Impacts of phosphorus and nitrogen absence on microbial diversity and sulfate removal in anaerobic batch reactors.

Sulfate-rich effluents have been successfully treated in anaerobic reactors using sulfate-reducing bacteria (SRB). Many authors have demonstrated that these systems require nitrogen and phosphorous supplementation to achieve high sulfate removal rates. However, the resource ratio theory assumes that some species can be dominant according to the nutritional relations used or even without external nutrient supplementation. Thus, this study evaluated the SRB communities in batch reactors without external nitrogen and phosphorus sources based on most probable number (MPN) quantification, denaturing gradient gel electrophoresis (DGGE) analyses and sequencing. The sulfate and chemical oxygen demand (COD) removal and kinetic parameters were also determined. After 100 days of operation, the sulfate and COD removal achieved 71.8 ± 10% and 86.5 ± 10%, respectively. The SRB population increased from 8.106 to 4 × 1012 MPN 100 mL-1, and the richness of SRB bands was much higher at the end of the experiment compared to the inoculum. In addition, the sequenced bands from SRB-DGGE showed similarities to Desulfacinum infernum, Desulfobulbus sp, Syntrophobacter and Desulfomicrobium aestuarii-related sequences. Therefore, biological treatment of acid mine drainage wastewater was effective in the absence of nutrients, lowering costs and providing high sulfate removal efficiency.

Lactate as an effective electron donor in the sulfate reduction: impacts on the microbial diversity.

The competition between sulfate-reducing bacteria and methane-producing archaea has a major influence on organic matter removal, as well as the success of sulfidogenic systems. This study investigated the performance of six batch sulfidogenic reactors in response to different COD/sulfate ratios (1.0 and 2.0) and electron donors (cheese whey, ethanol, and sodium lactate) by evaluating the biochemical mechanisms of sulfate reduction, organic matter oxidation, and microbial structure modification. A COD/sulfate ratio of 1.0 resulted in high sulfidogenic activity for all electron donors, thereby achieving a nearly 80% sulfate removal. Lactate provided high sulfate removal rates at COD/sulfate ratios of 1.0 (80%) and 2.0 (90%). A COD/sulfate ratio of 2.0 decreased the sulfate removal rates by 25 and 28% when ethanol and cheese whey were used as substrates. The sulfate-reducing bacteria populations increased using ethanol and lactate at a COD/sulfate ratio of 1.0. Particularly, Desulfovibrio, Clostridium, and Syntrophobacter were predominant. Influent composition and COD/sulfate ratio influenced the relative abundance of the microbial communities. Therefore, controlling these parameters may facilitate the wastewater treatment with high sulfate levels through bacterial activity.

Real effluents and fractionation in the supply of COD: Rapid adaptation and high efficiency to treat mine drainage combined with industrial by-products.

The biological treatment of mine drainage (MD) using sulfate-reducing bacteria (SRB) is a technology in growing exploitation. The use of by-products as sources of electrons can make this treatment more environmentally and economically advantageous. However, the high chemical oxygen demand (COD) and the presence of recalcitrant molecules can lead to the accumulation of metabolic intermediates that acidify the system, thus interrupting the treatment. Besides, the adaptation of the inoculum to the establishment of sulfidogenesis with MD and by-product may be slow. This study aimed to investigate prompt adaptation and operation strategies that do not require additives to enable the sulfidogenic process to occur while maintaining a pH close to neutrality. The sources of electrons tested were trub (brewery residue) and crude glycerol - CG (residue from the biodiesel production). The inoculum from a methanogenic reactor was stored with a real MD for a month. The adapted inoculum was applied in a batch reactor for 168 h of hydraulic detention time, and promoted 75.8 ± 4.3% of sulfate removal from an MD with 3756.4 ± 258 mg.L-1 of sulfate using CG in a COD/SO42- ratio of 3 ratio. With higher initial substrate concentrations, acidification occurred and the treatment was interrupted. Using trub instead of CG, the acidification occurred at a COD/SO42- ratio of 3. Acidification was prevented and the best efficiencies in sulfate removal were obtained when the amount of substrate corresponding to COD/SO42- ratio of 3 was fractioned into equal parts and added over six days in the CG reactor. It was achieved 94.15 ± 1.76% of sulfate removal. With trub, the same procedure in which this COD was divided into seven parts, and resulted in a sulfate removal of 88.49 ± 1.02%. The removal of metals and metalloids were greater than 94.5% in all the systems in which the substrate supply was made fractionally, and the effluent generated presented alkalinity between 3370 and 4242 mg CaCO3.L-1, and pH between 6.8 and 7. The method of adaptation and operation applied allowed the realization of a MD treatment with quick establishment of sulfidogenesis and without the use of neutralizing additives. Finally, the effluent presented characteristics considered favorable for a later stage of post-treatment of the effluent with methane generation.

Sulfate and metals removal from acid mine drainage in a horizontal anaerobic immobilized biomass (HAIB) reactor.

The acid mine drainage (AMD) can causes negative impacts to the environment. Physico-chemical methods to treat AMD can have high operational costs. Through passive biological methods, such as anaerobic reactors, sulfate reduction, and recovery of metals are promoted. This study evaluated the performance of a horizontal anaerobic immobilized biomass (HAIB) reactor for the treatment of synthetic AMD using polyurethane foam as support material, and anaerobic sludge as inoculum. Ethanol was used as an electron donor for sulfate reduction, resulting in an influent chemical oxygen demand (COD) in the range of 500-1,500 mg/L and COD/sulfate ratio at 1. A gradual increase of sulfate and COD concentration was applied that resulted in COD removal efficiencies higher than 78%, and sulfate removal efficiencies of 80%. Higher sulfate and COD concentrations associated with higher hydraulic retention times (36 h) proved to be a better strategy for sulfate removal. The HAIB reactor was able to accommodate an increase in the SLR up to 2.25 g SO42-/L d-1 which achieved the greatest performance on the entire process. Moreover, the reactor proved a suitable alternative for reaching high levels of metal removal (86.95 for Zn, 98.79% for Fe, and 99.59% for Cu).

Long-term performance of a UASB reactor treating acid mine drainage: effects of sulfate loading rate, hydraulic retention time, and COD/SO42- ratio.

Acid mine drainage (AMD) is among the most serious threats to water and the typical alkali-based treatment costs are high. This study's main objective was the establishment of a highly efficient biological process using an upflow anaerobic sludge blanket (UASB) reactor to treat AMD based on a shorter hydraulic retention time (HRT) and lower organic matter input. The process was evaluated for a long-term operation (739 days) in terms of the influence of HRT (14-24 h), metal addition, sulfate loading rate (0.5-2.6 g SO42- l-1 d-1), and the COD/SO42- ratio (0.67-1.0) using ethanol as the only electron donor at a pH of 4.0. Neutral effluent pH was achieved throughout the time apart from operational modifications. The reduction in HRT from 24 to 16 h and an increase in the sulfate loading rate (SLR) up to 2.25 g SO42- l-1 d-1 improved the sulfate removal to (92.1 ± 1.8)% with 80% chemical oxygen demand (COD) removal. However, the sulfate reduction was less than 80% when the HRT and SLR was changed to 14 h and 2.6 g SO42- l-1 d-1, respectively. The oxidation of organic matter by sulfate reduction was greater than 50% regardless of the conditions imposed but the use of ethanol to treat AMD was more efficient when either the HRT was 16 h (1.5 g SO42- l-1 d-1) in the presence of Fe, Zn, and Cu or the HRT was 14 h (2.6 g SO42- l-1 d-1) but the COD/SO42- ratio was reduced to 0.67. The fully optimized conditions of the UASB reactor were set at an HRT of 16 h, SLR of 1.5 g SO42- l-1 d-1, and a COD/SO42- ratio of 1.0.