PCR-single-strand conformation polymorphism (SSCP) and Southern blotting tech-niques were adopted to investigate microbial community dynamics in a sulfate-reducing bioreactor caused by decreasing influent alkalinity. Experimental results indicated that the sulfate-removal rate approached 87% in 25 d under the conditions of influent alkalinity of 4000 mg/L (as CaCO3) and sul-fate-loading rate of 4.8 g/(L·d), which indicated that the bioreactor started up successfully. The analy-sis of microbial community structure in this stage showed that Lactococcus sp., Anaerofilum sp. and Kluyvera sp. were dominant populations. It was found that when influent alkalinity reduced to 1000 mg/L, sulfate-removal rate decreased rapidly to 35% in 3 d. Then influent alkalinity was increased to 3000 mg/L, the sulfate-removal rate rose to 55%. Under these conditions, the populations of Dysgo-nomonas sp., Sporobacte sp., Obesumbacterium sp. and Clostridium sp. got to rich, which predomi-nated in the community together with Lactococcus sp., Anaerofilum sp. and Kluyvera sp. However, when the alkalinity was decreased to 1500 mg/L, the sulfate-removal rate rose to and kept stable at 70% and populations of Dysgonomonas sp., Sporobacter sp. and Obesumbacterium sp. died out, while some strains of Desulfovibrio sp. and Clostridium sp. increased in concentration. In order to determine the minimum alkalinity value that the system could tolerate, the influent alkalinity was de-creased from 1500 to 400 mg/L secondly. This resulted in the sulfate-removal rate, pH value and ef-fluent alkalinity dropping quickly. The amount of Petrotoga sp., Prevotella sp., Kluyvera sp. and Neisseria sp. reduced obviously. The result data from Southern blotting indicated that the amount of sulfate-reducing bacteria (SRBs) decreased with influent alkalinity dropping. Analysis of the microbial community structure and diversity showed that the SRBs populations were very abundant in the in-oculated activated sludge and the alkalinity decrease caused the reduction of the p
REN Nanqi1, ZHAO Yangguo1, WANG Aijie1, GAO Chongyang2, SHANG Huaixiang1, LIU Yiwei1 & WAN Chunli1 1. School of Environmental and Municipal Engineering, Harbin Institute of Technology, Harbin 150090, China
Alkalinity is one of the most important parameters that influence microbial metabolism and activity during sulfate-laden wastewater biological treatment. To comprehensively understand the structure and dynamics of functional microbial community under alkalinity changes in sulfate-reducing continuous stirred tank reactor (CSTR), fluorescent in situ hybridization (FISH) technique was selected for qualitative and semi-quantitative analysis of functional microbial compositions in activated sludge. During 93d of bioreactor operation, the influent alkalinity was adjusted by adding sodium bicarbonate from 4000mg·L^-1 down to 3000mg·L^-1, then to 1500mg·L^-1, whereas other parameters, such as the loading rates of chenucal oxygen demand (COD) and sulfate (SO4^2-), hydraulic retention time (HRT), and pH value, were continuously maintained at 24g·L^-1·d^-1 and 4.8g·L^-1·d^-1, 10h,and about 6.7, respectively. Sludge samples were collected during diflerent alkalinity levels, and total Bacteria, tlae sulfate-reducing bacteria (SRB), and four SRB genera were demonstrated with 16S ribosomal .RNA-targeted oligonucleotide probes. The results indicated that bioreactor started-up successfully in 30d. The two instances ot drop in alkalinity resulted in the fluctuation of sulfate removal rate. The diversity of SRB community showed significant shift, and the alteration of microbial community directly resulted in the corresponding statuses of bioreactor. The dominant genera during the bioreactor start-up and alkalinity drops were Desulfovibrio, Desulfobacter, Desulfovibrio, Desulfobacter, and Desulfovibrio, respectively. In addition, the acetotrophic SRB sutterecl more trom me reduction of alkalinity than the non-acetotrophic SRB. This strategy can present the functional microbial community structure during start-up and alkalinity drop stages, and provides a powerful theoretical guideline for optimization and adjustment of bioreactor, as well.
In order to investigate the change of ecological characteristics due to the decrease of COD/SO_4^(2-) ratio during sulfate reduction, continuous-flew tests were conducted in an acidogenic sulfate-reducing reactor with molasses wastewater as sole organic carbon source and sodium snlfate as electron acceptor, and the change of pH value, oxidation reduction potential (ORP), volatile fat acids (VFAs), alkalinity (ALK) and the predominant populations with COD/SO_4^(2-) ratio decreasing from 4.2 to 2.0 were investigated. The experimental results demonstrated that, with decreasing COD/SO_4^(2-) ratio, ORP and ALK increased, pH value decreased, and the proportion of acetic acid in terminal products decreased significantly, and a stable -type microbial community with high COD/SO_4^(2-) ratio was converted into a sub -stable -type one with low COD/SO_4^(2-) ratio.