Reduced H2S Production Research Articles

P0149 Deep functional analysis of a defined sulphide-reducing diet (4-SURE diet) as a novel therapy for ulcerative colitis (UC) demonstrates all specific luminal micro environmental objectives were achieved

Abstract Background As a dietary approach to reducing inflammation in ulcerative colitis (UC), the 4-SURE diet was designed to correct pathogenic alterations of excessive protein fermentation and hydrogen sulphide (H2S) production in the distal colon. Specific dietary objectives were achieved (Day et al,J Nutr 2022;152:1690) but it is uncertain whether mechanistic objectives could be achieved with 8 weeks of diet. Therefore, we aimed to perform a deep functional analysis (microbial and metabolomic) of the faeces. Methods Faecal samples of 28 adults with mild-moderately active UC were collected at week 0 and 8 of diet intervention, subsampled and stored at−80 °C. Shotgun metagenomic sequencing was used to identify genes involved in H2S metabolism. Metagenomic reads were trimmed, reads mapping to the human genome removed. Gene identification was performed using Diamond v2.1.9 for alignment against a database of genes involved in global sulphur cycling. Gas-chromatography mass-spectrometry was used to characterise volatile organic compounds (VOCs) with specific analysis of products of protein fermentation. Microbiota capacity to produce H2S was assessed by anaerobic incubation of homogenates followed by spectrophotometric determination of total H2S production. Results Majority of microbiome members belonged to the Bacteroidota and Bacillota phyla, with no significant difference in bacterial diversity determined between time points (p=0.16). However, using a Random Forest Classifier, known H2S producers, Odoribacter and Peptostreptococcaceae, were identified as the most important taxa in discriminating between pre and post-diet samples, with abundances markedly lower after diet intervention. A shift in microbiota metagenomic profiles putatively involved in H2S metabolism was identified post-diet, with differences (p<0.05, Wilcoxon signed-rank with Benjamini-Hochberg correction) in 12 of 23 analysed genes involved in H2S cycling determined, including iscS, cysK (Fig), metC, luxS, asrC and sseA. 173 faecal VOCs were identified across all samples. Indole (a specific marker of protein fermentation) decreased from 0.42 [-0.25,0.61] at week 0 to -0.28 [-0.77,0.48] at week 8 (p=0.007, FDR correction; Fig). Faecal H2S reduced from median 2.61 (interquartile range 2–4.96)µmol to 1.41 (0.81-2.27)µmol of H2S/g stool (wet weight)(p=0.002)(Fig). Conclusion Deep functional analysis of the faeces demonstrated the micro-environmental objectives of the 4-SURE diet successfully reduced protein fermentation, changed microbial community sulphur-metabolic gene profile and reduced H2S production ex vivo. Applying functional analysis to a diet study is novel, highlighting exemplar framework for including biomarkers of pathogenic relevance in analysis of therapeutic diets.

Mechanistic insights into Fe3O4-mediated inhibition of H2S gas production in sludge anaerobic digestion

The addition of iron-based conductive materials has been extensively validated as a highly effective approach to augment methane generation from anaerobic digestion (AD) process. In this work, it was additionally discovered that Fe3O4 notably suppressed the production of hazardous H2S gas during sludge AD. As the addition of Fe3O4 increased from 0 to 20 g/L, the accumulative H2S yields decreased by 89.2 % while the content of element sulfur and acid volatile sulfide (AVS) respectively increased by 55.0 % and 30.4 %. Mechanism analyses showed that the added Fe3O4 facilitated sludge conductive capacity, and boosted the efficiency of extracellular electron transfer, which accelerated the bioprocess of sulfide oxidation. Although Fe3O4 can chemically oxidize sulfide to elemental sulfur, microbial oxidation plays a major role in reducing H2S accumulation. Moreover, the released iron ions reacted with soluble sulfide, which promoted the chemical equilibrium of sulfide species from H2S to metal sulfide. Microbial analysis showed that some SRBs (i.e., Desulfomicrobium and Defluviicoccus) and SOB (i.e., Sulfuritalea) changed into keystone taxa (i.e., connectors and module hubs) in the reactor with Fe3O4 addition, showing that the functions of sulfate reduction and sulfur oxidation may play important roles in Fe3O4-present system. Fe3O4 presence also increased the content of functional genes encoding sulfide quinone reductase and flavocytochrome c sulfidedehydrogenase (e.g., Sqr and Fcc) that could oxidize sulfide to sulfur. The impact of other iron-based conductive material (i.e., zero-valent iron) was also verified, and the results showed that it could also significantly reduce H2S production. These findings provide new insights into the effect of iron-based conductive materials on anaerobic process, especially sulfur conversion.

Deciphering the role of Moringa oleifera seeds and probiotic bacteria on mitigation of biogas production from ruminants

Maintaining cleaner and more sustainable ecosystems by mitigating greenhouse gas (GHG) emissions from livestock through dietary manipulation is in demand. This study was aimed to assess the effect of Moringa oleifera seeds and probiotics (Pediococcus acidilactici BX-B122 and Bacillus coagulans BX-B118) as feed supplements on GHG production and fermentation profile from steers and sheep. The treatments included diets containing 0, 6, 12, and 18% of M. oleifera seeds meal and a mixture of probiotic bacteria (0.2 ml/g of diet). Total biogas production, CH4, CO, and H2S emission from animals (up to 48 h), rumen fermentation profile, and CH4 conversion efficiency were recorded using standard protocols. Results showed interaction among M. oleifera seeds and probiotics on asymptotic biogas production and total biogas production up to 48 h (P < 0.05). The rate of CH4 emission in steers was reduced from 0.1694 to 0.0447 ml/h using 6 and 18% of M. oleifera seeds (P < 0.05). Asymptotic CO and the rate of CO production were increased (P < 0.05) by supplementing different doses of M. oleifera seeds and probiotics. Adding 12% of M. oleifera seeds and probiotics reduced H2S production from 0.0675 to 0.0112 ml H2S/g DM (at 48 h of fermentation) in steers. In sheep, the additives mitigated H2S production from 0.0364 to 0.0029 ml H2S/g DM (at 48 h of fermentation), however there were not interaction (P = 0.7744). In addition, M. oleifera seeds and probiotics reduced the pH level and dry matter degradability (DMD) in steers and sheep (P < 0.0001) showing a positive impact on CH4:ME and CH4:OM (in steers) and CH4:SCFA (in sheep), while the interaction was not significant (P > 0.05) for CH4:SCFA (in steers) and CH4:ME and CH4:OM (in sheep). In conclusion, the interaction of M. oleifera seeds and probiotics in the feeding diet reduced GHG emissions and affected the fermentation profile of steers and sheep.

Effect of sodium sulfate concentration in drinking water for beef heifers, and the in vitro effect of bismuth subsalicylate on H2S production and fiber disappearance

This study evaluated the effects of ( i) increasing the concentration of sulfate in drinking water on dry matter intake (DMI), water intake, ruminal fermentation, and apparent total-tract digestibility, and ( ii) water sulfate concentration and bismuth subsalicylate (BSS) dose on in vitro ruminal fermentation. Eight ruminally cannulated beef heifers (382 ± 45 kg) were used in a replicated incomplete 3 × 3 Latin square design. Water treatments contained low (342 ± 29 mg/L; LS), medium (2785 ± 72 mg/L; MS), or high (4948 ± 163 mg/L; HS) sulfate. Ruminal digesta was collected to evaluate water sulfate and BSS dose (0.0%, 0.2%, 0.4%, and 0.6% dry matter) on ruminal H2S production in vitro. Water intake increased linearly as water sulfate concentration increased ( p = 0.002), but DMI was not affected. Heifers drinking MS and HS had greater ruminal H2S at 10.5 h after water provision than LS (sulfate × time, p &lt; 0.001). In vitro H2S production (µg and µg/mL of rumen inoculum) increased and plateaued with increasing sulfate ( p &lt; 0.001) and was linearly reduced ( p &lt; 0.001) by increasing BSS dose. Increasing water sulfate concentrations did not negatively affect water or feed intake but increased ruminal H2S concentrations. Bismuth subsalicylate may reduce H2S production.

Microbial electrolysis cell simultaneously enhancing methanization and reducing hydrogen sulfide production in anaerobic digestion of sewage sludge

The effects of microbial electrolysis cells (MECs) at three applied voltages (0.8, 1.3, and 1.6 V) on simultaneously enhancing methanization and reducing hydrogen sulfide (H2S) production in the anaerobic digestion (AD) of sewage sludge were studied. The results showed that the MECs at 1.3 V and 1.6 V simultaneously enhanced the methane production by 57.02 and 12.70% and organic matter removal by 38.77 and 11.13%, and reduced H2S production by 94.8 and 98.2%, respectively. MECs at 1.3 V and 1.6 V created a micro-aerobic conditions for the digesters with oxidation-reduction potential as −178∼-232 mv, which enhanced methanization and reduced H2S production. Sulfur reduction, H2S and elemental sulfur oxidation occurred simultaneously in the ADs at 1.3 V and 1.6 V. The relative abundances of sulfur-oxidizing bacteria increased from 0.11% to 0.42% and those of sulfur-reducing bacteria decreased from 1.24% to 0.33% when the applied voltage of MEC increased from 0 V to 1.6 V. Hydrogen produced by electrolysis enhanced the abundance of Methanobacterium and changed the methanogenesis pathway.

Reduced hydrogen sulfide production contributes to adrenal insufficiency induced by hypoxia via modulation of NLRP3 inflammasome activation

ABSTRACTObjective: Adrenocortical responsiveness is critical for maintaining glucocorticoids production and homeostasis during stress. We sought to investigate adrenocortical responsiveness during hypoxia in mice and the mechanisms responsible for the regulation of adrenal responsiveness.Methods: (1) Adult male WT mice were randomly divided into four groups: normoxia, hypoxia (24h), hypoxia (72h), hypoxia (72h) + GYY4137(hydrogen sulfide (H2S) donor, 133mmol/kg/day); (2) WT mice were randomly divided into four groups: sham, adrenalectomy (ADX), sham+hypoxia, ADX+hypoxia; (3) Cse-/- mice were randomly divided into two groups: Cse-/-, Cse-/- +GYY4137.Results: The circulatory level of corticosteroid induced by ACTH stimulation was significantly reduced in the mice with hypoxia compared with control mice. The mortality rate induced by lipopolysaccharide (LPS) increased during hypoxia. Cystathionine-γ-lyase (CSE) expression was significantly reduced in adrenal glands during hypoxia. GYY4137 treatment significantly increased adrenal responsiveness and attenuated NLRP3 inflammasome activation in mice treated by hypoxia and Cse-/- mice. Furthermore, The sulfhydrated level of PSMA7 in adrenal gland was decreased in the mice with hypoxia and Cse-/- mice. PSMA7 was S-sulfhydrated at cysteine 70. Blockage of S-sulfhydration of PSMA7 increased NLRP3 expression in adrenocortical cells.Conclusion: Reduced H2S production mediated hypo-adrenocortical responsiveness and NLRP3 inflammasome activation via PAMA7 S-sulfhydration during hypoxia.

148 Effect of Increasing Sodium Sulfate in Water on Intake on Ruminal Fermentation and the in Vitro Effect of Bismuth Subsalicylate on gas Production and NDF Digestibility

Abstract This study evaluated in vivo effects of increasing sulfate concentrations of drinking water on dry matter intake (DMI), water intake, ruminal fermentation, and apparent total-tract digestibility, and in vitro effects of water sulfate and bismuth subsalicylate (BSS) dose. Eight ruminal-cannulated beef heifers (382 ± 45 kg) were stratified into two complete and one incomplete 3×3 Latin squares with 28 d periods. Water contained 341 ± 29 (LS), 2,785 ± 71 (MS), or 4,948 ± 163 mg/L (HS) sulfate. Feed and water intake, ruminal fermentation, and nutrient digestibility were evaluated. Ruminal gas samples were collected every 4 h on d 27 to analyze hydrogen sulfide (H2S) concentration. At the end of each period, digesta collected from heifers in the complete Latin squares were used for in vitro incubation to evaluate water sulfate and BSS dose (0.0, 0.2, 0.4, and 0.6% dry matter). Water intake was 7 L/d greater (P &amp;lt; 0.01) for HS than LS, while MS did not differ. Water sulfate did not affect DMI, but total sulfur intake increased with increasing water sulfate concentration (P &amp;lt; 0.01). Heifers drinking MS and HS had greater ruminal H2S for 10.5 h after water provision and HS continued to be greater for another 4 h relative to LS (sulfate hour, P &amp;lt; 0.01). Ruminal SCFA concentration, molar proportions of major acids, and nutrient digestibility were not affected (P≥0.19). There were no interactions between water sulfate and BSS in vitro. In vitro H2S production (µg/48 h) increased with increasing sulfate concentration (P &amp;lt; 0.01) and was reduced (P=0.05) by BSS inclusion. Neutral detergent fiber digestibility was not affected by sulfate or BSS. Increasing water sulfate concentrations did not negatively affect water or feed intake but did increase H2S concentration and production in vivo and in vitro, respectively. Bismuth subsalicylate may be a potential strategy to reduce H2S production.

Heart and kidney H2S production is reduced in hypertensive and older rats

The prevalence of hypertension increases with age, but the mechanisms linking this phenomenon are not well understood. Hydrogen sulfide (H2S) may be involved in this process, as it plays a role in the cardiovascular system, affecting blood pressure and heart and kidney functions. The aim of this study was to evaluate the influence of hypertension and aging on sulfur-containing compounds metabolism in the hearts and kidneys of Wistar Kyoto (WKY) and Spontaneously Hypertensive Rats (SHR) of different age groups. We determined the expression and activity of four enzymes participating in H2S production: cystathionine beta-synthase (CBS), cystathionine gamma-lyase (CTH), 3-mercaptopyruvate sulfurtransferase (MPST), and thiosulfate sulfurtransferase (TST). The levels of reduced/oxidized glutathione, cysteine, cystine, and cystathionine, and the ability of tissues to form hydrogen sulfide were also investigated. Tissues obtained from younger WKY rats produced the highest amounts of H2S. The effect of hypertension on the metabolism of sulfur-containing compounds was manifested by a decrease in sulfane sulfur concentrations in heart homogenates and a decrease in CTH activity in the kidneys. The hearts and kidneys of older WKY rats were characterized by lower MPST or CTH gene expression, respectively, compared to younger animals. Our study demonstrates that hypertension and aging influence cardiac and renal sulfur-containing compounds metabolism and reduce H2S production. Furthermore, we showed that MPST plays a major role in the production of hydrogen sulfide in the heart and CTH in the kidneys of rats.

Roles of a Cysteine Desulfhydrase LCD1 in Regulating Leaf Senescence in Tomato.

Hydrogen sulfide (H2S), a novel gasotransmitter in both mammals and plants, plays important roles in plant development and stress responses. Leaf senescence represents the final stage of leaf development. The role of H2S-producing enzyme L-cysteine desulfhydrase in regulating tomato leaf senescence is still unknown. In the present study, the effect of an L-cysteine desulfhydrase LCD1 on leaf senescence in tomato was explored by physiological analysis. LCD1 mutation caused earlier leaf senescence, whereas LCD1 overexpression significantly delayed leaf senescence compared with the wild type in 10-week tomato seedlings. Moreover, LCD1 overexpression was found to delay dark-induced senescence in detached tomato leaves, and the lcd1 mutant showed accelerated senescence. An increasing trend of H2S production was observed in leaves during storage in darkness, while LCD1 deletion reduced H2S production and LCD1 overexpression produced more H2S compared with the wild-type control. Further investigations showed that LCD1 overexpression delayed dark-triggered chlorophyll degradation and reactive oxygen species (ROS) accumulation in detached tomato leaves, and the increase in the expression of chlorophyll degradation genes NYC1, PAO, PPH, SGR1, and senescence-associated genes (SAGs) during senescence was attenuated by LCD1 overexpression, whereas lcd1 mutants showed enhanced senescence-related parameters. Moreover, a correlation analysis indicated that chlorophyll content was negatively correlated with H2O2 and malondialdehyde (MDA) content, and also negatively correlated with the expression of chlorophyll degradation-related genes and SAGs. Therefore, these findings increase our understanding of the physiological functions of the H2S-generating enzyme LCD1 in regulating leaf senescence in tomato.

A nuclear-localized cysteine desulfhydrase plays a role in fruit ripening in tomato

Hydrogen sulfide (H2S) is a gaseous signaling molecule that plays multiple roles in plant development. However, whether endogenous H2S plays a role in fruit ripening in tomato is still unknown. In this study, we show that the H2S-producing enzyme l-cysteine desulfhydrase SlLCD1 localizes to the nucleus. By constructing mutated forms of SlLCD1, we show that the amino acid residue K24 of SlLCD1 is the key amino acid that determines nuclear localization. Silencing of SlLCD1 by TRV-SlLCD1 accelerated fruit ripening and reduced H2S production compared with the control. A SlLCD1 gene-edited mutant obtained through CRISPR/Cas9 modification displayed a slightly dwarfed phenotype and accelerated fruit ripening. This mutant also showed increased cysteine content and produced less H2S, suggesting a role of SlLCD1 in H2S generation. Chlorophyll degradation and carotenoid accumulation were enhanced in the SlLCD1 mutant. Other ripening-related genes that play roles in chlorophyll degradation, carotenoid biosynthesis, cell wall degradation, ethylene biosynthesis, and the ethylene signaling pathway were enhanced at the transcriptional level in the lcd1 mutant. Total RNA was sequenced from unripe tomato fruit treated with exogenous H2S, and transcriptome analysis showed that ripening-related gene expression was suppressed. Based on the results for a SlLCD1 gene-edited mutant and exogenous H2S application, we propose that the nuclear-localized cysteine desulfhydrase SlLCD1 is required for endogenous H2S generation and participates in the regulation of tomato fruit ripening.

Magnetite-assisted in situ microbial oxidation of H2S to S0 during anaerobic digestion: A new potential for sulfide control

Direct interspecies electron transfer (DIET) between exoelectrogenic fatty acid-oxidizing bacteria and electrotrophic methanogens has recently been discovered, and studies have suggested that promoting DIET by adding electrically conductive material can effectively enhance the methanogenic performance and stability of anaerobic digestion (AD). This study investigated the effect of conductive magnetite (Fe3O4) addition on the AD of a sulfur-rich organic waste mixture, with an emphasis on the fate of sulfur and on H2S production. In contrast to previous findings, methanogenic performance under magnetite-added conditions was not significantly enhanced within the tested dose range of up to 20 mM Fe. When magnetite was added, H2S production decreased remarkably along with the extracellular accumulation of S0. Moreover, the H2S content in biogas was below 100 ppmv at magnetite doses of 8 mM Fe or higher (>6000 ppmv under control conditions, i.e., without magnetite). The reduced H2S production appears to be due to the anaerobic oxidation of sulfide to S0 because sulfate reduction remained active, whereas FeS was not produced under magnetite-added conditions. Based on microbial community analysis results and thermodynamic calculations, the electric syntrophy via DIET between exoelectrogenic anaerobic sulfide-oxidizing bacteria and electrotrophic methanogens is suggested to have been promoted by magnetite. To the best of our knowledge, this study is the first to propose an electro-syntrophic association that couples the oxidation of sulfide to S0 with the reduction of CO2 to CH4 in methanogenic environments. The present findings open a new possibility for in situ H2S control and sulfur recovery in AD processes for sulfur-rich waste treatment.

Biochar addition with Fe impregnation to reduce H2S production from anaerobic digestion

Corn stover biochar (CSB) and maple biochar (MB) were added into anaerobic digesters and evaluated for hydrogen sulfide (H2S) reductions. This was the first study to show Fe-impregnated biochar can eliminate H2S production. The novel study evaluated biochar addition on H2S reduction and nutrient concentrations using three experiments to test the effect of: 1) biochar concentration, 2) biochar particle size, and 3) Fe-impregnated biochar using triplicate lab-scale reactors. At the highest biochar dose (1.82 g biochar/g manure TS), H2S production was 90.5% less than the control treatment (351 mL H2S/kg VS). Biochar particle size did not significantly affect H2S concentration. The Fe-impregnated biochar (0.5 g biochar/g manure TS) reactors had no H2S detected in the CSB-Fe system. Methane (CH4) in the biochar and control treatments were not significantly different in all three experiments. The results show that biochar added to digesters can significantly reduce H2S production without affecting CH4 production.

The interventional effects of saturated hydrogen saline on lung injury in rats with cecal ligation and puncture operation

To investigate the hypothesis that hydrogen could ameliorate cecal ligation and puncture (CLP)-induced lung injury of rats by inhibiting cystathionine-gamma-lyase/hydrogen sulfide (CSE/H2S) system. A total number of 24 healthy male SD rats weighting 250～300 g were randomly divided into four groups (n=6 in each group): sham operation group(sham group), hydrogen-rich saline control group(H2 group), CLP group and hydrogen-rich saline treatment group(CLP+H2 group). The rats were treated with hydrogen-rich saline or saline 10 min before CLP or sham operation. At 8 h of sham or CLP operation, lung samples were obtained to detect the changes of the CSE/H2S system using biochemical and RT-PCR methods. In order to further confirm the role of H2S during hydrogen improve the lung injury of CLP rats, we also observed the effect of hydrogen-rich saline on the lung injury induced by H2S donor-sodium sodium hydrosulfide (NaHS). Thirty-two healthy male SD rats (250~300 g) were randomly divided into four groups (n=8 in each group): control group, H2S group, H2S+H2 group and H2 group. Saline(10 mg/kg) or NaHS(H2S donor, 56 μmol/kg) was injected intraperitoneally (10 mg/kg) respectively into rats in the control rats or H2S group. For rats in the H2S+H2 and H2 group, hydrogen-rich saline (10 mg/kg) was injected 10 min before saline or NaHS administration. Eight hours after the LPS saline or NaHS administration, lung coefficient, MDA content, and MPO activity were detected. The contents of TNF-α, IL-6 and IL-10 in lung tissue were measured, and the morphological changes of lung tissue were also observed. CSE/H2S system up-regulating were observed in animals exposed to CLP. Hydrogen-rich saline treatment significantly inhibited CSE/H2S system as indicated by significantly reduced H2S production in lung, along with a decreased CSE activity and CSE mRNA expression (all P＜0.05). Importantly, the results showed that lung injury and lung tissue inflammation were observed in animals exposed to NaHS. Hydrogen-rich saline treatment significantly attenuated lung injury as indicated by significantly improved histological changes in lung, significantly reduced index of quantitative assessment (IQA), MDA content and lung coefficient (all P＜0.05). MPO activity in lung tissue was significantly reduced along with decreased productions of TNF-α and IL-6, and an increased production of IL-10 in the presence of hydrogen (all P＜0.05), demonstrating antioxidant and anti-inflammatory effect of hydrogen in NaHS-induced ALI. These results indicate that hydrogen-rich saline peritoneal injection improves the lung injury induced by CLP operation. The therapeutic effects of hydrogen-rich saline may be related to suppressing the production of H2S.

Modulation of colonic hydrogen sulfide production by diet and mesalazine utilizing a novel gas-profiling technology

ABSTRACTExcessive hydrogen sulfide (H2S) production from gut microbial metabolism may have clinically important relevance in the pathogenesis of gut disorders, including ulcerative colitis. However, little is known regarding factors that alter its production. Using a newly-designed in vitro gas-profiling technology, the study aimed to verify real-time H2S measurement reproducibility and thereafter, assess its production following exposure to dietary factors and 5-aminosalicylate acid (5-ASA). Measurements of H2S, carbon dioxide, hydrogen and methane measurements were compared between gas-profiling systems. Homogenized slurries were prepared from freshly-passed healthy human feces. Fifty ml slurries were aliquoted into separate fermentation chambers and substrates added including 1 g highly fermentable fructo-oligosaccharides (FOS) or resistant starch Hi-Maize (RS), or minimally fermentable psyllium or sterculia, 1 g cysteine, 0.9 g sodium sulfate or 1.2 mL of 1 M 5-ASA alone or in combinations. H2S release was sampled every 5 mins over 4-h and expressed relative to unspiked controls. RS suppressed H2S production by a mean 89.0 (SEM 4.8)% and FOS by 82.2 (6.2)% compared to <35 (17)% by psyllium and sterculia (p<0.001, two-way ANOVA). Cysteine stimulated H2S production by 1557 (532)%. The addition of FOS to slurries containing cysteine significantly suppressed H2S by 90 (2)% over the addition of 5-ASA (0.3 (2)%, p<0.001). Sulfate and 5-ASA had minimal overall effects. In conclusion, the H2S-profiling technology is a reproducible tool. Production of H2S is greatly enhanced by sulfur-amino acids but not inorganic sulfate, and is effectively suppressed by readily fermentable fibers. These findings inform potential designs of dietary therapies to reduce H2S production in vivo.

Iron Oxide to Mitigate Hydrogen Sulfide Gas Release from Gypsum-Bedded Dairy Manure Storages

Abstract. To mitigate noxious hydrogen sulfide (H2S) gas release that has been observed from gypsum-laden dairy manure, three additives were studied in sequential investigations. Three trials with specific aims were conducted using experimental vessels containing 15 kg of dairy manure each. Trial 1 investigated two additives: iron oxide (specifically, iron oxide-hydroxide, FeOOH) and a proprietary gypsum-lime based product (DriMatt). Trial 2 investigated effective ratios of gypsum to iron oxide and a modified DriMatt additive, and trial 3 evaluated iron oxide at the most effective ratio. Manure agitation events were monitored in the first two trials, while gas releases were continuously monitored in trial 3 during and between agitations. Hydrogen sulfide concentrations were captured using electrochemical sensors or a Fourier transfer infrared (FTIR) gas analyzer assembly over an incubation period of two months for the first two trials and over 40 days for the third trial. Additionally, nutrient analyses were performed for each trial. Extremely high concentrations of H2S were observed during most manure agitation events (500 to 8000 ppm), while minimum releases (&amp;lt;10 ppm) were found when samples were static. Means of maximum concentrations of H2S were compared among treatments in each trial. Statistical tests showed that adding iron oxide to gypsum-laden manure reduced H2S production by an average of 94% compared to treatments without iron oxide. With a 1:1 molar ratio of iron oxide to gypsum, the level of H2S released was diminished to as low as the control manure (without gypsum). Therefore, iron oxide is a promising additive to mitigate H2S production in gypsum-laden dairy manure during agitation events. Keywords: Additive, Dairy, Gas, Gypsum bedding, Hydrogen sulfide, Iron oxide, Manure, Safety, Storage.

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.

Variation in properties of the sediment following electrokinetic treatments

ABSTRACT Many studies have reported variation in properties of the sediment within electrokinetic treatments (EKTs). However, we aim to reveal the variation in properties of the sediment following EKTs through laboratory experiments. We collected sewage-derived sediment from a littoral region, and passed it through a 2-mm sieve. We used a potentiostat to cause electrical current in EKT. We measured the sediment properties such as pH, redox potential (ORP), and hydrogen sulphide (H2S) concentration at the end of EKT and at 30 days following EKT. Results showed decreases in pH, increases in ORP, and decreases in H2S concentration at the end of EKT. Compared with the sediment without EKT, the decrease in ORP for the sediment within EKT was higher at 30 days following EKT. These suggest that anaerobic digestion of organic compounds occurs in the sediment following EKT, of which the oxidants produced by EKT serve as electron acceptors and organic compounds serve as electron donors. Furthermore, we found that EKT can remove H2S from the sediment and reduce H2S production in the sediment within EKT when compared to the case without EKT. These ensure that EKT can be used to remove H2S and control H2S production in the sediment.

Effects of L-tryptophan, Fructan, and Casein on Reducing Ammonia, Hydrogen Sulfide, and Skatole in Fermented Swine Manure.

The effects of daily dietary Bacillus subtilis (Bs), and adding L-tryptophan, fructan, or casein to fecal fermentation broths were investigated as means to reduce the production of noxious gas during manure fermentation caused by ammonia, hydrogen sulfide (H2S), and 3-methylindole (skatole). Eighty swine (50.0±0.5 kg) were equally apportioned to an experimental group given Bs in daily feed, or a control group without Bs. After 6 weeks, fresh manure was collected from both groups for fermentation studies using a 3×3 orthogonal array, in which tryptophan, casein, and fructan were added at various concentrations. After fermentation, the ammonia, H2S, L-tryptophan, skatole, and microflora were measured. In both groups, L-tryptophan was the principle additive increasing skatole production, with significant correlation (r = 0.9992). L-tryptophan had no effect on the production of ammonia, H2S, or skatole in animals fed Bs. In both groups, fructan was the principle additive that reduced H2S production (r = 0.9981). Fructan and Bs significantly interacted in H2S production (p = 0.014). Casein was the principle additive affecting the concentration of ammonia, only in the control group. Casein and Bs significantly interacted in ammonia production (p = 0.039). The predominant bacteria were Bacillus spp. CWBI B1434 (26%) in the control group, and Streptococcus alactolyticus AF201899 (36%) in the experimental group. In summary, daily dietary Bs reduced ammonia production during fecal fermentation. Lessening L-tryptophan and increasing fructan in the fermentation broth reduced skatole and H2S.

Endogenous Hydrogen Sulfide Production Is Essential for Dietary Restriction Benefits

Dietary restriction (DR) without malnutrition encompasses numerous regimens with overlapping benefits including longevity and stress resistance, but unifying nutritional and molecular mechanisms remain elusive. In a mouse model of DR-mediated stress resistance, we found that sulfur amino acid (SAA) restriction increased expression of the transsulfuration pathway (TSP) enzyme cystathionine γ-lyase (CGL),resulting in increased hydrogen sulfide (H2S) production and protection from hepatic ischemia reperfusion injury. SAA supplementation, mTORC1 activation, or chemical/genetic CGL inhibition reduced H2S production and blocked DR-mediated stress resistance. Invitro, the mitochondrial protein SQR was required for H2S-mediated protection during nutrient/oxygen deprivation. Finally, TSP-dependent H2S production was observed in yeast, worm, fruit fly, and rodent models of DR-mediated longevity. Together, these data are consistent with evolutionary conservation of TSP-mediated H2S as a mediator ofDR benefits with broad implications for clinical translation. PAPERFLICK:

MiRNA-30 family inhibition protects against cardiac ischemic injury by regulating cystathionine-γ-lyase expression.

Myocardial infarction (MI) is a leading cause of death globally. MicroRNAs (miRNAs) have been identified as a novel class of MI injury regulators. Hydrogen sulfide (H2S) is a gaseous signaling molecule that regulates cardiovascular function. The purpose of this study was to explore the role of the miR-30 family in protecting against MI injury by regulating H2S production. The expression of miR-30 family was upregulated in the murine MI model as well as in the primary cardiomyocyte hypoxic model. However, the cystathionine-γ-lyase (CSE) expression was significantly decreased. The overexpression of miR-30 family decreased CSE expression, reduced H2S production, and then aggravated hypoxic cardiomyocyte injury. In contrast, silencing the whole miR-30 family can protect against hypoxic cell injury by elevating CSE and H2S level. Nonetheless, the protective effect was abolished by cotransfecting with CSE-siRNA. Systemic delivery of a locked nucleic acid (LNA)-miR-30 family inhibitor correspondingly increased CSE and H2S level, then reduced infarct size, decreased apoptotic cell number in the peri-infarct region, and improved cardiac function in response to MI. However, these cardioprotective effects were absent in CSE knockout mice. MiR-30b overexpression in vivo aggravated MI injury because of H2S reduction, and this could be rescued by S-propargyl-cysteine (SPRC), which is a novel modulator of CSE, or further exacerbated by propargylglycine (PAG), which is a selective inhibitor of CSE. Our findings reveal a novel molecular mechanism for endogenous H2S production in the heart at the miRNA level and demonstrate the therapeutic potential of miR-30 family inhibition for ischemic heart diseases by increasing H2S production.