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1.
Most probable number (MPN) estimates indicated that a mean of 4.3×107 and 5×106 bacteria per ml of rumen fluid from a predominantly alfalfa hay-fed steer demethoxylated ferulate and syringate, respectively. After further enrichment from an MPN tube of the highest dilution showing demethoxylation of syringate, strain S195 was isolated using roll tubes with syringate as an added energy source. S195 was an anaerobic, Gram-negative, nonmotile coccus, 1 to 1.3 m in diameter, and was unique in using various carbohydrates as electron donor with acetate as the sole organic product. One of the following electron acceptor systems allowed growth (organic products in parentheses): Methanobrevibacter simithii (CH4), formate (acetate), 3,4,5-trimethoxybenzoate and syringate (acetate and gallate), vanillate (acetate and protocatechuate), vanillin (acetate, protocatechuic aldehyde and protocatechuate), ferulate (acetate, caffeate and hydrocaffeate), caffeate (hydrocaffeate). Strain S195 required 30% (v/v) rumen fluid in the medium for good growth. S195 was placed in a new genus and species, Syntrophococcus sucromutans, of the family Veillonellaceae.Abbreviations G+C Guanine plus cytosine - MPN most probable number - OD optical density  相似文献   

2.
Five strains of acetogenic bacteria were isolated by selective enrichment from the rumen of a mature Hereford crossbred steer fed a typical high forage diet. Suspensions of rumen bacteria, prepared from contents collected 7 h postfeeding, blended and strained through cheesecloth, were incubated in a minimal medium containing 10% clarified rumen fluid under either H2:CO2 (80:20) or N2:CO2 (80:20) headspace atmosphere. The selection criterion was an increment of acetate in the enrichments incubated under H2:CO2. Periodically, the enrichment broths were plated onto agar media and presumed acetogenic bacteria subsequently were screened for acetate production. Selected acetogenic bacteria utilized a pressurized atmosphere of H2:CO2 to form acetate in quantities 2 to 8-fold higher than when grown under N2:CO2. All presumptive acetogenic isolates were derived from either the 10-7 or 10-8 dilutions of rumen contents. All 5 strains were Gram-positive rods, and all utilized formate, glucose and CO. One strain required, and all were stimulated by, rumen fluid. No spores were observed with phase-contast microscopy and two strains were motile. No methane was detected in the headspace of pure cultures grown under either gas phase. The isolation of these bacteria indicates that acetogenic bacteria are inhabitants of the rumen of the bovine fed a typical diet and suggests that they may be participants in the utilization of hydrogen in the rumen ecosystem. Strain 139B (= ATCC 43876) is named Acetitomaculum ruminis gen. nov., sp. nov. and is the type strain of this new species. Portions of this work were presented previously (Greening RC, Leedle JAZ (1987) Abstr Annu Meet Am Soc Microbiol I 131, pp 194)  相似文献   

3.
A Synthrophomonas wolfei-Methanospirillum hungatei coculture was adapted to catabolize crotonate. S. wolfei was then isolated in axenic culture using agar spread plates and roll tubes with crotonate as the sole energy source. S. wolfei catabolized crotonate via a disproportionation mechanism similar to that of some Clostridium species. Growth on crotonate was very slow (specific growth rate of 0.029 h–1) but the conversion of energy into cell material was very efficient with cell yields of 14.6 g (dry wt.) per mol of crotonate. S. wolfei alone did not catabolize butyrate, but butyrate was stoichiometrically degraded to acetate and presumably methane when S. wolfei was reassociated with M. hungatei. S. wolfei-M. hungatei cocultures accumulated some butyrate during growth on crotonate indicating that protons were not the sole electron acceptors used for crotonate oxidation by the coculture.  相似文献   

4.
The paper deals with the microbiological characterization of water-saturated horizons in permafrost soils (cryopegs) found on the Varandei Peninsula (Barents Sea coast), 4–20 m deep. The total quantity of bacteria in the water of cryopegs was 3.5 × 108 cells/ml. The population of cultivated aerobic heterotrophic bacteria was 3–4 × 107 cells/ml and the number of anaerobic heterotrophic bacteria varied from 102 to 105 cells/ml depending on cultivation temperature and salinity. Sulfate-reducing bacteria and methanogenic archaea were found as hundreds and tens of cells per ml of water, respectively. A pure culture of a sulfate-reducing strain B15 was isolated from borehole 21 and characterized. Phylogenetic analysis has shown that the new bacterium is a member of the genus Desulfovibrio with Desulfovibrio mexicanus as its closest relative (96.5% similarity). However, the significant phenotypic differences suggest that strain B15 is a new species of sulfate-reducing bacteria.  相似文献   

5.
A new species of anaerobic bacterium that degrades the even-numbered carbon fatty acids, butyrate, caproate and caprylate, to acetate and H2 and the odd-numbered carbon fatty acids, valerate and heptanoate, to acetate, propionate and H2 was obtained in coculture with either an H2-utilizing methanogen or H2-utilizing desulfovibrio. The organism could be grown only in syntrophic association with the H2-utilizer and no other energy sources or combination of electron donor and acceptors were utilized. It was a Gram-negative helical rod with 2 to 8 flagella, about 20 nm in diameter, inserted in a linear fashion about 130 nm or more apart along the concave side of the cell. It grew with a generation time of 84 h in co-culture with Methanospirillum hungatii and was present in numbers of at least 4.5×10-6 per g of anaerobic digestor sludge.  相似文献   

6.
A saccharolytic spirochete that associated and interacted with cellulolytic bacteria was isolated from bovine rumen fluid. Isolation was accomplished by means of a procedure involving serial dilution of a sample of rumen fluid into a cellulose-containing agar medium. Clear zones appeared within the medium as a result of cellulose hydrolysis by rumen bacteria. The saccharolytic spirochete and a cellulolytic bacterium later identified as a strain of Bacteroides succinogenes were isolated from the clear zones. The spirochete did not utilize cellulose, but grew in coculture with the cellulolytic bacterium in cellulose-containing media. When cocultured in these media the spirochete used, as fermentable substrates, soluble sugars released from cellulose by the cellulolytic bacterium. In cellulosecontaining agar medium the spirochete enhanced cellulose breakdown by the B. succinogenes strain. Electron microscopy showed that the helical spirochete cells possessed an outer sheath, a protoplasmic cylinder, and two periplasmic fibrils. Under a CO2 atmosphere, in a reduced medium containing inorganic salts, rumen fluid, glucose, and NaHCO3, the spirochete grew to a final density of 1.9×109 cells/ml. Succinate, acetate, and formate were products of the fermentation of glucose by growing cells. CO2 (HCO3 -), branched short-chain fatty acids, folic acid, biotin, niacinamide, thiamine, pyridoxal, and a carbohydrate were required for growth of the spirochete. The results of this study indicated that the rumen spirochete represents a new species of Treponema. It is proposed that the new species be named Treponema bryantii.Abbreviations cpm counts per minute - GC guanine plus cytosine - Tm melting temperature - PC protoplasmic cylinder - PF pertplasmic fibrils (axial fibrils) - OS outer sheath - ID insertion disk  相似文献   

7.
From anoxic marine sediment samples, new anaerobic, microaerotolerant, Gram-negative, non-sporeforming bacteria were isolated which grew in mineral medium with malonate as sole source of carbon and energy. Cells were motile thin rods, often forming large aggregates. Malonate was decarboxylated to acetate with concomitant growth yields of 1.9–2.1 g dry cell matter per mol malonate degraded. Fumarate and malate were fermented to succinate and CO2. No other substrates were used. No inorganic electron acceptors were reduced. At least 150 mM NaCl was required for growth with either substrate. High amounts of a periplasmic cytochrome c were detected, as well as small amounts of a membrane-bound cytochrome b. All enzymes of the citric acid cycle were found to be present. The DNA base ratio was 48.3 mol% guanine plus cytosine. Since this new bacterium cannot be affiliated with any of the known genera and species, a new genus and species, Malonomonas rubra is proposed.  相似文献   

8.
A gram-positive, motile, rod-shaped, strictly anaerobic, sporulating bacterium was isolated from an enrichment initiated with mullet gut contents. The organism grew optimally at 30°C and pH6.5, and at a salinity of 1–103. Out of a variety of polysaccharides tested as growth substrates, only alginate supported growth in either semidefined or complex culture medium. The organism also grew on a variety of mono- and disaccharides. Moles product per 100mol of alginate monomer degraded were: acetate, 186; ethanol, 19; formate, 54; and CO2, 0.19. Moles product per 100mol of hexose in cellobiose or glucose degraded were: acetate, 135; ethanol,61; formate, 63: and CO2, 61. Hydrogen was not detectable during the incubations (detection limit, <10-5atm) and propionate, butyrate, lactate, or succinate were not produced as fermentation end products (<2 mol per 100 mol of monomer). The G+C content of DNA from the bacterium was 30.2±0.3 mol%, and the cell walls contained the peptidoglycan component meso-diaminopimelic acid. A phylogenetic analysis of the 16S rDNA sequence indicated that the organism grouped closely with members of the RNA-DNA homology group 1 of the genus Clostridium. However, it differed from other species of the genus with regard to morphology, growth temperature optimum, substrate range, and fermentation pattern and is therefore designated as a new species of Clostridium; the type strain is A-1 (DSM 8605).  相似文献   

9.
From anoxic sewage sludge a new strictly anaerobic, spore-forming bacterium was isolated with 2-hydroxybutyrate as sole substrate. 2-, 3-, and 4-hydroxybutyrate, 4-chlorobutyrate, crotonate, vinylacetate, and pyruvate were fermented to acetate and butyrate. Fructose was converted to acetate, butyrate, butanol, and H2. Lactate and acrylate were fermented to acetate and propionate. Cells pregrown with lactate fermented 2-hydroxybutyrate to butyrate, propionate and acetate. No inorganic electron acceptors were reduced. The DNA base ratio was 32.0±1.0 mol % and was similar to that of Clostridium propionicum, which was determined to be 35.3±0.5 mol %. Strain LuHBu1 is described as type strain of a new species, Clostridium homopropionicum sp. nov. Another isolate obtained from marine sediment degraded 2-and 3-hydroxybutyrate to acetate and butyrate and was in some respects similar to the known species Ilyobacter polytropus.  相似文献   

10.
Chopped maize stover, ammoniated at ambient and elevated temperatures or steam treated, was evaluated with four Cheviot crossbred wethers fitted with permanent rumen fistulae in a 4 × 4 Latin square design. Treatments were: (i) control, 60% H2O, ensiled for at least 40 days; (ii) 3% NH3, 60% H2O for 30 days at 21°C; (iii) 3% NH3, 60% H2O for 12 h at 90°C; (iv) steamed at 16.2 kg/cm2 and 213°C for 4 min. The sheep received two meals per day at a restricted level of 90% of lowest ad libitum intake. The markers 51Cr-EDTA (100 μ ci) and 103Ru-phenanthroline complex (10 μ ci) were used for the liquid and particulate phases, respectively. Samples for rumen metabolites were collected in each period at 0, 0.5, 1.0, 2.0, 3.0, 4.0, 6.0 and 8.0 h after the morning meal. On the following day, a mixture of the markers in 200 ml of demineralized water was infused into the rumen in a single dose and rumen fluid was sampled at specific times for 4 days.Rumen fluid pH tended to be lower with ammoniated stovers. High NH3-N levels were maintained with ammoniated stovers, indicating gradual release of nitrogen. Total VFA tended to increase more with the thermoammoniated stover. Molar proportions of propionate increased (P < 0.05) and butyrate decreased (P < 0.05) with ammoniation. In contrast, steam treatment increased (P < 0.05) molar proportions of acetate and butyrate. The largest decrease (P < 0.05) for C2 : C3 acids occurred with thermoammoniated stover. Treatments with NH3 or steam increased (P < 0.05) dilution rate. Mean retention time of particulate matter was decreased (P < 0.05) by treatment, being shortest with steamed material.  相似文献   

11.
A new strictly anaerobic bacterium was isolated from an enrichment culture with glutarate as sole substrate and freshwater sediment as inoculum, however, glutarate was not metabolized by the pure culture. The isolate was a mesophilic, spore-forming, Gram-negative, motile curved rod. It fermented various organic acids, alcohols, fructose, acetoin, and H2/CO2 to acetate, usually as the only product. Other acids were fermented to acetate and propionate or acetate and butyrate. Succinate and malonate were decarboxylated to propionate or acetate, respectively, and served as sole sources of carbon and energy for growth. No inorganic electron acceptors except CO2 were reduced. Yeast extract (0.05% w/v) was required for growth. Small amounts of cytochrome b were detected in membrane fractions. The guanine-plus-cytosine content of the DNA was 44.1±2 mol%. The isolate is described as a new species of the genus Sporomusa, S. malonica.  相似文献   

12.
An anaerobic, nonphototrophic bacterium that β-oxidizes saturated fatty acids (butyrate through octanoate) to acetate or acetate and propionate using protons as the electron acceptor (H2 as electron sink product) was isolated in coculture with either a non-fatty acid-degrading, H2-utilizing Desulfovibrio sp. or methanogens. Three strains of the bacterium were characterized and are described as a new genus and species, Syntrophomonas wolfei. S. wolfei is a gram-negative, slightly helical rod with round ends that possesses between two to eight flagella laterally inserted along the concave side of the cell. It has a multilayered cell wall of the gram-negative type. The presence of muramic acid, inhibition of growth by penicillin, and increased sensitivity of the cells to lysis after treatment with lysozyme indicate that peptidoglycan is present in the cell wall. Cells of S. wolfei contain poly-β-hydroxybutyrate. Isoheptanoate was degraded to acetate, isovalerate, and H2. Carbohydrates, proteinaceous materials, alcohols, or other tested organic compounds do not support growth. Common electron acceptors are not utilized with butyrate as the electron donor. Growth and degradation of fatty acids occur only in syntrophic association with H2-using bacteria. The most rapid generation time obtained by cocultures of S. wolfei with Desulfovibrio and Methanospirillum hungatei is 54 and 84 h, respectively. The addition of Casamino Acids but neither Trypticase nor yeast extract stimulated growth and resulted in a slight decrease in the generation time of S. wolfei cocultured with M. hungatei. The addition of H2 to the medium stopped growth and butyrate degradation by S. wolfei.  相似文献   

13.
Two types of new anaerobic bacteria were isolated from anoxic freshwater sediments. They grew in mineral medium with oxalate as sole energy source and with acetate as main carbon source. Oxalate as well as oxamate (after deamination) were decarboxylated to formate with growth yields of 1.2–1.4 g dry cell matter per mol oxalate degraded. No other organic or inorganic substrates were used, and no electron acceptors were reduced. Strain WoOx3 was a Gramnegative, non-sporeforming, motile vibrioid rod with a guanine-plus-cytosine content of the DNA of 51.6 mol%. It resembled the previously described genus Oxalobacter, and is described as a new species, O. vibrioformis. Strain AltOx1 was a Gram-positive, spore-forming, motile rod with a DNA base ratio of 36.3 mol% guanine-plus-cytosine. This isolate is described as a new species of the genus Clostridium, C. oxalicum.  相似文献   

14.
The marine green sulfur bacterium Chlorobium vibrioforme strain 1930 produced H2 and elemental sulfur from sulfide or thiosulfate under N limitation in the light. H2 production depended on nitrogenase and occurred only in the absence of ammonia. Methionine sulfoximine, an inhibitor of glutamine synthetase, prevented the switch-off by ammonia. In defined syntrophic cocultures of the acetate-oxidizing, sulfur-reducing bacterium Desulfuromonas acetoxidans with green sulfur bacteria, H2 was produced from acetate via a light-driven sulfur cycle. The sulfur-reducing bacterium could not be replaced by sulfate-reducing bacteria in these experiments. In a coculture of the marine Chlorobium vibrioforme strain 1930 and the sulfur-reducing bacterium Desulfuromonas acetoxidans strain 5071, optimum long-term H2 production from acetate was obtained with molecular nitrogen as N source, at low light intensity (110 mol · m-2 · s-1), in sulfide-reduced mineral medium (2 mM Na2S) at pH 6.8. Traces of sulfide (10 M) were sufficient to keep the sulfur cycle running. The coculture formed no poly--hydroxyalkanoates (PHA), but 20%–40% polysaccharide per cell dry mass. Per mol acetate added, the coculture formed 3.1 mol of H2 (78% of the theoretical maximum). Only 8% of the reducing equivalents was incorporated into biomass. The maximum rate of H2 production was 1300 ml H2 per day and g cell dry mass.Non-standard abbrevations MOPS 2-(N-morpholino) propane sulfonic acid - MSX Methionine sulfoximine - PHA poly--hydroxyalkanoates  相似文献   

15.
A gram positive, motile rod-shaped strictly anaerobic non sporulating bacterium was isolated from an enrichment initiated with mullet gut contents. The organism grew optimally at 30°C at pH 6.5 and at a salinity of 10/103. Out of a variety of mono-, di-, and polysaccharides tested only pectin, cellobiose and starch actively supported growth in either semi defined medium or peptone-yeast extract (PY) medium. Galacturonic acid and maltose were less effective as substrates. Mol product per 100 mol of pectin monomer degraded were: acetate, 163; ethanol, 30; methanol, 88 and formate, 48. Per 100 mol of hexose in cellobiose or starch degraded, the amounts were acetate, 39; ethanol, 128 and formate, 41. Hydrogen was not detectable in the incubations (detection limit, <10-5 atm) and propionate, butyrate, lactate or succinate were not produced as fermentation end-products (<2 mol per 100 mol monomer). The guanine plus cytosine content of DNA from the bacterium was 31 mol%, and the cell walls contained meso-diaminopimelic acid. A phylogenetic analysis of the organism by 16S rDNA sequencing and DNA-DNA homology indicated that the organism grouped more closely with several species of Clostridium than with Eubacterium. The phenotypic characteristics of the organism indicated that it did not fit within the genus Clostridium and more closely resembled Eubacterium. The organism is therefore designated as a species of Eubacterium; the type strain is P-1 (DSM 6788).  相似文献   

16.
Methanogenesis by a Syntrophomonas wolfei/ Methanospirillum hungatei coculture was inhibited in presence of ethylene and the hydrogenation catalyst Pd-BaSO4. However, butyrate oxidation by S. wolfei continued and ethylene was reduced to ethane. Per mol of butyrate oxidized, 2.4 mol acetate was produced and 0.8 mol ethylene was reduced. Acetylene, propylene and butene were less effective as H2 acceptors than ethylene, and addition of bromoethanesulfonic acid was necessary to inhibit methanogenesis in the presence of the two longer-chain olefins. Other hydrogenation catalysts were less effective in the order Pd-charcoal < PE-asbestos < Pd-PEI beads < Pt-Al2O3, Pd-CaCO3. Optimal ethylene hydrogenation was achieved with still incubation in presence of 7.2 mg Pd-BaSO4 and 0.7 g sand per ml medium. The higher catabolic rate of S. wolfei in presence of the methanogen indicated that the biological H2 removal mechanism was more efficient than the catalytic olefin reduction.Abbreviations BES bromoethane sulfonic acid - VFA volatile fatty acid  相似文献   

17.
The determination of enzymatic activities in cell-free extracts of Acidaminococcus fermentans and Peptostreptococcus asaccharolyticus led to a refined scheme for the pathway of glutamate fermentation via (R)-2-hydroxyglutarate to acetate and butyrate. From the ratio of these products the amount of ATP generated by substrate level phosphorylation was calculated. Growth experiments with the organisms including Clostridium symbiosum and Clostridium tetanomorphum indicated that a sodium gradient contributed additional energy for growth. The high growth yields found in organisms containing the biotin dependent sodium pump glutaconyl-CoA decarboxylase could be reduced by the sodium ionophor monensin. In P. asaccharolyticus energy equivalent up to 0.6 mol ATP per mol of glutaconyl-CoA decarboxylated was conserved via the Na+ gradient. The data may explain the growth promoting effects of monensin in cattle.  相似文献   

18.
Ergot alkaloids in endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum) have been shown to cause a reduction in blood flow to the rumen epithelium as well as a decrease in volatile fatty acids (VFA) absorption from the washed rumen of steers. Previous data also indicates that incubating an extract of endophyte-infected tall fescue seed causes an increase in the amount of VFA absorbed per unit of blood flow, which could result from an alteration in the absorptive or barrier function of the rumen epithelium. An experiment was conducted to determine the acute effects of an endophyte-infected tall fescue seed extract (EXT) on total, passive or facilitated acetate and butyrate flux across the isolated bovine rumen as well as the barrier function measured by inulin flux and tissue conductance (Gt). Flux of ergovaline across the rumen epithelium was also evaluated. Rumen tissue from the caudal dorsal sac of Holstein steers (n=6), fed a common diet, was collected and isolated shortly after slaughter and mounted between two halves of Ussing chambers. In vitro treatments included vehicle control (80% methanol, 0.5% of total volume), Low EXT (50 ng ergovaline/ml) and High EXT (250 ng ergovaline/ml). Results indicate that there is no effect of acute exposure to ergot alkaloids on total, passive or facilitated flux of acetate or butyrate across the isolate bovine rumen epithelium (P>0.51). Inulin flux (P=0.16) and Gt (P>0.17) were not affected by EXT treatment, indicating no alteration in barrier function due to acute ergot alkaloid exposure. Ergovaline was detected in the serosal buffer of the High EXT treatment indicating that the flux rate is ~0.25 to 0.44 ng/cm2 per hour. Data indicate that specific pathways for VFA absorption and barrier function of the rumen epithelium are not affected by acute exposure to ergot alkaloids from tall fescue at the concentrations tested. Ergovaline has the potential to be absorbed from the rumen of cattle that could contribute to reduced blood flow and motility and lead to reduced growth rates of cattle.  相似文献   

19.
The number and diversity of culturable microorganisms involved in sulfur oxidation and sulfate reduction were investigated in the oxidized sediments of gold mine tailings, Kuznetsk Basin, Russia. The sediments had a low pH (2.4–2.8), high SO 4 2? content (up to 22 g/l), and high concentrations of dissolved metals. The arsenic content was as high as 1.9 g/l. Bacterial phylogeny in microcosms was investigated by amplification of 16S rRNA gene fragments with subsequent denaturing gradient gel electrophoresis (DGGE). Spore-forming bacteria Desulfosporosinus were the only bacteria revealed for which the capacity for dissimilatory sulfate reduction is known. Strain Desulfosporosinus sp. DB was obtained in pure culture, and it was phylogenetically remote from other cultured and uncultured members of the genus. No sulfate-reducing members of the Deltaproteobacteria were detected. The Firmicutes members were the most numerous phylotypes in the microcosms, including a separate cluster with the similarity to Pelotomaculum not exceeding 94%. Acidithiobacillus ferrooxidans and A. caldus were found in anaerobic and microaerophilic microcosms. The number of sulfate reducers did not exceed 9.5 × 102 cells/ml.  相似文献   

20.
Nitrogenase activity (acetylene reduction activity) was found to occur universally in the Cyperus papyrus swamp in Lake Naivasha. Low rates of acetylene reduction activity (0.9–104.9 nmol C2H4 g d.wt. roots-1 h-1) were associated with excised roots of C. papyrus but higher rates of activity (89.0–280.4 nmol C2H4 g d.wt. roots-1 h-1) were associated with intact root systems of the plant. It was estimated that nitrogen fixation associated with young roots alone could supply about 26% of the nitrogen requirements of growing papyrus plants. Acetylene reduction activity in the lake bottom sediments was generally low and associated with adjacent papyrus stands. Plate counts of putative aerobic and facultatively anaerobic N2-fixing bacteria associated with papyrus roots showed the presence of high numbers of diazotrophs (5.4 × 106 CFU g d.wt. roots-1). Fewer numbers of N2-fixing bacteria were detected in the sediments (1.9 × 103-3.2 × 104 CFU g d.wt. sediment-1).  相似文献   

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