Qianwen Sui

Distribution of antibiotic resistance genes (ARGs) in anaerobic digestion and land application of swine wastewater

Swine farm and the adjacent farmland are hot spots of ARGs. However, few studies have investigated the on-site occurrence of ARGs distributed in the process of anaerobic digestion (AD) followed by land application of swine wastewater. Two typical swine farms, in southern and northern China respectively, with AD along with land application were explored on ARG distributions. ARGs were highly abundant in raw swine wastewater, AD effectively reduced the copy number of all detected ARGs (0.21-1.34 logs removal), but the relative abundance with different resistance mechanisms showed distinctive variation trends. The reduction efficiency of ARGs was improved by stable operational temperature and longer solid retention time (SRT) of AD. ARGs in soil characterized the contamination from the irrigation of the digested liquor. The total ARGs quantity in soil fell down by 1.66 logs in idle period of winter compared to application period of summer in the northern region, whereas the total amount was steady with whole-year application in south. Some persistent (sul1 and sul2) and elevated ARGs (tetG and ereA) in AD and land application need more attention.

Effect of red mud addition on tetracycline and copper resistance genes and microbial community during the full scale swine manure composting.

Swine manure has been considered as the reservoir of antibiotic resistance genes (ARGs). Composting is one of the most suitable technologies for treating livestock manures, and red mud was proved to have a positive effect on nitrogen conservation during composting. This study investigated the abundance of eight tetracycline and three copper resistance genes, the bacterial community during the full scale swine manure composting with or without addition of red mud. The results showed that ARGs in swine manure could be effectively removed through composting (reduced by 2.4log copies/g TS), especially during the thermophilic phase (reduced by 1.5log copies/g TS), which the main contributor might be temperature. Additionally, evolution of bacterial community could also have a great influence on ARGs. Although addition of red mud could enhance nitrogen conservation, it obviously hindered removal of ARGs (reduced by 1.7log copies/g TS) and affected shaping of bacterial community during composting.

Fate of antibiotic resistance genes and its drivers during anaerobic co-digestion of food waste and sewage sludge based on microwave pretreatment

In this study, anaerobic digestion of mono-SS, MW-SS:FW and SS:MW-FW was investigated to understand the fate of ARGs and its drivers. Anaerobic digestion was effective for the reduction of metal resistance genes (MRGs), and could reduce the abundance of blaOXA-1, sulI and tetG, while sulII in co-digestion and blaTEM and ereA only in MW-SS. ARGs reduction could be partly attributed to the reduction of co-selective pressure from heavy metals reflected by MRGs. However, the abundance of mefA/E, ermB, ermF, tetM and tetX increased significantly. Anaerobic co-digestion, especially for MW-SS, could reduce total ARGs abundance compared with mono-SS, and evolution of bacterial community was the main driver for the fate of ARGs.

Sludge bio-drying: Effective to reduce both antibiotic resistance genes and mobile genetic elements

Sewage sludge is considered as one of major contributors to the increased environmental burden of ARGs. Sludge bio-drying was increasingly adopted due to its faster sludge reduction compared with composting. The fate of ARGs during full-scale sludge bio-drying was investigated to determine whether it could effectively reduce ARGs, and the contributions of bacterial community, horizontal gene transfer (HGT) through mobile genetic elements (MGEs) and co-selection from heavy metals to ARGs profiles were discussed in detail. Two piles with different aeration strategies (Pile I, the improved and Pile II, the control) were operated to elucidate effects of aeration strategy on ARGs profiles. Results showed that sludge bio-drying could effectively reduce both most of targeted ARGs (0.4-3.1 logs) and MGEs (0.8-3.3 logs) by the improved aeration strategy, which also enhanced both the sludge bio-drying performance and ARGs reduction. The enrichment of ARGs including ermF, tetX and sulII could be well explained by the evolution of bioavailable heavy metals, not HGT through MGEs, and their potential host bacteria mainly existed in Bacteroidetes. Although changes of bacterial community contributed the most to ARGs profiles, HGT through MGEs should be paid more attention especially in the thermophilic stage of sludge bio-drying.

Effects of chlortetracycline and copper on tetracyclines and copper resistance genes and microbial community during swine manure anaerobic digestion

As antibiotic and heavy metals are over used in the livestock industry, animal manure is a reservoir of antibiotic resistance genes (ARGs). Anaerobic digestion has been reported to have the potential to reduce ARGs. However, few studies investigated whether reduction of ARGs would be affected by different external pressures including antibiotics and heavy metals during anaerobic digestion. The purpose of this study was thus to investigate effects of both chlortetracycline (CTC) and Cu on reduction of ARGs, heavy metal resistance genes (HMRGs) and mobile genetic elements (MGEs) during the swine manure anaerobic digestion. The results showed that the predominant ARGs (tetO, tetW, tetX, tetL) could be effectively reduced (approximately 1.00 log copies/g TS) through mesophilic anaerobic digestion. Microbial community evolution was the main driver. It was interesting that Treponema might indicate the termination of anaerobic digestion and compete with ARGs host bacteria. Addition of CTC, Cu and CTC+Cu affected microbial community change and hindered removal of ARGs, especially, CTC+Cu seriously affected Treponema and ARGs during anaerobic digestion.

Influence of natural zeolite and nitrification inhibitor on organics degradation and nitrogen transformation during sludge composting

Sludge composting is one of the most widely used treatments for sewage sludge resource utilization. Natural zeolite and nitrification inhibitor (NI) are widely used during composting and land application for nitrogen conservation, respectively. Three composting reactors (A—the control, B—natural zeolite addition, and C—3,4-dimethylpyrazole phosphate (DMPP) addition) were established to investigate the influence of NI and natural zeolite addition on organics degradation and nitrogen transformation during sludge composting conducted at the lab scale. The results showed that, in comparison with the control, natural zeolite addition accelerated organics degradation and the maturity of sludge compost was higher, while the DMPP addition slowed down the degradation of organic matters. Meanwhile, the nitrogen transformation functional genes including those responses for nitrification (amoA and nxrA) and denitrification (narG, nirS, nirK, and nosZ) were quantified through quantitative PCR (qPCR) to investigate the effects of natural zeolites and DMPP addition on nitrogen transformation. Although no significant difference in the abundance of nitrogen transformation functional genes was observed between treatments, addition of both natural zeolite and DMPP increases the final total nitrogen content by 48.6 % and 23.1 %, respectively. The ability of natural zeolite for nitrogen conservation was due to the absorption of NH3 by compost, and nitrogen conservation by DMPP was achieved by the source reduction of denitrification. Besides, it was assumed that the addition of natural zeolite and DMPP may affect the activity of these genes instead of the abundance.

Seasonal variation and removal efficiency of antibiotic resistance genes during wastewater treatment of swine farms.

The seasonal variation and removal efficiency of antibiotic resistance genes (ARGs), including tetracycline resistance genes (tetG, tetM, and tetX) and macrolide (ermB, ermF, ereA, and mefA), were investigated in two typical swine wastewater treatment systems in both winter and summer. ARGs, class 1 integron gene, and 16S rRNA gene were quantified using real-time polymerase chain reaction assays. There was a 0.31–3.52 log variation in ARGs in raw swine wastewater, and the abundance of ARGs in winter was higher than in summer. tetM, tetX, ermB, ermF, and mefA were highly abundant. The abundance of ARGs was effectively reduced by most individual treatment process and the removal efficiencies of ARGs were higher in winter than in summer. However, when examining relative abundance, the fate of ARGs was quite variable. Anaerobic digestion reduced the relative abundance of tetX, ermB, ermF, and mefA, while lagoon treatment decreased tetM, ermB, ermF, and mefA. Sequencing batch reactor (SBR) decreased tetM, ermB, and ermF, but biofilters and wetlands did not display consistent removal efficiency on ARGs in two sampling seasons. As far as the entire treatment system is concerned, ermB and mefA were effectively reduced in both winter and summer in both total and relative abundance. The relative abundances of tetG and ereA were significantly correlated with intI1 (p < 0.01), and both tetG and ereA increased after wastewater treatment. This may pose a great threat to public health.

Performance of a sequencing-batch membrane bioreactor (SMBR) with an automatic control strategy treating high-strength swine wastewater

Due to high-strength of organic matters, nutrients and pathogen, swine wastewater is a major source of pollution to rural environment and surface water. A sequencing-batch membrane bioreactor (SMBR) system with an automatic control strategy was developed for high-strength swine wastewater treatment. Short-cut nitrification and denitrification (SND) was achieved at nitrite accumulation rate of 83.6%, with removal rates of COD, NH4+-N and TN at 95%, 99% and 93%, respectively, at reduced HRT of 6.0 d and TN loading rate of 0.02kgN/(kgVSS d). With effective membrane separation, the reduction of total bacteria (TB) and putative pathogen were 2.77 logs and 1%, respectively. The shift of microbial community was well responded to controlling parameters. During the SND process, ammonia oxidizing bacteria (AOB) (Nitrosomonas, Nitrosospira) and nitrite oxidizing bacteria (NOB) (Nitrospira) were enriched by 52 times and reduced by 2 times, respectively. The denitrifiers (Thauera) were well enriched and the diversity was enhanced.

Who contributes more to N2O emission during sludge bio-drying with two different aeration strategies, nitrifiers or denitrifiers?

Global warming effects have drawn more and more attention to studying all sources and sinks of nitrous oxide (N2O). Sludge bio-drying, as an effective sludge treatment technology, is being adopted worldwide. In this study, two aeration strategies (piles I and II) were compared to investigate the primary contributors to N2O emission during sludge bio-drying through studying the evolution of functional genes involved in nitrification (amoA, hao, and nxrA) and denitrification (narG, nirS, nirK, norB, and nosZ) by quantitative PCR (qPCR). Results showed that the profile of N2O emission can be divided into three stages, traditional denitrification contributed largely to N2O emission at stage I (days 1–5), but N2O emission mainly happened at stage II (days 5–14) due to nitrifier denitrification and NH2OH accumulation by ammonia-oxidizing bacteria (AOB), accounting for 51.4% and 58.2% of total N2O emission for piles I and II, respectively. At stage III (days 14–21), nitrifier denitrification was inhibited because sludge bio-drying proceeded mainly by the physical aeration, thus N2O emission decreased and changed little. The improved aeration strategy availed pile I to reduce N2O emission much especially at stages II and III, respectively. These results indicated that nitrifier denitrification by AOB and biological NH2OH oxidation due to AOB made more contribution to N2O emission, and aeration strategy was crucial to mitigate N2O emission during sludge bio-drying.

Profiles and drivers of antibiotic resistance genes distribution in one-stage and two-stage sludge anaerobic digestion based on microwave-H2O2 pretreatment.

Three anaerobic digestion (AD) processes of waste activated sludge (WAS) were established including the control (mono-WAS), one-stage AD and two-stage AD along with microwave-H2O2 pre-treatment (MW-H2O2) to investigate the profiles and drivers of antibiotic resistance genes (ARGs) distribution concerning co-selection from heavy metals, intI1 and microbial community through qPCR and high-throughput sequencing method. Results showed that MW-H2O2 could reduce the absolute gene copies of all ARGs while increased the relative abundance of most ARGs. After subsequent AD, both total ARGs quantities and relative abundance were enriched while two-stage AD showed some advantages over ARGs abundance reduction. Besides, AD was more effective on the potential pathogens reduction than MW-H2O2. AD could reduce the role of intI1 on the spread of ARGs, while mantel test and procrustes analysis indicated that the variation of ARGs abundance was closely associated with the discrepancy of bacterial community.