TABLE 3 - uploaded by Meyer Wolin
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except fortheaddition ofa fewvitamins. It consisted ofNH4Cl,0.1%;KH2PO4,0.04%; MgCl26H20,0.02%;sodiumformate, 0.5%; FeSO4, 0.001%; phenol red, 0.0003%; NaHCO3, 0.078%; Na2S-9H20, 0.02%; vitamin B12, 0.02 ,g/ml;andp-aminobenzoic acid, 0.30,ug/ml. Themediumwasprepared using tapwaterand placed inserumbottles. Bicarbonate andsulfide weresterilized byfi...
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... of nitrate. Although nitrite accumu- lates in the medium when the vibrio is grown with H2 or formate and nitrate, the stoichiometry of the reduction of nitrate by H2 obtained with resting cells indicates that the nitrate is reduced beyond the nitrite stage ( Table 3). The stoichi- ometry suggests almost a complete reduction of nitrate to NH3. ...
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Osnabrück, Univ., Diss., 2005 (Nicht für den Austausch).
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... Fibrobacter succinogenes (Hungate, 1947;Stewart and Flint, 1989), Butyrivibrio fibrisolvens (Hungate, 1950), Bacillus licheniformis (Fujimoto et al., 2011), Ruminococcus flavefaciens (Sijpesteijn, 1951), R. albus (Hungate, 1957), Clostridium cellobioparum (Hungate, 1944), C. chartadabidum (Kelly et al., 1987), C. longisporum (Hungate, 1957), C. lochheadii (Hungate, 1957), Eubacterium cellulosolvens (Taguchi et al., 2008) Hemicellulolytic Eubacterium xylanophilum (Johns, 1951), E. uniformis (Johns, 1951) Amylolytic Streptococcus bovis (Hungate et al., 1952), Ruminobacter amylophilus (Hamlin and Hungate, 1956) Lipolytic Anaerovibrio lipolytica (Hobson and Mann, 1961) Proteolytic Prevotella ruminicola (Wallace, 1996), Clostridium bifermentans (Clarke, 1961) Saccharolytic Succinivibrio dextrinosolvens (Bryant and Small, 1956), S. amylolytica, Bacteroides ruminocola (Bryant et al., 1958), Selenomonas ruminantium (Bryant, 1956), Lactobacillus acidophilus, L. casei, L. fermentum, L. plantarum (Perry and Briggs, 1957), L. brevis, L. helveticus (Briges, 1953), Bifidobacterium globosum, B. longum, B. thermophilum, B. ruminale, B. ruminantium (Scardovi et al., 1969;Trovatelli and Matteuzzi, 1976) Pectinolytic Treponema saccharophilum (Paster and Canale-Parola, 1985), Lachnospira multiparus (Bryant and Small, 1956) Acid utilizers Megasphaera elsdeni (Elsden et al., 1956), Wolinella succinogenes (Wolin et al., 1961), Veillonella gazogene (Johns, 1951), Micrococcus lactolytica, Oxalobacter formigenes (Allison et al., 1985), Desulfovibrio desulfuricans (Howard and Hungate, 1976), Desulfotomaculum ruminis (Coleman, 1960b), Succiniclasticum ruminis (Van Gylswyk, 1995) ...
Over the last two decades, biotechnology has advanced at a rapid pace, propelled by the incorporation of bio-products into various aspects of pharmaceuticals, industry, and the environment. These developments have sparked interest in the bioprospecting of microorganisms and their products in a variety of niche environments. Furthermore, the use of omics technologies has greatly aided our analyses of environmental samples by elucidating the microbial ecological framework, biochemical pathways, and bio-products. However, the more often overemphasis on taxonomic identification in most research publications, as well as the data associated with such studies, is detrimental to immediate industrial and commercial applications. This review identifies several factors that contribute to the complexity of sequence data analysis as potential barriers to the pragmatic application of functional genomics, utilizing recent research on ruminants to demonstrate these limitations in the hopes of broadening our horizons and drawing attention to this gap in bioprospecting studies for other niche environments as well. The review also aims to emphasize the importance of routinely incorporating functional genomics into environmental metagenomics analyses in order to improve solutions that drive rapid industrial biocatalysis developments from derived outputs with the aim of achieving potential benefits in energy-use reduction and environmental considerations for current and future applications.
... Two samples from gastric biopsies showed a 100% identity of the 16S rRNA gene sequence with W. succinogenes species. W. succinogenes was initially isolated from cattle rumen (Wolin et al. 1961). Phylogenetic analyses of one sample from the camel revealed that this sample was bootstrapped to H. cinaedi. ...
Helicobacter species have been reported in animals, some of which are of zoonotic importance. This study aimed to detect Helicobacter species among human and animal samples using conventional PCR assays and to identify their zoonotic potentials. Helicobacter species was identified in human and animal samples by genus-specific PCR assays and phylogenetic analysis of partial sequencing of the 16S ribosomal RNA gene. The results revealed that Helicobacter species DNA was detected in 13 of 29 (44.83%) of the human samples. H. pylori was identified in 2 (15.38%), and H. bovis was detected in 4 (30.77%), whereas 7 (53.85%) were unidentified. H. bovis and H. heilmannii were prevalent among the animal samples. Phylogenetic analysis revealed bootstrapping of sequences with H. cinaedi in camel, H. rappini in sheep and humans, and Wollinella succinogenes in humans. In conclusion, the occurrence of non-H. pylori infections among human and animal samples suggested zoonotic potentials.
... Despite this, it has been proposed that stimulation of acetogens may be an effective strategy for methane mitigation in methanogen-inhibited scenarios [14,20,48,49]. Various microorganisms have also been isolated from cows and sheep that support anaerobic hydrogenotrophic respiration, including dissimilatory sulfate reduction (e.g., Desulfovibrio desulfuricans) [50,51], fumarate and nitrate reduction (e.g., Selenomonas ruminantium, Wolinella succinogenes) [52][53][54][55][56][57][58][59] and trimethylamine N-oxide reduction (e.g., Denitrobacterium detoxificans) [60]. The first described and most comprehensively studied of these hydrogen oxidisers is W. succinogenes, which mediates interspecies hydrogen transfer with R. albus [25]. ...
... We demonstrate that ruminants harbour a diverse community of hydrogenogenic fermenters and hydrogenotrophic methanogens, acetogens and sulfate, fumarate and nitrate reducers. Second, we used the model system of the H 2 -producing carbohydrate fermenter R. albus 7 and the H 2 -utilising fumarate-reducing syntrophic partner Wolinella succinogenes DSM 1740 [25,54,55,68] to gain a deeper mechanistic understanding of how and why ruminant bacteria regulate H 2 metabolism. We observed significant differences in the growth, transcriptome and metabolite profiles of these bacteria in co-culture compared with pure culture. ...
... The bovine rumen isolates R. albus 7 [68] and Wolinella succinogenes DSM 1740 [54] were cultured anaerobically at 37 °C in modified Balch medium [79] (Table S6). Precultures were grown in Balch tubes (18 × 150 mm; Chemglass Life Sciences, Vineland, NJ) containing 20% v/v culture medium and sealed with butyl rubber stoppers crimped with aluminium caps. ...
Farmed ruminants are the largest source of anthropogenic methane emissions globally. The methanogenic archaea responsible for these emissions use molecular hydrogen (H 2), produced during bacterial and eukaryotic carbohydrate fermentation, as their primary energy source. In this work, we used comparative genomic, metatranscriptomic and co-culture-based approaches to gain a system-wide understanding of the organisms and pathways responsible for ruminal H 2 metabolism. Two-thirds of sequenced rumen bacterial and archaeal genomes encode enzymes that catalyse H 2 production or consumption, including 26 distinct hydrogenase subgroups. Metatranscriptomic analysis confirmed that these hydrogenases are differentially expressed in sheep rumen. Electron-bifurcating [FeFe]-hydrogenases from carbohydrate-fermenting Clostridia (e.g., Ruminococcus) accounted for half of all hydrogenase transcripts. Various H 2 uptake pathways were also expressed, including methanogenesis (Methanobrevibacter), fumarate and nitrite reduction (Selenomonas), and acetogenesis (Blautia). Whereas methanogenesis-related transcripts predominated in high methane yield sheep, alternative uptake pathways were significantly upregulated in low methane yield sheep. Complementing these findings, we observed significant differential expression and activity of the hydrogenases of the hydrogenogenic cellulose fermenter Ruminococcus albus and the hydrogenotrophic fumarate reducer Wolinella succinogenes in co-culture compared with pure culture. We conclude that H 2 metabolism is a more complex and widespread trait among rumen microorganisms than previously recognised. There is evidence that alternative hydrogenotrophs, including acetogenic and respiratory bacteria, can prosper in the rumen and effectively compete with methanogens for H 2. These findings may help to inform ongoing strategies to mitigate methane emissions by increasing flux through alternative H 2 uptake pathways, including through animal selection, dietary supplementation and methanogenesis inhibitors.
... Representatives of the Epsilonproteobacteria inhabit a broad spectrum of environments like mammalian digestive systems (Wolin et al., 1961;Engberg et al., 2000), brackish water (Brettar et al., 2006), hydrothermal sediments , or subsurface systems (Watanabe et al., 2000;Gittel et al., 2012;Hubert et al., 2012;Handley et al., 2013). Previously obtained isolates have been described as chemolithoautotrophs Sievert et al., 2008) fixing carbon via the reductive tricarboxylic acid (rTCA) cycle (Hügler et al., 2005). ...
... A model organism representing the metabolic versatility of this proteobacterial class is Wolinella succinogenes. It couples anaerobic fumarate or nitrate respiration with hydrogen, sulfide, or formate oxidation (Kröger et al., 2002;Kern and Simon, 2009) but can also grow under limited oxic conditions (Wolin et al., 1961). During the last decade, research focused on Epsilonproteobacteria thriving in marine systems such as hydrothermal vents (Miroshnichenko et al., 2004;Nakagawa et al., 2007;Schauer et al., 2011;Stokke et al., 2015) or pelagic oxic-anoxic interfaces (Grote et al., 2012) as well as in terrestrial sulfidic caves and springs (Engel et al., 2004;Porter and Engel, 2008;Jones et al., 2010;Rossmassler et al., 2012;Hamilton et al., 2015), or mud volcanos (Green-Saxena et al., 2012) where they are thought to be mainly involved in the oxidation or reduction of sulfur compounds. ...
The population genome of an uncultured bacterium assigned to the Campylobacterales (Epsilonproteobacteria) was reconstructed from a metagenome dataset obtained by whole-genome shotgun pyrosequencing. Genomic DNA was extracted from a sulfate-reducing, m-xylene-mineralizing enrichment culture isolated from groundwater of a benzene-contaminated sulfidic aquifer. The identical epsilonproteobacterial phylotype has previously been detected in toluene- or benzene-mineralizing, sulfate-reducing consortia enriched from the same site. Previous stable isotope probing (SIP) experiments with 13C6-labeled benzene suggested that this phylotype assimilates benzene-derived carbon in a syntrophic benzene-mineralizing consortium that uses sulfate as terminal electron acceptor. However, the type of energy metabolism and the ecophysiological function of this epsilonproteobacterium within aromatic hydrocarbon-degrading consortia and in the sulfidic aquifer are poorly understood. Annotation of the epsilonproteobacterial population genome suggests that the bacterium plays a key role in sulfur cycling as indicated by the presence of an sqr gene encoding a sulfide quinone oxidoreductase and psr genes encoding a polysulfide reductase. It may gain energy by using sulfide or hydrogen/formate as electron donors. Polysulfide, fumarate, as well as oxygen are potential electron acceptors. Auto- or mixotrophic carbon metabolism seems plausible since a complete reductive citric acid cycle was detected. Thus the bacterium can thrive in pristine groundwater as well as in hydrocarbon-contaminated aquifers. In hydrocarbon-contaminated sulfidic habitats, the epsilonproteobacterium may generate energy by coupling the oxidation of hydrogen or formate and highly abundant sulfide with the reduction of fumarate and/or polysulfide, accompanied by efficient assimilation of acetate produced during fermentation or incomplete oxidation of hydrocarbons. The highly efficient assimilation of acetate was recently demonstrated by a pulsed 13C2-acetate protein SIP experiment. The capability of nitrogen fixation as indicated by the presence of nif genes may provide a selective advantage in nitrogen-depleted habitats. Based on this metabolic reconstruction, we propose acetate capture and sulfur cycling as key functions of Epsilonproteobacteria within the intermediary ecosystem metabolism of hydrocarbon-rich sulfidic sediments.
... Supporting this, even epsilonproteobacterial genera that associate with terrestrial animals, such as Campylobacter, Helicobacter and Wolinella, retain physiological characteristics reflective of a hydrothermal environment where they most likely evolved. These include a need for high CO 2 (Al-Haideri et al., 2016), intolerance to high levels of O 2 (Kendall et al., 2014), and the ability to use hydrogen as an electron donor (Wolin et al., 1961). Below, the underlying geochemistry of deep-sea vents and its effect on the growth conditions of the microbes inhabiting this habitat are discussed. ...
Chemoautotrophic ecosystems at deep-sea hydrothermal vents were discovered in 1977, but not until 1995 were free-living autotrophic Epsilonproteobacteria identified as important microbial community members. Because the deep-sea is food-starved, the autotrophic metabolism of hydrothermal vent Epsilonproteobacteria may be very important for deep-sea consumers. However, quantifying their metabolic activities in situ has remained difficult, and biochemical mechanisms underlying their autotrophic physiology are poorly described. To gain insight into environmental processes, an approach was developed for incubations of microbes at in situ pressure and temperature (25 MPa, 24°C) with various combinations of electron donors/acceptors (H₂ , O₂ and NO₃- and ¹³HCO₃-) as a tracer to track carbon fixation. During short (18-24 h) incubations of low-temperature vent fluids from Crab Spa (9°N East Pacific Rise), the concentration of electron donors/acceptors and cell numbers were monitored to quantify microbial processes. Measured rates were generally higher than previous studies, and the stoichiometry of microbially-catalyzed redox reactions revealed new insights into sulfur and nitrogen cycling. Single-cell, taxonomically-resolved tracer incorporation showed Epsilonproteobacteria dominated carbon fixation, and their growth efficiency was calculated based on electron acceptor consumption. Using these data, in situ primary productivity, microbial standing stock, and average biomass residence time of the deep-sea vent subseafloor biosphere were estimated. Finally, the population structures of the most abundant genera Sulfurimonas and Thioreductor were shown to be strongly influenced by pO₂ and temperature respectively, providing a mechanism for niche differentiation in situ. To gain insights into the core biochemical reactions underlying autotrophy in Epsilonprotebacteria, a theoretical metabolic model of Sulfurimonas denitrificans was developed. Validated iteratively by comparing in silico yields with data from chemostat experiments, the model generated hypotheses explaining critical, yet so far unresolved reactions supporting chemoautotrophy in Epsilonproteo bacteria. For example, it provides insight into how energy is conserved during sulfur oxidation coupled to denitrification, how reverse electron transport produces ferredoxin for carbon fixation, and why aerobic growth yields are only slightly higher compared to denitrification. As a whole, this thesis provides important contributions towards understanding core mechanisms of chemoautrophy, as well as the in situ productivity, physiology and ecology of autotrophic Epsilonproteobacteria.
... With the exception of H. pylori, no investigation on the presence of other Helicobacteraceae in the milk has been so 2 BioMed Research International far performed and very few studies researched Helicobacteraceae in gastrointestinal tract of cattle. Wolinella succinogenes, which belongs to the family Helicobacteraceae, was originally isolated from cattle rumen [12], and "Candidatus Helicobacter bovis" was described in the pyloric portion of the abomasum [13]. The study of Helicobacteraceae other than H. pylori in animals is valuable because some Helicobacter spp. ...
... As "Candidatus Helicobacter bovis" is present at high levels in cattle abomasa [13], one might expect a transit into the intestinal tract and fecal excretion. A similar situation is expected for Wolinella spp., which are found in cattle rumen [12]. Personal observations confirm that such bacteria can be frequently found in the gastrointestinal tract of cattle of our region [20]. ...
Helicobacter pylori is responsible for gastritis and gastric adenocarcinoma in humans, but the routes of transmission of this bacterium have not been clearly defined. Few studies led to supposing that H. pylori could be transmitted through raw milk, and no one investigated the presence of other Helicobacteraceae in milk. In the current work, the presence of Helicobacteraceae was investigated in the bulk tank milk of dairy cattle herds located in northern Italy both by direct plating onto H. pylori selective medium and by screening PCR for Helicobacteraceae, followed by specific PCRs for H. pylori, Wolinella spp., and “Candidatus Helicobacter bovis.” Three out of 163 bulk milk samples tested positive for Helicobacteraceae, but not for the subsequent PCRs. H. pylori was not isolated in any case. However, given similar growth conditions, Arcobacter butzleri, A. cryaerophilus, and A. skirrowii were recovered. In conclusion, the prevalence of Helicobacteraceae in raw milk was negligible (1.8%), and H. pylori was not identified in any of the positive samples, suggesting that, at least in the farming conditions of the investigated area, bovine milk does not represent a potential source of infection.
... In the rumen, CH 4 is produced by methanogenic archaea from CO 2 and H 2 (Madigan et al. 2000). As some ruminal bacteria are able to reduce FA to propionate using 2H as reducing equivalents (Wolin et al. 1961), the addition of FA serves as an alternative hydrogen sink diverting 2H from methanogenesis towards propionate formation (Demeyer and Henderickx 1967;López et al. 1999;Bayaru et al. 2001). The availability of 2H can influence microbial metabolism, growth and gene expression of microorganisms (Wolin et al. 1961;Morgan et al. 1997;Hendrickson et al. 2007). ...
... As some ruminal bacteria are able to reduce FA to propionate using 2H as reducing equivalents (Wolin et al. 1961), the addition of FA serves as an alternative hydrogen sink diverting 2H from methanogenesis towards propionate formation (Demeyer and Henderickx 1967;López et al. 1999;Bayaru et al. 2001). The availability of 2H can influence microbial metabolism, growth and gene expression of microorganisms (Wolin et al. 1961;Morgan et al. 1997;Hendrickson et al. 2007). Since the community of methanogenic archaea was affected by hydrogen concentrations in enrichment cultures (Leybo et al. 2006), it is conceivable that the addition of 2H acceptors like FA might result in changes of the microbial population. ...
The greenhouse gas methane (CH4) contributes substantially to global climate change. As a potential approach to decrease ruminal methanogenesis, the effects of different dosages of fumaric acid (FA) on ruminal microbial metabolism and on the microbial community (archaea, bacteria) were studied using a rumen simulation technique (RUSITEC). FA acts as alternative hydrogen acceptor diverting 2H from methanogenesis of archaea towards propionate formation of bacteria. Three identical trials were conducted with 12 fermentation vessels over a period of 14 days. In each trial, four fermentation vessels were assigned to one of the three treatment groups differing in FA dosage: low fumaric acid (LFA), high fumaric acid (HFA) and without FA (control). FA was continuously infused with the buffer. Grass silage and concentrate served as substrate. FA led to decreases in pH and to higher production rates of total short chain fatty acids (SCFA) mediated by increases in propionate for LFA of 1.69 mmol d(-1) and in propionate and acetate production for HFA of 4.49 and 1.10 mmol d(-1), respectively. Concentrations of NH3-N, microbial crude protein synthesis, their efficiency, degradation of crude nutrients and detergent fibre fraction were unchanged. Total gas and CH4 production were not affected by FA. Effects of FA on structure of microbial community by means of single strand conformation polymorphism (SSCP) analyses could not be detected. Given the observed increase in propionate production and the unaffected CH4 production it can be supposed that the availability of reduction equivalents like 2H was not limited by the addition of FA in this study. It has to be concluded from the present study that the application of FA is not an appropriate approach to decrease the ruminal CH4 production.
... Morphological classifications were drawn from multiple chapters of The Prokaryotes [7,8] and Bergey's Manual of Systematic Bacteriology [9,10]. We consulted additional primary research articles for the following species: Arcobacter butzleri [11,12], Arcobacter nitrofigilis [13], Campylobacter concisus and Campylobacter curvus [14], Campylobacter hominis [15], Nautilia profundicola [16], Sulfuricurvum kujiense [17], Sulfurimonas dentrificans [18], Sulfurospirillum deleyianum [19], and Wolinella succinogenes [20,21]. mutant is on the left, wild-type on the right. ...
Supporting information. This file contains supplemental materials and methods, four supplemental figures, five supplemental tables, and references for the supporting information. Supplemental materials and methods describe genetic manipulations, Csd4 purification, statistical analysis of cell shape distributions, and bioinformatics analyses. Supplemental figures include Figure S1 Phylogenetic relatedness of Csd4 and Csd5 homologues and morphological complementation of their respective mutant strains, Figure S2 Growth of wild-type, csd4, and csd5 mutant strains independently and in co-culture, Figure S3 Prediction of Csd4 functional residues through structural threading analysis, and Figure S4 Morphological characterization of cross-shape class and straight rod double mutants. Supplemental tables include Table S1 Muropeptide composition of Csd4 treated Δcsd4 mutant sacculi, Table S2 Muropeptide composition of wild-type, mutant, and complemented mutant strains, Table S3 Bacterial strains, Table S4 Primers, and Table S5 Plasmids.
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... Sulfur-reduction pathways can be classified into two types based on the substrates: (1) sulfur-reduction (sulfur respiration), and (2) sulfate reduction (sulfate respiration). Sulfur-reduction pathways have been intensively studied in gastrointestinal epsilon- Proteobacterium, Wolinella succinogenes (Wolin et al., 1961; Hedderich et al., 1999). Sulfur-reduction is catalyzed by polysulfide reductase (Psr), in which the actual substrate of the reaction is polysulfide but not elemental sulfur (Pfennig and Biebel, 1986). ...
... They are heterotrophic and do not have inorganic sulfur metabolic pathways . The genus Wolinella isolated from cattle rumen can reduce elemental sulfur to hydrogen sulfide (Wolin et al., 1961). There are many reports that chemoautotrophic epsilon- Proteobacteria are predominant in sulfidic environments such as deep-sea hydrothermal and subsurface environments (Takai et al., 2004; Campbell et al., 2006; Nakagawa and Takai, 2008). ...
In deep-sea hydrothermal systems, super hot and reduced vent fluids from the subseafloor blend with cold and oxidized seawater. Very unique and dense ecosystems are formed within these environments. Many molecular ecological studies showed that chemoautotrophic epsilon- and gamma-Proteobacteria are predominant primary producers in both free-living and symbiotic microbial communities in global deep-sea hydrothermal fields. Inorganic sulfur compounds are important substrates for the energy conservative metabolic pathways in these microorganisms. Recent genomic and metagenomic analyses and biochemical studies have contributed to the understanding of potential sulfur metabolic pathways for these chemoautotrophs. Epsilon-Proteobacteria use sulfur compounds for both electron-donors and -acceptors. On the other hand, gamma-Proteobacteria utilize two different sulfur-oxidizing pathways. It is hypothesized that differences between the metabolic pathways used by these two predominant proteobacterial phyla are associated with different ecophysiological strategies; extending the energetically feasible habitats with versatile energy metabolisms in the epsilon-Proteobacteria and optimizing energy production rate and yield for relatively narrow habitable zones in the gamma-Proteobacteria.
... A second nonenzymatic mechanism for the production of elemental selenium could involve a reaction between H2S and s~o:-, since W. succinogenes has been reported to produce hydrogen sulfide from cysteine (Wolin et al. 1961). This mechanism could not explain most of the formation that occurred because there was not enough cysteine to account for all of the reductive activity. ...
Cultures of Wolinella succinogenes were adapted to grow in the presence of 1 mM or 10 mM . Both selenium salts were reduced to red, amorphous, elemental selenium but only after the culture reached the stationary growth phase. Bacterial cells taken from a culture actively reducing selenium were examined by transmission electron microscopy and were found to have large, electron-dense granules in the cytoplasm. These granules were verified by energy-dispersive X-ray spectroscopy to consist of selenium. Wolinella succinogenes was unable to grow with or as the final electron acceptor. Key words: Wolinella, selenium, cytology, selenate.
