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Supercooled water brines within permafrost-an unknown ecological niche for microorganisms: a model for astrobiology.

This study describes brine lenses (cryopegs) found in Siberian permafrost derived from ancient marine sediment layers of the Arctic Ocean. The cryopegs were formed and isolated from sediment ~100,000-120,000 years ago. They remain liquid at the in situ temperature of -10 degrees C as a result of their high salt content (170-300 g/L). [(14)C] Glucose is taken up by the cryopeg biomass at -15 degrees C, indicating microbial metabolism at low temperatures in this habitat. Furthermore, aerobic, anaerobic heterotrophs, sulfate reducers, acetogens, and methanogens were detected by most probable number analysis. Two psychrophilic microbes were isolated from the cryopegs, a Clostridium and a Psychrobacter. The closest relatives of each were previously isolated from Antarctica. The cryopeg econiche might serve as a model for extraterrestrial life, and hence is of particular interest to astrobiology.

Antarctic Regions↗

Multiple linear regression analysis of bacterial deposition to polyurethane coatings after conditioning film formation in the marine environment.

Many studies have shown relationships of substratum hydrophobicity, charge or roughness with bacterial adhesion, although bacterial adhesion is governed by interplay of different physico-chemical properties and multiple regression analysis would be more suitable to reveal mechanisms of bacterial adhesion. The formation of a conditioning film of organic compounds adsorbed from seawater affects the properties of substratum surfaces prior to bacterial adhesion, which is a complicating factor in studying the mechanism of bacterial adhesion. In this paper, the impact of conditioning films adsorbed from natural seawater to four polyurethane coatings with different hydrophobicity, elasticity and roughness was studied for three different marine bacterial strains in a multiple linear regression analysis. The water contact angle on hydrophobic coatings decreased on average by 8 degrees and increased on average by the same amount on hydrophilic coatings. These changes were accompanied by increased concentrations of oxygen and nitrogen on the surface as determined by X-ray photoelectron spectroscopy, indicative of adsorption of proteinaceous material. Furthermore, the mean surface roughness increased on average by 4 nm after conditioning film formation. Multiple linear regression analysis revealed that changes in deposition due to conditioning film formation of Marinobacter hydrocarbonoclasticus, Psychrobacter sp. SW5H and Halomonas pacifica in a stagnation-point flow chamber could be explained in a model comprising hydrophobicity and the prevalence of nitrogen-rich components on the surface for the most hydrophobic strain. For the two more hydrophilic strains, deposition was governed by a combination of surface roughness and hydrophobicity. Elasticity was not a factor in bacterial adhesion to conditioning films.

Bacterial Adhesion↗

Recombinant protein production in Antarctic Gram-negative bacteria.

This review reports some results from our laboratory on the setting up of a psychrophilic expression system for the homologous/heterologous protein production in cold-adapted bacteria by using natural plasmids as cloning vectors. By screening some Antarctic bacteria for the presence of extrachromosomal elements, we identified three new plasmids, pMtBL from Pseudoalteromonas haloplanktis TAC125, and pTAUp and pTADw, from Psychrobacter sp. TA144. The latter autoreplicating elements were isolated, cloned, and fully sequenced and their molecular characterisation was carried out; however, we focused our attention on the small multicopy plasmid, pMtBL, from the Gram-negative P. haloplanktis TAC125 strain. This episome turned out to be an interesting extrachromosomal element, since it displays unique molecular features as its transcriptional inactivity. Being cryptic, the inheritance of pMtBL totally relied on the efficiency of its replication function. This function was bound to a region of about 850 bp, identified by an in vivo assay based on the possibility to efficiently mobilize plasmidic DNA from a mesophilic donor (Escherichia coli) to psychrophilic recipient by intergeneric conjugation. This information was instrumental in the construction of a shuttle vector, able to replicate either in E. coli or in several cold-adapted hosts (clone Q). Since the conversion of a cloning system into an expression vector requires the insertion of transcription and translation regulative sequences, the corresponding signals from the aspartate aminotransferase gene isolated from P. haloplanktis TAC125 were inserted, generating the pFF vector. To investigate the possibility of obtaining recombinant proteins in this cold-adapted host, we used the psychrophilic alpha-amylase from the Antarctic bacterium P. haloplanktis TAB23 (previously known as Alteromonas haloplanktis A23) as a model enzyme to be produced. Our results demonstrate that the cold-adapted enzyme was not only produced but also efficiently secreted by the recombinant PhTAC125 cells. The described expression system represents the first example of heterologous protein production based on a true cold-adapted replicon.

Calcium Chloride↗

Marine bacteria associated with the Korean brown alga, Undaria pinnatifida.

Several marine bacterial strains were isolated from Undaria pinnatifida (Miyok in Korean). Sixty-six strains were isolated on R2A agar media at 10 degrees and identified by a phylogenetic analysis of the 16S rRNA gene sequences. They were grouped into 10 different sequence types based on the initial sequence analysis of the 5' domain of the gene (approximately 500 bp). Full sequences of 16S rRNA gene were obtained from one strain in each sequence type and the species-affiliation was determined using phylogenetic and sequence similarity analyses. The results of the analyses indicated that they were closely related to Psychrobacter aquimaris, P. celer, P. nivimaris, P. pulmonis, Psychromonas arctica or Bacillus psychrodurans. These bacteria are marine or psychrotrophic bacteria. Because the sporophytes of U.pinnatifida are cultured on the costal area during winter, the U. pinnatifida-associated bacteria appeared to grow at low temperatures. U. pinnatifida sporophytes can be a good source for the isolation of psychrotrophic bacteria.

Alteromonadaceae↗

Degradation of crude oil by an arctic microbial consortium.

The ability of a psychrotolerant microbial consortium to degrade crude oil at low temperatures was investigated. The enriched arctic microbial community was also tested for its ability to utilize various hydrocarbons, such as long-chain alkanes (n-C24 to n-C34), pristane, (methyl-)naphthalenes, and xylenes, as sole carbon and energy sources. Except for o-xylene and methylnaphthalenes, all tested compounds were metabolized under conditions that are typical for contaminated marine liquid sites, namely at pH 6-9 and at 4-27 degrees C. By applying molecular biological techniques (16S rDNA sequencing, DGGE) nine strains could be identified in the consortium. Five of these strains could be isolated in pure cultures. The involved strains were closely related to the following genera: Pseudoalteromonas (two species), Pseudomonas (two species), Shewanella (two species), Marinobacter (one species), Psychrobacter (one species), and Agreia (one species). Interestingly, the five isolated strains in different combinations were unable to degrade crude oil or its components significantly, indicating the importance of the four unculturable microorganisms in the degradation of single or of complex mixtures of hydrocarbons. The obtained mixed culture showed obvious advantages including stability of the consortium, wide range adaptability for crude oil degradation, and strong degradation ability of crude oil.

Arctic Regions↗

Changes in aerobic microflora of skin and gills of Mediterranean sardines (Sardina pilchardus) during storage in ice.

Sardines from the Adriatic Sea were examined fresh and after 4 and 8 days of storage in ice. A total of 1500 strains isolated were identified from the gills and the surface of the fish. Pseudomonadaceae, Neisseriaceae, Flavobacterium/Cytophaga, Enterobacteriaceae, coryneform bacteria and Micrococcaceae were the most common bacteria in fresh fish. During storage the pseudomonads (mainly the non-fluorescent strains) increased and became the dominating microflora; the Neisseriaceae (Moraxella, Psychrobacter and Acinetobacter) showed a distinct increase during the first 4 days in ice; the percentage of the other bacterial groups clearly decreased. On the gills the quantitative changes in the microflora were less pronounced than on the surface.

Animals↗

Comparison of antibacterial activity in the hemocytes of different crustacean species.

Antibacterial activity in hemocytes of the squat lobster, Galathea strigosa, the Norway lobster, Nephrops norvegicus, the common shrimp, Crangon crangon, and the giant Antarctic isopod, Glyptonotus antarcticus, was investigated in vitro. For all species, the marine bacterium, Psychrobacter immobilis, was used as the test organism, although with G. antarcticus, the Gram positive bacteria, Planococcus citreus and BS 68 (an isolate from Antarctic waters), were also used. Hemocyte lysate supernatants (HLS) from all four species reduced the viable count of test bacteria over a period of 4 hr showing that their hemocytes contain factors able to neutralize bacteria in vitro. However, comparison of responses produced by serially diluted samples of HLS from G. strigosa, N. norvegicus and C. crangon, revealed that activity (per unit protein) is weaker than for Carcinus maenas. Using G. antarcticus, positive activity was also observed against P. citreus and BS 68; with the response effective against all of the bacteria at both 0 degree C and 20 degrees C. These results show that: (1) the hemocytes from a range of crustacean species contain factor(s) able to neutralize bacteria in vitro; (2) antibacterial potency varies from species to species; and (3) antibacterial immunity in at least one polar invertebrate functions at low temperature.

Animals↗

Structural organization of a Bacillus subtilis operon encoding menaquinone biosynthetic enzymes.

Menaquinone (MK) is a non-protein component of the Bacillus subtilis (Bs) electron transport chain synthesized from chorismate through a series of MK-specific reactions. The genes encoding biosynthesis of the naphthoquinone ring of MK are clustered at 273 degrees on the Bs chromosome. A 3.9-kb region capable of rescuing men mutants blocked in the early stages of MK biosynthesis was sequenced and found to contain three major open reading frames (ORFs). The first ORF (menF) has a predicted size of 51.8 kDa and 34% amino-acid identity with the isochorismate synthases of Escherichia coli (EntC) and Aeromonas hydrophila (AmoA), ORF2 (menD) a predicted size of 60.2 kDa and 21% identity with MenD of E. coli. ORF3 has a predicted size of 21.4 kDa and 29% identity to triacylglycerol lipase of Psychrobacter immobilis. No sequence corresponding to menC was identified. Plasmid integrational studies of the men gene cluster had suggested the presence of promoters secondary to the previously identified p1 men promoter. Sequence analysis revealed a putative promoter region upstream from ORF3.

Bacillus subtilis↗

Evaluation of pyrrolidonyl arylamidase for the identification of nonfermenting Gram-negative rods.

To evaluate the activity of pyrrolidonyl arylamidase (PYR) for the differentiation and identification of nonfermenting gram negative rods (NFGNR), 293 isolates were tested. A 24 h culture of each test organism was prepared. From this a 108-109 cfu/mL suspension was added to 0.25 mL of sterile physiologic solution. A PYR disk was then added and the test was incubated for 30 minutes at 35-37 degrees C, at environmental atmosphere. Reading was done by adding 1 drop of cinnamaldehyde reagent. Strains of Acinetobacter baumannii, Acinetobacter haemolyticus, Alcaligenes faecalis, Bergeyella zoohelcum, Bordetella bronchiseptica, Bordetella hinzii, Brevundimonas diminuta, Brevundimonas vesicularis, Brucella ovis, Brucella spp., Brucella suis, Burkholderia cepacia complex, Moraxella catarrhalis, Moraxella lacunata, Moraxella nonliquefaciens, Moraxella osloensis, Oligella ureolytica, Pseudomonas alcaligenes, Pseudomonas mendocina, Pseudomonas pseudoalcaligenes, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas Vb3, Psychrobacter phenylpyruvicus, and Stenotrophomonas maltophilia were PYR negative. On the other hand Achromobacter piechaudii, Achromobacter denitrificans, Achromobacter xylosoxidans, Burkholderia gladioli, Chryseobacterium gleum-indologenes, Comamonas testosroni, Cupriavidus pauculus, Delftia acidovorans, Elizabethkingia meningoseptica, Myroides spp., Ochrobactrum anthropi, Pseudomonas oryzihabitans, Ralstonia pickettii, Rhizobium radiobacter, Shewanella spp., Sphingobacterium multivorum, Sphingobacterium spiritivorum, and Weeksella virosa were PYR positive. Finally, Acinetobacter lwoffii, Pseudomonas aeruginosa, Pseudomonas fluorescens, Roseomonas spp., and Sphingomonas paucimobilis-parapaucimobilis were PYR variable. PYR testing should be considered as a useful tool to facilitate the identification of NFGNR.

Aminopeptidases↗

Dynamics of antibiotic resistance genes during manure composting: Reduction in herbivores manure and accumulation in carnivores.

The elevated levels of antibiotic resistance genes (ARGs) in livestock manure represent a significant threat to both the environment and human health. Composting has been recognized as an effective strategy to mitigate the abundance of ARGs in manure. However, notable rebounds in ARGs abundance have been observed during this process. This study explored the changes in ARGs abundance and the underlying influencing factors during the composting of carnivore (chicken and pig) and herbivore (sheep and cow) manures, along with mushroom residues. The findings revealed that the total relative abundance of ARGs increased by 6.96 and 10.94 folds in chicken and pig manure composts, respectively, whereas it decreased by a remarkable 91.72% and 98.37% in sheep and cow manure composts. Nitrogen content emerged as the primary physicochemical factors governing the abundance of ARGs in chicken and pig manure composts. Conversely, carbon content played a pivotal role in determining ARGs abundance in chicken and pig manure composts. Furthermore, the presence of dominant hosts, such as Corynebacterium, Bacillus, and Clostridium, along with emerging bacteria like Thermobifida, Saccharomonospora, and Actinomadura, contributed significantly to the enrichment of total ARGs, including tetG, tetO, tetX, and sul2, in chicken and pig manure composts. The coexistence of these genes with mobile genetic elements and a plethora of host bacteria, coupled with their high abundance, renders them particularly high-risk ARGs. On the other hand, the observed decrease in the abundance of total ARGs in sheep and cow manure composts can be attributed to the decline in the population of host bacteria, specifically Atopostipes, Psychrobacter, and Corynebacterium. Collectively, these results provide crucial insights into the management of ARGs risks and offer essential theoretical support for enhancing the safe utilization of organic fertilizer in agriculture.

Manure↗

Aerobic bacterial flora of the vagina and prepuce of California sea lions (Zalophus californianus) and investigation of associations with urogenital carcinoma.

To investigate the association between genital bacterial infection and urogenital carcinoma in California sea lions (Zalophus californianus), vaginal and preputial swabs for bacterial isolation were taken from 148 free-ranging and 51 stranded California sea lions including 16 animals with urogenital carcinoma. Cytological examination of vaginal or preputial smears showed a majority (65.5%, 57/87) of animals examined had mild or no inflammation. Aerobic bacteria were isolated from 116 (78.4%) wild sea lions and 100% of stranded animals. A total of 403 isolates were identified representing 51 unique bacterial species. The median number of isolates per animal increased with age in the wild group, but there was no difference in the number of isolates per animal between wild and stranded adults. The most common bacteria isolated from the wild sea lions were Psychrobacter phenylpyruvicus (39 isolates), non-hemolytic Streptococcus (35 isolates), Corynebacterium spp. (30 isolates), and Escherichia coli (20 isolates). More bacterial species were isolated from stranded animals than wild animals (33 versus 26) and there was significantly less growth of P. phenylpyruvicus, Corynebacterium spp., and Moraxella-like spp. in the stranded animals. Beta-hemolytic Streptococcus was the only bacterium significantly associated with urogenital carcinomas in California sea lions, but only in females.

Age Factors↗

Change in the composition of the microflora on vacuum-packaged beef during chiller storage.

Five fresh beefcut samples were divided into 10 pieces, respectively, vacuum-packaged and stored at 2 degrees C for up to 6 weeks. The average pH values of five pieces from five beef samples were 5.62 +/- 0.04 at the start of storage and 5.12 +/- 0.07 after 6 weeks of storage. The pieces were homogenized, diluted and cultivated at weekly intervals on glucose-blood-liver (BL), de Man Rogosa and Sharpe (MRS) and trypticase soya (TS) agars at 7 degrees C for 10-14 days. From plates with 30-300 colonies, or the highest number if below 30, 15 colonies were randomly picked from each of the three media used for each beef sample. Based on morphologies, SDS-PAGE whole-cell protein profiles and physio-chemical characteristics, a total of 1493 strains isolated were identified as Brochothrix thermosphacta (64), Carnobacterium divergens (79), Carnobacterium piscicola (27), Lactobacillus algidus (637), Lactobacillus sp. (4), Lactococcus piscium (270), Leuconostoc gelidum (375), Acinetobacter (3), Aeromonas (1), Bacillus (10), Corynebacterium (3), Enterobacteriaceae (1), Pseudomonas (13) or Psychrobacter (6). A wider range of organisms was isolated from TS (13 organisms) and BL (7) agars than from MRS agar (3). Leuc. gelidum, Lc. piscium and L. algidas increased in numbers during the first 3 weeks of storage from about 5 x 10(3) cfu/g to the level of about 10(8) cfu/g and persisted at this level thereafter. C. divergens and C. piscicola were inconsistently detected, but seemed to persist at the relatively low level of about 5 x 10(7) cfu/g during the last 3 weeks of storage. B. thermosphacta increased to the level of about 7 x 10(5) cfu/g during the first 2 weeks of storage, but was not detected thereafter. Remaining organisms were detected sporadically at levels of <3.5 x 10(2) cfu/g during the first 2 weeks of storage.

Agar↗

The microbial ecology of processing equipment in different fish industries-analysis of the microflora during processing and following cleaning and disinfection.

The microflora adhering to the processing equipment during production and after cleaning and disinfecting procedures was identified in four different processing plants. A total of 1009 microorganisms was isolated from various-agar plates and identified. A stepwise procedure using simple phenotypic tests was used to identify the isolates and proved a fast way to group a large collection of microorganisms. Pseudomonas, Neisseriaceae, Enterobactericeae, Coryneform, Acinetobacter and lactic acid bacteria dominated the microflora of cold-smoked salmon plants, whereas the microflora in a plant processing semi-preserved herring consisted of Pseudomonas, Alcaligenes and Enterobactericeae. Psychrobacter, Staphylococcus and yeasts were found in a caviar processing plant. Overall, many microorganisms that are often isolated from fish were also isolated from the fish processing plants. However, some selection depending on processing parameters occurred, since halo- and osmo-tolerant organisms dominated in the caviar processing. After cleaning and disinfection, yeasts, Pseudomonas, Neisseriaceae and Alcaligenes remained in smokehouses, yeasts and Pseudomonas in the herring plant and Pseudomonas, Staphylococcus and yeasts in the caviar plant. The dominant adhering organisms after cleaning and disinfection were pseudomonads and yeasts independently of the microflora during processing. Knowledge of the adhering microflora is essential in the Good Hygienic Practises programme of food processing plants, as the development and design of improved cleaning and disinfecting procedures should target the microorganisms persisting and potentially contaminating the product.

Animals↗

Predictive models as means to quantify the interactions of spoilage organisms.

The purpose of this paper is to quantify the interactions of some groups of spoilage organisms that can be usually found in refrigerated meat stored in air, such as: Enterobacteriaceae, Pseudomonas, Acinetobacter, Psychrobacter, Shewanella, Carnobacterium, Lactobacillus, Leuconostoc, Brochothrix and Kurthia spp. The growth of these organisms was studied in the range of temperature 2-11 degrees C and pH 5.2-6.4, which is characteristic of refrigerated meat. The main growth parameters (maximum specific growth rate and lag time) were modelled by multivariate quadratic polynomials of temperature and pH. The interactions of the organisms were analyzed by comparing their growth models obtained in isolation with those obtained in mixture. The difference between the models was quantified by statistical F-values which were used to measure how much the growth of an organism or group of organisms was affected by others and which of them dominated their joint growth.

Bacteria↗

Styelins, broad-spectrum antimicrobial peptides from the solitary tunicate, Styela clava.

Two novel phenylalanine-rich antimicrobial peptides, Styelin A and Styelin B, were purified from the hemocytes of Styela clava. The peptides had very similar masses (Styelin A, 3685.8; Styelin B, 3700.6) and amino acid compositions, and at least 17 of their first 20 N-terminal residues were identical. Both Styelins were effective against a panel of gram negative and gram positive bacterial pathogens of humans, usually acting with minimal inhibitory concentrations < 1.5 microgram/ml (< 0.5 microM), even in the presence of 100 mM NaCl. Styelins also killed marine bacteria, Psychrobacter immobilis and Planococcus citreus, in media containing 0.4 M NaCl. The presence of antimicrobial peptides (Styelins) in tunicate hemocytes is evidence that such molecules are ancient mediators of host defense within the vertebrate lineage. Peptide antibiotics from marine organisms could afford design template for the development of topical microbicides that manifest broad-spectrum antibacterial activity in the presence of physiological or elevated NaCl concentrations.

Amino Acids↗

Reversed unidirectional transport in a Janus polyurethane/alginate dressing for directional postbiotic delivery to infected wounds.

Probiotic-derived postbiotics exhibit significant potential for infected wound control; however, their effective and localized delivery at wound sites remains a challenge. This study developed a polyurethane/alginate composite nonwoven via electrospinning to establish a postbiotic delivery platform for Bifidobacterium bifidum BD-1 (PU/Alg/BD-1). The beaded fibrous hydrophobic PU layer and hydrophilic Alg layer form a wettability gradient, enabling reversed unidirectional fluid transport toward the wound interface while inhibiting backflow. In vitro results showed that PU/Alg/BD-1 exhibited significant antibacterial activity against Staphylococcus aureus and Escherichia coli and good cytocompatibility with a hemolysis rate of <5%. Targeted metabolomic analysis revealed multiple organic acids in the BD-1 metabolites, which contributed to its antibacterial activity. In vivo microbial analysis verified that PU/Alg/BD-1 effectively reduced the relative abundance of Staphylococcus at the wound site while increasing the proportions of Corynebacterium and Psychrobacter. This microbial modulation contributed to infection control in a rat full-thickness infected wound model, accompanied by a shift in the macrophage phenotype and the downregulation of inflammatory factors including IL-6, TNF-&#x3b1;, and TGF-&#x3b2; in the PU/Alg/BD-1 group. Compared with the blank control, conventional gauze, PU/Alg, and BD-1 groups, PU/Alg/BD-1 significantly promoted wound contraction and re-epithelialization and enhanced collagen deposition. Hence, this study provides an effective material construction strategy for the application of probiotic-derived postbiotics to promote wound healing, demonstrates the potential of BD-1 to regulate the wound microenvironment and accelerate healing, and thereby offers a novel approach for the treatment of infected wounds.

Journal Article↗

The effect of dissolved organic carbon on bacterial adhesion to conditioning films adsorbed on glass from natural seawater collected during different seasons.

Adhesion of three marine bacterial strains, i.e. Marinobacter hydrocarbonoclasticus, Psychrobacter sp. and Halomonas pacifica with different cell surface hydrophobicities was measured on glass in a stagnation point flow chamber. Prior to bacterial adhesion, the glass surface was conditioned for 1 h with natural seawater collected at different seasons in order to determine the effect of seawater composition on the conditioning film and bacterial adhesion to it. The presence of a conditioning film was demonstrated by an increase in water contact angle from 15 degrees on bare glass to 50 degrees on the conditioned glass, concurrent with an increase in the amount of adsorbed organic carbon and nitrogen, as measured by X-ray photoelectron spectroscopy. Multiple linear regression analysis on initial deposition rates, with as explanatory variables the temperature, salinity, pH and concentration of dissolved organic carbon (DOC) of the seawater at the time of collection, showed that the concentration of DOC was most strongly associated with the initial deposition rates of the three strains. Initial deposition rates of the two most hydrophilic strains to a conditioning film, increased with the concentration of DOC in the seawater, whereas the initial deposition rate of the most hydrophobic strain decreased with an increasing concentration of DOC.

Adhesiveness↗

Bacterial community in ancient Siberian permafrost as characterized by culture and culture-independent methods.

The microbial composition of ancient permafrost sediments from the Kolyma lowland of Northeast Eurasia was examined through culture and culture-independent approaches. These sediments have been continuously frozen for 5,000 to 2-3 million years. A total of 265 Bacteria 16S rRNA gene sequences were amplified from the permafrost total-community genomic DNA and screened by amplified ribosomal 16S rRNA restriction analysis. Members of three major lineages were found: gamma-Proteobacteria (mostly Xanthomonadaceae), Actinobacteria, and Firmicutes. We also determined partial 16S rRNA gene sequences of 49 isolates from a collection of 462 aerobes isolated from these sediments. The bacteria included Actinomycetales (Arthrobacter and Microbacteriaceae); followed by the Firmicutes (Exiguobacterium and Planomicrobium); the Bacteroidetes (Flavobacterium); the gamma-Proteobacteria (Psychrobacter); and the alpha-Proteobacteria (Sphingomonas). Both culture and culture-independent approaches showed the presence of high and low G+C Gram-positive bacteria and gamma-Proteobacteria. Some of the 16S rRNA gene sequences of environmental clones matched those of Arthrobacter isolates. Two-thirds of the isolates grew at -2.5 degrees C, indicating that they are psychroactive, and all are closely related to phylogenetic groups with strains from other cold environments, mostly commonly from Antarctica. The culturable and non-culturable microorganisms found in the terrestrial permafrost provide a prototype for possible life on the cryogenic planets of the Solar System.

Bacteria↗