Xun Qian

Variable effects of oxytetracycline on antibiotic resistance gene abundance and the bacterial community during aerobic composting of cow manure

Livestock manure is often subjected to aerobic composting but little is known about the variation in antibiotic resistance genes (ARGs) during the composting process under different concentrations of antibiotics. This study compared the effects of three concentrations of oxytetracycline (OTC; 10, 60, and 200mg/kg) on ARGs and the succession of the bacterial community during composting. Very similar trends were observed in the relative abundances (RAs) of each ARG among the OTC treatments and the control during composting. After composting, the RAs of tetC, tetX, sul1, sul2, and intI1 increased 2-43 times, whereas those of tetQ, tetM, and tetW declined by 44-99%. OTC addition significantly increased the absolute abundances and RAs of tetC and intI1, while 200mg/kg OTC also enhanced those of tetM, tetQ, and drfA7. The bacterial community could be grouped according to the composting time under different treatments. The highest concentration of OTC had a more persistent effect on the bacterial community. In the present study, the succession of the bacterial community appeared to have a greater influence on the variation of ARGs during composting than the presence of antibiotics. Aerobic composting was not effective in reducing most of the ARGs, and thus the compost product should be considered as an important reservoir for ARGs.

Reducing antibiotic resistance genes, integrons, and pathogens in dairy manure by continuous thermophilic composting

This study explored the effects of composting using three temperature regimes, namely, insufficient thermophilic composting (ITC), normal thermophilic composting (NTC), and continuous thermophilic composting (CTC), on antibiotic resistance genes (ARGs), integrons, and human pathogenic bacteria (HPB), as well as the mechanisms involved. The NTC and CTC treatments led to greater decreases in 5/10 ARGs and two integrons than ITC, and the abundances of ARGs (tetC, tetG, and tetQ) and int1 only declined in the NTC and CTC treatments. The abundances of HPB decreased by 82.8%, 76.9%, and 96.9% under ITC, NTC, CTC, respectively. Redundancy analysis showed that both bacterial succession and horizontal gene transfer play important roles in the variation of ARGs, and the changes in different ARGs were due to diverse mechanisms. CTC performed significantly better at reducing ARGs, integrons, and HPB, thus it may be used to manage the public health risks of ARGs in animal manure.

Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure

Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

Effects of bamboo charcoal on antibiotic resistance genes during chicken manure composting

Composting is widely used for animal waste disposal, and bamboo charcoal (BC) can be used for nitrogen conservation during composting. However, the effects of BC on antibiotic resistance genes (ARGs) during chicken manure composting are still unclear. This study investigated the effects on ARGs of adding different proportions of BC (0%, 5%, 10%, and 20% w/w) to chicken manure compost. After 26 days, the relative abundances (RAs) of most ARGs (tetC, tetG, tetW, tetX, sul2, drfA1, drfA7, ermB, ermF, ermQ, and ermX) and intI1 declined by 21.6-99.5%, whereas sul1 increased by 7.5-17.7 times. The average RAs reductions with 0%, 5%, 10%, and 20% BC were 0.85, 1.05, 1.08, and 1.15 logs, respectively. The most important environmental factor for the ARG profiles was temperature according to redundancy analysis. Furthermore, BC significantly decreased the bio-Cu and bio-Zn levels, thereby reducing the co-selection pressure from heavy metals. Different proportions of BC had no significant effects on the removal of tetG, tetW, tetX, sul2, drfA1, and ermB. Supplementation with 10% BC was more effective at removing tetC and drfA7 compared with the other treatments. The results suggested that 10% BC supplementation is appropriate for reducing ARGs in chicken manure compost.

Effects of adding different surfactants on antibiotic resistance genes and intI1 during chicken manure composting.

Aerobic composting is usually employed to treat livestock manure. In this study, a bio-surfactant (rhamnolipid, RL) and chemical surfactant (Tween 80, Tw) were added to chicken manure during composting and their effects were determined on the variations in ARGs and intI1. After composting, the reductions in the RAs of ARGs and intI1 with the addition of Tw (1-4.7logs) were generally greater than that with the addition of RL (0.8-3.7logs) and the control (CK) (0.3-2.6logs), and the enrichment of ARGs was higher with CK (0.9-1.8logs). The ARG profiles were affected significantly by temperature and the water-soluble carbon contents. RL and Tw effectively reduced the concentrations of bio-available Cu and Zn, thereby hindering the co-selection of ARGs by heavy metals. The effects of RL and Tw on ARGs and intI1 indicate that the addition of Tw was slightly more effective than RL after composting.

Mechanisms and effects of arsanilic acid on antibiotic resistance genes and microbial communities during pig manure digestion

High concentrations of residual arsanilic acid occur in pig manure due to its use in feed to promote growth and control diseases. This study compared the effects of arsanilic acid at three concentrations (0, 325, and 650mg/kg dry pig manure) on the abundance of antibiotic resistance genes (ARGs) and the microbial community during anaerobic digestion. Addition of 650mg/kg arsanilic acid enhanced the absolute abundances of tetC, sul2, ermB, and gyrA more than twofold in the digestion product. Redundancy analysis indicated that the change in the microbial community structure was the main driver of variation in the ARGs profile. The As resistance gene arsC co-occurred with four ARGs and intI1, possibly causing the increase in ARGs under pressure by arsanilic acid. High arsanilic acid concentrations can increase the risk of ARGs occurring in anaerobic digestion products. The amount of arsanilic acid used as a feed additive should be controlled.

Relationships between sulfachloropyridazine sodium, zinc, and sulfonamide resistance genes during the anaerobic digestion of swine manure.

In this study, swine manure containing sulfachloropyridazine sodium (SCPS) and zinc was subjected to mesophilic (37°C) anaerobic digestion (AD). The absolute abundances (AAs) of antibiotic resistance genes (ARGs) were evaluated, as well as intI1 and intI2, and the degradation of SCPS according to variation in the amount of bio-available zinc (bio-Zn). In digester that only contained SCPS, the concentrations of SCPS were lower than that digesters both contain SCPS and Zn. Compared with the control digester, the addition of SCPS increased the AAs of sul1, sul3, drfA1, and drfA7 by 1.3-13.1 times. However, compared with the digester with SCPS but no added Zn, the AAs of sul3, drfA1, and drfA7 were decreased by 21.4-70.3% in the presence of SCPS and Zn, whereas sul1 and sul2 increased 1.3-10.7 times. There were significant positive correlations (P<0.05) between the concentrations of SCPS with several ARGs and bio-Zn.

Impacts of biochar on the environmental risk of antibiotic resistance genes and mobile genetic elements during anaerobic digestion of cattle farm wastewater

Biochar has positive effects on nitrogen conservation during anaerobic digestion, but its impacts on antibiotic resistance genes (ARGs) are unclear. Therefore, the effect of biochar (0, 5, 20, and 50 g/L) on the environmental risk of ARGs during cattle manure wastewater anaerobic digestion were investigated. The results showed that 5 g/L biochar reduced the relative abundances (RAs) of 5/13 ARGs while 20 g/L biochar significantly reduced the total RAs of ARGs in the digestion products, where the RA of ISCR1 was 0.89 log lower than the control. Biochar mainly affected the distribution of ARGs by influencing the RAs of Firmicutes and Proteobacteria, and the influence of 20 g/L biochar was greater than that of 5 g/L. Mobile genetic elements also influenced the ARG profiles, especially intI2 and ISCR1. The addition of 20 g/L biochar to cattle farm wastewater anaerobic digestion systems could reduce the environmental risk of ARGs.

Effects of oxytetracycline on archaeal community, and tetracycline resistance genes in anaerobic co-digestion of pig manure and wheat straw

ABSTRACT In this study, the effects of different concentrations of oxytetracycline (OTC) on biogas production, archaeal community structure, and the levels of tetracycline resistance genes (TRGs) were investigated in the anaerobic co-digestion products of pig manure and wheat straw. PCR denaturing gradient gel electrophoresis analysis and real-time quantitative polymerase chain reaction (RT-qPCR) (PCR) were used to detect the archaeal community structure and the levels of four TRGs: tet(M), tet(Q), tet(W), and tet(C). The results showed that anaerobic co-digestion with OTC at concentrations of 60, 100, and 140 mg/kg (dry weight of pig manure) reduced the cumulative biogas production levels by 9.9%, 10.4%, and 14.1%, respectively, compared with that produced by the control, which lacked the antibiotic. The addition of OTC substantially modified the structure of the archaeal community. Two orders were identified by phylogenetic analysis, that is, Pseudomonadales and Methanomicrobiales, and the methanogen present during anaerobic co-digestion with OTC may have been resistant to OTC. The abundances of tet(Q) and tet(W) genes increased as the OTC concentration increased, whereas the abundances of tet(M) and tet(C) genes decreased as the OTC concentration increased.

Effects of sulphamethazine and zinc on the functional diversity of microbial communities during composting

ABSTRACT The changes in the functional diversity of the microbial community in a compost matrix with a single or compound addition of zinc (Zn; 0, 600, and 1800 mg/kg) and sulphamethazine (SM2; 0, 1, and 25 mg/kg) were studied with the Biolog method during composting. The microbial community was extracted from the compost matrix comprising swine manure and wheat straw at day 6 (themophilic period) and day 25 (mature period) of composting. Results proved that the Shannon index, average well-colour development, and substrate utilization significantly decreased as the concentrations of SM2 and Zn increased on day 6. The negative effect of the combined addition of SM2 and Zn was lower than that of the individual addition of SM2 and Zn. On day 25, the inhibition effect disappeared, and microbial metabolic activities were higher than those on day 6. The effects of SM2 and Zn could be further differentiated via the principal component analysis (PCA) and cluster analysis. On day 6, the treatments were divided into three groups by PC1 and PC2. The separation of the different treatments in the PCA plots became increasingly apparent on day 25. In conclusion, the effects of SM2 and Zn on the microbial community during composting became evident in the themophilic period and that the microbial activity recovered in the mature period. The combination of SM2 and Zn decreased the inhibition with the addition of individual additive. GRAPHICAL ABSTRACT