The effects of Fe2+ on sulfur-oxidizing bacteria (SOB) driven autotrophic denitrification.

With the short-term exposure to Fe2+, the mechanism of autotrophic denitrification and sulfide oxidation and the correlation between microbial community changes and environmental factors have been explored in the ADSOB process. RSM was used to optimize conditions for the maximum nitrate reduction and sulfide oxidation. About 88% of nitrate could be autotrophically denitrified to nitrogen by utilizing sulfide as the electron donor with the molar ratio C/N of 1.14 and S/N of 0.99 at pH 7.1. Lower Fe2+ additions can reduce TDS inhibition with dissolved sulfide to form FeS precipitates, while high amount of Fe2+ limited the mass transfer of NO3- and intermediate products such as S0 may be generated. High-throughput sequencing and RDA analysis revealed the correlation between ferrous iron, environmental factors and microorganisms. Sulfurospirillum, Rhodanobacter, Thauera and Thiobacillus were all slightly promoted at NFL level and inhibited at NFH level. And the narrow angles of the arrows indicated that Thauera, Sulfurospirillum and Thiobacillus were positively correlated with SO42- concentrations, while large angles indicated these bacteria were inversely related with TDS and NO3- arrows, which further confirmed that these bacteria played a dominant role in the ADSOB process, and can reduce NO3- by the oxidation of TDS. The correlation further indicated that lower Fe2+ additions have a promoting effect, while high concentrations have an inhibiting effect.

Inherent humic substance promotes microbial denitrification of landfill leachate via shifting bacterial community, improving enzyme activity and up-regulating gene.

Microbial denitrification is the main pathway for nitrogen removal of landfill leachate. Although humic substances (HSs) have been reported in landfill leachate, the effects of HS on denitrification process of activated sludge for leachate treatment are still unknown. In this study, we adopted SAHA as the model HS to study the effects of HS on the denitrification of landfill leachate. After long-term operation at 10 mg/L of Shanghai Aladdin Humic Acid (SAHA), the final nitrate concentration and nitrite accumulation were much lower than the control (5.2 versus 96.2 mg/L; 0.5 versus 34.7 mg/L), and the final N2O emission was 13.1% of the control. The mechanistic study unveiled that SAHA substantially changed the activated sludge community structure and resulted in the dominance of Thauera after long-term exposure to SAHA. Thauera could be able to utilize HSs as electron shuttle to improve denitrificattion performance, especially for nitrite reduction. Moreover, SAHA significantly upregulated the gene expressions and catalytic activities of the key enzymes related to denitrification, the reducing power (NADH) generation, and the electron transport system activity, which accelerated nitrogen oxide reduction. The positive effects of HS on denitrification performance were confirmed by the addition of SAHA into real leachate.

Denitrifying sulfide removal process on high-tetracycline wastewater.

Antibiotics wastewater from tetracycline (TC) production unit can have high levels of chemical oxygen demand, ammonium and sulfate and up to a few hundreds of milligrams per liter of TC. Denitrifying sulfide removal (DSR) process is set up for simultaneously removal of sulfur, carbon and nitrogen from waters. The DSR process was for the first time studied for treating TC wastewaters. The TC stress has no adverse effects on removal rates of nitrate and acetate; however, it moderately deteriorated sulfide removal rates and S(0) accumulation rates when the concentration is higher than 100mgL(-1) TC. The Thauera sp., and Pseudomonas sp. present the heterotrophs and Sulfurovum sp. presented the autotroph for the present DSR reactions. The high tolerance of TC stress by the tested consortium was explained by the excess production of extracellular polymeric substances at high TC concentration, which can bind with TC for minimizing its inhibition effects.

Effects of high CO2 concentrations on ecophysiologically different microorganisms.

We investigated the effect of increasing CO(2) concentrations on the growth and viability of ecophysiologically different microorganisms to obtain information for a leakage scenario of CO(2) into shallow aquifers related to the capture and storage of CO(2) in deep geological sections. CO(2) concentrations in the gas phase varied between atmospheric conditions and 80% CO(2) for the aerobic strains Pseudomonas putida F1 and Bacillus subtilis 168 and up to 100% CO(2) for the anaerobic strains Thauera aromatica K172 and Desulfovibrio vulgaris Hildenborough. Increased CO(2) concentrations caused prolonged lag-phases, and reduced growth rates and cell yields; the extent of this effect was proportional to the CO(2) concentration. Additional experiments with increasing CO(2) concentrations and increasing pressure (1-5000 kPa) simulated situations occurring in deep CO(2) storage sites. Living cell numbers decreased significantly within 24 h at pressures ≥1000 kPa, demonstrating a severe lethal effect for the combination of high pressure and CO(2).

Adaptation of anaerobically grown Thauera aromatica, Geobacter sulfurreducens and Desulfococcus multivorans to organic solvents on the level of membrane fatty acid composition.

The effect of different solvents and pollutants on the cellular fatty acid composition of three bacterial strains: Thauera aromatica, Geobacter sulfurreducens and Desulfococcus multivorans, representatives of diverse predominant anaerobic metabolisms was investigated. As the prevailing adaptive mechanism in cells of T. aromatica and G. sulfurreducens whose cellular fatty acids patterns were dominated by palmitic acid (C16:0) and palmitoleic acid (C16:1cis), the cells reacted by an increase in the degree of saturation of their membrane fatty acids when grown in the presence of sublethal concentrations of the chemicals. Next to palmitic acid C16:0, the fatty acid pattern of D. multivorans was dominated by anteiso-branched fatty acids which are characteristic for several sulfate-reducing bacteria. The cells responded to the solvents with an increase in the ratio of straight-chain saturated (C14:0, C16:0, C18:0) to anteiso-branched fatty acids (C15:0anteiso, C17:0anteiso, C17:1anteisoΔ9cis). The results show that anaerobic bacteria react with similar mechanisms like aerobic bacteria in order to adapt their membrane to toxic organic solvents. The observed adaptive modifications on the level of membrane fatty acid composition can only be carried out with de novo synthesis of the fatty acids which is strictly related to cell growth. As the growth rates of anaerobic bacteria are generally much lower than in the so far investigated aerobic bacteria, this adaptive response needs more time in anaerobic bacteria. This might be one explanation for the previously observed higher sensitivity of anaerobic bacteria when compared with aerobic ones.

Anaerobically grown Thauera aromatica, Desulfococcus multivorans, Geobacter sulfurreducens are more sensitive towards organic solvents than aerobic bacteria.

The effect of seven important pollutants and three representative organic solvents on growth of Thauera aromatica K172, as reference strain for nitrate-reducing anaerobic bacteria, was investigated. Toxicity in form of the effective concentrations (EC50) that led to 50% growth inhibition of potential organic pollutants such as BTEX (benzene, toluene, ethylbenzene, and xylene), chlorinated phenols and aliphatic alcohols on cells was tested under various anaerobic conditions. Similar results were obtained for Geobacter sulfurreducens and Desulfococcus multivorans as representative for Fe(3+)-reducing and sulphate-reducing bacteria, respectively, leading to a conclusion that anaerobic bacteria are far more sensitive to organic pollutants than aerobic ones. Like for previous studies for aerobic bacteria, yeast and animal cell cultures, a correlation between toxicity and hydrophobicity (log P values) of organic compounds for different anaerobic bacteria was ascertained. However, compared to aerobic bacteria, all three tested anaerobic bacteria were shown to be about three times more sensitive to the tested substances.

Role of benzylsuccinate in the induction of the tutE tutFDGH gene complex of T. aromatica strain T1.

Expression of the tutE tutFDGH gene cluster of Thauera aromatica strain T1 was examined by Northern and Western analysis in a wild-type strain and chromosomally deleted strains with or without in-frame deletion plasmids. While expression was observed when the wild-type strain was induced with toluene, various chromosomally deleted strains exhibited little or no expression of the tut genes. In contrast, both wild-type and chromosomally deleted strains expressed the tut genes when induced with benzylsuccinate. We conclude that benzylsuccinate is required for the full induction of the tutE tutFDGH gene cluster of T. aromatica strain T1.