Prevention of formation of acid drainage from high-sulfur coal refuse by inhibition of iron- and sulfur-oxidizing microorganisms. II. Inhibition in "run of mine" refuse under simulated field conditions

The combination of sodium lauryl sulfate and benzoic acid effectively inhibits iron- and sulfur-oxidizing bacteria in coal refuse and prevents the conversion of iron pyrite to sulfate, ferric iron, and sulfuric acid, thereby significantly reducing the formation of acidic drainage from coal refuse. The inhibitors were effective in a concentration of 1.1 mg/kg refuse, and data indicate that the SLS was in excess of the concentration required. The treatment was compatible with the use of lime for neutralization of acid present prior to inhibition of its formation.     

Bacteria and Acidic Drainage from Coal Refuse: Inhibition by Sodium Lauryl Sulfate and Sodium Benzoate

The application of an aqueous solution of sodium lauryl sulfate and sodium benzoate to the surface of high-sulfur coal refuse resulted in the inhibition of iron-and sulfur-oxidizing chemoautotrophic bacteria and in the decrease of acidic drainage from the refuse, suggesting that acid drainage can be abated in the field by inhibiting iron- and sulfur-oxidizing bacteria.     

矿区高硫煤覆盖对土壤细菌群落组成和多样性的影响

【背景】土壤微生物生态研究对于矿区污染生态影响调查和矿区生态恢复都具有非常重要的意义,高硫煤覆盖对其下方土壤微生物群落的影响目前尚不清楚。【目的】探究煤矿区长期高硫煤覆盖对其下方土壤细菌群落组成和多样性产生的影响。【方法】从高硫煤矿附近煤场采集3种类型土壤样品(高硫煤覆盖层、高硫煤下方土壤、对照土壤),通过测定土壤理化性质和运用高通量测序技术研究高硫煤覆盖对土壤性质和细菌群落产生的影响。【结果】与对照土壤相比,高硫煤覆盖使其下方土壤pH值降低,硫酸盐、有机质、有效氮、有效磷含量升高。高硫煤覆盖使其下方土壤中细菌的多样性指数降低,细菌群落组成发生了较大改变。在属水平上,与对照土壤相比,高硫煤下方土壤中Bacillus的相对丰度较高;Acidiphilium与Sulfobacillus在3个样本组中的相对丰度呈现为高硫煤覆盖层>高硫煤下方土壤>对照土壤,硫氧化菌在高硫煤中的优势地位及其对下方土壤的影响由此可以得到证实。共现性网络分析表明,Acidiphilium和Sulfobacillus在高硫煤覆盖区域的土壤细菌群落中占有重要地位,对其他细菌类群影响非常大。【结论】高硫煤覆盖及硫氧化菌的优势存在,对高硫煤下方土壤的理化性质及细菌群落产生明显影响。研究结果有助于增进对矿区土壤微生物生态的认识,为高硫煤矿区生态恢复治理提供微生物学理论基础。     

Relative role of eukaryotic and prokaryotic microorganisms in phenanthrene transformation in coastal sediments

THE RELATIVE ROLE OF EUKARYOTIC VERSUS PROKARYOTIC MICROORGANISMS IN PHENANTHRENE TRANSFORMATION WAS MEASURED IN SLURRIES OF COASTAL SEDIMENT BY TWO DIFFERENT APPROACHES: detection of marker metabolites and use of selective inhibitors on phenanthrene biotransformation. Phenanthrene biotransformation was measured by polar metabolite formation and CO(2) evolution from [9-C]phenanthrene. Radiolabeled metabolites were tentatively identified by high-performance liquid chromatography (HPLC) separation combined with UV/visible spectral analysis of HPLC peaks and comparison to authentic standards. Both yeasts and bacteria transformed phenanthrene in slurries of coastal sediment. Two products of phenanthrene oxidation by fungi, phenanthrene trans-3,4-dihydrodiol and 3-phenanthrol, were produced in yeast-inoculated sterile sediment. However, only products of phenanthrene oxidation typical of bacterial transformation, 1-hydroxy-2-naphthoic acid and phenanthrene cis-3,4-dihydrodiol, were isolated from slurries of coastal sediment with natural microbial populations. Phenanthrene trans-dihydrodiols or other products of fungal oxidation of phenanthrene were not detected in the slurry containing a natural microbial population. A predominant role for bacterial transformation of phenanthrene was also suggested from selective inhibitor experiments. Addition of streptomycin to slurries, at a concentration which suppressed bacterial viable counts and rates of [methyl-H]thymidine uptake, completely inhibited phenanthrene transformation. Treatment with colchicine, at a concentration which suppressed yeast viable counts, depressed phenanthrene transformation by 40%, and this was likely due to nontarget inhibition of bacterial activity. The relative contribution of eukaryotic microorganisms to phenanthrene transformation in inoculated sterile sediment was estimated to be less than 3% of the total activity. We conclude that the predominant degraders of phenanthrene in muddy coastal sediments are bacteria and not eukaryotic microorganisms.     

Biodegradation of Benzene in a Laboratory-Scale Biobarrier at Low Dissolved Oxygen Concentrations

A laboratory-scale permeable biobarrier exhibited high removal efficiencies of benzene at inlet concentrations of 0.4 to 35.1?mg/L and with a limited supply of dissolved oxygen. The supplied oxygen was less than the demand for a complete aerobic oxidation of benzene. Stainless steel pieces or granulated peat moss were used as packing material for microbial support in the biobarrier. Removal efficiencies ranged from 63.9% to 99.9% in the stainless steel-packed biobarrier and from 70.4% to 97.2% in the peat moss-packed biobarrier, while benzene elimination rate changed from 0.2 to 10.4?mg/L-d and from 0.1 to 3.7?mg/L-d in the two biobarriers, respectively. The consumption of sulfate and the presence of sulfate-reducing bacteria suggested the contribution of anaerobic metabolism in the biodegradation of benzene. The biodegradation of benzene under microaerophilic conditions (defined as dissolved oxygen concentrations <2?mg/L) was demonstrated during independent batch experiments. The maximum specific rate of benzene biodegradation with concentrations of 22.0 to 65.9?mg/L under microaero-philic conditions was 2.6 mg/mg biomass-d.     

Development of a large-scale continuous substrate feed process for the biotransformation of simvastatin by Nocardia s.p

This article describes a process for microbial hydroxylation of simvastatin by a Nocardia sp. Simvastatin (Zocor) belongs to the family of HMGCoA reductase inhibitors used as cholesterol-lowering drugs. Studies at 14 L scale showed that high substrate (simvastatin) concentrations inhibited product formation; consequently, continuous slow feeding of the substrate was introduced to maintain low residual simvastatin concentrations. Dissolved oxygen levels above 50% air saturation were desirable for the biotransformation. The process was scaled up to 19,000-L fermentors using an on-line filter sterilization system for substrate feeding. The feed rate was regulated by off-line high-pressure liquid chromatography (HPLC) assays to keep the substrate concentration below 20 mg/L. Intermittent addition of nutrients helped to boost the bioconversion rate to give final titers of 400 mg/L 6-beta-hydroxymethyl simvastatin. Enrichment of the nutrient medium led to bioconversion titers of 800 mg/L 6-beta-hydroxymethyl simvastatin. Bioconversion efficiencies (desired product/substrate) of 22-25% with a ratio of desired product/side products of 0.7 were obtained by this process.     

锁阳原花青素对葡萄糖/甘氨酸模拟体系糖基化终产物的抑制效果

为探究锁阳原花青素对葡萄糖/甘氨酸模拟体系中形成的晚期糖基化终产物(advanced glycation end-product,AGEs)的抑制效果。本实验考察了不同温度,不同反应时间下锁阳原花青素对体系中AGEs抑制的影响,同时探讨了多种金属离子对抑制效果的作用。结果表明:在80℃下加热75 min,锁阳原花青素的质量浓度为1 mg/mL时,对AGEs的抑制效果可达85.73%±1.57%,而100℃下加热30 min时抑制率达到74.01%±1.45%。当加入金属离子(Al³⁺、Cu²⁺、Fe²⁺、Mg²⁺、Ca²⁺、Zn²⁺)时,对AGEs的形成既有抑制作用也有促进作用;与仅有锁阳原花青素存在时相比,金属离子在低浓度时减弱了锁阳原花青素对AGEs的抑制作用,高浓度差异显著。此结果可为天然AGEs抑制剂的开发和在食品中应用提供参考价值。     

Increases in microbial nitrogen production and efficiency in vitro with three inhibitors of ruminal methanogenesis

It was hypothesized that the addition of crotonic acid or 3-butenoic acid would relieve constraints in digestibility observed when methane formation is inhibited by lumazine, propynoic acid, or ethyl 2-butynoate. In six incubations, one of the three methanogenesis inhibitors, at three different concentrations, was combined with either crotonic acid or 3-butenoic acid at two different concentrations. A mixture of buffer and ruminal fluid (4:1) was incubated with grass hay in Erlenmeyer flasks for 72 h. Initial concentrations were 0, 0.6, and 1.2 mmol/L for lumazine; 0, 2, and 4 mmol/L for propynoic acid; and 0, 4, and 8 mmol/L for ethyl 2-butynoate. 15Nitrogen (N) incorporation was used as a microbial marker. All three methanogenesis inhibitors decreased proteolysis. Propynoic acid and ethyl 2-butynoate at 8 mmol/L also decreased the digestibility of organic matter and neutral detergent fibre. However, all three inhibitors of methanogenesis increased the production of microbial N through an improvement of synthetic efficiency. Crotonic acid and 3-butenoic acid were generally ineffective in compensating digestibility decreases caused by the inhibitors of methanogenesis. It is of interest to elucidate the mechanisms by which these compounds increased the efficiency of microbial N production. Lumazine and the addition of low levels of ethyl 2-butynoate could potentially benefit animal production by lowering methane emissions, decreasing ruminal proteolysis, and increasing microbial N production without affecting organic matter digestibility.     

Effects of imposed salinity gradients on dissimilatory arsenate reduction, sulfate reduction, and other microbial processes in sediments from two California soda lakes

Salinity effects on microbial community structure and on potential rates of arsenate reduction, arsenite oxidation, sulfate reduction, denitrification, and methanogenesis were examined in sediment slurries from two California soda lakes. We conducted experiments with Mono Lake and Searles Lake sediments over a wide range of salt concentrations (25 to 346 g liter(-1)). With the exception of sulfate reduction, rates of all processes demonstrated an inverse relationship to total salinity. However, each of these processes persisted at low but detectable rates at salt saturation. Denaturing gradient gel electrophoresis analysis of partial 16S rRNA genes amplified from As(V) reduction slurries revealed that distinct microbial populations grew at low (25 to 50 g liter(-1)), intermediate (100 to 200 g liter(-1)), and high (>300 g liter(-1)) salinity. At intermediate and high salinities, a close relative of a cultivated As-respiring halophile was present. These results suggest that organisms adapted to more dilute conditions can remain viable at high salinity and rapidly repopulate the lake during periods of rising lake level. In contrast to As reduction, sulfate reduction in Mono Lake slurries was undetectable at salt saturation. Furthermore, sulfate reduction was excluded from Searles Lake sediments at any salinity despite the presence of abundant sulfate. Sulfate reduction occurred in Searles Lake sediment slurries only following inoculation with Mono Lake sediment, indicating the absence of sulfate-reducing flora. Experiments with borate-amended Mono Lake slurries suggest that the notably high (0.46 molal) concentration of borate in the Searles Lake brine was responsible for the exclusion of sulfate reducers from that ecosystem.     

Biochemical effects of the hypoglycaemic compound pent-4-enoic acid and related non-hypoglycaemic fatty acids. Effects of the free acids and their carnitine esters on coenzyme A-dependent oxidations in rat liver mitochondria

1. The synthesis of pent-4-enoyl-l-carnitine, cyclopropanecarbonyl-l-carnitine and cyclobutanecarbonyl-l-carnitine is described. 2. Pent-4-enoate strongly inhibits palmitoyl-l-carnitine oxidation in coupled but not in uncoupled mitochondria. Pent-4-enoyl-l-carnitine strongly inhibits palmitoyl-l-carnitine oxidation in uncoupled mitochondria. Prior intramitochondrial formation of pent-4-enoyl-CoA is therefore necessary for inhibition. 3. There was a small self-limiting pulse of oxidation of pent-4-enoyl-l-carnitine during which the ability to inhibit the oxidation of subsequently added palmitoyl-l-carnitine developed. 4. Pent-4-enoate and pent-4-enoyl-l-carnitine are equally effective inhibitors of the oxidation of all even-chain acylcarnitines of chain length C(4)-C(16). Pent-4-enoyl-l-carnitine also inhibits the oxidation of pyruvate and of 2-oxoglutarate. 5. Pent-4-enoate strongly inhibits the oxidation of palmitate but not that of octanoate. This is presumably due to competition between octanoate and pent-4-enoate for medium-chain acyl-CoA ligase. 6. There was less inhibition of the oxidation of pyruvate by pent-4-enoyl-l-carnitine, and of palmitoyl-l-carnitine by cyclopropanecarbonyl-l-carnitine, after pre-incubation with 10mm-arsenate. This suggests that these inhibitions were caused either by depletion of free CoA or by increase of acyl-CoA concentrations, since arsenate deacylates intramitochondrial acyl-CoA. There was little effect on the inhibition of palmitoyl-l-carnitine oxidation by pent-4-enoyl-l-carnitine. 7. Penta-2,4-dienoate strongly inhibited palmitoyl-l-carnitine oxidation in coupled mitochondria; acrylate only inhibited slightly. 8. Pent-4-enoate (0.1mm) caused a rapid and almost complete decrease in free CoA and a large increase in acid-soluble acyl-CoA when incubated with coupled mitochondria. Cyclopropanecarboxylate caused a similar decrease in CoA, with an equivalent rise in acid-soluble acyl-CoA concentrations. n-Pentanoate caused extensive lowering of CoA and a large increase in acid-soluble acyl-CoA and acetyl-CoA concentrations. Octanoate caused a 50% lowering of CoA and an increase in acid-soluble acyl-CoA and acetyl-CoA concentrations. 9. Cyclopropanecarboxylate and n-pentanoate were less potent inhibitors of palmitate oxidation than was pent-4-enoate. 10. It is concluded that pent-4-enoate causes a specific inhibition of beta-oxidation after the formation intramitochondrially of its metabolites.     

Role of methanogens and other bacteria in degradation of dimethyl sulfide and methanethiol in anoxic freshwater sediments

The roles of several trophic groups of organisms (methanogens and sulfate- and nitrate-reducing bacteria) in the microbial degradation of methanethiol (MT) and dimethyl sulfide (DMS) were studied in freshwater sediments. The incubation of DMS- and MT-amended slurries revealed that methanogens are the dominant DMS and MT utilizers in sulfate-poor freshwater systems. In sediment slurries, which were depleted of sulfate, 75 micromol of DMS was stoichiometrically converted into 112 micromol of methane. The addition of methanol or MT to DMS-degrading slurries at concentrations similar to that of DMS reduced DMS degradation rates. This indicates that the methanogens in freshwater sediments, which degrade DMS, are also consumers of methanol and MT. To verify whether a competition between sulfate-reducing and methanogenic bacteria for DMS or MT takes place in sulfate-rich freshwater systems, the effects of sulfate and inhibitors, like bromoethanesulfonic acid, molybdate, and tungstate, on the degradation of MT and DMS were studied. The results for these sulfate-rich and sulfate-amended slurry incubations clearly demonstrated that besides methanogens, sulfate-reducing bacteria take part in MT and DMS degradation in freshwater sediments, provided that sulfate is available. The possible involvement of an interspecies hydrogen transfer in these processes is discussed. In general, our study provides evidence for methanogenesis as a major sink for MT and DMS in freshwater sediments.     

Inhibition of Auxin Polar Transport Affecting the Model of Leaf Growth and Development

Tobacco (Nicotiana tabacum L. cv. Gexin No. 1) leaf slices were cultured in MS medium with different concentrations of auxin polar transport inhibitors (2, 3, 5-triiodobenzoic acid (TIBA), trans-cinnamic acid (CA), and 9-hydooxyflurence-9-carboxylic acid (HFCA)) and their effects on bud formation were observed. Although the effective concentrations vary with different inhibitors, all of them induced the formation of trumpet-shaped leaves. The frequencies of trumpet-shaped leaves were increased with the concentrations of inhibitors in media, and it was up to 82.1% when cultured in the medium containing 7.5 mg/L TIBA. The trumpet-shaped leaves were formed in different sites of the adventitious buds. These results indicated that inhibition of auxin polar transport could affect the morphogenesis of leaves, so the polar transport of auxin is essential for the bilateral symmetry of leaf growth.     

Somatic embryogenesis in cotton (Gossypium) I. Effects of source of explant and hormone regime

Optimal media for induction of somatic embryogenesis from mature and immature tissues ofG. hirsutum L. cv Coker 312 were determined. Explants of three-day-old seedlings form somatic embryos in 100% of cultures when treated with 0.1 mg/1 2,4-dichlorophenoxyacetic acid plus 0.5 mg/1 kinetin. Mature tissues are more recalcitrant than immature tissues and formed somatic embryos on a limited number of hormone treatments. Stem tissue is most readily induced to form somatic embryos by 2 mg/1 napthaleneacetic acid plus 0.1 mg/1 kinetin, whereas leaf tissue formed embryos best when treated with 0.1 mg/1 2,4-dichlorophenoxyacetic acid plus 1.0 mg/1 (2-isopentyl)-adenine, or 1.0 mg/1 napthaleneacetic acid plus 0.5 mg/1 (2-isopentyl)-adenine.     

Ferrous Iron Oxidation by Thiobacillus ferrooxidans: Inhibition with Benzoic Acid, Sorbic Acid, and Sodium Lauryl Sulfate

Benzoic acid, sorbic acid, and sodium lauryl sulfate at low concentrations (5 to 10 mg/liter) each effectively inhibited bacterial oxidation of ferrous iron in batch cultures of Thiobacillus ferrooxidans. The rate of chemical oxidation of ferrous iron in low-pH, sterile batch reactors was not substantially affected at the tested concentrations (5 to 50 mg/liter) of any of the compounds.     

Role of Methanogens and Other Bacteria in Degradation of Dimethyl Sulfide and Methanethiol in Anoxic Freshwater Sediments

The roles of several trophic groups of organisms (methanogens and sulfate- and nitrate-reducing bacteria) in the microbial degradation of methanethiol (MT) and dimethyl sulfide (DMS) were studied in freshwater sediments. The incubation of DMS- and MT-amended slurries revealed that methanogens are the dominant DMS and MT utilizers in sulfate-poor freshwater systems. In sediment slurries, which were depleted of sulfate, 75 μmol of DMS was stoichiometrically converted into 112 μmol of methane. The addition of methanol or MT to DMS-degrading slurries at concentrations similar to that of DMS reduced DMS degradation rates. This indicates that the methanogens in freshwater sediments, which degrade DMS, are also consumers of methanol and MT. To verify whether a competition between sulfate-reducing and methanogenic bacteria for DMS or MT takes place in sulfate-rich freshwater systems, the effects of sulfate and inhibitors, like bromoethanesulfonic acid, molybdate, and tungstate, on the degradation of MT and DMS were studied. The results for these sulfate-rich and sulfate-amended slurry incubations clearly demonstrated that besides methanogens, sulfate-reducing bacteria take part in MT and DMS degradation in freshwater sediments, provided that sulfate is available. The possible involvement of an interspecies hydrogen transfer in these processes is discussed. In general, our study provides evidence for methanogenesis as a major sink for MT and DMS in freshwater sediments.     

生长素极性运输的抑制对叶生长发育模式的影响

以烟草 ( Nicotiana tabacum L.品种“革新一号”)无菌幼苗叶片为材料 ,在 MS培养基中分别加不同浓度的生长素极性运输抑制剂 (三碘苯甲酸、反式肉桂酸、9-羟基芴 - 9-羧酸 )和 2 mg/ L BA ,探讨不定芽分化的情况 ,在这些培养基上都观察到不同形态的喇叭状叶片形成。结果表明 ,再生喇叭叶的发生频率与培养基中抑制剂一定范围内的浓度密切有关。当三碘苯甲酸浓度为 7.5 mg/ L 时 ,喇叭叶的发生频率最高可达到 82 .1 % ,在再生不定芽的不同位置均观察到有喇叭叶的发生。实验证明 ,抑制生长素的极性运输可导致叶形态发生改变 ,说明生长素的极性运输在叶片两侧对称性生长中有重要作用     

多酚氧化酶抑制剂对蝴蝶兰叶外植体褐变的影响

将多酚氧化酶(PPO)抑制剂添加到酶反应液中,抗坏血酸和半胱氨酸在0.5mmol/L就完全抑制蝴蝶兰PPO活性。300mg/L柠檬酸和100~200mg/L亚硫酸氢钠分别添加到培养基中,可使蝴蝶兰外植体褐变程度降低;采用抑制剂浸泡处理外植体,对外植体褐变抑制效果最好的为50mg/L抗坏血酸,外植体在褐变发生前PPO活性低于对照。     

The susceptibility of the giant freshwater prawn Macrobrachium rosenbergii to Lactococcus garvieae and its resistance under copper sulfate stress.

Addition of copper sulfate (0.1 to 0.4 mg l(-1)) to tryptic soy broth (TSB) had no effect on growth rate of the bacterial pathogen Lactococcus garvieae. Giant freshwater prawns Macrobrachium rosenbergii were injected with L. garvieae (4 x 10(6) colony-forming units [cfu] prawn(-1)) grown in TSB or TSB containing copper sulfate at 0.1, 0.2, 0.3 or 0.4 mg l(-1). After 48 h, the cumulative mortality was significantly (p < 0.05) higher for prawns exposed to L. garvieae grown in 0.4 mg l(-1) copper sulfate than at the lower concentrations examined. In other experiments, prawns were injected with TSB-grown L. garvieae (4 x 10(6) and 2 x 10(5) cfu prawn(-1)), then held in water containing copper sulfate. After 8 h the mortality of L. garvieae-exposed prawns held in water containing 0.4 mg l(-1) copper sulfate was significantly higher than prawns held in water containing 0.2 and 0.3 mg l(-1) copper sulfate. At the lower L. garvieae density, cumulative mortality of prawns increased directly with ambient copper sulfate concentrations in the range of 0.2 to 0.4 mg l(-1). All prawns survived a 168 h exposure to 0.1 mg l(-1) copper sulfate. Prawns exposed to different concentrations of copper sulfate were examined for hemocyte density, phenoloxidase activity and respiratory burst. No significant differences in hemocyte density were observed among treatments. In prawns following a 48 h exposure to 0.1 mg l(-1) copper sulfate, phenoloxidase activity was decreased, but respiratory burst was increased. In conclusion, copper sulfate increased the virulence of L. garvieae to M. rosenbergii and modulated its immune system. Copper sulfate at 0.1 mg l(-1) decreased susceptibility of M. rosenbergii to L garvieae infection, whereas at 0.2 mg l(-1) the susceptibility was increased. The generation of superoxide anion by M. rosenbergii exposed to copper sulfate at a concentration higher than 0.2 mg l(-1) was considered to be cytoxic.     

Organogenesis in leafy spurge (Euphorbia esula L.)

Summary All parts of leafy spurge seedlings can be regenerated when isolated and placed onto B5 medium. One-centimeter isolated hypocotyl segments were tested successfully for their usefulness as a bioassay system by comparing the response of auxins, herbicides, and cytokinins. Indole-3-acetic acid (IAA) was the most effective auxin to stimulate root formation. IAA was effective whether the hypocotyl segments remained on the same medium up to 60 days, or the segments were transferred to basal media after 2 or 5 days (pulse treatment). Pulse treatments with the other auxins resulted in stimulation of root formation; continuous or 5-day pulses of higher concentrations of indole-3-butyric acid,&#x03B1;-naphthaleneacetic acid and especially 2,4-dichlorophenoxyacetic acid and picloram formed excessive callus instead of roots. Picloram did not stimulate root formation, whether the treatment was continuous or pulse-treated. No roots formed with continuous picloram at 0.1 mg/liter or greater, but transfer to basal media did result in root and shoot formation at about 50% of the number formed on the controls. Lesser picloram concentrations had no effect. Shoots formed readily on untreated (control) segments, but continuous treatment with all three cytokinins, kinetin, zeatin, and zeatin riboside, increased the numbers of shoots about equally. Root formation was inhibited by the cytokinins at the higher concentrations (0.1 to 0.2 mg/liter). With the exception of a 5-day pulse of 0.04 mg/liter IAA, the auxins did not stimulate shoot formation, but generally inhibited shoot formation, even in pulse-treated cultures.     

Microbial sulfate reduction in acidic (pH 3) strip-mine lakes

Abstract ³⁵SO₄ reduction was detected in slurries of sediments obtained from Reservoir 29 (pH 3.8) and Lake B (pH 6.2), two acid strip-mine lakes in Indiana. The rates varied seasonally and were higher in summer and fall than in the spring. The optimal pH for sulfate reduction in Reservoir 29 sediments was 5, but samples had increased activity at pH 7 within 24 h after adjusting the pH to this value. In Lake B, the optimal pH for sulfate reduction was the in situ pH (6.2). Sulfate reduction in both lakes was stimulated 2–3-fold by increasing p _H2. High concentrations (5 mM) of organic acids inhibited sulfate reduction at pH 3.8, but stimulation was observed at concentrations of 0.1 mM. Acid-volatile sulfides accounted for about 70% of the products of ³⁵SO₄ reduction.