[Show abstract][Hide abstract] ABSTRACT: Strain DVT, a halotolerant, Gram-negative, facultatively anaerobic bacterium, was isolated from a hypersaline pond located in Death Valley, California. The cells were non-spore-forming, motile, curved rods (1.0-1.8 x 0.5-0.6 microns) and occurred singly, in pairs or rarely in chains. Strain DVT was oxidase-, catalase-, Voges-Proskauer-, amylase-, gelatinase- and lipase-positive and indole-negative. Nitrate, sulfate and fumarate were not used as electron acceptors. Carbohydrates served as energy sources both aerobically and anaerobically. Strain DVT grew optimally at 37 degrees C (temperature range 20-50 degrees C) with 2.5% NaCl (NaCl range 0-12.5%) and pH 7.3 (pH range of 5.5-8.5) in a glucose/yeast extract medium with a doubling time of 20 min (aerobically) or 41 min (anaerobically). The end products of glucose fermentation were ethanol, isobutyrate, propionate, lactate, formate and CO2. Strain DVT was resistant to penicillin, D-cycloserine, streptomycin and tetracycline (200 micrograms ml-1). The G + C content was 50 mol%. 16S rRNA gene sequence analysis indicated that it was closely related to Salinivibrio costicola (97.7%) and this was confirmed by DNA-DNA hybridization (93% relatedness). However, phenotypic characteristics such as halotolerance, gas production, growth at 50 degrees C, antibiotic resistance, sugar-utilization spectrum and phylogenetic signatures are sufficiently different from Salinivibrio costicola to warrant designating strain DVT as a new subspecies of Salinivibrio costicola, Salinivibrio costicola subsp. vallismortis subsp. nov. (= DSM 8285T).
International journal of systematic and evolutionary microbiology 04/2000; 50 Pt 2:615-22. · 2.11 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A new halotolerant Desulfovibrio, strain CVLT (T = type strain), was isolated from a solar saltern in California. The curved, gram-negative, nonsporeforming cells (0.3 x 1.0-1.3 &mgr;m) occurred singly, in pairs, or in chains, were motile by a single polar flagellum and tolerated up to 12.5% NaCl. Strain CVLT had a generation time of 60 min when grown in lactate-yeast extract medium under optimal conditions (37 degreesC, pH 7.6, 2.5% NaCl). It used lactate, pyruvate, cysteine, or H2/CO2 + acetate as electron donors, and sulfate, sulfite, thiosulfate, or fumarate as electron acceptors. Elemental sulfur, nitrate, or oxygen were not used. Sulfite and thiosulfate were disproportionated to sulfate and sulfide. The G+C content of the DNA was 62 mol%. Phylogenetic analysis revealed that Desulfovibrio fructosovorans was the nearest relative. Strain CVLT is clearly different from other Desulfovibrio species, and is designated Desulfovibrio senezii sp. nov. (DSM 8436).
Archives of Microbiology 10/1998; 170(4):313-7. · 1.91 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A new halotolerant Desulfovibrio, strain CVLT (T = type strain), was isolated from a solar saltern in California. The curved, gram-negative, nonsporeforming cells (0.3
× 1.0–1.3 μm) occurred singly, in pairs, or in chains, were motile by a single polar flagellum and tolerated up to 12.5% NaCl.
Strain CVLT had a generation time of 60 min when grown in lactate-yeast extract medium under optimal conditions (37°C, pH 7.6, 2.5% NaCl).
It used lactate, pyruvate, cysteine, or H2/CO2 + acetate as electron donors, and sulfate, sulfite, thiosulfate, or fumarate as electron acceptors. Elemental sulfur, nitrate,
or oxygen were not used. Sulfite and thiosulfate were disproportionated to sulfate and sulfide. The G+C content of the DNA
was 62 mol%. Phylogenetic analysis revealed that Desulfovibrio fructosovorans was the nearest relative. Strain CVLT is clearly different from other Desulfovibrio species, and is designated Desulfovibrio senezii sp. nov. (DSM 8436).
Archives of Microbiology 08/1998; 170(4):313-317. · 1.91 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: An anaerobic, extremely thermophilic, xylanolytic, non-spore-forming bacterium was isolated from a sediment sample taken from Owens Lake, California, and designated strain OLT (T = type strain). Strain OLT had a Gramnegative reaction and occurred as short rods which sometimes formed long chains containing a few coccoid cells. It grew at 50-80 degrees C, with an optimum at 75 degrees C. The pH range for growth was 5.5-9.0 with an optimum at about pH 7.5. When grown on glucose at optimal conditions, its doubling time was 7.3 h. In addition to glucose, the isolate utilized sucrose, xylose, fructose, ribose, xylan, starch, pectin and cellulose. Yeast extract stimulated growth on carbohydrates but was not obligately required. The end products from glucose fermentation were lactate, acetate, ethanol, H2 and CO2. The G + C content of strain OLT was 36.6 mol%. The 16S rDNA sequence analysis indicated that strain OLT was a member of the subdivision containing Gram-positive bacteria with DNA G + C content of less than 55 mol% and clustered with members of the genus Caldicellulosiruptor. Because strain OLT is phylogenetically and phenotypically different from other members of this genus, it is proposed to designate this isolate Caldicellulosiruptor owensensis sp. nov. Strain OLT is the type strain (= ATCC 700167T).
International journal of systematic bacteriology 01/1998; 48 Pt 1:91-7. · 2.27 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The 16S rRNA gene (rDNA) sequence analysis of four halophilic anaerobes: Halobacteroides halobius, H. lacunaris. Haloanaerobacter (Hb.) chitinovorans and H. acetoethylicus confirmed that they were all members of the family Haloanaerobiaceae. H. lacunaris and H. halobius were found to be more closely related to each other and were distantly related to Sporohalobacter lortetti and the members of the genera Haloanaerobium and Halothermothrix. These data are in agreement with their assignment to the genus Halobacteroides. Further analysis indicated that Hb. chitinovorans was closely affiliated to members of the genus Halobacteroides, and therefore we propose to transfer it to the genus Halobacteroides as H. chitinovorans comb. nov. This transfer would invalidate the genus Haloanaerobacter, as Hb. chitinovorans is the only member of this genus. The 16S rDNA sequence analysis of H. acetoethylicum indicated that it was very closely related to members of the genus Haloanaerobium, viz. Haloanaerobium (Ha.) praevalens, Ha. salsugo, and Ha. alcaliphilum, and hence we propose to transfer it to the genus Haloanaerobium as Ha. acetoethylicus comb. nov.
[Show abstract][Hide abstract] ABSTRACT: A strictly anaerobic, moderately halophilic, gram-negative, rod-shaped bacterium was isolated from Great Salt Lake, Utah, sediments and designated GSLST (T = type strain). Strain GSLST grew optimally at pH 6.7 to 7.0 but had a very broad pH range for growth (pH 5.8 to 10.0). The optimum temperature for growth was 37 degrees C, and no growth occurred at 15 or 55 degrees C. The optimum salt concentration for growth was 10%. Strain GSLST required yeast extract and Trypticase peptone to ferment carbohydrates, pyruvate, and glycine betaine. Strain GSLST was resistant to penicillin, D-cycloserine, tetracycline, and streptomycin. The G + C content of this isolate was 31 mol%. The fermentation products from glucose utilization were acetate, butyrate, lactate, CO2, and H2, and in addition strain GSLST fermented glycine betaine to acetate and trimethylamine. All of these traits distinguish this organism from all previously described halophilic anaerobes. The 16S rRNA gene sequence of strain GSLST was found to be similar to, but also significantly different from, the 16S rRNA sequences of Haloanaerobium salsugo and Haloanaerobium praevalens. Therefore, strain GSLST (= DSM 8275T) is described as a new species, Haloanaerobium alcaliphilum.
International journal of systematic bacteriology 05/1995; 45(2):301-7. · 2.27 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Strictly anaerobic halophiles, namely fermentative, sulfate-reducing, homoacetogenic, phototrophic, and methanogenic bacteria are involved in the oxidation of organic carbon in hypersaline environments. To date, six anaerobic fermentative genera, containing nine species, have been described. Two of them are homoacetogens. Six species belong to the family Haloanaerobiaceae, as indicated by their unique 16S rRNA oligonucleotide sequences. Desulfohalobium retbaense and Desulfovibrio halophilus represent the only two moderately halophilic sulfate reducers so far reported. Among anoxygenic phototrophic anaerobes, a few purple bacteria with optimal growth at salinities between 6 and 11% NaCl have been isolated from hypersaline habitats. They belong to the genera Rhodospirillum, Chromatium, Thiocapsa, and Ectothiorhodospira. The commonest organisms isolated so far are Chromatium salexigens, Thiocapsa halophila, and Rhodospirillum salinarum. Extremely halophilic purple bacteria have most commonly been isolated from alkaline brines and require about 20 to 25% NaCl for optimal growth. They belong to the family Ectothiorodhospiraceae. Their osmoregulation involves synthesis or uptake of compatible solutes such as glycine-betaine that accumulate in their cytoplasm. The existence of methanogens in hypersaline environments is related to the presence of noncompetitive substrates such as methylamines, which originate mainly from the breakdown of osmoregulatory amines. Methanogenesis probably does not contribute to the mineralization of carbohydrates at NaCl concentrations higher than 15%. Above this concentration, sulfate reduction is probably the main way to oxidize H2 (although at rates too low to use up all the H2 formed) and occupies a terminal function kn the degradation of carbohydrates. Three genera and five species of halophilic methylotrophic methanogens have been reported. A bloom of phototrophic bacteria in the marine salterns of Salins-de-Giraud, located on the Mediterranean French coast in the Rhone Delta, is also described.
[Show abstract][Hide abstract] ABSTRACT: The biodegradation of organic matter to form methane and carbon dioxide requires the interactions of diverse populations of bacteria. The roles of each of these organisms in the process and how they interact with each other is understood only in a rudimentary way. This paper describes the investigation of the microbial ecology of the anaerobic degradation of biomass feedstocks.
[Show abstract][Hide abstract] ABSTRACT: Two halophilic anaerobic bacteria, one of which had chitinolytic activity, were isolated from a solar saltern in southern California. These organisms were long, gram-negative, motile, flexible rods. The biochemical and physiological characteristics of these bacteria were very similar but were different from the characteristics of other haloanaerobic bacteria. Both grew at salt concentrations ranging from 0.5 to 5 M and at temperatures ranging from 23 to 50 degrees C. They were sensitive to chloramphenicol but resistant to penicillin, carbenicillin, d-cycloserine, streptomycin, and tetracycline. An analysis of DNAs and whole-cell proteins showed that they were closely related taxonomically and distinguishable from other halophilic anaerobic bacteria. They exhibited 92.3 to 100% DNA homology as determined by DNA-DNA hybridization. The guanine-plus-cytosine contents of their DNAs were 34.8+/-1 mol%. The two isolates, strains W5C8 and W3C1, differed from other halophilic anaerobic bacteria sufficiently to support establishment of a new genus and species, Haloanaerobacter chitinovorans. Strain W5C8 exhibited chitinolytic activity and is designated the type strain. Two chitin-induced extracellular proteins with molecular weights of 38 x 10 and 40 x 10 were detected in strain W5C8.
Applied and Environmental Microbiology 02/1992; 58(1):260-6. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Methanosarcina thermophila TM-1 produced hydrogen during growth on acetate, maintaining a concentration of approx. 0.3 M in the culture, corresponding to a hydrogen partial pressure of approx. 40 Pa. Increasing the partial pressure of hydrogen to 250 Pa and more led to a gradually increasing inhibition of acetate metabolism. No growth was observed when the gas phase contained 2000 Pa or more and acetate metabolism did not occur even after prolonged incubation (more than 2 weeks). M. thermophila was capable of limited consumption of hydrogen. Consumption of low concentrations of hydrogen proceeded simultaneously with acetate utilization. The affinity for hydrogen (K
m=5 M) was within the range normally found for hydrogenutilizing methanogens, while the corresponding V
max (1.2 mol hydrogen consumed mg-1 cells h-1) was orders of magnitude lower.
Archives of Microbiology 11/1991; 157(1):38-42. · 1.91 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The effects of different concentrations of Mg2+, Ca2+, or Na+ on the morphology and growth of Methanosarcina thermophila TM-1 growing on acetate at concentrations comparable with those found in anaerobic digestors was studied. At 30 mm Mg2+ or less, M. thermophila grew as large aggregates that settled rapidly. At 100 mm Mg2+ or more, the bacteria grew as single cells or a mixture of single cells and small aggregates is suspended culture. Mg2+ was necessary for growth and could not be substituted by addition of either Ca2+ or Na+. The optimal Mg2+ concentration was 30 mm and no growth was observed at 400 mm Mg2+. Cultures could be adapted to 300 mm Mg2+ without a change in growth rate. Added Ca2+ was not required for growth and had no effect on cell morphology. Inhibition by Na+ was directly related to the Mg2+ concentration. When the Mg2+ was 0.05 mm or less, 0.35 m Na+ completely inhibited growth. However, more Na+ was required for inhibition at higher Mg2+ concentrations. The same inhibitory effect of Na+ was observed when the temperature was 52C or 45C. The potential for disaggregation of Methanosarcina aggregates in anaerobic digestor environments was discussed.
Applied Microbiology and Biotechnology 01/1991; 35(5):686-689. · 3.69 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: At certain stages in its growth cycle, Methanosarcina mazei produces an enzyme (disaggregatase) that causes aggregates to separate into single cells. M. mazei S-6 and LYC both produce this enzymatic activity, although the specificities of activities differ. The disaggregatase of M. mazei S-6 had little effect on strain LYC cells, but the disaggregatase of M. mazei LYC disaggregated both strain LYC and strain S-6 cells. The disaggregatase of M. mazei LYC was purified by column chromatography, and it apparently consisted of two similar subunits with a combined molecular size of about 180,000 Da. Strain S-6 culture supernatants contained 14 U of activity per liter when activity was measured as uronic acids released from purified cell wall material. When the activity was quantified as the release of uronic acids from boiled M. mazei S-6 cells, the highest activity was found at pH 4.7 and at 35 degrees C.
Applied and Environmental Microbiology 01/1991; 56(12):3693-8. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Methanosarcina barkeri MS and 227 and Methanosarcina mazei S-6 produced acetate when grown on H(2)-CO(2), methanol, or trimethylamine. Marked differences in acetate production by the two bacterial species were found, even though methane and cell yields were nearly the same. M. barkeri produced 30 to 75 mumol of acetate per mmol of CH(4) formed, but M. mazei produced only 8 to 9 mumol of acetate per mmol of CH(4).
Applied and Environmental Microbiology 10/1989; 55(9):2257-61. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The affinity of Methanosarcina barkeri 227 for acetate and hydrogen at different incubation temperatures was investigated. Increasing the temperature from 20 to 37 degrees C resulted in a 4.5-fold increase in K(m) for acetate and a 4.8-fold increase for hydrogen. The corresponding increase in V(max) for acetate was 8.3-fold (5.4-fold for hydrogen). This response implied a decrease in the temperature coefficient (Q(10)) and hence a decrease in the temperature dependency as a function of decreasing substrate concentration.
Applied and Environmental Microbiology 06/1989; 55(5):1262-6. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Marked differences were found for minimum threshold concentrations of acetate catabolism by Methanosarcina barkeri 227 (1.180 mM), Methanosarcina mazei S-6 (0.396 mM), and a Methanothrix sp. (0.069 mM). This indicates that the aceticlastic methanogens responsible for the conversion of acetate to methane in various ecosystems might be different, depending on the prevailing in situ acetate concentrations.
Applied and Environmental Microbiology 03/1989; 55(2):514-5. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The morphology of Methanosarcina mazei was controlled by magnesium, calcium, and substrate concentrations and by inoculum size; these factors allowed manipulation of the morphology and interconversions between pseudosarcinal aggregates and individual, coccoid cells. M. mazei grew as aggregates in medium with a low concentration of catabolic substrate (either 50 mM acetate, 50 mM methanol, or 10 mM trimethylamine) unless Ca and Mg concentrations were high. Growth in medium high in Ca, Mg, and substrate (i.e., 150 mM acetate, 150 mM methanol, or 40 mM trimethylamine) converted pseudosarcinal aggregates to individual cocci. In such media, aggregates separated into individual cells which continued to grow exclusively as single cells during subsequent transfers. Conversion of single cells back to aggregates was complicated, because conditions which supported the aggregated morphology (e.g., low calcium or magnesium concentration) caused lysis of coccoid inocula. We recovered aggregates from coccoid cells by inoculating serial dilutions into medium high in calcium and magnesium. Cells from very dilute inocula grew into aggregates which disaggregated on continued incubation. However, timely transfer of the aggregates to medium low in calcium, magnesium, and catabolic substrates allowed continued growth as aggregates. We demonstrated the activity of the enzyme (disaggregatase) which caused the dispersion of aggregates into individual cells; disaggregatase was produced not only during disaggregation but also in growing cultures of single cells. Uronic acids, the monomeric constituents of the Methanosarcina matrix, were also produced during disaggregation and during growth as coccoids.
Applied and Environmental Microbiology 09/1988; 54(8):2064-8. · 3.95 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Methanohalophilus zhilinae, a new alkaliphilic, halophilic, methylotrophic species of methanogenic bacteria, is described. Strain WeN5T (T = type strain) from Bosa Lake of the Wadi el Natrun in Egypt was designated the type strain and was further characterized. This strain was nonmotile, able to catabolize dimethylsulfide, and able to grow in medium with a methyl group-containing substrate (such as methanol or trimethylamine) as the sole organic compound added. Sulfide (21 mM) inhibited cultures growing on trimethylamine. The antibiotic susceptibility pattern of strain WeN5T was typical of the pattern for archaeobacteria, and the guanine-plus-cytosine content of the deoxyribonucleic acid was 38 mol%. Characterization of the 16S ribosomal ribonucleic acid sequence indicated that strain WeN5T is phylogenetically distinct from members of previously described genera other than Methanohalophilus and supported the partition of halophilic methanogens into their own genus.
International journal of systematic bacteriology 05/1988; 38(2):139-42. · 2.27 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Methanosarcina mazei S-6 grew faster and its morphology changed to individual coccoid cells in medium with elevated concentrations of divalent cations and a large amount of catabolic substrate.
Applied and Environmental Microbiology 08/1987; 53(7):1699-700. · 3.95 Impact Factor