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The effect of microplastics (MPs) retained in waste activated sludge (WAS) on anaerobic digestion (AD) performance has attracted more and more attention. However, their effect on thermophilic AD remains unclear. Here, the influence of polyvinyl chloride (PVC) MPs on methanogenesis and active microbial communities in mesophilic (37 °C) and thermophilic (55 °C) AD was investigated. The results showed that 1, 5, and 10 mg/L PVC MPs significantly promoted the cumulative methane yield in mesophilic AD by 5.62%, 7.36%, and 8.87%, respectively, while PVC MPs reduced that in thermophilic AD by 13.30%, 18.82%, and 19.99%, respectively. Moreover, propionate accumulation was only detected at the end of thermophilic AD with PVC MPs. Microbial community analysis indicated that PVC MPs in mesophilic AD enriched hydrolytic and acidifying bacteria (Candidatus Competibacter, Lentimicrobium, Romboutsia, etc.) together with acetoclastic methanogens (Methanosarcina, Methanosaeta). By contrast, most carbohydrate-hydrolyzing bacteria, propionate-oxidizing bacterium (Pelotomaculum), and Methanosarcina were inhibited by PVC MPs in thermophilic AD. Network analysis further suggested that PVC MPs significantly changed the relationship of key microorganisms in the AD process. A stronger correlation among the above genera occurred in mesophilic AD, which may promote the methanogenic performance. These results suggested that PVC MPs affected mesophilic and thermophilic AD of WAS via changing microbial activities and interaction. [ABSTRACT FROM AUTHOR]

The metagenomics-based behavior and risk of antibiotic resistance genes (ARGs) were investigated during cattle manure thermophilic anaerobic digestion with tetracycline and copper, namely, bulk-copper oxide, nano-copper oxide, and copper sulfate, which are common feed additives. Although bulk-copper oxide reduced ARGs' diversity, it enriched high-risk ARGs the most than the other two copper species, while copper sulfate could strongly mitigate the ARG risk by decreasing their abundances. Compared to corresponding individual effects, copper and tetracycline combinations may decrease ARGs' co-occurrence potential by 22.0%, and particularly, tetracycline combined separately with copper sulfate and nano-copper oxide reduces the ARGs' risk in abundance (by 7.2%) and human health (by 4.0%). These were mainly driven by bioavailable copper, volatile fatty acids, and pH, as well as the main potential hosts in phyla Firmicutes, Coprothermobacterota, and Euryarchaeota. Notably, the twin risks of pathogenicity and ARGs should be emphasized due to the ARGs' positive correlation with human pathogens of Clostridioides difficile and Arcobacter peruensis. These findings are important for understanding the potential ARGs' risk in treatments of livestock wastes containing feed additives of different sizes and speciation. [ABSTRACT FROM AUTHOR]

Syntrophic oxidization of acetate and propionate are both critical steps of methanogenesis during thermophilic anaerobic digestion. However, knowledge on syntrophic acetate-oxidizing bacteria (SAOB) and syntrophic propionate-oxidizing bacteria (SPOB) is limited because of the difficulty in pure culture isolation due to symbiotic relationship. In this study, two thermophilic acetate-fed anaerobic chemostats, ATL (dilution rate of 0.025 day−1) and ATH (0.05 day−1) and one thermophilic propionate-fed anaerobic chemostat PTL (0.025 day−1) were constructed, AOB and POB in these chemostats were studied via microbial community analysis and DNA stable-isotope probing (SIP). The results showed that, in addition to Tepidanaerobacter, a known SAOB, species of Thauera, Thermodesulfovibrio, Anaerobaculum, Ruminiclostridium, Comamonas, and uncultured bacteria belonging to Lentimicrobiaceae, o_MBA03, Thermoanaerobacteraceae, Anaerolineaceae, Clostridiales, and Ruminococcaceae were determined to be potential AOB in chemostats. Pelotomaculum was the key SPOB detected in the propionate-fed chemostat. Based on the intense fluorescence of coenzyme F420, majority of Methanosarcina cells in acetate-fed chemostats were involved in hydrogenotrophic methanogenesis, suggesting the existence of highly active SAOB among the detected AOB. In the propionate-fed chemostat, most of the species detected as AOB were similar to those detected in the acetate-fed chemostats, suggesting the contribution of the syntrophic acetate oxidization pathway for methane generation. These results revealed the existence of previously unknown AOB with high diversity in thermophilic chemostats and suggested that methanogenesis from acetate via the syntrophic oxidization pathway is relevant for thermophilic anaerobic digestion. [ABSTRACT FROM AUTHOR]

• Anaerobic VFA production from fruit waste at high OLRs of 8–24 g VTS/(L·d). • An OLR of 14 g VTS/(L·d) yielded the highest VFA concentration (61.90 g COD/L). • OLR increase affected the VFA profile and facilitated butyrate-type fermentation. • Firmicutes and Actinobacteriota were predominant in the VFA production reactor. This study examined the performance and microbial community dynamics of an anaerobic volatile fatty acid (VFA) production reactor for treating fruit waste by stepwise increasing organic loading rates (OLRs) from 8 to 24 g volatile total solids (VTS)/(L·d). Results showed that higher VFA concentrations of 52.25–61.90 g chemical oxygen demand (COD)/L can be maintained at each OLR, thereby resulting to a production of 0.70–0.76 g chemical oxygen demand (COD) VFA /g VTS. Notably, an increase in OLR from 8 to 14 g VTS/(L·d) was beneficial for achieving higher VFA concentrations and yields. Moreover, an increase in OLR affected the VFA distribution significantly; acetate and butyrate became dominant in the fermentation liquid at OLRs ≥ 14 g VTS/(L·d). Microbial community dynamics analysis revealed that phyla Firmicutes and Actinobacteriota were predominant at each OLR, and the genera Lactobacillus , Clostridium_sensu_stricto_ 12, and Caproiciproducens were closely related to anaerobic VFA production. [ABSTRACT FROM AUTHOR]

Fluctuation disturbance of organic loading rate (OLR) is common in actual anaerobic digestion (AD), but its effects on AD of municipal sludge gets little attention. This study investigated the responses of reactor performance and active microbial community in mesophilic and thermophilic AD of municipal sludge before, during and after OLR periodic fluctuation disturbance. The performance of both reactors were similar before and after disturbance although some parameter values changed during the disturbance, which indicated their enough buffer capacity to OLR periodic fluctuation. Different microbial community at RNA level was observed in the two reactors. When the OLR disturbance commenced, the microbial community changed greatly in thermophilic AD. Error and attack tolerance of the microbial network was analyzed in order to learn the response mechanisms to OLR disturbance. The results assisted that the thermophilic microbial community was more vulnerable, but the reactor performance of which could be maintained using the functional redundancy strategy under OLR fluctuation disturbance. • Thermophilic AD of sludge showed higher performance than mesophilic AD. • Both reactors showed enough buffer capacity during the OLR disturbance period. • AD performance could recover to former situation after the OLR disturbance finished. • Microbial network of thermophilic AD was more susceptible to node removal. • Functional redundancy of thermophilic community could keep the reactor stable. [ABSTRACT FROM AUTHOR]

Ammonia inhibition easily affects the performance of anaerobic digestion (AD) for municipal sludge and the oxidization of volatile fatty acids (VFAs) is the rate-limiting step of this process. Bioaugmentation is considered to be an effective method to alleviate ammonia inhibition of AD, but most study used the hydrogenotrophic methanogens as the bioaugmentation culture. In this study, bioaugmentation of mesophilic AD (MAD) and thermophilic AD (TAD) under ammonia inhibition with syntrophic acetate and propionate oxidizing consortia was investigated. The results showed that the bioaugmented reactors recovered earlier than control reactors with 20 (MAD) and 8 (TAD) days, respectively. The high-throughput 16S rRNA gene sequencing indicated that the relative abundance of carbohydrates fermenter (Lentimicrobium), syntrophic VFAs-oxidizing bacteria (Rikenellaceae_ DMER64, Smithella and Syntrophobacter) and acetoclastic and hydrogenotrophic methanogens (Methanosaeta , Methanolinea and Methanospirillum) increased in MAD after bioaugmentation. However, part of the bioaugmentation culture could not adapt to the high free ammonia (FAN) concentration in MAD and the effect was weakened. In TAD, proteolytic bacteria (Keratinibaculum and Tepidimicrobium), syntrophic VFAs-oxidizing bacteria (Syntrophomonas) and hydrogenotrophic methanogen (Methanosarcina) were strengthened. The effect of bioaugmentation in TAD was durable even at higher organic loading rate (OLR), due to its positive influence on microbial community. These results suggested that the different bioaugmentation mechanism occurred in MAD and TAD, which are derived from the synergetic effects of ammonia tolerance and microbial interactions. Our study revealed the VFAs-oxidizing consortia as bioaugmented culture could be the potential strategy to alleviate the ammonia stress of AD. [Display omitted] • Bioaugmentation recovered MAD and TAD performance from the ammonia inhibition. • Bioaugmentation increased the robustness of thermophilic community at higher OLR. • Enrichment of hydrolysis, syntrophic VFAs-oxidation and methanogenesis is crucial. • The bioaugmentation mechanisms were different between MAD and TAD. • Syntrophic VFAs-oxidizing consortia was effective as the bioaugmented culture. [ABSTRACT FROM AUTHOR]

• MAD and TAD of high-solids containing sludge were comprehensively compared. • The response of microbial community composition to sludge properties was monitored. • TAD performed better than MAD in biogas production and VTS reduction. • Ammonia inhibition was the main risk for high-solids TAD of sludge. • Microbial analysis based on RNA datasets more accurately reflected the AD process. This study involved a comparison between mesophilic (MAD) and thermophilic anaerobic digestion (TAD) of municipal sludge with high (10%) solids content; the reactor performance and the response of total and active microbial communities to changes in sludge properties were monitored. Both TAD and MAD were stably maintained. TAD performed better than MAD in biogas production and volatile total solids reduction upon feeding sludge 1. TAD was slightly inhibited by ammonia, whereas the performance of MAD was improved when sludge 2 was used as the feedstock. Alpha- and beta-diversity analyses revealed significant differences in the microbial community based on DNA and RNA datasets, indicating that not all microbes function in AD. The active microbial community diversity and composition in MAD and TAD were also driven by sludge properties. Moreover, MAD showed significantly higher richness and diversity of the active microbial community compared with TAD, regardless of changes in sludge properties. [ABSTRACT FROM AUTHOR]