BPAC-UF对二级出水中抗生素抗性基因的去除及膜污染缓解机制

采用生物粉末活性炭(BPAC)-超滤(UF)组合工艺去除控制二级出水中抗生素抗性基因(ARGs),并对ARGs的去除和BPAC缓解膜污染机制进行了探讨。结果表明:与直接超滤工艺相比,组合工艺对水中四环素类抗性基因(tetA、tetW)、磺胺类抗性基因(sulⅠ、sulⅡ)以及溶解性有机碳(DOC)的去除效果均有较大的改善,这主要是由于BPAC对ARGs的吸附降解作用所致;水中16S rDNA、intⅠ1和DOC含量与不同种类ARGs浓度具有显著相关性,强化上述指标的去除可有效促进ARGs的削减;在BPAC投加量较低时,组合工艺的膜比通量较直接UF有所提高,膜污染状况明显改善;直接UF时,膜污染状况与滤饼层过滤模型的拟合度最好,而组合工艺的膜污染状况与标准膜孔堵塞模型和滤饼层过滤模型拟合度均较好。BPAC-UF组合工艺是一种较好的去除ARGs的工艺。     

喹诺酮抗性基因在城市污水处理系统中的分布及去除

城市污水处理厂是耐药细菌和抗生素抗性基因进入自然环境的主要途径之一。对污水处理厂不同工艺采集7组样品,重点考察污水处理系统中喹诺酮类耐药细菌和7种喹诺酮类抗性基因污染分布及其去除效率。结果显示,在不同处理工艺阶段均检测到耐环丙沙星细菌,最高浓度为(6.65±0.19)×106CFU/m L,耐药率为0.87%~7.24%。污水处理系统受抗生素抗性基因严重污染,入水样品中抗性基因总浓度为(1.28±0.12)×107copies/m L。污水处理工艺对抗环丙沙星细菌和喹诺酮类抗性基因具有有效的去除作用,除qnr C去除效率为75%,其余均在98%以上,但是仍有浓度为2.77×104CFU/m L的耐环丙沙星细菌和3.98×104copies/m L的抗性基因进入自然环境。表明污水处理系统已成为耐药细菌和抗生素抗性基因的储存库和污染源。     

混凝法去除城市生活污水中抗性基因

采用聚合氯化铝(PACl)和聚合硫酸铁(PFS)混凝处理城市生活污水(格栅出水和二级出水),考察混凝剂投加量、p H值对污水中抗生素抗性基因(ARGs)的去除率影响。结果表明,随着PACl和PFS投加量的增加,污水中目标基因的去除率先升高后降低;在p H 3~10的范围内,PACl对目标基因的去除效果随p H值的增加而降低,PFS对目标基因的去除随着p H值的增加先升高后降低。对于格栅出水,PACl和PFS的最佳投加量为400 mg/L,基因的去除率分别达到2.33~2.97 log和0.98~2.11 log,对于二级出水,PACl和PFS的最佳投加量分别为200 mg/L和150 mg/L,基因的去除率分别为1.85~2.64log、1.81~2.46 log;混凝去除最优p H条件分别为p H=3(投加PACl)及p H=5~6(投加PFS)。PACl的处理效果优于PFS。     

龙葵-镉抗性菌株修复镉污染土壤

为研究镉抗性菌株对龙葵修复镉污染土壤的效应,开展龙葵室外盆栽试验。从镉污染土壤、龙葵根际土壤和标准菌株中共筛选出3株抗镉产铁载体菌株,经16S rRNA基因序列对比分析,以上3株菌株分别被鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)、苍黄假棍状杆菌(Pseudoclavibacter helvolus)和荧光假单胞菌(Pseudomonas aeruginosa)。试验中,研究了此3株菌株单种及其复合接菌处理对龙葵生长和富集镉能力以及土壤修复效率的强化作用。结果表明,与空白对照相比,复合接种苍黄假棍状杆菌和荧光假单胞菌显著(P<0.05)提高龙葵地上部分干质量(提高31.07%)、地下部分干质量(提升75.59%)、龙葵镉总积累量(提高30.28%)、地上部分镉积累量(提高31.53%)、转运系数(提高39.47%)和土壤修复效率(提高67.83%)。此外,接种以上复合菌株后土壤镉去除量的86.16%随淋溶流失,13.84%由龙葵吸收。     

人工湿地中氨化细菌去除有机氮的效果

人工湿地去除有机氮主要由于氨化细菌的作用.为了了解人工湿地中氨化细菌去除有机氮的效果,对人工湿地基质中5株氮化细菌进行了初步鉴定,比较了不同氨化细菌去除有机氮的效果,氨化细菌去除有机氮的量通过其生成的NH4< >-N来衡量.结果表明,芽孢杆菌属(Bacillus)、假单胞菌属(Pseudomonas)为人工湿地中氧化细菌的优势菌属;氨化细蒲-1、氨化细菌-2及氨化细菌-5对有机氮的去除效果相对较好,分别达到46.2%、49.4%和52.6%.添加沸石对玄除氨氮有明显效果,从而能够提高有机氮的去除率.     

固定化反硝化菌联合固体碳源小球处理低碳氮比污水的性能研究

以谷壳为固体碳源,制备固定化反硝化菌联合固体碳源小球(简称固定化小球)投加至人工湿地基质层中,并对人工湿地在不同条件下处理低碳比污水的性能进行了研究。结果表明,在相同条件下,投加了固定化小球的人工湿地对COD、氨氮和TN的去除性能更好,其平均去除率分别达到了78.98%、85.46%和78.33%,并能有效解决人工湿地处理低碳氮比污水时反硝化阶段碳源不足的问题。     

种植不同植物的表面流人工湿地净化效果和微生物群落差异分析

为了解植物种类对表面流人工湿地的净化效果的影响及其与微生物群落的关系,研究了4种植物条件下表面流人工湿地的氮磷平衡以及微生物群落结构。结果表明,各组人工湿地对氨氮(45.53%～80.95%)、总氮(53.67%～80.30%)和总磷(32.97%～55.77%)都有较好的处理效果,种植植物的人工湿地比未种植的人工湿地具有更高的氨氮、总氮和总磷去除效果,其中黄菖蒲组对氮的去除效果最好,美人蕉组对磷的去除效果最好。在表面流人工湿地中,微生物作用(34.84%～45.44%)是人工湿地氮去除的主要途径,基质吸附(20.90%～23.91%)是人工湿地磷去除的主要途径,但是种植植物的人工湿地的氮磷通过微生物去除的量更高。高通量测序分析表明,相较于未种植植物的人工湿地,种植植物的人工湿地显示出更高的微生物丰富度、多样性和更高的脱氮除磷功能微生物的丰度。假单胞菌属、不动杆菌属、芽孢杆菌属和硝化螺菌属是人工湿地中主要的脱氮菌属,也是种植植物的人工湿地高生物脱氮的原因。假单胞菌属和不动杆菌属丰度增加是种植植物的人工湿地高生物除磷的原因。     

模块化人工湿地基质筛选与除污性能评价

为进一步解决农村污水处理问题,在利用单一基质传统人工湿地筛选出最佳COD、TP和TN去除基质的基础上,比较了最佳基质组成的组合基质模块化人工湿地和传统人工湿地对COD、TP和TN的去除效果。结果表明,无烟煤、铝污泥和沸石分别是去除COD、TP和TN的最佳基质。综合考虑COD、TP和TN的去除效果,确定运行时间为4个月。组合基质模块化人工湿地对COD、TP和TN的综合效果最佳,第4个月的平均去除率分别为82.26%、92.16%、76.50%,相应出水平均质量浓度分别为26.29、0.49、7.11mg/L,达到了《城镇污水处理厂污染物排放标准》(GB 18918—2002)的一级A标准,满足城镇景观用水和一般回用水的要求。     

回流立式组合人工湿地处理生活污水的效果

针对目前我国农村存在的生活污水随意排放问题,构建了一种安装方便、工艺简单的回流立式组合人工湿地系统用于处理生活污水,研究其对污染物质的去除效果和过程,及不同基质(砾石、陶粒、火山石)间污染物质去除的差异。结果显示,回流立式组合人工湿地系统对COD、NH+4-N、TN、TP的去除率分别高达95%、99%、53%、57%,其中COD、NH+4-N的去除过程符合一级动力学模型。不同基质组湿地对生活污水处理效果存在显著差异(P0.05),砾石组对NH+4-N、TN的去除效果较好,火山石组对COD、TP去除效果较好,可分别作为人工湿地脱氮除磷的优先选择。结果表明,回流立式组合人工湿地去除能力好、占地面积小、操作方便,是一种具有广阔发展前景的农村污水处理技术。     

复合人工湿地对城市受污染河水的净化效果

采用2种复合人工湿地系统处理昆明市城市河流污水,为期1年的运行结果表明:2种复合人工湿地系统对城市河流污水具有良好的污染物去除性能,TN、TP、COD和SS的月平均去除率分别达到50%、85%、85%和80%以上;2种复合人工湿地系统具有良好的耐冲击性,季节变化对出水水质不存在显著影响。其中,垂直上行流复合人工湿地系统性能更优于垂直上行流与水平流复合人工湿地系统;强降雨会对复合人工湿地出水水质造成一定的影响。     

Diversity of antibiotic resistance genes and encoding ribosomal protection proteins gene in livestock waste polluted environment

The rapid development and increase of antibiotic resistance are global phenomena resulting from the extensive use of antibiotics in human clinics and animal feeding operations. Antibiotics can promote the occurrence of antibiotic resistance genes (ARGs), which can be transferred horizontally to humans and animals through water and the food chain. In this study, the presence and abundance of ARGs in livestock waste was monitored by quantitative PCR. A diverse set of bacteria and tetracycline resistance genes encoding ribosomal protection proteins (RPPs) from three livestock farms and a river were analyzed through denaturing gradient gel electrophoresis (DGGE). The abundance of sul(I) was 10³ to 10⁵ orders of magnitude higher than that of sul(II). Among 11 tet-ARGs, the most abundant was tet(O). The results regarding bacterial diversity indicated that the presence of antibiotics might have an evident impact on bacterial diversity at every site, particularly at the investigated swine producer. The effect of livestock waste on the bacterial diversity of soil was stronger than that of water. Furthermore, a sequencing analysis showed that tet(M) exhibited two genotypes, while the other RPPs-encoding genes exhibited at least three genotypes. This study showed that various ARGs and RPPs-encoding genes are particularly widespread among livestock.     

Antibiotic resistance and genetic diversity of Escherichia coli isolates from traditional and integrated aquaculture in South China

This study investigated antibiotic resistance profiles including antibiotic resistance frequencies, resistance genes and resistance patterns in Escherichia coli strains isolated from traditional and integrated aquaculture systems in South China by using antibiotic susceptibility testing and real time polymerase chain reaction (PCR) technique. The E. coli isolates were found to be resistant to at least one antibiotic among 12 antibiotics. Higher resistance frequencies to ampicillin, sulfamethoxazole, trimethoprime, streptomycin and tetracycline were found compared to the rest antibiotics. Among the 10 tetracycline resistance genes detected in the resistant isolates, the most prevalent tetracycline resistance genes were tetA, tetW and tetB with the frequency of 69.7%, 63.5% and 21.9%, respectively. Three sulfonamide resistance genes were detected in these resistant isolates, with their detection frequencies in the following order: sul2 (55.3%) > sul3 (28.2%) > sul1 (6.2%). Four resistance genes mainly encoding extended-spectrum β-lactamases (ESBLs) were detected in these resistant isolates, with the detection frequencies of blaTEM (28.4%) > blaOXA (9.7%) > blaCTX (9.3%) > blaCARB (5.2%) > blaSHV (0.0%). It was found that the integrated aquaculture system exhibited generally higher prevalence of antibiotic resistance than the traditional aquaculture system. An integrated aquaculture system could facilitate development of bacterial resistance and spread of the antibiotic resistance genes, and consequently become an important reservoir of resistance genes.     

A comparison of antibiotics,antibiotic resistance genes,and bacterial community in broiler and layer manure following composting

Animal manure is an important source of antibiotics and antibiotic resistance genes (ARGs) in the environment. However, the difference of antibiotic residues and ARG profiles in layer and broiler manure as well as their compost remains unexplored. In this study, we investigated the profiles of twelve antibiotics, seventeen ARGs, and class 1 integrase gene (intI1) in layer and broiler manure, and the corresponding compost at large-scale. Compared with layer manure, broiler manure exhibited approximately six times more residual tetracyclines, especially chlortetracycline. The relative abundances of qnrS and ermA genes in broiler manure were significantly higher than those in layer manure. The concentration of tetracyclines not only had a significantly positive correlation with tetracycline resistance genes (tetA and tetC) but was also positively correlated with quinolone resistance (qepA, qnrB, and qnrS) and macrolide resistance (ermA and ermT). Most ARGs in manure were reduced after composting. However, the relative abundance of sulfonamide resistance gene sul1 increased up to 2.41% after composting, which was significantly higher than that of broiler (0.41%) and layer (0.62%) manure. The associated bacterial community was characterized by high-throughput 16S rRNA gene sequencing. The relative abundances of thermophilic bacteria had significant positive correlations with the abundance of sul1 in compost. The composting has a significant impact on the ARG-associated gut microbes in poultry manure. Variation partitioning analysis indicated that the change of bacterial community compositions and antibiotics contributed partially to the shift in ARG profiles. The results indicate that at industry-scale production broiler manure had more antibiotics and ARGs than layer manure did, and composting decreased most ARG abundances in poultry manure except for sulfonamide resistance genes.

     

The effect of wastewater treatment on antibiotic resistance in Escherichia coli and Enterococcus sp.

The effects of wastewater treatment on the proportion of Escherichia coli and Enterococcus sp. resistant to specific antibiotics were investigated at two facilities in Davis County, Utah, one of which received hospital waste. Samples were taken from the influent, effluent before disinfection, and secondary anaerobic sludge digester effluent. There was very little difference in antibiotic resistance among E. coli in the inflow waters of the plants but the plant receiving hospital waste had a significantly higher proportion of antibiotic resistant Enterococcus. The effect of wastewater treatment on antibiotic resistance was more pronounced on enterococci than E. coli. Although some increases in antibiotic resistance were observed, the general trend seemed to be a decrease in resistance, especially in the proportion of multidrug resistant Enterococcus sp.     

Dynamics of antibiotic resistance genes and microbial community in shortcut nitrification–denitrification process under antibiotic stresses

In this study, the performance of shortcut nitrification–denitrification (SCND) at different TC and SD stress conditions (0 μg/L, 1–97 days; 100 μg/L, 98–138 days; 500 μg/L, 139–175 days) was investigated. Higher level antibiotic stress (500 μg/L) led to the serious deterioration of nitrogen removal, and denitrification was more sensitive to antibiotic stress than nitrification. The dynamics of antibiotic resistance genes (ARGs) and microbial community were revealed by quantitative real-time PCR and 16S rDNA high-throughput sequencing, respectively. Tet-genes (tetA, tetQ, tetW), sul-genes (sulI, sulII), and mobile genetic element (intI1) in activated sludge increased by 1.2?~?2.5 logs with long-term exposure of antibiotic stress, and sulI, tetA, tetQ, and tetW were significantly positively correlated with intI1. Long-term antibiotics stress caused the decrease of most denitrifiers, and five genera were identified as the potential host of ARGs. The key impact factors of SCND drove the dynamics of ARGs and microbial community. Except for sulII gene, DO and FA were significantly positively correlated with ARGs, while FNA, NAR, and NO₂^?-N showed opposite effects to ARGs. Overall, maintaining relative lower DO, higher FNA, NAR, and NO₂^?-N conditions are not only benefit to the stable operation of SCND, but may also conducive to the control of ARG dissemination. This study provides theoretical basis on the control of ARGs in the SCND process.

     

High-throughput analysis of the effects of different fish culture methods on antibiotic resistance gene abundances in a lake

Freshwater lakes are important reservoirs for antibiotic resistance genes (ARGs). In this study, we determined the ARG profiles in water samples from Ying Lake, China, using high-throughput quantitative PCR. The high prevalence of ARGs suggested significant pollution with ARGs in the study area, where the ARG diversity and abundance were greater in an area with box-type fish farming than an area with fenced fish farming. Network analysis indicated the widespread co-occurrence of ARGs and mobile genetic elements. cphA-01, blalMP02, and blaCMY202 were identified as adequate indicator genes for estimating the total ARG abundances. Redundancy analysis indicated that changes in the microbial communities caused by variations in the physicochemical parameters with different fish culture methods mainly determined the ARGs in the lake system. Thus, analyzing the factors that affect ARGs provided novel insights into the mechanisms responsible for the maintenance and propagation of ARGs in a lake.

     

Abundance and removal of antibiotic resistance genes (ARGs) in the rearing environments of intensive shrimp aquaculture in South China

Although research regarding antibiotic resistance genes (ARGs) in aquaculture environments has gained increasing scientific interest, further studies are required to understand the abundances and removal mechanisms of ARGs during the entire rearing period of shrimp aquaculture. Thus, in this study, abundances, distributions and removal rates of ARGs in different environmental compartments of intensive shrimp farms in South China were investigated during the entire rearing period. The results indicated that sul1 and cmlA were the predominant ARGs in the water and sediment samples. Additionally, the total abundance of ARGs was higher in shrimp pond water than in the source water and farm effluent. Moreover, sediment samples indicated significantly higher ARG abundances than water samples from the shrimp ponds (P?<?0.05). Environmental factors were found to significantly affect the distribution of ARGs in shrimp rearing environments. Furthermore, stable ponds aided the removal of ARGs from shrimp pond water. This study accounted for temporal variations in ARG abundances as well as removal of ARGs in different environmental compartments during the entire shrimp rearing period. However, additional research is required to optimize the water treatment process for removal of ARGs from the aquaculture.     

Antibiotic resistant bacteria/genes dissemination in lacustrine sediments highly increased following cultural eutrophication of Lake Geneva (Switzerland)

This study investigates faecal indicator bacteria (FIB), multiple antibiotic resistant (MAR), and antibiotic resistance genes (ARGs), of sediment profiles from different parts of Lake Geneva (Switzerland) over the last decades. MARs consist to expose culturable Escherichia coli (EC) and Enterococcus (ENT) to mixed five antibiotics including Ampicillin, Tetracycline, Amoxicillin, Chloramphenicol and Erythromycin. Culture-independent is performed to assess the distribution of ARGs responsible for, β-lactams (blaTEM; Amoxicillin/Ampicillin), Streptomycin/Spectinomycin (aadA), Tetracycline (tet) Chloramphenicol (cmlA) and Vancomycin (van). Bacterial cultures reveal that in the sediments deposited following eutrophication of Lake Geneva in the 1970s, the percentage of MARs to five antibiotics varied from 0.12% to 4.6% and 0.016% to 11.6% of total culturable EC and ENT, respectively. In these organic-rich bacteria-contaminated sediments, the blaTEM resistant of FIB varied from 22% to 48% and 16% to 37% for EC and ENT respectively, whereas the positive PCR assays responsible for tested ARGs were observed for EC, ENT, and total DNA from all samples. The aadA resistance gene was amplified for all the sediment samples, including those not influenced by WWTP effluent water. Our results demonstrate that bacteria MARs and ARGs highly increased in the sediments contaminated with WWTP effluent following the cultural eutrophication of Lake Geneva. Hence, the human-induced changing limnological conditions highly enhanced the sediment microbial activity, and therein the spreading of antibiotic resistant bacteria and genes in this aquatic environment used to supply drinking water in a highly populated area. Furthermore, the presence of the antibiotic resistance gene aadA in all the studied samples points out a regional dissemination of this emerging contaminant in freshwater sediments since at least the late nineteenth century.     

Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China

This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta.     

Antibiotic resistance and virulence factors of Escherichia coli from eagles and goshawks

One of the major global problems in medicine is microbial resistance to antibiotics (antimicrobial resistance) and this has become an increasingly frequent research topic. This study focuses on antimicrobial resistance, phylogenetic and genetic characterization of Escherichia coli from wild birds: ten isolates from eagles (Aquila chrysaetos), nine from goshawks (Accipiter gentilis) and 24 from broilers in the Slovak Republic. Twenty-two strains with presence of int1 gene were selected and examined for the presence or absence of transposon gene (tn3), genes of antibiotic resistance and virulence factors. We detected sequence type (ST) in eagles ST 442 with genes iss, papC, iutA, cvaC, tsh, fyuA, iroN, kps, feoB, sitA, irp2, ireA for virulence factors and tetA, sul1, sul2, dfrA, aadA for antibiotic resistance; in goshawks ST 1011 with iss, papC, fyuA, iroN, feoB, sitA and qnrS1, tetA, sul1, sul2, dfrA, aadA, respectively. These ST types have been found in humans too and should be evaluated further for possible zoonotic potential and transfer of resistance genes from the environment.