Thermophilic Digestion Research Articles

One-step acquirement of superior microbial communities from mesophilic digested sludge to upgrade anaerobic digestion

Anaerobic digestion is a promising waste-to-energy alternative technology. However, the efficiency upgrading for conventional mesophilic digestion of organic solid waste is always indispensable. Employing hyperthermophilic or thermophilic microbial community is one of the viable upgrading alternatives. Given the unavailability of the superior microbial communities, mesophilic digested sludge was used as inoculum, and instantly controlled at 70 °C and 55 °C for acclimation of hyperthermophilic and thermophilic inocula, respectively. Waste activated sludge was continuously and synchronously fed into two digesters. After one round, thermophilic digester achieved stable biogas production rate at 0.22 L L−1 d−1, with a methane proportion over 60%, whereas fluctuation was observed in the hyperthermophilic digester, and approximately triple time was needed to reach a relatively stable biogas production rate 0.12 L L−1 d−1. Nevertheless, higher hydrolysis ratio 24.4% was observed in the hyperthermophilic digester despite the lower biogas production. Therefore, methanogenesis step limited the whole anaerobic process for the hyperthermophilic digestion, and digestion at 70 °C was appropriate as a pre-fermentation stage to enhanced hydrolysis. The genus Methanothrix proportion in the thermophilic digester gradually decreased, while another acetoclastic genus Methanosarcina ultimately was acclimated to the dominant methanogen. In addition to Methanothrix, hydrogenotrophic archaea became competitive in the hyperthermophilic digester, with Methanothermobacter dominant at 22.6%. The genus Psychrobacter, affiliated to the phylum Proteobacteria could survive better than the others at 70 °C, with a final proportion of 62.5%.

Establishment and differential performance of hyperthermophilic microbial community during anaerobic self-degradation of waste activated sludge

Hyperthermophilic anaerobic digestion, especially at 70 °C, has drawn wide attention. In order to acquire the inoculum and digestion characteristics, batch acclimation and continuous operation experiments were conducted under hyperthermophilic (70 °C), thermophilic (55 °C) and mesophilic (35 °C) conditions, respectively. Archaea at each temperature was successfully enriched from the sole-source waste activated sludge (WAS). Hyperthermophilic digestion achieved higher archaea diversity, close to the Shannon index 2.23 for the thermophilic digestion, but the population were not improved, at a 16S rRNA genes 5.99 × 105 copies mL−1. Hydrogenotrophic methanogens, Methanospirillum and Methanothermobacter, dominated in the hyperthermophilic digester, accounting for 27.15%, while the primary phylum Firmicutes was promoted to 36.31%, with the proteolytic genus Coprothermobacter in Firmicutes at 19.50%. Refractory organic fractions were converted more with a higher digestion temperature, which was demonstrated by the fact that the COD/VS increased to 5.8, 5.2 and 4.2 at 70 °C, 55 °C and 35 °C, respectively, at the end of batch acclimation. In addition, the most solubilization for the dominant fraction protein in the WAS occurred at 70 °C as well. Similar hydrolysis ratio, over 10%, and specific hydrolysis rate, around 0.025 g COD (g VSS·d)−1, were achieved at 70 °C and 55 °C. The higher hydrolysis for hyperthermophilic digestion even resulted in a higher methane yield than that for the mesophilic digestion. Nevertheless, contrary to higher hydrolysis, methanogenesis limited hyperthermophilic digestion in WAS degradation, with an ultimate methane yield 71.2 mL g−1 VSadded, despite an almost complete VFA conversion through the continuous operation.

Potentiality of municipal sludge for biological gas production at Soba Station South of Khartoum (Sudan)

Potentiality of municipal sludge for biological gas production at Soba Station South of Khartoum (Sudan)

Metagenomic approach reveals the fate of antibiotic resistance genes in a temperature-raising anaerobic digester treating municipal sewage sludge

The fate of diverse antibiotic resistance genes (ARGs) in anaerobic digestion (AD) system under different temperature remains largely unknow. This study employed the shotgun metagenomic sequencing and 16S rRNA amplicon sequencing approaches and investigated the shifts of composition and abundance of more than 1360 ARGs, as well as microbial community, in a temperature-raising AD system (from 35 °C to 55 °C) treating municipal sewage sludge. Results suggested that the six ARGs, inlcuding macB, tetA(58), bcrA, evgS, mtrA, and oleC, were predominated in digested sludge. More than 407ARG subtypes (30%) could be largely reduced under high temperature condition. Thermophilic AD (55 °C) provided a better removal for tetracycline, macrolide, penam, fluoroquinolone, acridine dye, and peptide resistance genes when compared to mesophilic AD (35 °C). The removal rates of these genes reached 29.7%–32.3%. In addition, at the end of thermophilic digestion process, the relative abundance of Firmicutes increased (from 18.22% to 74.89%), while Proteobacteria and Chloroflexi both decreased, from 36.39% to 6.80% and 20.29%–2.64%. This study further proposed the underlying mechanisms of effective elimination of ARGs under high temperature by: (1) the reduction of genes encoding antibiotic efflux pump, (2) the promotion of sludge biomass degradation rate, (3) the loss of microbial community diversity in AD systems, and (4) the inactivation of multidrug-resistant bacteria. The significance of current study is in characterizing the fate of ARGs across AD process, which allows to correlate diverse genetic properties with digesters’ operational condition. Results provided an insight into the manipulation of AD technology (e.g. temperature) to efficiently remove ARGs from municipal sewage sludge.

English

The current practice of using stanchion breeding cow houses results in a semi-dry manure that ideally should be used as the substrate for the anaerobic fermentation. The conventional liquid-slurry biogas fermentation is disadvantageous with high running costs due to requirement such as water addition and running efficiency, among others. This study aimed to establish a high-speed semi-dry methane fermentation system for cow manure, by comparing productivity in wet and semi-dry conditions (> 90 and 83-85% water content, respectively), and mesophilic and thermophilic conditions (38 and 55°C, respectively). The highest methane productivity was obtained for 30 days in semi-dry or wet, thermophilic conditions (0.4-16.2 L‧Kg-1VS‧day-1). The highest accumulated production was 60 days (192.1 L‧Kg-1VS) in semi-dry thermophilic conditions. The pronounced difference in methane production at different temperatures in semi-dry digesters was studied by characterizing the microbial community changes in mesophilic and thermophilic digesters. For bacteria, the class Clostridia (including family Clostridiaceae and the genus Clostridium) was dominant in the thermophilic digester. In the thermophilic digester, a succession of taxa from the genus Methanobrevibacter and family Methanobacteriaceae to genus Methanoculleus in archaea was observed. The convergence of archaeal microbiota in thermophilic conditions was more pronounced than in mesophilic conditions. These results indicate that the proposed procedure is capable of improving methane productivity per scale and lowering the running costs concomitant and downsizing the fermentation scale. Key words: Semi-dry fermentation, methane yield, anaerobic digestion, cow manure, biogas.

Heat integration of two-stage autothermal thermophilic aerobic digestion system for reducing the impact of uncertainty

The operation of Autothermal Thermophilic Aerobic Digestion (ATAD) wastewater treatment systems is subjected to daily uncertainties regarding the parameters of the inlet streams, which lead to prolongation of the process and increase the energy consumption. This, together with the heat loss in the environment from the product tank, makes the ATAD system energy inefficient. Heat integration plays an important role in improving the sustainability of ATAD systems. In this study an approach for optimal redesign of heat integrated two-stage ATAD system operating under uncertainties is presented. It includes a two-stage stochastic optimization model for heat integration with an optimization criterion - the annual capital cost for redesign of the heat integrated system and its operating costs. Artificial Neural Network (ANN) models of the bioreactors are used for simulation of the system operation. They are combined with the heat integration model for the obtained with the stochastic approach values of design parameters of the heat integration equipment. The simulation results show a thermal shock reduction of 5–6 °C, an increase of the inlet sludge temperature of 8–10 °C, achievement of close to normal operating temperatures in the ATAD system and reduction of solids in both bioreactors by about 2 wt%.

Syntrophic acetate oxidation replaces acetoclastic methanogenesis during thermophilic digestion of biowaste

BackgroundAnaerobic digestion (AD) is a globally important technology for effective waste and wastewater management. In AD, microorganisms interact in a complex food web for the production of biogas. Here, acetoclastic methanogens and syntrophic acetate-oxidizing bacteria (SAOB) compete for acetate, a major intermediate in the mineralization of organic matter. Although evidence is emerging that syntrophic acetate oxidation is an important pathway for methane production, knowledge about the SAOB is still very limited.ResultsA metabolic reconstruction of metagenome-assembled genomes (MAGs) from a thermophilic solid state biowaste digester covered the basic functions of the biogas microbial community. Firmicutes was the most abundant phylum in the metagenome (53%) harboring species that take place in various functions ranging from the hydrolysis of polymers to syntrophic acetate oxidation. The Wood-Ljungdahl pathway for syntrophic acetate oxidation and corresponding genes for energy conservation were identified in a Dethiobacteraceae MAG that is phylogenetically related to known SAOB. 16S rRNA gene amplicon sequencing and enrichment cultivation consistently identified the uncultured Dethiobacteraceae together with Syntrophaceticus, Tepidanaerobacter, and unclassified Clostridia as members of a potential acetate-oxidizing core community in nine full-scare digesters, whereas acetoclastic methanogens were barely detected.ConclusionsResults presented here provide new insights into a remarkable anaerobic digestion ecosystem where acetate catabolism is mainly realized by Bacteria. Metagenomics and enrichment cultivation revealed a core community of diverse and novel uncultured acetate-oxidizing bacteria and point to a particular niche for them in dry fermentation of biowaste. Their genomic repertoire suggests metabolic plasticity besides the potential for syntrophic acetate oxidation.1dV765cjiwNJNY-1M7VNFLVideo

SARS-CoV-2 from faeces to wastewater treatment: What do we know? A review

SARS-CoV-2, the virus that causes COVID-19, has been found in the faeces of infected patients in numerous studies. Stool may remain positive for SARS-CoV-2, even when the respiratory tract becomes negative, and the interaction with the gastrointestinal tract poses a series of questions about wastewater and its treatments. This review aims to understand the viral load of SARS-CoV-2 in faeces and sewage and its fate in wastewater treatment plants (WWTPs).The viral load in the faeces of persons testing positive for SARS-CoV-2 was estimated at between 5·103 to 107.6 copies/mL, depending on the infection course. In the sewerage, faeces undergo dilution and viral load decreases considerably in the wastewater entering a WWTP with a range from 2 copies/100 mL to 3·103 copies/mL, depending on the level of the epidemic. Monitoring of SARS-CoV-2 in sewage, although no evidence of COVID-19 transmission has been found via this route, could be advantageously exploited as an early warning of outbreaks. Preliminary studies on WBE seem promising; but high uncertainty of viral loads in wastewater and faeces remains, and further research is needed.The detection of SARS-CoV-2 in sewage, based on RNA sequences and RT-PCR, requires a shared approach on sample pre-treatment and on-site collection to ensure comparable results. The finding of viral RNA in stools does not imply that the virus is viable and infectious. Viability of CoVs such as SARS-CoV-2 decreases in wastewater - due to temperature, pH, solids, micropollutants - but high inactivation in WWTPs can be obtained only by using disinfection (free chlorine, UVC light). A reduction in the quantity of disinfectants can be obtained by implementing Membrane-Bioreactors with ultrafiltration to separate SARS-CoV-2 virions with a size of 60–140 nm. In sludge treatment, thermophilic digestion is effective, based on the general consensus that CoVs are highly sensitive to increased temperatures.

Comparative response of thermophilic and mesophilic sludge digesters to zinc oxide nanoparticles.

The inevitable discharge of zinc oxide nanoparticles (ZnO NPs), from consumer and industrial products, into wastewater treatment plants (WWTPs) has created a need to determine their effect on sludge digestion. In this study, the effect of particle size (30 nm and 100 nm), type (coated and non-coated), and dose (6, 75, and 150 mg/g feed total solids (TS)) of ZnO NPs on anaerobic sludge digestion was studied under mesophilic (35 °C) and thermophilic (55 °C) conditions. The effect was investigated in two stages with different digester feeding regime: (1) batch biochemical methane potential (BMP) assays, and (2) semi-continuously fed reactors. Results showed that ZnO NPs were inhibitory at medium and high levels (75 and 100 mg ZnO/g TS, respectively). Coated NPs created less inhibition than non-coated NPs. Thermophilic bacteria were more sensitive to ZnO NPs compared with mesophilic bacteria. For the non-coated ZnO NPs, only the mesophilic batch assays were able to recover at the medium concentration and the thermophilic reactors presented chronic inhibition and could not recover. As a beneficial outcome, coated ZnO NPs significantly reduced odor-causing volatile sulfur compounds in digester headspace in comparison with the non-coated NPs. In summary, the condition in which ZnO NPs would have little to no effect would be 6 mg/g TS-coated ZnO NPs under mesophilic conditions.

Dry mesophilic and thermophilic semi-continuous anaerobic digestion of cow manure: effects of solid loading rate on the process performance

Four solid loading rates (SLRs) of 3-9 kg TS/m3 day at fixed 20% total solids (TS) content were chosen in this study and assessed under two distinct circumstances: mesophilic and thermophilic under dry semi-continuous cow manure (CM) digestion. This research aimed to investigate the production of biogas, volatile solid reductions and volatile fatty acids (VFAs) concentration effects on process efficiency. This was done in 100 days of operation. The findings indicated that the production of biogas and volatile solid reduction in both digesters showed similar patterns of decrements alongside increasing SLRs. In all test cases, the best performance of the digesters was found at the SLR of 3 kg TS /m3 day, which was under thermophilic conditions. The methane yielded 0.39 m3 CH4/kg VS (65 % CH4). Meanwhile, the mesophilic reactor recorded the production of 0.25 m3 CH4/kg VS (60 % CH4). A 35.89 % increment in methane production was observed in the thermophilic state. About 67 % and 62 % of volatile solids reduction were observed in the thermophilic and mesophilic digesters, respectively. In contrast, it was observed that volatile fatty acids showed inhibitory effects at the SLR of 9 kg TS/m3 day, which in turn decreased the digesters’ performance.

Effects of reactive oxygen species scavengers on thermophilic micro-aerobic digestion for sludge stabilization

In thermophilic digestion systems, sludge stabilization may be adversely impacted by high concentrations of ammonia nitrogen or the rapid accumulation of fatty acids; however, few studies have focused on the mitigation of the inhibition of reactive oxygen species (ROS). In this study, fulvic acids or tea polyphenols were introduced to a thermophilic digestion system and the effects of ROS scavengers on sludge stabilization were investigated. As fulvic acids or tea polyphenols were added to sludge, they reacted with metal cations, such as Cu2+ and Zn2+, to form stable complexes that enabled active metal ions to be transported into cells to enhance the oxidase activities. Therefore, the digestion systems presented a lower O2•- content compared with that of a control group with no additive. Both fulvic acids and tea polyphenols mitigated the adverse effects of the ROS and enhanced the reduction of volatile solids (VS), however, fulvic acids better facilitated the sludge stabilization. The optimal dosage was 0.3% of the total solids as fulvic acids were added to the sludge every 48 h. The VS reduction in the digester reached 38.2% at 14 d, which was clearly higher than that in the control group. The pathway of ROS scavengers affecting sludge stabilization was proposed, and it may be helpful to gain deeper insight into the characteristics of thermophilic digestion processes as well as the mechanism of sludge stabilization.

Effect of sulfate addition on carbon flow and microbial community composition during thermophilic digestion of cellulose

Substrates with high sulfate levels pose problems for biogas production as they allow sulfate reducing bacteria to compete with syntrophic and methanogenic members of the community. In addition, the end product of sulfate reduction, hydrogen sulfide, is toxic and corrosive. Here we show how sulfate addition affects physiological processes in a thermophilic methanogenic system by analyzing the carbon flow and the microbial community with quantitative PCR and amplicon sequencing of the 16s rRNA gene. A sulfate addition of 0.5 to 3 g/L caused a decline in methane production by 73–92%, while higher sulfate concentrations had no additional inhibitory effect. Generally, sulfate addition induced a shift in the composition of the microbial community towards a higher dominance of Firmicutes and decreasing abundances of Bacteroidetes and Euryarchaeota. The abundance of methanogens (e.g., Methanoculleus and Methanosarcina) was reduced, while sulfate reducing bacteria (especially Candidatus Desulforudis and Desulfotomaculum) increased significantly in presence of sulfate. The sulfate addition had a significant impact on the carbon flow within the system, shifting the end product from methane and carbon dioxide to acetate and carbon dioxide. Interestingly, methane production quickly resumed, when sulfate was no longer present in the system. Despite the strong impact of sulfate addition on the carbon flow and the microbial community structure during thermophilic biogas production, short-term process disturbances caused by unexpected introduction of sulfate may be overcome due to the high resilience of the engaged microorganisms.

Comparative Analysis of Bacterial and Archaeal Community Structure in Microwave Pretreated Thermophilic and Mesophilic Anaerobic Digesters Utilizing Mixed Sludge under Organic Overloading

The effects of microwave (MW) pretreatment were investigated by six anaerobic digesters operated under thermophilic and mesophilic conditions at high organic loading rates (4.9–5.7 g volatile solids/L/d). The experiments and analyses were mainly designed to reveal the impact of MW pretreatment and digester temperatures on the process stability and microbial community structure by correlating the composition of microbial populations with volatile fatty acid (VFA) concentrations. A slight shift from biogas production (with a reasonable methane content) to VFA accumulation was observed in the thermophilic digesters, especially in the MW-irradiated reactors. Microbial population structure was assessed using a high-throughput sequencing of 16S rRNA gene on the MiSeq platform. Microbial community structure was slightly affected by different MW pretreatment conditions, while substantially affected by the digester temperature. The phylum Bacteroidetes proliferated in the MW-irradiated mesophilic digesters by resisting high-temperature MW (at 160 °C). Hydrogenotrophic methanogenesis (mostly the genus of Methanothermobacter) was found to be a key route of methane production in the thermophilic digesters, whereas aceticlastic methanogenesis (mostly the genus of Methanosaeta) was the main pathway in the mesophilic digesters.

Insights into Thermophilic Plant Biomass Hydrolysis from Caldicellulosiruptor Systems Biology.

Plant polysaccharides continue to serve as a promising feedstock for bioproduct fermentation. However, the recalcitrant nature of plant biomass requires certain key enzymes, including cellobiohydrolases, for efficient solubilization of polysaccharides. Thermostable carbohydrate-active enzymes are sought for their stability and tolerance to other process parameters. Plant biomass degrading microbes found in biotopes like geothermally heated water sources, compost piles, and thermophilic digesters are a common source of thermostable enzymes. While traditional thermophilic enzyme discovery first focused on microbe isolation followed by functional characterization, metagenomic sequences are negating the initial need for species isolation. Here, we summarize the current state of knowledge about the extremely thermophilic genus Caldicellulosiruptor, including genomic and metagenomic analyses in addition to recent breakthroughs in enzymology and genetic manipulation of the genus. Ten years after completing the first Caldicellulosiruptor genome sequence, the tools required for systems biology of this non-model environmental microorganism are in place.

Anaerobic digestion of purple phototrophic bacteria – The release step of the partition-release-recover concept

Purple phototrophic bacteria (PPB) have been proposed as a high-growth, assimilative option for wastewater treatment. The original partition-release-recover concept proposal requires their near complete digestion and release (and subsequent recovery) of energy and nutrients in an anaerobic digester. While the growth (partition) step has been extensively assessed, no work has been done on their anaerobic digestion characteristics (release). Continuous mesophilic (20d) and thermophilic (10d) digestion could achieve around 55% volatile solids degradation (VSD), with 35% (mesophilic) and 20% (thermophilic) nitrogen solubilisation. Post digestion (with/without pretreatment) could increase the VSD to 70% and nitrogen solubilisation to 43%. A number of pretreatment options were tested, with high temperature and sonication being relatively effective, and chemical treatment, and temperature phased digestion being relatively ineffective vs controls. Overall, anaerobic digestion of PPB results in substantial residual particulate material, with an increased nitrogen content, and avenues to effectively utilise this residue should be identified.

PH and Phosphate Induced Shifts in Carbon Flow and Microbial Community during Thermophilic Anaerobic Digestion.

pH is a central environmental factor influencing CH4 production from organic substrates, as every member of the complex microbial community has specific pH requirements. Here, we show how varying pH conditions (5.0–8.5, phosphate buffered) and the application of a phosphate buffer per se induce shifts in the microbial community composition and the carbon flow during nine weeks of thermophilic batch digestion. Beside monitoring the methane production as well as volatile fatty acid concentrations, amplicon sequencing of the 16S rRNA gene was conducted. The presence of 100 mM phosphate resulted in reduced CH4 production during the initial phase of the incubation, which was characterized by a shift in the dominant methanogenic genera from a mixed Methanosarcina and Methanoculleus to a pure Methanoculleus system. In buffered samples, acetate strongly accumulated in the beginning of the batch digestion and subsequently served as a substrate for methanogens. Methanogenesis was permanently inhibited at pH values ≤5.5, with the maximum CH4 production occurring at pH 7.5. Adaptations of the microbial community to the pH variations included shifts in the archaeal and bacterial composition, as less competitive organisms with a broad pH range were able to occupy metabolic niches at unfavorable pH conditions.

Comparative study of industrial-scale high-solid biogas production from food waste: Process operation and microbiology

Anaerobic high-solid treatment (HST) for processing food waste and biogas production is a viable technology with considerable commercial potential. In this study, we examined and compared mesophilic and thermophilic industrial-scale plug-flow digesters. The HSTs demonstrated reasonable biogas yields from food waste (0.4–0.6 Nm3 CH4/kg volatile solids). However, during operation at thermophilic conditions ammonia inhibition (~2 g NH3-N/L) and acid accumulation (6–14 g/L) caused severe process disturbance. Microbial community structures diverged between the processes, with temperature appearing to be a strong driver. A unique feature of the thermophilic HSTs was high abundance of the uncultivated Clostridia group MBA03 and temperature fluctuations in one mesophilic HST were linked to drastically decreased abundance of methanogens and relative abundance of Cloacimonetes. The process data obtained in this study clearly demonstrate both potential and challenges in HST of food waste but also possibilities for management approaches to tackle process imbalance and restore process function.

Methane yield enhancement of mesophilic and thermophilic anaerobic co-digestion of algal biomass and food waste using algal biochar: Semi-continuous operation and microbial community analysis

The impact of algal biochar addition on mesophilic and thermophilic anaerobic co-digestion of algal biomass and food waste was investigated with a focus on semi-continuous operations and functional microbial communities. Under batch co-digestion, the highest co-digestion synergy was observed for a mixture of 25% food waste and 75% algal biomass. During semi-continuous co-digestion of 25% food waste-75% algal biomass mixture, biochar amended digesters exhibited a 12–54% increase in average methane yield (275.8–394.6 mL/gVS) compared to the controls. Elevated temperature induced narrow distributions of volatile fatty acids (VFAs) by inhibiting the production of branched VFAs. Genus Proteiniphilum was selectively enriched by 3.2 folds in mesophilic digesters with biochar amendment while genus Defluviitoga was selectively enriched in thermophilic digesters due to elevated temperature. Methanogenic communities were significantly different in mesophilic and thermophilic digesters. Biochar amendment contributed to shifts in the predominant methanogens leading to a more balanced state of two methanogenic pathways.

Influence of Anaerobic Mesophilic and Thermophilic Digestion on Cytotoxicity of Swine Wastewaters.

Recycling wastewater from animal production for fertilizers using anaerobic digestion (AD) is a common method to recover the nutrients in the digestate. However, the digestate toxicity is not well understood because AD is mainly designed for chemical oxygen demand reduction. This study determined the toxicity during AD and the controlling factors with the goal to improve digestate safety during farmer handling to reuse the nutrients. Thermophilic and mesophilic AD of two swine wastewater sources were studied. Mammalian cell cytotoxicity revealed that the effluent after thermophilic digestion was at least 69% more toxic than the mesophilic effluent, owing to higher ammonia and total organic carbon in the former. Ammonia accounted for >55% total cytotoxicity, and the organics of the thermophilic digestate were twice more toxic than those in the mesophilic digestate. Despite less toxicity contribution than the ammonia, the organics did demonstrate significant adverse effects on the thiol-mediated cellular protection mechanism. For swine wastewater nutrient recovery, converting ammonia to less toxic nitrogen forms could lower the toxic hazard of the AD digestate. With much less ammonia, the organics would be the remaining decisive factor for toxicity, which is favorably reduced using thermophilic AD over mesophilic. If the ammonia is not reduced, mesophilic AD would generate a less toxic digestate.

Occurrence of the Persistent Antimicrobial Triclosan in Microwave Pretreated and Anaerobically Digested Municipal Sludges under Various Process Conditions.

Treatment of emerging contaminants, such as antimicrobials, has become a priority topic for environmental protection. As a persistent, toxic, and bioaccumulative antimicrobial, the accumulation of triclosan (TCS) in wastewater sludge is creating a potential risk to human and ecosystem health via the agricultural use of biosolids. The impact of microwave (MW) pretreatment on TCS levels in municipal sludge is unknown. This study, for the first time, evaluated how MW pretreatment (80 and 160 °C) itself and together with anaerobic digestion (AD) under various sludge retention times (SRTs: 20, 12, and 6 days) and temperatures (35 and 55 °C) can affect the levels of TCS in municipal sludge. TCS and its potential transformation products were analyzed with ultra-high-performance liquid chromatography and tandem mass spectrometry. Significantly higher TCS concentrations were detected in sludge sampled from the plant in colder compared to those in warmer temperatures. MW temperature did not have a discernible impact on TCS reduction from undigested sludge. However, AD studies indicated that compared to controls (no pretreatment), MW irradiation could make TCS more amenable to biodegradation (up to 46%), especially at the elevated pretreatment and digester temperatures. At different SRTs studied, TCS levels in the thermophilic digesters were considerably lower than that of in the mesophilic digesters.