Anaerobic digestion of sludge by different pretreatments: Changes of amino acids and microbial community

•Tryptophan protein, and aromatic protein I/II were the key identified proteins. •Cysteine was more correlated with methane production than other amino acids. •The presence of cysteine can promote methane production and degradation of VFAs. •The presence of cysteine can lower ORP and increase biomass activity. •Predominant Tissierella and Proteiniphilum were noted in pretreated sludge samples. Many studies have investigated the effects of different pretreatments on the performance of anaerobic digestion of sludge. However, the detailed changes of dissolved organic nitrogen, particularly the release behavior of proteins and the byproducts of protein hydrolysis-amino acids, are rarely known during anaerobic digestion of sludge by different pretreatments. Here we quantified the changes of three types of proteins and 17 types of amino acids in sludge samples solubilized by ultrasonic, thermal, and acid/alkaline pretreatments and their transformation during anaerobic digestion of sludge. Tryptophan protein, aromatic protein I, aromatic protein II, and cysteine were identified as the key dissolved organic nitrogen responsible for methane production during anaerobic digestion of sludge, regardless of the different pretreatment methods. Different from the depletion of other amino acids, cysteine was resistant to degradation after an incubation period of 30 days in all sludge samples. Meanwhile, the “cysteine and methionine metabolism (K00270)” was absent in all sludge samples by identifying 6755 Kyoto Encyclopedia of Genes and Genomes assignments of genes hits. Cysteine contributed to the generation of methane and the degradation of acetic, propionic, and n-butyric acids through decreasing oxidation-reduction potential and enhancing biomass activity. This study provided an alternative strategy to enhance anaerobic digestion of sludge through in situ production of cysteine.     

Influencing mechanism of high solid concentration on anaerobic mono-digestion of sewage sludge without agitation

High-solids anaerobic digestion of sewage sludge was a promising process, but high solid concentration negatively influenced methane production. The influencing mechanism was systematically analyzed in this study through a series of static anaerobic digestion experiments at total solids (TS) contents of 3%–15%. The results showed that TS 6% was the boundary between low-solids and high-solids anaerobic digestion, and the accumulative methane yield decreased exponentially when TS increased from 6% to 15%. The performance of anaerobic digestion was directly determined by the efficiency of mass transfer, and the relation between methane yield and sludge diffusive coefficients was well described by a power function. Thus, the increasing TS resulted in an exponential increase in sludge viscosity but an exponential decrease in diffusive coefficient. The blocked mass transfer led to the accumulation of volatile fatty acids (VFAs) and free ammonia. Acetic metabolism was the main process, whereas butyric and propionic metabolisms occurred at the initial stage of high-solids anaerobic digestion. The concentration of VFAs reached the maximum at the initial stage, which were still lower than the threshold influencing methanogens. The concentration of free ammonia increased gradually, and the methanogenesis was inhibited when free ammonia nitrogen exceeded 50 mg·L⁻¹. Consequently, the deterioration of high-solids anaerobic digestion was related to the blocked mass transfer and the resulting ammonia accumulation.     

微波碱解处理剩余污泥的厌氧消化性能

为了研究微波碱解预处理（microwave/NaOH pretreatment,MNP）剩余污泥在高温和中温条件下厌氧消化的性能,采用半连续完全混合式反应器分别研究了MNP处理后的剩余污泥在高温厌氧消化工艺（thermophilic anaerobic di-gestion process,TADP）和中温厌氧消化工艺...     

Enhanced 4-chlorophenol biodegradation by integrating Fe2O3 nanoparticles into an anaerobic reactor: Long-term performance and underlying mechanism

• 4-chlorophenol biodegradation could be enhanced in Fe₂O₃ coupled anaerobic system. • Metabolic activity and electron transport could be improved by Fe₂O₃ nanoparticles. • Functional microbial communities could be enriched in coupled anaerobic system. • Possible synergistic mechanism involved in enhanced dechlorination was proposed. Fe₂O₃ nanoparticles have been reported to enhance the dechlorination performance of anaerobic systems, but the underlying mechanism has not been clarified. This study evaluated the technical feasibility, system stability, microbial biodiversity and the underlying mechanism involved in a Fe₂O₃ nanoparticle-coupled anaerobic system treating 4-chlorophenol (4-CP) wastewater. The results demonstrated that the 4-CP and total organic carbon (TOC) removal efficiencies in the Fe₂O₃-coupled up-flow anaerobic sludge blanket (UASB) were always higher than 97% and 90% during long-term operation, verifying the long-term stability of the Fe₂O₃-coupled UASB. The 4-CP and TOC removal efficiencies in the coupled UASB increased by 42.9±0.4% and 27.5±0.7% compared to the control UASB system. Adding Fe₂O₃ nanoparticles promoted the enrichment of species involved in dechlorination, fermentation, electron transfer and acetoclastic methanogenesis, and significantly enhanced the extracellular electron transfer ability, electron transport activity and conductivity of anaerobic sludge, leading to enhanced 4-CP biodegradation performance. A possible synergistic mechanism involved in enhanced anaerobic 4-CP biodegradation by Fe₂O₃ nanoparticles was proposed.     

Environmental effects and risk control of antibiotic resistance genes in the organic solid waste aerobic composting system: A review

• ARGs were detected in livestock manure, sludge, food waste and fermentation dregs. • The succession of microbial community is an important factor affecting ARGs. • Horizontal transfer mechanism of ARGs during composting should be further studied. Antibiotic resistance genes (ARGs) have been diffusely detected in several kinds of organic solid waste, such as livestock manure, sludge, antibiotic fermentation residues, and food waste, thus attracting great attention. Aerobic composting, which is an effective, harmless treatment method for organic solid waste to promote recycling, has been identified to also aid in ARG reduction. However, the effect of composting in removing ARGs from organic solid waste has recently become controversial. Thus, this article summarizes and reviews the research on ARGs in relation to composting in the past 5 years. ARGs in organic solid waste could spread in different environmental media, including soil and the atmosphere, which could widen environmental risks. However, the conventional composting technology had limited effect on ARGs removal from organic solid waste. Improved composting processes, such as hyperthermophilic temperature composting, could effectively remove ARGs, and the HGT of ARGs and the microbial communities are identified as vital influencing factors. Currently, during the composting process, ARGs were mainly affected by three response pathways, (I) “Microenvironment-ARGs”; (II) “Microenvironment-microorganisms-ARGs”; (III) “Microorganisms-horizontal gene transfer-ARGs”, respectively. Response pathway II had been studied the most which was believed that microbial community was an important factor affecting ARGs. In response pathway III, mainly believed that MGEs played an important role and paid less attention to eARGs. Further research on the role and impact of eARGs in ARGs may be considered in the future. It aims to provide support for further research on environmental risk control of ARGs in organic solid waste.     

A syntrophic propionate-oxidizing microflora and its bioaugmentation on anaerobic wastewater treatment for enhancing methane production and COD removal

Methane fermentation process can be restricted and even destroyed by the accumulation of propionate because it is the most difficult to be anaerobically oxidized among the volatile fatty acids produced by acetogenesis. To enhance anaerobic wastewater treatment process for methane production and COD removal, a syntrophic propionate-oxidizing microflora B83 was obtained from an anaerobic activated sludge by enrichment with propionate. The inoculation of microflora B83, with a 1:9 ratio of bacteria number to that of the activated sludge, could enhance the methane production from glucose by 2.5 times. With the same inoculation dosage of the microflora B83, COD removal in organic wastewater treatment process was improved from 75.6% to 86.6%, while the specific methane production by COD removal was increased by 2.7 times. Hydrogen-producing acetogenesis appeared to be a rate-limiting step in methane fermentation, and the enhancement of hydrogen-producing acetogens in the anaerobic wastewater treatment process had improved not only the hydrogen-producing acetogenesis but also the acidogenesis and methanogenesis.

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城市有机生活垃圾溶胞处理对其厌氧消化的影响机理

由于城市有机生活垃圾成分的复杂性和厌氧消化的限速步骤的影响,导致厌氧发酵的速度比较缓慢、产气量较少和工艺不稳定等问题。文章综述了国内外有机生活垃圾的各种溶胞处理技术,如物理法、化学法、生物法及其联合处理等方法,以便改善发酵物料的性质,消除厌氧发酵的限速步骤。研究认为,通过溶胞处理能够改善有机垃圾的物理化学性质如发酵物料的溶解度、酸碱度等,提高微生物对难降解有机物的分解,增加可溶性COD和挥发性酸的浓度,优化发酵细菌的代谢途径以及产物的组成等,从而增加生物气产量,缩短水力停留时间,强化厌氧发酵过程,减轻了后续处理的负担。     

Anaerobic digestion and recycling of kitchen waste: a review

About 1.6 billion tons of food are wasted worldwide annually, calling for advanced methods to recycle food waste into energy and materials. Anaerobic digestion of kitchen waste allows the efficient recovery of energy, and induces low-carbon emissions. Nonetheless, digestion stability and biogas production are variables, due to dietary habits and seasonal diet variations that modify the components of kitchen waste. Another challenge is the recycling of the digestate, which could be partly solved by more efficient reactors of anaerobic digestion. Here, we review the bottlenecks of anaerobic digestion treatment of kitchen waste, with focus on components inhibition, and energy recovery from biogas slurry and residue. We provide rules for the optimal treatment of the organic fraction of kitchen waste, and guidelines to upgrade the anaerobic digestion processes. We propose a strategy using an anaerobic dynamic membrane bioreactor to improve anaerobic digestion of kitchen waste, and a model for the complete transformation and recycling of kitchen waste, based on component properties.

     

Influence of arsanilic acid,Cu2+, PO43− and their interaction on anaerobic digestion of pig manure

The methanogenesis was severely inhibited with 0.46 mM ASA addition.PO₄³⁻ didn’t attenuate the methanogenesis inhibition in the existence of ASA.ASA was transformed to As(III), As(V), MMA and DMA in anaerobic digestion.Cu²⁺ mitigated the methanogenesis inhibition via impeding the degradation of ASA.Arsanilic acid (ASA), copper ion (Cu²⁺) and phosphate (PO₄³⁻) are widely used as feed additives for pigs. Most of these three supplemented feed additives were excreted in feces and urine. Anaerobic digestion is often used for the management of pig manure. However, the interaction of ASA with Cu²⁺ or PO₄³⁻ on anaerobic digestion is still not clear. In this study, the influence of ASA, Cu²⁺, PO₄³⁻ and their interaction on anaerobic digestion of pig manure and the possible mechanisms were investigated. The initial concentrations of ASA, Cu²⁺ and PO₄³⁻ were 0.46 mM, 2 mM and 2 mM in the anaerobic digester, respectively. The methanogenesis was severely inhibited in the assays with only ASA addition, only Cu²⁺ addition and ASA+ PO₄³⁻ addition with the inhibition index of 97.8%, 46.6% and 82.6%, respectively, but the methanogenesis inhibition in the assay with ASA+ Cu²⁺ addition was mitigated with the inhibition index of 39.4%. PO₄³⁻ had no obvious impacts on the degradation of ASA. However, Cu²⁺ addition inhibited the degradation of ASA, mitigating the methanogenesis inhibition. The existence of ASA would inhibit methanogenesis and generate more toxic inorganic arsenic compounds during anaerobic digestion, implying the limitation of anaerobic digestion for ASA- contaminated animal manure. However, the co-existence of ASA and Cu²⁺ could mitigate the inhibition. These results could provide useful information for the management of anaerobic digestion of pig manure containing ASA with Cu²⁺.     

影响厌氧氨氧化与甲烷化反硝化耦合的因素

氨氮、氮氧化物对产甲烷菌有一定的抑制作用,但可以通过驯化去除毒性.亚硝酸盐在厌氧氨氧化菌作用下与氨发生厌氧氨氧化反应.虽然厌氧氨氧化菌是自养菌,但具有异养代谢能力,并且NO2可提高厌氧氨氧化菌的活性.因此,通过特殊的反应器技术,将厌氧氨氧化菌与甲烷菌、反硝化菌复合在一个有利的微生态环境中,充分发挥它们之间的协同耦合作用,把有机物转化为清洁能源又同时脱氮,是极有前景的废水厌氧(缺氧)处理研究新方向.表1参31     

Impact of Ni(II), Zn(II) and Cd(II) on biogassification of potato waste

A study was conducted on anaerobic digestion of potato waste and cattle manure mixture, inoculated with 12% inoculum and diluted to 1:1 substrate water ratio at 37 +/- 1 degrees C. Initially pH of substrate was found to be 4.5 to 5.0. Lime and sodium bicarbonate solutions were employed to adjust the pH to 7.5. Biogas production continued up to 10 and 7 days, when lime and sodium bicarbonate solutions were used to adjust the pH, respectively. Biogassification potential was studied in response to different ratio of waste and cattle manure. Biogas production rate was higher when potato waste and cattle manure were used in 50:50 ratio. Effect of two different concentrations (2.5 and 5.0 ppm) of three heavy metals viz. (Ni (II), Zn (II) and Cd (II)) on anaerobic digestion of substrate (potato waste--cattle manure, 50:50) was studied. At 2.5 ppm, all the three heavy metals increased biogas production rate over the control value. The percentage increase in biogas production over the control was highest by Cd, followed by Ni and Zn. In all the treatments, methane content of biogas increased with increase in time after feeding. Various physico-chemical parameters viz. total solids, total volatile solids, total organic carbon and chemical oxygen demand considerably declined after 7 days of digestion and decline was greater in presence of heavy metals as compared to control. The physico-chemical parameters revealed maximum decrease in the presence of 2.5-ppm concentrations of heavy metals with the substrate. Among all the three heavy metals employed in the study, Cd++ at 2.5 ppm was found to produce maximum biogas production rate. The use of three heavy metals to enhance biogas production from potato and other horticultural waste is discussed.     

四环素类抗生素和铜复合污染对猪粪厌氧消化的影响

兽用抗生素和矿物元素添加剂可起到预防动物疾病、促进动物生长、提高饲料转化率等作用,因此被广泛应用于畜禽养殖业。本研究以猪粪中温厌氧消化为研究目标,采用全自动甲烷潜势测试系统,考察了一定浓度的四环素(TC:30 mg·kg~(-1)dry weight,DW)、土霉素(OTC:50 mg·kg~(-1)DW)和金霉素(CTC:15 mg·kg~(-1)DW)对厌氧累积产甲烷量和日产甲烷速率的影响。结果表明,TC、OTC和CTC对猪粪中温厌氧消化累积产甲烷量和日产甲烷速率均有促进作用(累积产甲烷总量提高比例分别为7.9%、0.4%和5.4%)。另外,采用超高效液相色谱-四极杆串联质谱对猪粪厌氧消化前后样品中四环素类抗生素及其代谢产物进行了分析。结果表明,液相中的四环素类抗生素在猪粪厌氧消化过程中得到了明显的去除,去除率达到90%~100%;而固相中只有金霉素和差向异构金霉素有明显的去除效果,去除率分别为41.69%和41.58%。采用Tessier连续提取法对猪粪厌氧消化前后样品中5种形态的铜包括可交换态、碳酸盐结合态、铁锰氧化结合态、有机物结合态、残渣态进行了分析,结果表明,猪粪厌氧消化后,可交换态、碳酸盐结合态和铁锰结合态的铜浓度比厌氧消化前分别降低了1%~9%、0.1%~3%、12%~19%,而有机态和残渣态的铜浓度却在厌氧消化后分别增加了15%~35%、1%~2%。厌氧消化后,70%~80%的铜都是以有机铜的形态存在。铜逐渐从不稳定态转化为相对稳定的有机态和残渣态铜,因此,厌氧消化过程使铜从可生物利用态转变为不可生物利用态,趋于稳定化。     

Cytoenzymatic investigation of intracellular digestion in the symbiont-bearing hydrothermal bivalve <Emphasis Type="Italic">Bathymodiolus azoricus</Emphasis>

Invertebrates harbouring endosymbiotic chemoautotrophic bacteria are widely distributed in a variety of reducing marine habitats, including deep-sea hydrothermal vents. Bathymodiolids are dominants of the biomass at geochemically distinct vent sites of the Mid Atlantic Ridge (MAR) and thus are good candidates to study biological processes in response to site-specific conditions. To satisfy their nutritional requirements, these organisms depend to varying extent on two types of chemoautotrophic symbionts and on filterfeeding. The quantitative relationships of the nutritional modes are poorly understood. Using enzyme cytochemistry, electron microscopy and X-ray microanalysis, the structural and functional aspects of the cellular equipment necessary for lysosomal digestion was studied. We provide evidence for the following: (1) the basis of intracellular digestion of symbionts in Bathymodiolus azoricus from two geochemically distinct vent sites was not mainly in the large lysosomal bodies as previously thought (based on the membranous content resembling bacteria); (2) senescent bacteria are autolysed, possibly by bacterial acid phosphatase, that is more likely a cell cycling of the symbionts rather than an active lysosomal digestion by the host; (3) the consistent absence of hydrolases may indicate the improper use of the name “lysosome” for large vesicles at the base of the gill bacteriocytes (4) nutrient transfer in B. azoricus, therefore, may more likely be accomplished through leaking of metabolites from the symbiont to the host, not excluding lysosomal resorption of dead bacteria as an auxiliary strategy for organic molecule transfer; (5) evidence is provided for microvillar transfer of substances from the seawater that may indicate filter-feeding, in non-symbiotic ciliated gill cells of mussels from Lucky Strike; (6) two types of lysosomal vesicles can be distinguished in digestive cells based on their enzymatic content and their elemental composition.     

Increasing the performance of anaerobic digestion: Pilot scale experimental study for thermal hydrolysis of mixed sludge

The performance of a pilot plant operation combining thermal hydrolysis (170°C, 30 min) and anaerobic digestion (AD) was studied, determining the main properties for samples of fresh mixed sludge, hydrolyzed sludge, and digested sludge, in order to quantify the thermal pretreatment performance (disintegration, solubilisation, and dewaterability) and its impact on the anaerobic digestion performance (biodegradability, volatile solids reduction, and digester rheology) and end product characteristics (dewaterability, sanitation, organic and nitrogen content). The disintegration achieved during the thermal treatment enhances the sludge centrifugation, allowing a 70% higher total solids concentration in the feed to anaerobic digestion. The digestion of this sludge generates 40% more biogas in half the time, due to the higher solids removal compared to a conventional digester. The waste generated can be dewatered by centrifugation to 7% dry solids without polymer addition, and is pathogen free.     

Enhanced methane recovery and exoelectrogen-methanogen evolution from low-strength wastewater in an up-flow biofilm reactor with conductive granular graphite fillers

Methane production from low-strength wastewater (LSWW) is generally difficult because of the low metabolism rate of methanogens. Here, an up-flow biofilm reactor equipped with conductive granular graphite (GG) as fillers was developed to enhance direct interspecies electron transfer (DIET) between syntrophic electroactive bacteria and methanogens to stimulate methanogenesis process. Compared to quartz sand fillers, using conductive fillers significantly enhanced methane production and accelerated the start-up stage of biofilm reactor. At HRT of 6 h, the average methane production rate and methane yield of reactor with GG were 0.106 m³/(m³·d) and 74.5 L/kg COD, which increased by 34.3 times and 22.4 times respectively compared with the reactor with common quartz sand fillers. The microbial community analysis revealed that methanogens structure was significantly altered and the archaea that are involved in DIET (such as Methanobacterium) were enriched in GG filler. The beneficial effects of conductive fillers on methane production implied a practical strategy for efficient methane recovery from LSWW.

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添加剂对污泥厌氧消化性能的影响

在间歇培养条件下,研究了还原型辅酶Ⅱ（NADPH）、乙酰辅酶A（Acetyl Co A）和对氨基苯甲酸（PABA）3种添加剂对污泥厌氧消化性能的影响.结果表明,3种微生物活性促进剂均能促进污泥厌氧消化产气.其中,NADPH的促进效果最为显著,消化第35 d,产甲烷量比对照组高15.90%.在污泥含固率为3%、未调初始pH（pH=6.7）和温度35℃的厌氧消化条件下,NADPH的最佳添加量为50 mg.L-1,消化第36 d,污泥累积产甲烷量127.13 mL.g-1VSS.在含固率3%、初始pH=8.5、温度55℃和NADPH添加量为50 mg.L-1的工艺条件下,污泥厌氧产气效果最佳,消化第30 d时累积产甲烷量达158.02 mL.g-1VSS.     

猪场废水厌氧消化过程中的除磷效果

采用序批式半连续厌氧消化试验方法,研究猪场废水厌氧消化过程中磷的去除情况。结果表明,水力停留时间（HRT）为1、3、6和9 d的厌氧反应器平均除磷率分别为65.0%、81.1%、82.7%和83.0%,而COD平均去除率分别为54.5%、82.3%、87.0%和85.9%。厌氧反应器除磷能力随沼气产量的增加而增加,说明厌氧反应器中磷的去除与产甲烷过程密切相关。对厌氧消化前后的污泥进行浸提后发现,厌氧消化过程中,化学反应生成磷酸盐沉淀的除磷作用十分显著,污泥中正磷酸盐,与铁结合的磷化合物（Fe-RP）,还原可溶性磷,与钙、镁离子结合的磷化合物（Ca-RP、Mg-RP）以及无机或有机聚合磷增加量分别为0.027 8～0.101 5、0.013 5～0.081 0、0.2165～0.430 5、23.4～54.8和7.2～21.5 mg.g-1;且总体而言,HRT越长,污泥中与不同金属结合的磷增加量就越大。从HRT、磷和有机物的去除效果以及沼气产气速率3个方面综合考虑,猪场废水厌氧消化反应器的HRT控制在3 d为宜。     

Equation for isotope accumulation in products and biomass as a way to reveal the pathways in mesophilic methanol methanization by microbial community

Isotope-labelling of substrate is used to reveal the methabolic pathways of substrate transformation by microbial community. In this paper, in order to describe the batch mesophilic anaerobic methanization of ¹³C-labelled methanol and microbial ecology analysis (Li et al., 2008), an equation for the isotope accumulation in products and biomass was included into the basic mathematical model based on stoichiometric chemical reactions. The higher was the isotope level in substrate, the larger fraction of ¹³C accumulated in products and biomass. Acetate, total organic and inorganic carbon (TOC, TIC) concentrations and methane production were used for the model calibration, whereas ¹³C enrichment of acetate, TIC and biomass were used for model validation. In the model, chemical transformations including methanol and acetate oxidation, homoacetogenesis, hydrogenotrophic and aceticlastic methanogenesis were considered. The rate-limiting reactions were methanol and acetate consumption. According to the model, homoacetogens performing acetate formation and oxidation were competed with hydrogenotrophic methanogens for hydrogen. Biphasic methane production was due to hydrogenotrophic methanogenesis in the first phase and due to acetiticlastic and hydrogenotrophic methanogenesis following acetate oxidation in the second phase.     

Feasibility assessment of up-flow anaerobic sludge blanket treatment of sulfamethoxazole pharmaceutical wastewater

The UASB system successfully treated sulfamethoxazole pharmaceutical wastewater. High concentration sulfate of this wastewater was the main refractory factor. UASB recovery performance after a few days of inflow arrest was studied. The optimal UASB operating conditions for practical application were determined. Treatment of sulfamethoxazole pharmaceutical wastewater is a big challenge. In this study, a series of anaerobic evaluation tests on pharmaceutical wastewater from different operating units was conducted to evaluate the feasibility of using anaerobic digestion, and the results indicated that the key refractory factor for anaerobic treatment of this wastewater was the high sulfate concentration. A laboratory-scale up-flow anaerobic sludge blanket (UASB) reactor was operated for 195 days to investigate the effects of the influent chemical oxygen demand (COD), organic loading rate (OLR), and COD/SO₄^2? ratio on the biodegradation of sulfamethoxazole in pharmaceutical wastewater and the process performance. The electron flow indicated that methanogenesis was still the dominant reaction although sulfidogenesis was enhanced with a stepwise decrease in the influent COD/SO₄^2? ratio. For the treated sulfamethoxazole pharmaceutical wastewater, a COD of 4983 mg/L (diluted by 50%), OLR of 2.5 kg COD/(m³·d), and COD/SO₄^2? ratio of more than 5 were suitable for practical applications. The recovery performance indicated that the system could resume operation quickly even if production was halted for a few days.     

Evaluating the impact of sulfamethoxazole on hydrogen production during dark anaerobic sludge fermentation

● SMX promotes hydrogen production from dark anaerobic sludge fermentation. ● SMX significantly enhances the hydrolysis and acidification processes. ● SMX suppresses the methanogenesis process in order to reduce hydrogen consumption. ● SMX enhances the relative abundance of hydrogen-VFAs producers. ● SMX brings possible environmental risks due to the enrichment of ARGs. The impact of antibiotics on the environmental protection and sludge treatment fields has been widely studied. The recovery of hydrogen from waste activated sludge (WAS) has become an issue of great interest. Nevertheless, few studies have focused on the impact of antibiotics present in WAS on hydrogen production during dark anaerobic fermentation. To explore the mechanisms, sulfamethoxazole (SMX) was chosen as a representative antibiotic to evaluate how SMX influenced hydrogen production during dark anaerobic fermentation of WAS. The results demonstrated SMX promoted hydrogen production. With increasing additions of SMX from 0 to 500 mg/kg TSS, the cumulative hydrogen production elevated from 8.07 ± 0.37 to 11.89 ± 0.19 mL/g VSS. A modified Gompertz model further verified that both the maximum potential of hydrogen production (P_m) and the maximum rate of hydrogen production (R_m) were promoted. SMX did not affected sludge solubilization, but promoted hydrolysis and acidification processes to produce more hydrogen. Moreover, the methanogenesis process was inhibited so that hydrogen consumption was reduced. Microbial community analysis further demonstrated that the introduction of SMX improved the abundance of hydrolysis bacteria and hydrogen-volatile fatty acids (VFAs) producers. SMX synergistically influenced hydrolysis, acidification and acetogenesis to facilitate the hydrogen production.