Assessing Methanobrevibacter smithii and Clostridium difficile as not conventional faecal indicators in effluents of a wastewater treatment plant integrated with sludge anaerobic digestion.

Wastewater treatment plants (WWTP) are an important source of surface water contamination by enteric pathogens, affecting the role of environmental water as a microbial reservoir. We describe the release to the environment of certain anaerobes of human and environmental concern. The work was focused on emerging microbial targets. They are tracing, by RT-qPCR, on WWTP effluents, both liquid and solid, when an anaerobic digestion step is included. The focus is placed on Clostridium spp. with the specific quantification of Clostridium perfringens, as typical bioindicator, and Clostridium difficile, as emerging pathogen not only confined into nosocomial infection. Moreover methanogens were quantified for their involvement in the anaerobic digestion, and in particular on Methanobrevibacter smithii as major methanogenic component of the human gut microbiome and as not conventional faecal indicator. In the water samples, a reduction, statistically significant, in all microbial targets was observed (p < 0.01), 2 log for the total bacteria, 1.4 log for the Clostridium spp. and M. smithii, 1 log for total methanogens, C. perfringens and C. difficile. The AD process contribute to a significant change in microbial levels into the sludge for total bacteria and total methanogens (p < 0.01), both when the input sludge are primary and secondary, while for the presence of Clostridium spp. and C. difficile there was not a significant change. The produced data are innovative showing which is the diffusion of such anaerobic microorganisms throughout the WWTP and opening a discussion on the implementation of possible techniques for a more efficient microbial removal from effluents, particularly bio-solids, to reduce the potential release of pathogens into the environment.

Chlorination in a wastewater treatment plant: acute toxicity effects of the effluent and of the recipient water body.

This study investigates the impact of wastewater treatment plant (WWTP) effluent on the toxicity of the recipient water body and the effectiveness of the disinfection treatment applied (sodium hypochloride) to assure the compliance of both microbiological and toxicological emission limits. No toxicity was found in the majority of samples collected from the recipient river, upstream and downstream of the WWTP, using three different toxicity tests (Vibrio fischeri, Daphnia magna, and Pseudokirchneriella subcapitata). Only three samples presented toxic unit (TU) values with V. fischeri, and one presented TU with P. subcapitata. The influent toxicity ranged from slightly toxic to toxic (TU = 0.68-4.47) with V. fischeri, while only three samples presented TU values with the other tests. No toxicity was found in the absence of chlorination, while the mean toxicity was 3.42 ± 4.12 TU with chlorination in the effluent. Although no toxicity or very slight toxicity was found in the receiving water, its residual toxicity was higher than the US EPA Quality Standard in two samples. Escherichia coli concentration had a lower mean value in the chlorinated effluent: 13,993 ± 12,037 CFU/100 mL vs. 62,857 ± 80,526 CFU/100 mL for the not chlorinated effluent. This difference was shown to be significant (p < 0.05). E. coli in ten chlorinated samples was higher than the limit established by European and Italian Legislation. The mean highest trihalomethanes (THMs) value was found in the influent samples (2.79 ± 1.40 µg/L), while the mean highest disinfection by-products (DBPs) was found in the effluent samples (1.85 ± 2.25 µg/L). Significant correlations were found between toxicity, sodium hypochlorite, THMs, DBPs, E. coli, and residual chlorine. In conclusion, this study highlighted that the disinfection of wastewater effluents with sodium hypochlorite determines the increase of the toxicity, and sometimes is not enough to control the E. coli contamination.