农田生态系统中抗生素抗性基因迁移扩散的研究进展

农田生态系统是抗生素抗性基因(antibiotic resistance genes, ARGs)向人体传播的重要环境,其中农产品的食用是抗性基因暴露的主要途径之一。施肥等农业活动改变了农田中抗性基因及其宿主菌的组成,而复杂的微生物活动使抗性基因及其宿主菌进一步转移到农作物体内。近年来,PCR、宏基因组测序和外源基因标记等方法的进步不断拓宽了ARGs的研究思路。水平基因转移可促进ARGs快速向更广泛的宿主菌进行迁移,尤其是存在迁移到人类致病菌中的风险。为了深入探明ARGs在农田生态系统中的迁移途径和优势宿主菌,文中结合国内外研究进展,阐述了农田生态系统中ARGs的来源、分布传播,介绍了农田生态系统抗性菌和抗性基因的主要研究手段,总结了抗性菌和抗性基因在农田生态系统中的传播途径和扩散机制。基于当下研究的不足展望了继续深入探索的方向,为今后进一步探索ARGs在农田生态系统中的迁移机制提出了设想,以期降低潜在的食品安全和人体健康风险。     

城市社区农贸市场空气微生物及抗生素抗性基因研究

社区农贸市场活禽交易区是城市重要的人畜交叉感染区域,区内高存量的微生物和抗生素抗性基因(antibiotic resistance genes,ARGs)可通过粪便、冲洗水、空气等介质传播扩散。而空气介质中通过微生物气溶胶形式的传播途径,因其隐秘性、持久性的特点而对社区居民健康构成严重威胁。本文研究了深圳市某典型社区农贸市场内空气微生物及抗生素抗性基因。结果表明,活禽交易区可培养细菌浓度高达105CFU·m-3,远高于一般室内区域(103CFU·m-3),其中PM2.5精细颗粒物(0.65~3.3μm)中所含菌量占总菌量42%以上;活禽交易区空气介质中,抗生素抗性基因tet G、tet W、sul1和sul2检出率达70%以上,其绝对浓度在10~4~10~9copies·m-3之间;周边环境空气样品中,随着与活禽交易区距离的增加,空气微生物及抗生素抗性基因含量呈显著下降趋势。结果表明,农贸市场活禽交易区是微生物和抗生素抗性基因的一个重要储存库,活禽交易区空气会严重影响农贸市场及其外周边空气质量。     

抗生素抗性基因在土壤中积累、转移与消减的研究进展

细菌耐药性给人类健康及公共卫生带来巨大的威胁。土壤尤其是农业土壤是环境中抗生素抗性重要的源库。为减少抗生素抗性基因(ARGs)的传播风险,了解其在土壤中的传播规律非常重要。通过总结分析国内外发表的相关文献,对目前ARGs在土壤中的积累、转移情况及消减特征进行了综述。已有调查结果发现,农业发达及经济发地区土壤是ARGs积累的热区。有机肥施用及污水灌溉等原因导致ARGs在土壤中持续积累,其丰度可达10² gene copies/16S rRNA gene copies。胞内抗生素抗性基因(i ARGs)、胞外游离抗生素抗性基因(eARGs)是ARGs的两种赋存形态,其中,i ARGs是主要的赋存形态。i ARGs通过接合转移、转导在土壤中传播,其中接合转移是目前研究最多及最主要的水平转移方式。eARGs通过转化在土壤中传播。胞外DNA可以在土壤中留存几个月甚至一年以上,由于检测方法的限制eARGs在土壤中的自然转化并不经常被发现,因此,对土壤eARGs的风险研究有所忽略。外源ARGs进入土壤后的命运受到ARGs种类、形态、土壤特性、污染物等因素的影响。ARB进入土壤后...     

微塑料对农田土壤理化性质、土壤微生物群落结构与功能的影响

微塑料作为一种新污染物普遍存在于各类环境介质中,土壤环境中的微塑料污染已受到全球的广泛关注。该研究围绕农田土壤中微塑料污染这一主题,在总结分析国内外最新研究进展的基础上,综述了微塑料对农田土壤理化性质、土壤微生物生物量以及微生物群落结构与功能的影响。通过农业活动等途径进入农田土壤的微塑料会在非生物和生物作用下发生风化和降解,并对土壤理化性质、养分循环和污染物相互作用产生影响,进而影响微生物生物量、微生物群落结构与多样性、土壤酶活性,以及碳、氮循环和污染物降解等土壤生物地球化学过程,且微塑料对上述指标的影响与微塑料自身性质、土壤类型和暴露条件等多种因素有关。最后,对未来土壤微塑料的研究方向做了展望,以期为后续研究提供参考和思路。     

兽药抗生素对土壤微生物群落的影响

兽药抗生素广泛应用于畜禽养殖业,用于预防、治疗动物疾病及促进动物生长。然而,养殖过程中使用的抗生素不能被动物完全吸收,其中40%～90%以母体或其代谢物的形式排出动物体外并随畜禽粪便进入土壤环境,对土壤微生物群落结构和功能产生影响。在汇总了畜禽粪便和土壤中兽药抗生素的残留特征之后,概述了抗生素对土壤微生物群落结构和功能的影响以及微生物产生的污染诱导群落耐性。重点探讨了抗生素微生物毒性的影响因素和近年来对微生物共耐性方面的研究,并对未来的研究方向及目标提出了建议。     

污水处理厂空气介质抗生素抗性基因的分布

考察了污水处理厂空气介质中典型的抗性基因(antibiotic resistance genes,ARGs)污染水平和浓度分布,并通过16S r RNA高通量技术对样品进行亲缘性及溯源研究。结果表明,在污水厂空气样品中8种抗生素抗性基因的检出率均超过50%,其中tet C、sul1、sul2和erm B检出率为100%。在曝气池和污泥脱水车间空气样品中8种抗性基因检出率均为100%。对其中的sul1、sul2、tet G和tet X共4种ARGs的定量分析结果表明,以上4种基因的相对浓度范围在102~105copies·ng~(-1)DNA之间,与邻近居民区空气样品抗性基因浓度处于同一水平;空气样品16S r RNA高通量测序聚类分析结果显示,居民区空气与污水厂园区内空气有较高的种群相似度,污水厂处理单元对其邻近区域的空气介质微生物组成影响较大。     

我国土壤中抗生素抗性基因污染的消减策略

近年来,土壤环境的抗生素耐药性问题日益严重并逐渐成为威胁人类和动植物健康的全球性挑战。抗生素抗性基因(ARGs)是耐药性在土壤中传播的重要载体和指示物质,寻找合适的策略以降低其在土壤中的丰度及传播风险是当前和今后一个时期深入推进生命-生态一体化健康的主攻方向。ARGs进入土壤的途径主要包括粪肥施用和废水灌溉。针对不同来源,利用好氧堆肥和厌氧消化等阻控技术可从源头有效阻止ARGs进入土壤。当ARGs不可避免地进入土壤后,则仍需前瞻性研究和使用噬菌体、生物炭等ARGs污染土壤原位修复技术并深入开发植物间作等调控技术以构建免疫型土壤微生态环境。该文从阻控、修复和调控3个技术层面对ARGs去除效果和技术优劣势进行归纳,以期为土壤ARGs消减的技术开发和实际应用提供参考。     

生物炭添加对土壤中抗生素和抗性基因的环境行为影响研究进展

近年来,土壤抗生素和抗性基因污染已成为我国新兴的环境问题,生物炭作为土壤改良剂施用到土壤后会影响抗生素和抗性基因的环境行为.本文从我国土壤中抗生素和抗性基因污染现状和潜在风险出发,概述了生物炭添加土壤对抗生素的吸附、解吸及老化的影响,分析了生物炭特性、土壤类型、抗生素种类,和温度、pH值、共存物质等吸附条件对生物炭添加土壤吸附抗生素的影响,阐述了生物炭添加对土壤中抗生素和抗性基因迁移、消散、生物有效性,以及酶和微生物的影响,并对生物炭控制土壤中抗生素和抗性基因的研究前景进行了展望,拟为土壤中生物炭调控技术的发展提供参考.     

畜牧场土壤中重金属与抗生素抗性基因的分布规律研究

为研究畜牧场土壤中重金属和抗生素抗性基因(ARGs)的空间分布特点,揭示二者之间的相关性,为后续特色黑土的修复治理提供基础理论依据.结合GIS优化布点,在黑龙江省大庆市杜尔伯特蒙古族自治县某畜牧场园区分别采集内部10 cm和50 cm深度的土壤.采用原子吸收光谱仪和原子荧光分光光度计检测8种重金属含量,用单因子污染指数...     

抗生素及其抗性基因在环境中的污染、降解和去除研究进展

随着抗生素及其抗菌产品的广泛应用,自然和人工环境中的抗生素残留带来的危害引起人们关注.本文基于最新文献,综述了国内外抗生素及其抗性基因的污染水平和来源、它们之间的关系和传播机理以及这类污染物的降解和去除技术.现有研究表明,抗生素及其抗性基因的污染已遍布水、土壤、大气等介质,而在以污水处理厂和固废填埋场为代表的人工环境中,其污染水平更高.抗生素残留诱导产生抗性基因,其在环境中传播扩散与水平基因转移(Horizontal Gene Transfer,HGT)和微生物群落结构组成有关.抗生素和抗性基因在环境中自然降解过程受基质类型、光照、温度和微生物种群等因素的影响,其中光照是影响其降解的重要因子;而在人工处理系统中,紫外消毒和生化降解对抗生素及其抗性基因有较好的去除效果,但并非全部有效.建议今后加强对特定环境中抗生素和抗生素抗性基因的扩散规律和高效降解去除等方面的机理和工艺研究,进而有效控制其环境含量,降低其污染水平.     

Cu、抗生素协同污染对土壤微生物活性的影响

随着畜禽养殖业的规模化发展,重金属和抗生素在土壤环境中协同污染的几率不断升高。为分析和评价重金属、抗生素协同污染对土壤微生物生态系统的影响,以Cu、磺胺甲基嘧啶为添加毒物,其中,Cu的添加质量分数为0、100、500mg·kg-1;磺胺甲基嘧啶的添加质量分数为0、5、25、50、100mg·kg-1,采用室内培养试验的方法研究分析了cu、磺胺甲基嘧啶协同污染对土壤微生物微生物基础呼吸、微生物量碳、微生物量氮、硝化势、尿酶以及脱氢酶等土壤微生物指标的影响。结果表明,与磺胺甲基嘧啶单独污染处理相比,在Cu的质量分数为100mg·kg。协同污染污染下土壤微生物呼吸,土壤微生物量碳、氮以及土壤尿酶等指标的活性均明显增加;即表明它们对Cu与磺胺甲基嘧啶的协同污染表现出不同程度的交互抗性;在cu的协同污染质量分数为500mg·kg-1,Cu与磺胺甲基嘧啶对土壤各微生物指标则基本表现为协同抑制作用;在不同剂量Cu的协同污染处理下,当磺胺甲基嘧啶处理质量分数≥10mg·kg-1时,各土壤微生物指标的活性随着其处理质量分数的增加而显著降低,表现出很好的剂量依赖效应。因此,低剂量的Cu．磺胺甲基嘧啶协同污染可能会诱导土壤微生物对二者产生交互抗性;而高剂量协同污染则对土壤微生物生态功能产生较为严重的协同抑制作用。     

Tropical forest soil microbial communities couple iron and carbon biogeochemistry

We report that iron-reducing bacteria are primary mediators of anaerobic carbon oxidation in upland tropical soils spanning a rainfall gradient (3500-5000 mm/yr) in northeast Puerto Rico. The abundant rainfall and high net primary productivity of these tropical forests provide optimal soil habitat for iron-reducing and iron-oxidizing bacteria. Spatially and temporally dynamic redox conditions make iron-transforming microbial communities central to the belowground carbon cycle in these wet tropical forests. The exceedingly high abundance of iron-reducing bacteria (up to 1.2 x 10(9) cells per gram soil) indicated that they possess extensive metabolic capacity to catalyze the reduction of iron minerals. In soils from the higher rainfall sites, measured rates of ferric iron reduction could account for up to 44% of organic carbon oxidation. Iron reducers appeared to compete with methanogens when labile carbon availability was limited. We found large numbers of bacteria that oxidize reduced iron at sites with high rates of iron reduction and large numbers of iron reducers. The coexistence of large populations of iron-reducing and iron-oxidizing bacteria is evidence for rapid iron cycling between its reduced and oxidized states and suggests that mutualistic interactions among these bacteria ultimately fuel organic carbon oxidation and inhibit CH4 production in these upland tropical forests.     

Effects of cotton straw amendment on soil fertility and microbial communities

Maintaining soil fertility, while controlling pollution from excessive chemical fertilizer application is important for keeping soil productivity of sustainable agriculture. Variety of straws have been used and proven to be good soil amendments for increasing soil organic matter (OM) and a range of additional soil nutrients. However, little is known about the utilization of cotton straw for soil amendment. To better understand the mechanism behind cotton straw soil amendments, investigations were performed upon cucumber seedlings, where changes to soil nutrients and microbial communities were investigated. The results revealed that the cotton straw application promoted the cucumber seedling growth by significantly increasing the soil OM, available nitrogen, available phosphorus, and available potassium. The concentration of cotton straw was positively correlated to both the number of the culturable microorganisms and also the total microbial biomass within soil. Furthermore, assessment of cotton straw application using Biolog metabolic profiling and phospholipid fatty acid analysis revealed that such application increased the microbial community metabolic activity, and markedly changed the structure of microbial community. 16S rRNA gene clone library construction and phylogenetic analysis of soil bacteria revealed γ- Proteobacteria sequences dominated the cotton straw amendment soil, comprising 27.8% of the total number of analyzed sequences, while they were less represented in control soil (13.4%). On the contrary, the Sphingobacteria (7.8%) and Verrucomicrobia (2.4%) in the cotton straw amendment soil decreased after application when compared to the control soil 15.2% and 15.2%.     

Assessment of antibiotic resistance genes in dialysis water treatment processes

• Quantitative global ARGs profile in dialysis water was investigated. • Totally 35 ARGs were found in the dialysis treatment train. • 29 ARGs (highest) were found in carbon filtration effluent. • erm and mtrD-02 occurred in the final effluent. • The effluent was associated with health risks even after RO treatment. Dialysis water is directly related to the safety of hemodialysis patients, thus its quality is generally ensured by a stepwise water purification cascade. To study the effect of water treatment on the presence of antibiotic resistance genes (ARGs) in dialysis water, this study used propidium monoazide (PMA) in conjunction with high throughput quantitative PCR to analyze the diversity and abundance of ARGs found in viable bacteria from water having undergone various water treatment processes. The results indicated the presence of 35 ARGs in the effluents from the different water treatment steps. Twenty-nine ARGs were found in viable bacteria from the effluent following carbon filtration, the highest among all of the treatment processes, and at 6.96 Log (copies/L) the absolute abundance of the cphA gene was the highest. Two resistance genes, erm (36) and mtrD-02, which belong to the resistance categories macrolides-lincosamides-streptogramin B (MLSB) and other/efflux pump, respectively, were detected in the effluent following reverse osmosis treatment. Both of these genes have demonstrated the potential for horizontal gene transfer. These results indicated that the treated effluent from reverse osmosis, the final treatment step in dialysis-water production, was associated with potential health risks.     

Effect of the ultraviolet/chlorine process on microbial community structure,typical pathogens,and antibiotic resistance genes in reclaimed water

• UV/chlorine can effectively remove VBNC pathogens, ARGs and MGEs in reclaimed water. • Microbial community was changed with reduced diversity during UV/chlorine process. • CRBs-carried MGEswere the predominant groups during UV/chlorine process. • No direct co-selection strategy was shared between UV/chlorine and resistome. Urban wastewater contains a wide range of pathogens and antibiotic resistance genes (ARGs), which are a serious concern if reusing treated wastewater. However, few studies have explored the microbial communities in reclaimed water using ultraviolet (UV)/chlorine treatment and assessed the changes of the resistome. This study investigated the occurrence of typical pathogens, ARGs, and bacterial communities in UV/chlorine-treated reclaimed water samples. The numbers of culturable and viable but non-culturable pathogens were effectively reduced to 0 CFU/mL within 1–10 and 10–30 min after UV/chlorine treatment, respectively. Meanwhile, the physicochemical indices of water quality were not affected. UV/chlorine treatment could significantly change the bacterial community structure of reclaimed water, showing a decrease in bacterial abundance and diversity. Chlorine-resistant Acinetobacter and Mycobacterium were the dominant bacterial genera (>50%) after UV/chlorine treatment. Moreover, the number of ARGs and mobile genetic elements (MGEs) decreased with an increase in UV/chlorine exposure. However, eight ARGs and three MGEs were consistently detected in more than three seasons, making these major concerns because of their potential role in the persistence and dissemination of antibiotic resistance. Overall, the results of this study suggest that UV/chlorine treatment can potentially improve the microbiological safety of reclaimed water. And more attention should be paid to the pathogens that are both chlorine-resistant and carry MGEs because of their potential for resistance transmission.     

Mesophilic and thermophilic anaerobic digestion of swine manure with sulfamethoxazole and norfloxacin: Dynamics of microbial communities and evolution of resistance genes

• SMX addition had negative effect on acetoclastic methanogens in mesophilic AD. • Thermophilic AD was more effective in eliminating resistance genes than mesophilic. • ARGs variations in AD were mainly affected by succession of microbial community. • Methane production was significant associated to ARGs reduction. The role of norfloxacin (NOR) and sulfamethoxazole (SMX) in mesophilic and thermophilic anaerobic digestion (AD) of pig manure, with respect to methane production and variations in the microbial community and resistance genes, including antibiotic resistance genes (ARGs), class I integrase (intI1), and heavy metal resistance genes (MRGs), was investigated. The results indicated that NOR exerted little influence on the microbial community, whereas SMX negatively affected the acetoclastic methanogens. The abundance of two sulfonamide resistance genes (sul1 and sul2), three quinolone resistance genes (qnrS, parC, and aac(6’)-Ib-cr), and intI1 decreased by 2‒3 orders of magnitude at the end of thermophilic AD. In contrast, mesophilic AD was generally ineffective in reducing the abundance of resistance genes. According to the results of redundancy analysis, the abundance of ARGs was affected primarily by microbial community dynamics (68.5%), rather than the selective pressure due to antibiotic addition (13.3%). Horizontal gene transfer (HGT) through intI1 contributed to 26.4% of the ARG variation. The archaeal community also influenced the changes in the resistance genes, and ARG reduction was significantly correlated with enhanced methane production. Thermophilic AD presented a higher methane production potential and greater reduction in resistance gene abundance.     

黄浦江水域抗生素及抗性基因污染初步研究

采用高效液相色谱-质谱法,调查了黄浦江江水及底泥中6种常见兽用抗生素的含量特征,并采用实时荧光定量PCR方法,分别研究了该水域相应的8种代表性抗生素抗性基因(Antibiotics Resistance Genes,ARGs)的存在及丰度水平。结果显示,调查的四环素类、磺胺类及氯霉素抗生素在江水中的质量浓度范围分别在0.44～2.69、0.97～1.96、0.03～0.26μg·L-1之间,在底泥中的质量分数则分别为22.10～72.74、25.98～117.29、nd～50.47μg·kg-1。所有样品中除tet(O)、tet(B/P)未检出外,其他6种抗性基因均有检出。与四环素类抗性基因相比,磺胺类抗性基因的绝对拷贝数及相对丰度较高,为黄浦江水域中的优势抗性基因。相关性分析表明:江水中sul(Ⅲ)、tet(W),以及底泥中的sul(Ⅱ)丰度与对应相中磺胺类抗生素含量显著正相关,江水中sul(A)则与水相中氯霉素、四环素含量显著相关,其他几种抗生素与抗性基因间未见相关性存在。除抗生素外,黄浦江水域ARGs的相对丰度还可能与抗性基因种类及其环境因素(如光照、温度、pH和重金属等)有关。     

枯草芽孢杆菌Bs-15对板栗土壤微生物种群数量及功能多样性的影响

土壤微生物是维持土壤质量的重要组成部分,是土壤中生物活性的具体体现。土壤微生物多样性的变异可反映其对环境的响应与适应,能敏感反映生态系统的功能演变和生态环境的变化。本研究采用固体平板法研究了枯草芽孢杆菌（Bacillus subtilis）Bs-15对板栗（Castanea mollissimaBL）土壤微生物种群数量的影响,并通过BIOLOG ECO微孔板法分析Bs-15对板栗土壤功能多样性的影响。结果表明,接种Bs-15后,土壤中细菌数量比对照略有增加,但差异不显著;接种后放线菌的数量与对照相比有所减少,第7天达到极显著（p〈0.01）差异,之后差异变小,第14 d开始,处理与对照之间放线菌数量基本持平;真菌数量则先增加后减少,第7天开始接种后的真菌数量与对照相比达极显著（p〈0.01）差异。BIOLOG ECO微孔板分析显示,Bs-15使得土壤中微生物的AWCD值变大,72小时以后,AWCD值与对照相比差异达到极显著（p〈0.01）水平;接种Bs-15后增大了土壤微生物多样性指数,其中Shannon多样性指数、Simpson多样性指数和McIntosh多样性指数分别增加了4.09%、6.01%和7.20%,对对照相比差异均达到极显著水平（p〈0.01）,Simpson均匀度和McIntosh均匀度分别增加了2.07%和2.53%,与对照相比差异均达到显著水平（p〈0.05）。本研究结果表明,Bs-15不但提高了板栗土壤微生物的整体活性,丰富了土壤微生物种群,有利于保持和促进土壤肥力和健康状况;还提高了板栗土壤微生物功能多样性,使板栗土壤微生态系统功能更加稳定。     

长春市土壤重金属化学形态与土壤微生物群落结构的关系

采用样品采集、室内分析和多种数据统计方法相结合的方法,研究了长春市土壤重金属化学形态与土壤微生物群落结构的关系.结果表明:长春市土壤重金属(Pb、Cd、Cu、Zn和Ni)不同化学形态对土壤微生物群落结构影响明显不同,碳酸盐结合态的Pb和Ni对放线菌的生长繁殖具有刺激作用,碳酸盐结合态Zn对细菌的生长繁殖具有刺激作用,铁锰氧化物结合态和有机结合态的Zn与有机结合态Cu对细菌、放线菌的生长繁殖具有刺激作用.土壤理化性质对重金属化学形态与微生物群落结构之间的关系影响很大,其掩盖了土壤重金属化学形态对土壤微生物群落结构特征的影响,有机质含量、电导率、阳离子交换量掩盖了重金属化学形态对细菌、放线菌和微生物总数的影响.自然含水率掩盖了重金属化学形态对多样性指数的影响.采用重金属的化学形态评价重金属对土壤微生物参数的影响更合理,而且在确定评价长春市土壤重金属污染状况指标时,应该剔除土壤理化性质的影响.选择有机结合态Cu、铁锰氧化物结合态Zn与微生物群落多样性指数相结合来评价长春城市土壤重金属Cu和Zn的污染状况.     

Removal of trimethoprim and sulfamethoxazole in artificial composite soil treatment systems and diversity of microbial communities

• Novel ACST allowed biodegradation to effectively remove adsorbed SMX and TMP. • Ammonia and nitrite were efficiently removed in ACSTs and water quality was improved. Four artificial composite soil treatment systems (ACSTs) fed with reclaimed water containing trimethoprim (TMP) and sulfamethoxazole (SMX) were constructed to investigate SMX and TMP biodegradation efficiency, ammonia and nitrite removal conditions and the microbial community within ACST layers. Results showed SMX and TMP removal rates could reach 80% and 95%, respectively, and removal rates of ammonia and nitrite could reach 80% and 90%, respectively, in ACSTs. The MiSeq sequencing results showed that microbial community structures of the ACSTs were similar. The dominant microbial community in the adsorption and biodegradation layers of the ACSTs contained Proteobacteria, Chloroflexi, Acidobacteria, Firmicutes, Actinobacteria and Nitrospirae. Firmicutes and Proteobacteria were considerably dominant in the ACST biodegradation layers. The entire experimental results indicated that Nitrosomonadaceae_uncultured, Nitrospira and Bacillus were associated with nitrification processes, while Bacillus and Lactococcus were associated with SMX and TMP removal processes. The findings suggest that ACSTs are appropriate for engineering applications.