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Biological sulfate (SO42−) reduction was examined in anaerobic sequential batch reactors (ASBRs) operated under different hydraulic retention times (HRTs) ranging from 12 to 36 h and COD (Chemical Oxygen Demand)/SO42−) ratios of 2.4, 1.6 and 0.8. Competition between SO42− reducing bacteria (SRBs), methane producing archaea (MPAs) and homoacetogens (HACs) was examined in controls and cultures treated with linoleic acid (LA). The ASBR performance was influenced by the COD/SO42− ratio in control cultures with a SO42− reduction of 87% at a COD/SO42− ratio of 0.8. At a 12 h HRT, in both control and LA treated cultures, greater than 75% SO42− removal was observed under all the conditions examined. In control reactors operating at a 36 h HRT, high levels of MPAs belonging to Methanobacteriales and Methanosarcinales were detected; however, in comparison, under low COD/SO42− ratio and with decreasing HRT conditions, a relative increase in SRBs belonging to Desulfovibrio and Desulfatibacillum was observed. Adding 0.5 g·L−1 LA suppressed Methanobacteriales, while increasing the LA concentration to 1 g·L−1 completely suppressed MPAs with a relative increase in SRBs. HACs belonging to Bacteroidetes were observed in the control and in cultures operated at 12 h HRT with a COD/SO42− ratio of 1.6 and fed 0.5 g·L−1 LA; however, with all other LA levels (0.5 and 1.0 g·L−1) and HRTs (12, 24 and 36 h), HACs were not detected.

Taguchi statistical design, an orthogonal array (OA) method, was used to study the impact of the COD/SO42− ratio, hydraulic retention time (HRT) and linoleic acid (LA) concentration on sulfate (SO42−) reduction in an anaerobic sequencing batch reactor using glucose as the electron donor. Based on the OA, optimum condition for maximum SO42− reduction was evaluated. Increasing the COD/SO42− ratio and HRT caused decreasing SO42− reduction while increased SO42− reduction was observed with increasing LA concentration (1 g L−1). In control (not fed LA) cultures, higher SO42− reduction (87% ± 3%) was observed at a low COD/SO42− ratio of 0.8. This indicates that increasing SO42− reduction was observed at increasing SO42− loading rates. In general, results from this study reveal that limiting the substrate concentration with high SO42− levels (low COD/SO42− ratio) favors high SO42− removal. Surface plots were used to evaluate the significant interactions between the experimental factors. Accuracy of the model was verified using an analysis of residuals. Optimum conditions for maximum SO42− reduction (97.61%) were observed at a COD/SO42− ratio of 0.8 (level 1), 12 h HRT (level 1) together with 1000 mg L−1 LA addition (level 3). In general, the Taguchi OA provided a useful approach for predicting the percent SO42− reduction in inhibited mixed anaerobic cultures within the factor levels investigated.