Domain Archaea Research Articles

Factors influencing Euryarchaeal gut methanogens distribution in dairy cattle in smallholding farms

Guts of ruminants contain symbiotic domains (Eubacteria, Archaea and Eukarya) that aid in the breakdown of consumed carbohydrates from plants to simple molecules that can be absorbed into the ruminant’s bloodstream. Methanogenesis occurs during the gut fermentation and methane gas is released in the final step of biomass degradation from the fermentation chambers. The Archaea that play a major role critical for methane emissions are methanogens and are found freely in the ruminants’ gut. Methane production from ruminants has attracted global attention due to their input on the Green House Gases effect, contribution to global warming and negative effects on farmers’ productivity. The objective of this study was to determine the factors contributing to the methanogens’ gut distribution in dairy cows from smallholder farms using next generation sequencing techniques. A total of 48 samples from smallholding dairy farms were used during this study and were collected from Kenya (Kiambu county) and Tanzania (Lushoto and Rungwe). The collected data samples from the experimental animals were from both the rumen fluid (6) and fecal (42). Samples were analyzed using metagenomic approaches and statistical analysis was undertaken using IBM SPSS statistics software version 28.0.0.0. Results showed that the gut site along the gastrointestinal tract and the feeding regime significantly contributed to the distribution and presence of various methanogenic species (P<0.1). The herd and the genotype had no statistical effect. A total of 12 families were identified. The family Methanobacteriaceae was identified with the leading number (8) of the methanogenic species. A third of the identified families showed presence for at least two methanogenic species with Methanobrevibacter ruminantium being abundant. For proper curbing mechanisms, efforts to reduce methane release should be channeled to the whole gastrointestinal tract and advanced studies carried out on any potential interspecies presence facilitation and/or elimination.

Influence of cosubstrate and hydraulic retention time on the removal of drugs and hygiene products in sanitary sewage in an anaerobic Expanded Granular Sludge Bed reactor

Diclofenac (DCF), ibuprofen (IBU), propranolol (PRO), triclosan (TCS) and linear alkylbenzene sulfonate (LAS) can be recalcitrant in Wastewater Treatment Plants (WWTP). The removal of these compounds was investigated in scale-up (69 L) Expanded Granular Sludge Bed (EGSB) reactor, fed with sanitary sewage from the São Carlos-SP (Brazil) WWTP and 200 mg L−1 of ethanol. The EGSB was operated in three phases: (I) hydraulic retention time (HRT) of 36±4 h; (II) HRT of 20±2 h and (III) HRT of 20±2 h with ethanol. Phases I and II showed no significant difference in the removal of LAS (63 ± 11–65 ± 12 %), DCF (37 ± 18–35 ± 11 %), IBU (43 ± 18–44 ± 16 %) and PRO (46 ± 25–51 ± 23 %) for 13±2–15 ± 2 mg L−1, 106 ± 32–462 ± 294 μg L−1, 166 ± 55–462 ± 213 μg L−1 and 201 ± 113–250 ± 141 μg L−1 influent, respectively. Higher TCS removal was obtained in phase I (72 ± 17 % for 127 ± 120 μg L−1 influent) when compared to phase II (51 ± 13 % for 135 ± 119 μg L−1 influent). This was due to its greater adsorption (40 %) in the initial phase. Phase III had higher removal of DCF (42 ± 10 % for 107 ± 26 μg L−1 influent), IBU (50 ± 15 % for 164 ± 47 μg L−1 influent) and TCS (85 ± 15 % for 185 ± 148 μg L−1 influent) and lower removal of LAS (35 ± 14 % for 12 ± 3 mg L−1 influent) and PRO (−142 ± 177 % for 188 ± 88 μg L−1 influent). Bacteria similar to Syntrophobacter, Smithella, Macellibacteroides, Syntrophus, Blvii28_wastewater-sludge_group and Bacteroides were identified in phase I with relative abundance of 3.1 %–4.7 %. Syntrophobacter was more abundant (15.4 %) in phase II, while in phase III, it was Smithella (12.7 %) and Caldisericum (15.1 %). Regarding the Archaea Domain, Methanosaeta was more abundant in phases I (84 %) and II (67 %), while in phase III it was Methanobacterium (86 %).

Metabolically Active Prokaryotic Complex in Grassland and Forests’ Sod-Podzol under Polycyclic Aromatic Hydrocarbon Influence

Molecular genetic techniques (FISH, RT-PCR, and metagenomic analysis) were used to investigate the comparative functional biodiversity in the prokaryotic complex in grassland and forests’ sod-podzol under polycyclic aromatic hydrocarbon influence. The polluted samples showed a decrease in the biomass of the prokaryotic community representatives and a change in the metabolically active dominants–representatives of the Bacteria and Archaea domains compared to the control samples. The suppression of the metabolic activity of prokaryote cells under the influence of PAHs in sod-podzolic soil under meadow vegetation was more pronounced compared to soils under forest vegetation. The representatives of prokaryotes that are sensitive and resistant to the studied PAHs were identified. The representatives of the phylogenetic groups from the bacterial complex resistant to PAH pollution were Proteobacteria (Alphaproteobacteria), Bacteroidetes, Firmicutes, Chloroflexi, and Thaumarhaeota in the archaeal complex. Representatives of the phylum Acidobacteria and Actinobacteria (Streptosporangiales) are noted among those sensitive to PAH contamination. The presence and expression of the functional alkane monooxygenase (alkB) gene were established in all the experimental variants studied. In the plant variants, the number of copies of alkB genes increased by an order of magnitude and the biomass of metabolically active prokaryotic representatives with the functional alkB gene doubled compared to the unpolluted territories. The copy number index of the alkB gene can be used as one of the parameters when characterizing an ecosystem for the presence of PAH pollutants.

Diverse methanogens, bacteria and tannase genes in the feces of the endangered volcano rabbit (Romerolagus diazi).

BackgroundThe volcano rabbit is the smallest lagomorph in Mexico, it is monotypic and endemic to the Trans-Mexican Volcanic Belt. It is classified as endangered by Mexican legislation and as critically endangered by the IUCN, in the Red List. Romerolagus diazi consumes large amounts of grasses, seedlings, shrubs, and trees. Pines and oaks contain tannins that can be toxic to the organisms which consume them. The volcano rabbit microbiota may be rich in bacteria capable of degrading fiber and phenolic compounds.MethodsWe obtained the fecal microbiome of three adults and one young rabbit collected in Coajomulco, Morelos, Mexico. Taxonomic assignments and gene annotation revealed the possible roles of different bacteria in the rabbit gut. We searched for sequences encoding tannase enzymes and enzymes associated with digestion of plant fibers such as cellulose and hemicellulose.ResultsThe most representative phyla within the Bacteria domain were: Proteobacteria, Firmicutes and Actinobacteria for the young rabbit sample (S1) and adult rabbit sample (S2), which was the only sample not confirmed by sequencing to correspond to the volcano rabbit. Firmicutes, Actinobacteria and Cyanobacteria were found in adult rabbit samples S3 and S4. The most abundant phylum within the Archaea domain was Euryarchaeota. The most abundant genera of the Bacteria domain were Lachnoclostridium (Firmicutes) and Acinetobacter (Proteobacteria), while Methanosarcina predominated from the Archaea. In addition, the potential functions of metagenomic sequences were identified, which include carbohydrate and amino acid metabolism. We obtained genes encoding enzymes for plant fiber degradation such as endo 1,4 β-xylanases, arabinofuranosidases, endoglucanases and β-glucosidases. We also found 18 bacterial tannase sequences.

Microbial analysis of sewage sludge shows closer monitoring needed for landfill waste

A method of high-throughput sequencing of gene libraries was used to identify the microbial communities in samples of sewage sludge at waste treatment facilities in Saint Petersburg, Russia. It was established that the waste had a complicated taxonomic structure and consisted of two main domains: Archaea (8.4% average) and Bacteria (77.6% average). The Archaea domain included three phyla (methanogens): Euryarchaeota, Crenarchaeota and Parvarchaeota. The Bacteria domain was represented by 49 phyla, including Proteobacteria, Firmicutes, Bacteroidetes, Chloroflexi, Cyanobacteria, Planctomycetes, Gemmatimonadetes, Acidobacteria and Verrucomicrobia. Up to 2.8% share of methanogens in sewage sludge deposited in landfills for around 15 years indicated the ongoing destruction of organic matter. In addition, ten pathogenic microorganisms were detected, including Clostridium and Mycobacterium. The results indicated that the national regulatory requirements for waste in Russia need to be reviewed in terms of compiling a list of standardised indicators for environmental monitoring of landfill sites.

Expanding Asgard members in the domain of Archaea sheds new light on the origin of eukaryotes.

The hypothesis that eukaryotes originated from within the domain Archaea has been strongly supported by recent phylogenomic analyses placing Heimdallarchaeota-Wukongarchaeota branch from the Asgard superphylum as the closest known archaeal sister-group to eukaryotes. However, our understanding is still limited in terms of the relationship between eukaryotes and archaea, as well as the evolution and ecological functions of the Asgard archaea. Here, we describe three previously unknown phylum-level Asgard archaeal lineages, tentatively named Sigyn-, Freyr- and Njordarchaeota. Additional members in Wukongarchaeota and Baldrarchaeota from distinct environments are also reported here, further expanding their ecological roles and metabolic capacities. Comprehensive phylogenomic analyses further supported the origin of eukaryotes within Asgard archaea and a new lineage Njordarchaeota was supposed as the known closest branch with the eukaryotic nuclear host lineage. Metabolic reconstruction suggests that Njordarchaeota may have a heterotrophic lifestyle with capability of peptides and amino acids utilization, while Sigynarchaeota and Freyrarchaeota also have the potentials to fix inorganic carbon via the Wood-Ljungdahl pathway and degrade organic matters. Additionally, the Ack/Pta pathway for homoacetogenesis and de novo anaerobic cobalamin biosynthesis pathway were found in Freyrarchaeota and Wukongrarchaeota, respectively. Some previously unidentified eukaryotic signature proteins for intracellular membrane trafficking system, and the homologue of mu/sigma subunit of adaptor protein complex, were identified in Freyrarchaeota. This study expands the Asgard superphylum, sheds new light on the evolution of eukaryotes and improves our understanding of ecological functions of the Asgard archaea.

Novel Glutamate-Putrescine Ligase Activity in Haloferax mediterranei: A New Function for glnA-2 Gene.

The genome of the halophilic archaea Haloferax mediterranei contains three ORFs that show homology with glutamine synthetase (GS) (glnA-1, glnA-2, and glnA-3). Previous studies have focused on the role of GlnA-1, suggesting that proteins GlnA-2 and GlnA-3 could play a different role to that of GS. Glutamine synthetase (EC 6.3.1.2) belongs to the class of ligases, including 20 subclasses of other different enzymes, such as aspartate–ammonia ligase (EC 6.3.1.1), glutamate–ethylamine ligase (EC 6.3.1.6), and glutamate–putrescine ligase (EC 6.3.1.11). The reaction catalyzed by glutamate–putrescine ligase is comparable to the reaction catalyzed by glutamine synthetase (GS). Both enzymes can bind a glutamate molecule to an amino group: ammonium (GS) or putrescine (glutamate–putrescine ligase). In addition, they present the characteristic catalytic domain of GS, showing significant similarities in their structure. Although these proteins are annotated as GS, the bioinformatics and experimental results obtained in this work indicate that the GlnA-2 protein (HFX_1688) is a glutamate–putrescine ligase, involved in polyamine catabolism. The most significant results are those related to glutamate–putrescine ligase’s activity and the analysis of the transcriptional and translational expression of the glnA-2 gene in the presence of different nitrogen sources. This work confirms a new metabolic pathway in the Archaea domain which extends the knowledge regarding the utilization of alternative nitrogen sources in this domain.

Natural attenuation of legacy hydrocarbon spills in pristine soils is feasible despite difficult environmental conditions in the monsoon tropics

The Kimberley region of Western Australia is a National Heritage listed region that is internationally recognised for its environmental and cultural significance. However, petroleum spills have been reported at a number of sites across the region, representing an environmental concern. The region is also characterised as having low soil nutrients, high temperatures and monsoonal rain – all of which may limit the potential for natural biodegradation of petroleum. Therefore, this work evaluated the effect of legacy petroleum hydrocarbons on the indigenous soil microbial community (across the domains Archaea, Bacteria and Fungi) at three sites in the Kimberley region. At each site, soil cores were removed from contaminated and control areas and analysed for total petroleum hydrocarbons, soil nutrients, pH and microbial community profiling (using16S rRNA and ITS sequencing on the Illumina MiSeq Platform). The presence of petroleum hydrocarbons decreased microbial diversity across all kingdoms, altered the structure of microbial communities and increased the abundance of putative hydrocarbon degraders (e.g. Mycobacterium, Acremonium, Penicillium, Bjerkandera and Candida). Microbial community shifts from contaminated soils were also associated with an increase in soil nutrients (notably Colwell P and S). Our study highlights the long-term effect of legacy hydrocarbon spills on soil microbial communities and their diversity in remote, infertile monsoonal soils, but also highlights the potential for natural attenuation to occur in these environments.

Analysis of Carotenoids in Haloarchaea Species from Atacama Saline Lakes by High Resolution UHPLC-Q-Orbitrap-Mass Spectrometry: Antioxidant Potential and Biological Effect on Cell Viability.

Haloarchaea are extreme halophilic microorganisms belonging to the domain Archaea, phylum Euryarchaeota, and are producers of interesting antioxidant carotenoid compounds. In this study, four new strains of Haloarcula sp., isolated from saline lakes of the Atacama Desert, are reported and studied by high-resolution mass spectrometry (UHPLC-Q-Orbitrap-MS/MS) for the first time. In addition, determination of the carotenoid pigment profile from the new strains of Haloarcula sp., plus two strains of Halorubrum tebenquichense, and their antioxidant activity by means of several methods is reported. The effect of biomass on cellular viability in skin cell lines was also evaluated by MTT assay. The cholinesterase inhibition capacity of six haloarchaea (Haloarcula sp. ALT-23; Haloarcula sp. TeSe-41; Haloarcula sp. TeSe-51; Haloarcula sp. Te Se-89 and Halorubrum tebenquichense strains TeSe-85 and Te Se-86) is also reported for the first time. AChE inhibition IC50 was 2.96 ± 0.08 μg/mL and BuChE inhibition IC50 was 2.39 ± 0.09 μg/mL for the most active strain, Halorubrum tebenquichense Te Se-85, respectively, which is more active in BuCHe than that of the standard galantamine. Docking calculation showed that carotenoids can exert their inhibitory activity fitting into the enzyme pocket by their halves, in the presence of cholinesterase dimers.

Functional expression of the eukaryotic proton pump rhodopsin OmR2 in Escherichia coli and its photochemical characterization

Microbial rhodopsins are photoswitchable seven-transmembrane proteins that are widely distributed in three domains of life, archaea, bacteria and eukarya. Rhodopsins allow the transport of protons outwardly across the membrane and are indispensable for light-energy conversion in microorganisms. Archaeal and bacterial proton pump rhodopsins have been characterized using an Escherichia coli expression system because that enables the rapid production of large amounts of recombinant proteins, whereas no success has been reported for eukaryotic rhodopsins. Here, we report a phylogenetically distinct eukaryotic rhodopsin from the dinoflagellate Oxyrrhis marina (O. marina rhodopsin-2, OmR2) that can be expressed in E. coli cells. E. coli cells harboring the OmR2 gene showed an outward proton-pumping activity, indicating its functional expression. Spectroscopic characterization of the purified OmR2 protein revealed several features as follows: (1) an absorption maximum at 533 nm with all-trans retinal chromophore, (2) the possession of the deprotonated counterion (pKa = 3.0) of the protonated Schiff base and (3) a rapid photocycle through several distinct photointermediates. Those features are similar to those of known eukaryotic proton pump rhodopsins. Our successful characterization of OmR2 expressed in E. coli cells could build a basis for understanding and utilizing eukaryotic rhodopsins.

Comparative Genomics Provides Insights into the Genetic Diversity and Evolution of the DPANN Superphylum.

ABSTRACTDPANN is known as highly diverse, globally widespread, and mostly ectosymbiotic archaeal superphylum. However, this group of archaea was overlooked for a long time, and there were limited in-depth studies reported. In this investigation, 41 metagenome-assembled genomes (MAGs) belonging to the DPANN superphylum were recovered (18 MAGs had average nucleotide identity [ANI] values of <95% and a percentage of conserved proteins [POCP] of >50%, while 14 MAGs showed a POCP of <50%), which were analyzed comparatively with 515 other published DPANN genomes. Mismatches to known 16S rRNA gene primers were identified among 16S rRNA genes of DPANN archaea. Numbers of gene families lost (mostly related to energy and amino acid metabolism) were over three times greater than those gained in the evolution of DPANN archaea. Lateral gene transfer (LGT; ∼45.5% was cross-domain) had facilitated niche adaption of the DPANN archaea, ensuring a delicate equilibrium of streamlined genomes with efficient niche-adaptive strategies. For instance, LGT-derived cytochrome bd ubiquinol oxidase and arginine deiminase in the genomes of “Candidatus Micrarchaeota” could help them better adapt to aerobic acidic mine drainage habitats. In addition, most DPANN archaea acquired enzymes for biosynthesis of extracellular polymeric substances (EPS) and transketolase/transaldolase for the pentose phosphate pathway from Bacteria.IMPORTANCE The domain Archaea is a key research model for gaining insights into the origin and evolution of life, as well as the relevant biogeochemical processes. The discovery of nanosized DPANN archaea has overthrown many aspects of microbiology. However, the DPANN superphylum still contains a vast genetic novelty and diversity that need to be explored. Comprehensively comparative genomic analysis on the DPANN superphylum was performed in this study, with an attempt to illuminate its metabolic potential, ecological distribution and evolutionary history. Many interphylum differences within the DPANN superphylum were found. For example, Altiarchaeota had the biggest genome among DPANN phyla, possessing many pathways missing in other phyla, such as formaldehyde assimilation and the Wood-Ljungdahl pathway. In addition, LGT acted as an important force to provide DPANN archaeal genetic flexibility that permitted the occupation of diverse niches. This study has advanced our understanding of the diversity and genome evolution of archaea.

A methylotrophic origin of methanogenesis and early divergence of anaerobic multicarbon alkane metabolism.

Methanogens are considered as one of the earliest life forms on Earth, and together with anaerobic methane-oxidizing archaea, they have crucial effects on climate stability. However, the origin and evolution of anaerobic alkane metabolism in the domain Archaea remain controversial. Here, we present evidence that methylotrophic methanogenesis was the ancestral form of this metabolism. Carbon dioxide-reducing methanogenesis developed later through the evolution of tetrahydromethanopterin S-methyltransferase, which linked methanogenesis to the Wood-Ljungdahl pathway for energy conservation. Anaerobic multicarbon alkane metabolisms in Archaea also originated early, with genes coding for the activation of short-chain or even long-chain alkanes likely evolving from an ethane-metabolizing ancestor. These genes were likely horizontally transferred to multiple archaeal clades including Candidatus (Ca) Bathyarchaeia, Ca. Lokiarchaeia, Ca. Hadarchaeia, and the methanogenic Ca. Methanoliparia.

The Proposed Molecular Mechanisms Used by Archaea for Fe(III) Reduction and Fe(II) Oxidation.

Iron (Fe) is the fourth most abundant element in the Earth’s crust where ferrous Fe [Fe(II)] and ferric Fe [Fe(III)] can be used by archaea for energy conservation. In these archaea-Fe interactions, Fe(III) serves as terminal electron acceptor for anaerobic respiration by a variety of archaea, while Fe(II) serves as electron donor and/or energy sources for archaeal growth. As no Fe is incorporated into the archaeal cells, these redox reactions are referred to as dissimilatory Fe(III) reduction and Fe(II) oxidation, respectively. Dissimilatory Fe(III)-reducing archaea (FeRA) and Fe(II)-oxidizing archaea (FeOA) are widespread on Earth where they play crucial roles in biogeochemical cycling of not only Fe, but also carbon and sulfur. To reduce extracellular Fe(III) (oxyhydr)oxides, some FeRA transfer electrons directly to the Fe(III) (oxyhydr)oxides most likely via multiheme c-type cytochromes (c-Cyts). These multiheme c-Cyts may form the pathways similar to those found in bacteria for transferring electrons from the quinone/quinol pool in the cytoplasmic membrane to the Fe(III) (oxyhydr)oxides external to the archaeal cells. Use of multiheme c-Cyts for extracellular Fe(III) reduction by both Domains of Archaea and Bacteria emphasizes an ancient mechanism of extracellular electron transfer, which is well conserved. Other FeRA, however, reduce Fe(III) (oxyhydr)oxides indirectly via electron shuttles. Similarly, it is proposed that FeOA use pathways to oxidize Fe(II) on the surface of the cytoplasmic membrane and then to transfer the released electrons across the cytoplasmic membrane inward to the O2 and NAD+ in the cytoplasm. In this review, we focus on the latest understandings of the molecular mechanisms used by FeRA and FeOA for Fe(III) reduction and Fe(II) oxidation, respectively.

Lipoproteins Are Responsible for the Pro-Inflammatory Property of Staphylococcus aureus Extracellular Vesicles.

Staphylococcal aureus (S. aureus), a Gram-positive bacteria, is known to cause various infections. Extracellular vesicles (EVs) are a heterogeneous array of membranous structures secreted by cells from all three domains of life, i.e., eukaryotes, bacteria, and archaea. Bacterial EVs are implied to be involved in both bacteria–bacteria and bacteria–host interactions during infections. It is still unclear how S. aureus EVs interact with host cells and induce inflammatory responses. In this study, EVs were isolated from S. aureus and mutant strains deficient in either prelipoprotein lipidation (Δlgt) or major surface proteins (ΔsrtAB). Their immunostimulatory capacities were assessed both in vitro and in vivo. We found that S. aureus EVs induced pro-inflammatory responses both in vitro and in vivo. However, this activity was dependent on lipidated lipoproteins (Lpp), since EVs isolated from the Δlgt showed no stimulation. On the other hand, EVs isolated from the ΔsrtAB mutant showed full immune stimulation, indicating the cell wall anchoring of surface proteins did not play a role in immune stimulation. The immune stimulation of S. aureus EVs was mediated mainly by monocytes/macrophages and was TLR2 dependent. In this study, we demonstrated that not only free Lpp but also EV-imbedded Lpp had high pro-inflammatory activity.

Bacteria and Methanogens in the Human Microbiome: a Review of Syntrophic Interactions.

Methanogens are microorganisms belonging to the Archaea domain and represent the primary source of biotic methane. Methanogens encode a series of enzymes which can convert secondary substrates into methane following three major methanogenesis pathways. Initially recognized as environmental microorganisms, methanogens have more recently been acknowledged as host-associated microorganisms after their detection and initial isolation in ruminants in the 1950s. Methanogens have also been co-detected with bacteria in various pathological situations, bringing their role as pathogens into question. Here, we review reported associations between methanogens and bacteria in physiological and pathological situations in order to understand the metabolic interactions explaining these associations. To do so, we describe the origin of the metabolites used for methanogenesis and highlight the central role of methanogens in the syntrophic process during carbon cycling. We then focus on the metabolic abilities of co-detected bacterial species described in the literature and infer from their genomes the probable mechanisms of their association with methanogens. The syntrophic interactions between bacteria and methanogens are paramount to gut homeostasis. Therefore, any dysbiosis affecting methanogens might impact human health. Thus, the monitoring of methanogens may be used as a bio-indicator of dysbiosis. Moreover, new therapeutic approaches can be developed based on their administration as probiotics. We thus insist on the importance of investigating methanogens in clinical microbiology.

Evaluating the potential for Haloarchaea to serve as ice nucleating particles

Abstract. Aerosols play a crucial role in cloud formation. Biologically derived materials from bacteria, fungi, pollen, lichen, viruses, algae, and diatoms can serve as ice nucleating particles (INPs), some of which initiate glaciation in clouds at relatively warm freezing temperatures. However, determining the magnitude of the interactions between clouds and biologically derived INPs remains a significant challenge due to the diversity and complexity of bioaerosols and limited observations of such aerosols facilitating cloud ice formation. Additionally, microorganisms from the domain Archaea have, to date, not been evaluated as INPs. Here, we present the first results reporting the ice nucleation activity of four species in the class Haloarchaea. Intact cells of Halococcus morrhuae and Haloferax sulfurifontis demonstrated the ability to induce immersion freezing at temperatures up to −18 ∘C, while lysed cells of Haloquadratum walsbyi and Natronomonas pharaonis were unable to serve as immersion INPs. Exposure to heat and peroxide digestion indicated that the INPs of intact cells were driven by organic (H. morrhuae and H. sulfurifontis) and possibly also heat labile materials (H. sulfurifontis only). While halophiles are prominent in hypersaline environments such as the Great Salt Lake and the Dead Sea, other members of the Archaea, such as methanogens and thermophiles, are prevalent in anoxic systems in seawater, sea ice, marine sediments, glacial ice, permafrost, and other cold niches. Archaeal extremophiles are both diverse and highly abundant. Thus, it is important to assess their ability to serve as INPs as it may lead to an improved understanding of biological impacts on clouds.

Prokaryotic and eukaryotic diversity in hydrothermal continental systems.

The term extremophile was suggested more than 30years ago and represents microorganisms that are capable of developing and living under extreme conditions, these conditions being particularly hostile to other types of microorganisms and to humankind. In terrestrial hydrothermal sites, like hot springs, "mud pools", solfataras, and geysers, the dominant extreme conditions are high temperature, low or high pH, and high levels of salinity. The diversity of microorganisms inhabiting these sites is determined by the conditions of the environment. Organisms belonging to the domains Archaea and Bacteria are more represented than the one belonging to Eukarya. Eukarya members tend to be less present because of their lower tolerance to higher temperatures, however, they perform important ecosystem processes when present. Both prokaryotes and eukaryotes have morphological and physical adaptations that allow them to colonize extreme environments. Microbial mats are complex associations of microorganisms that help the colonization of more extreme systems. In this review, a characterization of prokaryotic and eukaryotic organisms that populate terrestrial hydrothermal systems are made.

The largest moss carpet transplant in Antarctica and its bryosphere cryptic biodiversity.

As part of the reconstruction of the Brazilian Antarctic Station on King George Island, three areas of moss carpet were transplanted to minimize the impact of the new facilities on the local biodiversity. A total of 650 m2 of moss carpet was transplanted to neighboring but previously uncolonized locations and has subsequently survived for the last 3years. Antarctic moss carpets typically comprise low moss species diversity and are often monospecific. We investigated the cryptic biodiversity that was transplanted along with the carpets using a metabarcoding approach through high throughput sequencing. We targeted 16S rRNA for Bacteria and Archaea, ITS for Fungi and Viridiplantae and Cox1 for Metazoa. We detected DNA representing 263 taxa from five Kingdoms (Chromista, Fungi, Metazoa, Protista and Viridiplantae), two Domains (Archaea and Bacteria) and 33 Phyla associated with the carpet. This diversity included one Archaea, 189 Bacteria, 24 Chromista, 19 Fungi, eight Metazoa, seven Protista and 16 Viridiplantae. Bacteria was the most abundant, rich and diverse group, with Chromista second in diversity and richness. Metazoa was less diverse but second highest in dominance. This is the first study to attempt transplanting a significant area of moss carpet to minimize anthropogenic environmental damage in Antarctica and to use metabarcoding as a proxy to assess diversity associated with Antarctic moss carpets, further highlighting the importance of such habitats for other organisms and their importance for conservation.

Methanotrophs: Discoveries, Environmental Relevance, and a Perspective on Current and Future Applications.

Methane is the final product of the anaerobic decomposition of organic matter. The conversion of organic matter to methane (methanogenesis) as a mechanism for energy conservation is exclusively attributed to the archaeal domain. Methane is oxidized by methanotrophic microorganisms using oxygen or alternative terminal electron acceptors. Aerobic methanotrophic bacteria belong to the phyla Proteobacteria and Verrucomicrobia, while anaerobic methane oxidation is also mediated by more recently discovered anaerobic methanotrophs with representatives in both the bacteria and the archaea domains. The anaerobic oxidation of methane is coupled to the reduction of nitrate, nitrite, iron, manganese, sulfate, and organic electron acceptors (e.g., humic substances) as terminal electron acceptors. This review highlights the relevance of methanotrophy in natural and anthropogenically influenced ecosystems, emphasizing the environmental conditions, distribution, function, co-existence, interactions, and the availability of electron acceptors that likely play a key role in regulating their function. A systematic overview of key aspects of ecology, physiology, metabolism, and genomics is crucial to understand the contribution of methanotrophs in the mitigation of methane efflux to the atmosphere. We give significance to the processes under microaerophilic and anaerobic conditions for both aerobic and anaerobic methane oxidizers. In the context of anthropogenically influenced ecosystems, we emphasize the current and potential future applications of methanotrophs from two different angles, namely methane mitigation in wastewater treatment through the application of anaerobic methanotrophs, and the biotechnological applications of aerobic methanotrophs in resource recovery from methane waste streams. Finally, we identify knowledge gaps that may lead to opportunities to harness further the biotechnological benefits of methanotrophs in methane mitigation and for the production of valuable bioproducts enabling a bio-based and circular economy.

THE MICROBIAL COMMUNITY COMPOSITION OF AN ANAEROBIC REACTOR IN A SUGAR INDUSTRY WASTEWATER TREATMENT PLANT-FROM CLASSICAL TO NEW APPROACHES

In this study, the microbial characteristics of the anaerobic reactor of a sugar industry wastewater treatment plant were analyzed using cloning, FISH (Fluoresan in situ hybridization) and metagenomic analysis. Samples were obtained from seven different ports of the reactor on the 148th day of operation. The temperature was maintained at mesophilic conditions. The system’s pH range was operated at 6.8. The cloning results showed that most of the bacterial clones belonged to uncultured members of the Bacteria domain. Many archaeal clones were related to uncultured Archaea and Methanosarcina. The FISH method was applied to determine the microbial composition of the samples, which showed that bacterial and archaeal species had nearly equal rates. Rod-shaped cells, long bacilli, coccus and long chains were detected in the samples. After metagenomic analysis, in all samples, Archaea domain members ranged between 60-36% and Bacteria domain members ranged between 58-31%. At the phylum level, in all samples, Euryarchaeota was the most dominant phylum. Proteobacteria (14.8-21.97%) and Actinobacteria (5.53-15.94%) phyla were high in rate. Furthermore, members of Spirochaeotes (0.63-4.82%) and Bacteroidetes (1.72-2.38%) were analyzed in the samples. This study revealed both bacterial and archaeal populations in the reactor of high-concentration organic sugar wastewater. These results will help in the development of more efficient anaerobic treatment systems.