Description of Paraclostridium bifermentans subsp. muricolitidis subsp. nov., emended description of Paraclostridium bifermentans (Sasi Jyothsna et al., 2016), and creation of Paraclostridium bifermentans subsp. bifermentans subsp. nov.

Taxonomic studies of strain PAGU 1678T , an obligately anaerobic, gram-positive, spore-forming bacterium isolated from biobreeding rat feces, were performed. This strain has been demonstrated to have the ability to exacerbate pathosis in a mouse model of dextran sulfate sodium-induced ulcerative colitis. Phylogenetic analysis based on the 16S rRNA gene showed high homology with Paraclostridium bifermentans. To clarify the correct taxonomic position of strain PAGU 1678T , a comparative taxonomic study using P. bifermentans PAGU 2008T (═JCM 1386T ) and the closely related bacterial species P. benzoelyticum PAGU 2068T (═LMG 28745T ) was carried out. Despite the close similarity of 16S rRNA gene sequences, DNA-DNA hybridization between strain PAGU 1678T and P. bifermentans PAGU 2008T was 60.03% on average, average nucleotide identity was 96.17%, and it was shown to have different genomic sequences. Biochemically, strain PAGU 1678T could be differentiated from P. bifermentans PAGU 2008T by H2 S production. Furthermore, strain PAGU 1678T was characterized by the presence of two phospholipids with different polarity on polar lipid analysis. In addition, strain PAGU 1678T differed from P. bifermentans PAGU 2008T in findings on whole-cell protein analysis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. On the basis of these biochemical and genetic characteristics, a novel subspecies of P. bifermentans with the name Paraclostridium bifermentans subsp. muricolitidis subsp. nov. is here proposed, with PAGU 1678T (═CCUG 72489T ═NBRC 113386T ) as the type strain, which automatically creates P. bifermentans subsp. bifermentans subsp. nov. JCM 1386T (═ATCC 638T ═DSM 14991T ).

Whole genome sequence analysis indicates recent diversification of mammal-associated Campylobacter fetus and implicates a genetic factor associated with H2S production.

BACKGROUND: Campylobacter fetus (C. fetus) can cause disease in both humans and animals. C. fetus has been divided into three subspecies: C. fetus subsp. fetus (Cff), C. fetus subsp. venerealis (Cfv) and C. fetus subsp. testudinum (Cft). Subspecies identification of mammal-associated C. fetus strains is crucial in the control of Bovine Genital Campylobacteriosis (BGC), a syndrome associated with Cfv. The prescribed methods for subspecies identification of the Cff and Cfv isolates are: tolerance to 1 % glycine and H2S production. RESULTS: In this study, we observed the deletion of a putative cysteine transporter in the Cfv strains, which are not able to produce H2S from L-cysteine. Phylogenetic reconstruction of the core genome single nucleotide polymorphisms (SNPs) within Cff and Cfv strains divided these strains into five different clades and showed that the Cfv clade and a Cff clade evolved from a single Cff ancestor. CONCLUSIONS: Multiple C. fetus clades were observed, which were not consistent with the biochemical differentiation of the strains. This suggests the need for a closer evaluation of the current C. fetus subspecies differentiation, considering that the phenotypic differentiation is still applied in BGC control programs.

CysB-dependent upregulation of the Salmonella Typhimurium cysJIH operon in response to antimicrobial compounds that induce oxidative stress.

It has been proposed that some antibiotics exert additional damage through reactive oxygen species (ROS) production. Since H2S protects neurons and cardiac muscle from oxidative stress, it has been hypothesized that bacterial H2S might, similarly, be a cellular protector against antibiotics. In Enterobacteriaceae, H2S can be produced by the cysJIH pathway, which uses sulfate as the sulfur source. CysB, in turn, is a positive regulator of cysJIH. At present, the role of S. Typhimurium cysJIH operon in the protection to reactive oxygen species (ROS) induced by antimicrobial compounds remains to be elucidated. In this work, we evaluated the role of cysJIH and cysB in ROS accumulation, superoxide dismutase (SOD) activity, reduced thiol accumulation, and H2S accumulation in S. Typhimurium, cultured in either sulfate or cysteine as the sole sulfur source. Furthermore, we assessed the effects of the addition of ceftriaxone (CEF) and menadione (MEN) in these same parameters. In sulfate as the sole sulfur source, we found that the cysJIH operon and the cysB gene were required to full growth in minimal media, independently on the addition of CEF or MEN. Most importantly, both cysJIH and cysB contributed to diminish ROS levels, increase the SOD activity, increase the reduced thiols, and increase the H2S levels in presence of CEF or MEN. Moreover, the cysJIH operon exhibited a CysB-dependent upregulation in presence of these two antimicrobials compounds. On the other hand, when cysteine was used as the sole sulfur source, we found that cysJIH operon was completely negligible, were only cysB exhibited similar phenotypes than the described for sulfate as sulfur source. Unexpectedly, CysB downregulated cysJIH operon when cysteine was used instead of sulfate, suggesting a complex regulation of this system.

Increasing prevalence of hydrogen sulfide negative Salmonella in retail meats.

Hydrogen sulfide (H2S) production is considered a typical characteristic of Salmonella and an important marker for Salmonella isolation. In this study, a total of 82 (26%) Salmonella strains were isolated from 113 chicken and 204 pork samples, within which 49 Salmonella strains were H2S positive and 33 were H2S negative. Salmonella enterica serovar Derby was most prevalent in both pork and chicken followed by S. Typhimurium in pork and S. Heidelberg in chicken. Salmonella isolated from pork exhibited a much higher H2S positive rate than those from chicken (68% versus 31%). The most prevalent H2S negative serotypes were S. Derby (40%) and S. Heidelberg (30%) in chicken, and S. Typhimurium (23%) and S. Enteritidis (23%) in pork. spvC, a plasmid-encoded virulence marker, was detected in 51% and 42% of the H2S positive and negative Salmonella respectively. The presence of the two most important serotypes, S. Enteritidis and S. Typhimurium, as well as a virulence plasmid in H2S negative Salmonella suggested that H2S negative Salmonella is also a significant public health concern. Such finding warrants the development of an improved method for effective coverage of H2S negative Salmonella.

The Role of the Transsulfuration Pathway in Non-Alcoholic Fatty Liver Disease.

The prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing and approximately 25% of the global population may have NAFLD. NAFLD is associated with obesity and metabolic syndrome, but its pathophysiology is complex and only partly understood. The transsulfuration pathway (TSP) is a metabolic pathway regulating homocysteine and cysteine metabolism and is vital in controlling sulfur balance in the organism. Precise control of this pathway is critical for maintenance of optimal cellular function. The TSP is closely linked to other pathways such as the folate and methionine cycles, hydrogen sulfide (H2S) and glutathione (GSH) production. Impaired activity of the TSP will cause an increase in homocysteine and a decrease in cysteine levels. Homocysteine will also be increased due to impairment of the folate and methionine cycles. The key enzymes of the TSP, cystathionine ß-synthase (CBS) and cystathionine γ-lyase (CSE), are highly expressed in the liver and deficient CBS and CSE expression causes hepatic steatosis, inflammation, and fibrosis in animal models. A causative link between the TSP and NAFLD has not been established. However, dysfunctions in the TSP and related pathways, in terms of enzyme expression and the plasma levels of the metabolites (e.g., homocysteine, cystathionine, and cysteine), have been reported in NAFLD and liver cirrhosis in both animal models and humans. Further investigation of the TSP in relation to NAFLD may reveal mechanisms involved in the development and progression of NAFLD.

Hydrogen Sulfide Alleviates Anxiety, Motor, and Cognitive Dysfunctions in Rats with Maternal Hyperhomocysteinemia via Mitigation of Oxidative Stress.

Hydrogen sulfide (H2S) is endogenously produced from sulfur containing amino acids, including homocysteine and exerts neuroprotective effects. An increase of homocysteine during pregnancy impairs fetal growth and development of the offspring due to severe oxidative stress. We analyzed the effects of the H2S donor-sodium hydrosulfide (NaHS) administered to female rats with hyperhomocysteinemia (hHcy) on behavioral impairments and levels of oxidative stress of their offspring. Rats born from females fed with control or high methionine diet, with or without H2S donor injections were investigated. Rats with maternal hHcy exhibit increased levels of total locomotor activity and anxiety, decreased muscle endurance and motor coordination, abnormalities of fine motor control, as well as reduced spatial memory and learning. Oxidative stress in brain tissues measured by activity of glutathione peroxidases and the level of malondialdehyde was higher in rats with maternal hHcy. Concentrations of H2S and the activity and expression of the H2S generating enzyme-cystathionine-beta synthase-were lower compared to the control group. Administration of the H2S donor to females with hHcy during pregnancy prevented behavioral alterations and oxidative stress of their offspring. The acquisition of behavioral together with biochemical studies will add to our knowledge about homocysteine neurotoxicity and proposes H2S as a potential agent for therapy of hHcy associated disorders.

Elemental sulfur reduction to H2S by Tetrahymena thermophila.

Eukaryotic nucleocytoplasm is believed to be descended from ancient Archaea that respired on elemental sulfur. If so, a vestige of sulfur reduction might persist in modern eukaryotic cells. That was tested in Tetrahymena thermophila, chosen as a model organism. When oxygenated, the cells consumed H2S rapidly, but when made anoxic they produced H2S mostly by amino acid catabolism. That could be inhibited by adding aminooxyacetic acid, and then H2S production from elemental sulfur became more evident. Anoxic cell lysates produced H2S when provided with sulfur and NADH, but not with either substrate alone. When lysates were fractionated by centrifugation, NADH-dependent H2S production was 83% in the soluble fraction. When intact cells that had just previously oxidized H2S were shifted to anoxia, the cells produced H2S evidently by re-using the oxidized sulfur. After aerobic H2S oxidation was stopped, the oxidation product remained available for H2S production for about 10 min. The observed H2S production is consistent with an evolutionary relationship of nucleocytoplasm to sulfur-reducing Archaea. Mitochondria often are the cellular site of H2S oxidation, suggesting that eukaryotic cells might have evolved from an ancient symbiosis that was based upon sulfur exchange.

A new method for the simultaneous enhancement of methane yield and reduction of hydrogen sulfide production in the anaerobic digestion of waste activated sludge.

The biogas generated from anaerobic digestion (AD) also includes undesirable by-product such as hydrogen sulfide (H2S), which must be removed before the biogas can be used as a clean energy source. Therefore, it is necessary to find an appropriate strategy to simultaneously enhance the methane yield and reduce H2S production. An efficient strategy-pretreating sludge at pH 10 for 8d and adjusting the system at neutral pH to produce methane for 20d-is reported for the synchronous enhancement of methane production and reduction of H2S production during AD. The experimental results showed that the cumulative methane yield was 861.2±6.1mL/g volatile solids (VS) of sludge pretreated at pH 10 in semi-continuous stirred anaerobic reactors for 84d, an increase of 49.6% over the yield in the control. Meanwhile, the cumulative production of H2S was 144.1×10-4mL/g VS, 54.2% lower than that in the control.