micrometer-scale aggregates of nanocrystalline Mg-clay around cyanobacterial cell sheaths and throughout alveolar networks of microbial extracellular polymeric substances. High-Mn calcite occupies pore spaces and surfaces within the clay matrix. We propose a sequence whereby extracellular polymeric substance networks serve as surfaces for precipitation of Mg-clay, which permineralizes the original microorganisms. Formation of the clay reduces kinetic barriers and provides a suitable substrate for the formation of carbonate minerals. We therefore suggest that authigenic clays are important in the preservation of microbial communities and the precipitation of carbonates in non-marine settings such as the saline lakes of the Atacama Desert.

Comparative computational study to augment UbiA prenyltransferases inherent in purple photosynthetic bacteria cultured from mangrove microbial mats in Qatar for coenzyme Q10 biosynthesis.

Coenzyme Q10 (CoQ10) is a powerful antioxidant with a myriad of applications in healthcare and cosmetic industries. The most effective route of CoQ10 production is microbial biosynthesis. In this study, four CoQ10 biosynthesizing purple photosynthetic bacteria: Rhodobacter blasticus, Rhodovulum adriaticum, Afifella pfennigii and Rhodovulum marinum, were identified using 16S rRNA sequencing of enriched microbial mat samples obtained from Purple Island mangroves (Qatar). The membrane bound enzyme 4-hydroxybenzoate octaprenyltransferase (UbiA) is pivotal for bacterial biosynthesis of CoQ10. The identified bacteria could be inducted as efficient industrial bio-synthesizers of CoQ10 by engineering their UbiA enzymes. Therefore, the mutation sites and substitution residues for potential functional enhancement were determined by comparative computational study. Two mutation sites were identified within the two conserved Asp-rich motifs, and the effect of proposed mutations in substrate binding affinity of the UbiA enzymes was assessed using multiple ligand simultaneous docking (MLSD) studies, as a groundwork for experimental studies.

Absence of canonical trophic levels in a microbial mat.

In modern ecosystems, the carbon stable isotope (δ13 C) ratios of consumers generally conform to the principle "you are what you eat, +1‰." However, this metric may not apply to microbial mat systems where diverse communities, using a variety of carbon substrates via multiple assimilation pathways, live in close physical association and phagocytosis is minimal or absent. To interpret the δ13 C record of the Proterozoic and early Paleozoic, when mat-based productivity likely was widespread, it is necessary to understand how a microbially driven producer-consumer structure affects the δ13 C compositions of biomass and preservable lipids. Protein Stable Isotope Fingerprinting (P-SIF) is a recently developed method that allows measurement of the δ13 C values of whole proteins, separated from environmental samples and identified taxonomically via proteomics. Here, we use P-SIF to determine the trophic relationships in a microbial mat sample from Chocolate Pots Hot Springs, Yellowstone National Park (YNP), USA. In this mat, proteins from heterotrophic bacteria are indistinguishable from cyanobacterial proteins, indicating that "you are what you eat, +1‰" is not applicable. To explain this finding, we hypothesize that sugar production and consumption dominate the net ecosystem metabolism, yielding a community in which producers and consumers share primary photosynthate as a common resource. This idea was validated by confirming that glucose moieties in exopolysaccharide were equal in δ13 C composition to both cyanobacterial and heterotrophic proteins, and by confirming that highly 13 C-depleted fatty acids (FAs) of Cyanobacteria dominate the lipid pool, consistent with flux-balance expectations for systems that overproduce primary photosynthate. Overall, the results confirm that the δ13 C composition of microbial biomass and lipids is tied to specific metabolites, rather than to autotrophy versus heterotrophy or to individual trophic levels. Therefore, we suggest that aerobic microbial heterotrophy is simply a case of "you are what you eat."

Draft Genome Sequence of the Cyanobacterium Synechococcus sp. Strain Nb3U1.

ABSTRACTWe report the 3.5-Mb draft genome sequence of the cyanobacterium Synechococcus sp. strain Nb3U1, which was isolated from a microbial mat sample collected from Nakabusa Hot Spring, Nagano, Japan.

Draft Genome Sequence of Clostridium bowmanii DSM 14206T, Isolated from an Antarctic Microbial Mat.

ABSTRACTClostridium bowmanii type strain DSM 14206 (ATCC BAA-581) was isolated from a microbial mat sample retrieved from Lake Fryxell, Antarctica. This report describes the generation and annotation of the 4.9-Mb draft genome sequence of C. bowmanii DSM 14206T.

Micromonospora tarapacensis sp. nov., a bacterium isolated from a hypersaline lake.

Strain Llam7T was isolated from microbial mat samples from the hypersaline lake Salar de Llamará, located in Taracapá region in the hyper-arid core of the Atacama Desert (Chile). Phenotypic, chemotaxonomic and genomic traits were studied. Phylogenetic analyses based on 16S rRNA gene sequences assigned the strain to the family Micromonosporaceae with affiliation to the genera Micromonospora and Salinispora. Major fatty acids were C17 : 1ω8c, iso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The cell walls contained meso-diaminopimelic acid and ll-2,6 diaminopimelic acid (ll-DAP), while major whole-cell sugars were glucose, mannose, xylose and ribose. The major menaquinones were MK-9(H4) and MK-9(H6). As polar lipids phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol and several unidentified lipids, i.e. two glycolipids, one aminolipid, three phospholipids, one aminoglycolipid and one phosphoglycolipid, were detected. Genome sequencing revealed a genome size of 6.894 Mb and a DNA G+C content of 71.4 mol%. Phylogenetic analyses with complete genome sequences positioned strain Llam7T within the family Micromonosporaceae forming a distinct cluster with Micromonospora (former Xiangella) phaseoli DSM 45730T. This cluster is related to Micromonospora pelagivivens KJ-029T, Micromonospora craterilacus NA12T, and Micromonospora craniellae LHW63014T as well as to all members of the former genera Verrucosispora and Jishengella, which were re-classified as members of the genus Micromonospora, forming a clade distinct from the genus Salinispora. Pairwise whole genome average nucleotide identity (ANI) values, digital DNA-DNA hybridization (dDDH) values, the presence of the diamino acid ll-DAP, and the composition of whole sugars and polar lipids indicate that Llam7T represents a novel species, for which the name Micromonospora tarapacensis sp. nov. is proposed, with Llam7T (=DSM 109510T,=LMG 31023T) as the type strain.

Benthic Bacterial Diversity of High-Altitude Athalassohaline Lakes of the Puna de Atacama (Central Andes)

High-altitude saline lakes (located more than 3000 m a.s.l.) provide multiple environmental extremities for polyextremophilic bacteria. In this study, lake sediment samples from Laguna Santa Rosa and microbial mat samples from the warm water pool of Laguna Verde (Puna de Atacama, Chile) were examined by cultivation and pyrosequencing. The 16S rRNA gene-based identification showed that most of the bacterial isolates belonged to different moderately halophilic Marinobacter species (Proteobacteria). Sample-specific occurrence was characteristic of genera Caenispirillum, Halomonas, Idiomarina, Nitratireductor and Salinarimonas (Proteobacteria), Planococcus, Jeotgalibacillus and Paraliobacillus (Firmicutes), Psychroflexus (Bacteroidetes) and Arthrobacter (Actinobacteria). Using cultivation-independent technique, the benthic communities of both lakes were dominated by sequences related to the phyla Proteobacteria and Bacteroidetes. In addition, representatives of phyla Actinobacteria, Chlorobi, Chloroflexi, Cloacimonetes, Cyanobacteria, Deferribacteres, Deinococcus-Thermus, Firmicutes, Gracilibacteria, Latescibacteria, Lentisphaerae, Parcubacteria, Planctomycetes, Spirochaetae, Tenericutes and Verrucomicrobia were present at least 1% relative abundance. Large differences were explored in the bacterial community composition of the sediment and mat samples at the level of the identified families. It is hypothesized that depending on the type of benthic habitat, different bacteria – involved in the phototrophy based primary production as well as the mainly anaerobic biodegradation of organic materials – can participate in the local carbon, nitrogen, and sulfur cycles.

Genomic and Phenotypic Characterization of Chloracidobacterium Isolates Provides Evidence for Multiple Species.

Chloracidobacterium is the first and until now the sole genus in the phylum Acidobacteriota (formerly Acidobacteria) whose members perform chlorophyll-dependent phototrophy (i.e., chlorophototrophy). An axenic isolate of Chloracidobacterium thermophilum (strain BT) was previously obtained by using the inferred genome sequence from an enrichment culture and diel metatranscriptomic profiling analyses in situ to direct adjustments to the growth medium and incubation conditions, and thereby a defined growth medium for Chloracidobacterium thermophilum was developed. These advances allowed eight additional strains of Chloracidobacterium spp. to be isolated from microbial mat samples collected from Mushroom Spring, Yellowstone National Park, United States, at temperatures of 41, 52, and 60°C; an axenic strain was also isolated from Rupite hot spring in Bulgaria. All isolates are obligately photoheterotrophic, microaerophilic, non-motile, thermophilic, rod-shaped bacteria. Chloracidobacterium spp. synthesize multiple types of (bacterio-)chlorophylls and have type-1 reaction centers like those of green sulfur bacteria. Light harvesting is accomplished by the bacteriochlorophyll a-binding, Fenna-Matthews-Olson protein and chlorosomes containing bacteriochlorophyll c. Their genomes are approximately 3.7 Mbp in size and comprise two circular chromosomes with sizes of approximately 2.7 Mbp and 1.0 Mbp. Comparative genomic studies and phenotypic properties indicate that the nine isolates represent three species within the genus Chloracidobacterium. In addition to C. thermophilum, the microbial mats at Mushroom Spring contain a second species, tentatively named Chloracidobacterium aggregatum, which grows as aggregates in liquid cultures. The Bulgarian isolate, tentatively named Chloracidobacterium validum, will be proposed as the type species of the genus, Chloracidobacterium. Additionally, Chloracidobacterium will be proposed as the type genus of a new family, Chloracidobacteriaceae, within the order Blastocatellales, the class Blastocatellia, and the phylum Acidobacteriota.

The Santorini Volcanic Complex as a Valuable Source of Enzymes for Bioenergy

Marine microbial communities are an untapped reservoir of genetic and metabolic diversity and a valuable source for the discovery of new natural products of biotechnological interest. The newly discovered hydrothermal vent field of Santorini volcanic complex located in the Aegean Sea is gaining increasing interest for potential biotechnological exploitation. The conditions in these environments, i.e., high temperatures, low pH values and high concentration of heavy metals, often resemble harsh industrial settings. Thus, these environments may serve as pools of enzymes of enhanced catalytic properties that may provide benefits to biotechnology. Here, we screened 11 metagenomic libraries previously constructed from microbial mat samples covering the seafloor and the polymetallic chimneys of Kolumbo volcano as well as mat samples from Santorini caldera, to mine, in silico, genes associated with bioenergy applications. We particularly focused on genes encoding biomass hydrolysis enzymes such as cellulases, hemicellulases and lignin-degrading enzymes. A total of 10,417 genes were found for three specific groups of enzymes—i.e., the endoglucanases, the three different beta-glucosidases BGL, bglX and bglB, and the alpha-galactosidases melA, and rafA. Overall, we concluded that the Santorini–Kolumbo volcanic ecosystems constitute a significant resource of novel genes with potential applications in bioenergy that deserve further investigation.

Phosphate-Arsenic Interactions in Halophilic Microorganisms of the Microbial Mat from Laguna Tebenquiche: from the Microenvironment to the Genomes.

Arsenic (As) is a metalloid present in the earth's crust and widely distributed in the environment. Due to its high concentrations in the Andean valleys and its chemical similarity with phosphorus (P), its biological role in Andean Microbial Ecosystems (AMEs) has begun to be studied. The AMEs are home to extremophilic microbial communities that form microbial mats, evaporites, and microbialites inhabiting Andean lakes, puquios, or salt flats. In this work, we characterize the biological role of As and the effect of phosphate in AMEs from the Laguna Tebenquiche (Atacama Desert, Chile). Using micro X-ray fluorescence, the distribution of As in microbial mat samples was mapped. Taxonomic and inferred functional profiles were obtained from enriched cultures of microbial mats incubated under As stress and different phosphate conditions. Additionally, representative microorganisms highly resistant to As and able to grow under low phosphate concentration were isolated and studied physiologically. Finally, the genomes of the isolated Salicola sp. and Halorubrum sp. were sequenced to analyze genes related to both phosphate metabolism and As resistance. The results revealed As as a key component of the microbial mat ecosystem: (i) As was distributed across all sections of the microbial mat and represented a significant weight percentage of the mat (0.17 %) in comparison with P (0.40%); (ii) Low phosphate concentration drastically changed the microbial community in microbial mat samples incubated under high salinity and high As concentrations; (iii) Archaea and Bacteria isolated from the microbial mat were highly resistant to arsenate (up to 500 mM), even under low phosphate concentration; (iv) The genomes of the two isolates were predicted to contain key genes in As metabolism (aioAB and arsC/acr3) and the genes predicted to encode the phosphate-specific transport operon (pstSCAB-phoU) are next to the arsC gene, suggesting a functional relationship between these two elements.

Application of δ15N to trace the impact of penguin guano on terrestrial and aquatic nitrogen cycles in Victoria Land, Ross Sea region, Antarctica

Penguin colonies in Antarctica offer an ideal “natural laboratory” to investigate ecosystem function and the nitrogen (N) cycle. This study assessed the spatial distribution of penguin-derived N from guano and quantitatively assessed its impact on plant N utilization strategies in Victoria Land, Ross Sea region, Antarctica. Soil, moss, and aquatic microbial mats were collected inside and outside an active Adélie penguin (Pygoscelis adeliae) colony and analyzed for δ15N of total and inorganic nitrogen (NH4+-N and NO3−-N). The soil total nitrogen (TN), NH4+-N, and NO3−-N concentrations, as well as their δ15N values were significantly higher in guano-impacted areas than those in guano-free areas, verifying that guano is an important N source at and near penguin colonies. However, even far from the penguin colonies, soil δ15N values resembled those in penguin colonies, suggesting strong spatial impacts of penguin-derived N. The moss impacted by guano was more enriched in δ15N than in guano-free areas. The δ15N values of NH4+-N and NO3−-N in soils covered with moss revealed that the moss might prefer inorganic N in the absence of guano, while the dissolved organic N would become an important source for moss growing on ornithogenic soils. Aquatic microbial mat samples near penguin colonies were 15N-enriched, but 15N-depleted at upland sites.

The Effect of Bacterial Sulfate Reduction Inhibition on the Production and Stable Isotopic Composition of Methane in Hypersaline Environments

The aim of this research was to investigate the competition between methanogens and sulfate-reducing bacteria in hypersaline environments. Samples of photosynthetic microbial mats, both soft mats (salinities of 55–126 ppt) and gypsum-hosted endoevaporite mats (salinities of 77–320 ppt), were obtained from hypersaline environments in California, USA, Mexico and Chile. Methane production was determined from the increase in headspace methane concentration within incubation vials containing mat samples. At the end of the incubation period, the δ13C values of produced methane were measured. Soft microbial mat vials containing molybdate, a specific inhibitor of bacterial sulfate reduction, exhibited dramatically higher methane production rates and higher (enriched in 13C) methane δ13C values than the controls. This suggests that the inhibition of sulfate reduction allowed the methanogens at these sites to use the competitive substrates (H2 and/or acetate) made available. Further, the higher δ13C values of the produced methane suggest that substrates (both competitive and non-competitive) were used to near completion. At the endoevaporite sites, which have much higher salinities than the soft mat sites, methane production was not significantly different and the methane δ13C values either remained the same or decreased (depleted in 13C) with added molybdate. We suggest that substrate availability increased enough to allow for somewhat greater isotopic fractionation resulting in the lower methane δ13C values that were observed, but not enough to significantly increase measured production rates. Where no changes in either methane production rates or δ13C values occurred, we hypothesize that salinity itself was inhibiting sulfate reduction and thus controlling microbe populations and rates of metabolism.

Arsenite Oxidation by a Newly Isolated Betaproteobacterium Possessing arx Genes and Diversity of the arx Gene Cluster in Bacterial Genomes.

Microbes play essential roles in arsenic transformation in the environment. Microbial arsenite oxidation is catalyzed by either of two distantly related arsenite oxidases, referred to as AIO and ARX. The arx genes encoding ARX and its regulatory proteins were originally defined in the genomes of gammaproteobacteria isolated from an alkaline soda lake. The arx gene cluster has been identified in a limited number of bacteria, predominantly in gammaproteobacteria isolated from lakes characterized by high pH and high salinity. In the present study, a novel arsenite-oxidizing betaproteobacterium, strain M52, was isolated from a hot spring microbial mat. The strain oxidized arsenite under both microaerophilic and nitrate-reducing conditions at nearly neutral pH. Genome analysis revealed that the strain possesses the arx gene cluster in its genome and lacks genes encoding AIO. Inspection of the bacterial genomes available in the GenBank database revealed that the presence of this gene cluster is restricted to genomes of Proteobacteria, mainly in the classes Gammaproteobacteria and Betaproteobacteria. In these genomes, the structure of the gene cluster was generally well-conserved, but genes for regulatory proteins were lacking in genomes of strains belonging to a specific lineage. Phylogenetic analysis suggested that ARX encoded in the genomes can be divided into three groups, and strain M52 belongs to a group specific for organisms living in low-salt environments. The ArxA protein encoded in the genome of strain M52 was characterized by the presence of a long insertion, which was specifically observed in the same group of ARX. In clone library analyses with a newly designed primer pair, a diverse ArxA sequence with a long insertion was detected in samples of lake water and hot spring microbial mat, characterized by low salinity and a nearly neutral pH. Among the isolated bacterial strains whose arsenite oxidation has been demonstrated, strain M52 is the first betaproteobacterium that possesses the arx genes, the first strain encoding ARX of the group specific for low-salt environments, and the first organism possessing the gene encoding ArxA with a long insertion.

Comparative metagenomic analyses of a high-altitude Himalayan geothermal spring revealed temperature-constrained habitat-specific microbial community and metabolic dynamics.

Metagenomic surveys across microbial mat (~ 55°C) samples of high-altitude (1760m above sea level) Himalayan geothermal springs have revealed specialized community enriched with niche-specific functions. In this study, we have performed metagenomic sequence-based analyses to get insights into taxonomic composition and functional potential of hyperthermophiles in water (~ 95°C) and sediment samples (78-98°C). Community analyses revealed predominance of thermophilic bacterial and archeal genera dwelling in water in contrast to microbial mats (55°C), namely Methylophilus, Methyloversatilis, Emticicia, Caulobacter, Thermus, Enhydrobacter and Pyrobaculum. Sediment samples having surface temperature (~ 78°C) were colonized by Pyrobaculum and Chloroflexus while genus Massilia was found to be inhabited in high-temperature sediments (~ 98°C). Functional analyses of metagenomic sequences revealed genetic enrichment of genes such as type IV secretion system, flagellar assembly and two-component system in contrast to mats. Furthermore, inter-sample comparison of enriched microbial diversity among water, sediment and microbial mats revealed habitat-specific clustering of the samples within same environment highlighting the role of temperature dynamics in modulating community structure across different habitats in same niche. However, function-based analysis demonstrated site-specific clustering among sediment, microbial mat and water samples. Furthermore, a novel thermophilic genotype of the genus Emticicia (designated as strain MM) was reconstructed from metagenome data. This is a correlative study between three major habitats present in geothermal spring environment, i.e., water, sediment and microbial mats revealing greater phylogenetic and functional dispersion emphasizing changing habitat-specific dynamics with temperature.

Bacterial and Archaeal Viruses of Himalayan Hot Springs at Manikaran Modulate Host Genomes.

Hot spring-associated viruses, particularly the archaeal viruses, remain under-examined compared to bacteriophages. Previous metagenomic studies of the Manikaran hot springs in India suggested an abundance of viral DNA, which prompted us to examine the virus–host (bacterial and archaeal) interactions in sediment and microbial mat samples collected from the thermal discharges. Here, we characterize the viruses (both bacterial and archaeal) from this Himalayan hot spring using both metagenomics assembly and electron microscopy. We utilized four shotgun samples from sediment (78–98°C) and two from microbial mats (50°C) to reconstruct 65 bacteriophage genomes (24–200 kb). We also identified 59 archaeal viruses that were notably abundant across the sediment samples. Whole-genome analyses of the reconstructed bacteriophage genomes revealed greater genomic conservation in sediments (65%) compared to microbial mats (49%). However, a minimal phage genome was still maintained across both sediment and microbial mats suggesting a common origin. To complement the metagenomic data, scanning-electron and helium-ion microscopy were used to reveal diverse morphotypes of Caudovirales and archaeal viruses. The genome level annotations provide further evidence for gene-level exchange between virus and host in these hot springs, and augments our knowledgebase for bacteriophages, archaeal viruses and Clustered Regularly Interspaced Short Palindromic Repeat cassettes, which provide a critical resource for studying viromes in extreme natural environments.

Bacterial Diversity of a Microbial Mat from Hot Spring at Wartawan Beach, Lampung and Its Potential as a Source of Hydrogenases

Biohydrogen produced from thermophilic hydrogenases is an ideal and clean energy sources. As the biggest tectonic area in the world, Indonesia is potential for thermophile isolation. The aims of this study were to analyze the bacterial diversity of a microbial mat from hot spring at Wartawan beach, Lampung and to analyze the potency of microbial mat for hydrogenases, using clone library method. The diversity of 16S rRNA showed that the microbial mat sample contained 9 phyla of bacteria, and dominated by Cyanobacteria and Proteobacteria. These phyla indicate that the bacterial community of the microbial mat consisted of phototrophic and heterotrophic groups. In addition, a microbial mat of Wartawan beach environment might be influenced by marine environment and hydrothermal vent which was indicated by detection of both associated bacteria. The diversity of hydrogenase genes using NiFe hydrogenase (NiFe) and FeFe hydrogenase (FeFe) genes showed that Cyanobacteria was specifically related to NiFe, while Firmicutes was associated with FeFe. Proteobacteria and Bacteroidetes, however, were detected for both genes. The detected hydrogenase genes indicate that the microbial mat from hot spring at Wartawan beach is a promising source for hydrogenases isolation and further applications for biohydrogen production as a renewable energy.

Toxic Cyanobacteria in Svalbard: Chemical Diversity of Microcystins Detected Using a Liquid Chromatography Mass Spectrometry Precursor Ion Screening Method.

Cyanobacteria synthesize a large variety of secondary metabolites including toxins. Microcystins (MCs) with hepato- and neurotoxic potential are well studied in bloom-forming planktonic species of temperate and tropical regions. Cyanobacterial biofilms thriving in the polar regions have recently emerged as a rich source for cyanobacterial secondary metabolites including previously undescribed congeners of microcystin. However, detection and detailed identification of these compounds is difficult due to unusual sample matrices and structural congeners produced. We here report a time-efficient liquid chromatography-mass spectrometry (LC-MS) precursor ion screening method that facilitates microcystin detection and identification. We applied this method to detect six different MC congeners in 8 out of 26 microbial mat samples of the Svalbard Archipelago in the Arctic. The congeners, of which [Asp3, ADMAdda5, Dhb7] MC-LR was most abundant, were similar to those reported in other polar habitats. Microcystins were also determined using an Adda-specific enzyme-linked immunosorbent assay (Adda-ELISA). Nostoc sp. was identified as a putative toxin producer using molecular methods that targeted 16S rRNA genes and genes involved in microcystin production. The mcy genes detected showed highest similarities to other Arctic or Antarctic sequences. The LC-MS precursor ion screening method could be useful for microcystin detection in unusual matrices such as benthic biofilms or lichen.