抗生素最小抑制浓度法(MICs)评估环境大肠杆菌抗生素抗性

2015年世界卫生组织将抗生素的滥用列为21世纪最大的挑战之一,全球范围内抗生素抗药性的散播已严重威胁人类的健康。如何检测环境中的细菌抗药性,并有效评估抗药性感染的风险,是环境微生物研究的一项重要课题。本文通过改进的培养基微量稀释(broth micro-dilution)法,确定了2个地区(中国成都和美国夏威夷)不同环境来源(天然水系和市政污水)的大肠杆菌的抗生素最小抑制浓度(minimum inhibitory concentrations,MICs)。计算所得的MIC分位数(MIC50和MIC90)、菌体抗性百分比及多抗药性指数(multiple antibiotic resistance indexes,MARIs)显示两地不同环境区划的抗生素抗性存在明显的差异。天然水系(成都锦江)中的抗生素抗性是随时间可变的,与当地的降雨事件相关。环境菌株的抗药性模式通过聚类和非度量多维测度(non-metric multidimensional scaling,NMDS)进行分析。广谱β-内酰胺酶基因筛查显示出抗性基因与抗性表型之间的正相关性。结合现有的两地抗生素的使用数据讨论了两地环境抗生素抗性与当地人类活动及抗生素的使用实践之间的紧密联系。采集环境菌株抗生素MIC数据的实验及数据分析方法实现了环境抗药性的跨时空对比,为规范抗生素的区域性使用提供了指导作用。     

Earthworms highly increase ciprofloxacin mineralization in soils

This report shows that earthworms increase up to eight times the mineralization of the antibiotic ciprofloxacin in soils. Antibiotics are extensively used and disseminated in environmental compartments. Antibiotics may enter food chains and thus induce resistance in environmental and human commensal bacteria. The antibiotic ciprofloxacin is suspected to induce significant adverse effects on soil microbial processes, with possible consequences on soil functions. Nevertheless, little is known concerning the fate of ciprofloxacin in soils. Here, we studied the mineralization and distribution of the [2-¹⁴C]-ciprofloxacin in soil–plant–water systems where ciprofloxacin was applied by amendment of spiked pig slurry. Results show that a very weak microbial mineralization of the antibiotic, lower than 0.01 %, occurred after 84 days of incubation. By contrast, the addition of earthworms increased from 5 to 8 times ciprofloxacin mineralization during the following 84 days incubation. In addition, earthworm activity induced the transfer of 40 % of radioactive compounds from the upper to the lower layer of soil, modifying the distribution of the antibiotic within the soil profile. We conclude that earthworms can be used efficiently to mineralize ciprofloxacin and modify its distribution in soils. As a consequence, earthworms change the exposure of soil organisms to ciprofloxacin, and, in turn, the eco-toxicological impact of the antibiotic.     

环境中抗药基因水平转移研究进展

抗药基因是一种新型环境污染物,一旦被致病菌获得,将导致抗生素的临床使用失效,从而危害公共健康。抗药基因可通过转移因子在细菌间传播,加剧抗药基因的扩散效应,提升了环境健康风险性,因此抗药基因水平转移受到了越来越多的关注。本文介绍了抗药基因水平转移的分子机制,总结了不同环境中各种抗药基因水平转移途径的研究现状,并对该领域将来的研究方向提出了展望,以期为环境中抗药基因的调查评估、污染控制及合理管理提供理论参考。     

土壤环境中四环素类抗生素残留及抗性基因污染的研究进展(Tetracycline Residues and Tetracycline Resistance Gene Pollution in Soil: A Review)

近年来,致病菌耐药性的增加和扩散已成为全世界关注的热点问题,而人类医疗和畜禽养殖业抗生素的滥用正不断加剧这一问题.众多研究表明土壤环境作为一个巨大的抗性基因储存库,在抗性微生物和抗性基因的传播中起重要的作用,具有潜在的生态与健康风险.在总结国内外最新研究基础上,对土壤环境中典型抗生素-四环素类抗生素的主要污染源以及其在土壤中的基本环境行为等进行了分析,并探讨了土壤中四环素类抗性基因的来源、迁移和扩散及分子检测手段等问题.我国作为抗生素的生产和消费大国,抗生素污染问题较其他国家更为严重,而国内相关研究才刚刚起步,迫切需要开展有关环境中抗生素和抗生素抗性基因污染的系统研究.     

抗生素类兽药对植物和土壤微生物的生态毒理学效应研究进展

规模化养殖快速发展,常使用兽药来防治各种禽畜病害,导致大量兽药随动物粪便排出体外.含有残留兽药的粪便作为有机肥施入农田而造成土壤污染,对人类健康和生态系统产生潜在危害.养殖业使用的主要兽药种类为抗生素类药物,且用量逐年增加,目前土壤中兽药残留浓度范围为μg·kg-1级到g·kg-1级.在总结国内外及本课题组相关研究的基础上,论文较为系统地概述了兽药对植物生长和土壤微生物群落功能和结构的影响,探讨了今后兽药生态毒理学研究的主要方向.抗生素类兽药对植物和土壤微生物群落的影响受兽药种类、土壤因子(如有机质含量、矿物类型等)的影响.植物吸收抗生素类兽药可能是主动吸收过程,且大量在植物根系内累积,同时也可在植物地上部累积.抗生素类兽药极易诱导产生大量抗药菌,并可能诱导产生群落抗性(Pollution-Induced Community Tolerance,PICT),将对包括人类在内的生态系统健康产生深远影响.     

抗生素、重金属和杀生剂抗性共选择机制

细菌抗生素抗性已被世界卫生组织认定为对公众健康的重要威胁。细菌抗性除了受到抗生素的直接选择压力,也受到其他物质如重金属和杀生剂的共同影响。研究发现,细菌可以通过外排泵系统、细胞膜通透性改变、作用靶标点的改变、酶修饰或降解作用、应激反应改变生理特性、金属螯合和生物转化等抗性机制对这些选择压力产生抗性,或者通过协同抗性、交叉抗性和共调控抗性的机制产生多重抗性。本文综述了细菌对抗生素、重金属和杀生剂的单一抗性机制、多重抗性机制,以及重金属和杀生剂对细菌抗生素抗性的影响机制,并分析了目前研究的不足之处。     

Xα21基因导入水稻及转基因植株的鉴定

以水稻愈伤组织为受体、质粒pCXK1301为Xα21基因供体,用农杆菌EHA105和PDS－1000／He基因枪转化受体,经Hyg浓度梯度筛选,获得一些转基因水稻植株,GUS检测、PCR扩增和Southern杂交分析表明,报告基因（GUS）、选择标记基因（Hyg^r）和抗白叶枯病基因Xα21都已整合入转化水稻基因组中,Southern杂分析显示了Xα21基因在转化水稻基因组中整合的拷贝数。田间接菌鉴定表现抗白叶枯病。     

气溶胶中抗生素抗性基因研究进展:以养殖场和医院为例

抗生素在医药和养殖业的大量使用导致耐药菌的出现,加速了抗生素抗性基因(antibiotic resistance genes,ARGs)在不同环境介质中的传播扩散,抗生素耐药性已成为目前全球卫生、食品安全和发展的最大威胁之一。气溶胶作为ARGs的潜在储存库缺乏系统的研究数据,而通过空气传播具有较高抗生素抗性水平的细菌可能是引起重要疾病的主要传播途径。本文针对养殖场和医院2个抗生素大量使用的典型场所,对气溶胶中ARGs的污染现状、样品采集与检测技术进行综述,并探讨了这一环境污染的潜在风险,表明开展气溶胶中ARGs研究的必要性,并为以后需开展的工作提出几点建议。     

Effects of adding biochar on tetracycline removal during anaerobic composting of swine manure

Residual antibiotics in manure pose a potential threat to public and ecological health when the manure is released to the environment via land application of manure as fertiliser. Anaerobic composting has the potential to decrease antibiotic concentrations prior to land application. We therefore investigated the degradation of the antibiotics tetracycline (TC) and oxytetracycline (OTC) during anaerobic composting under different conditions. All composting treatment conditions were able to effect significant decreases in the antibiotic concentrations, though without biochar antibiotic residues remained. TC and OTC were removed more efficiently when the compost was treated at 55°C than treatments at 35°C or 25°C. Adding biochar significantly and rapidly decreased the antibiotic concentrations in the compost samples. After 15 days of treatment with biochar at 55°C, 100% of TC and OTC had been removed from the manure. The marked ability of biochar to remove the antibiotics was primarily attributed to adsorption of TC and OTC by the biochar. However, the addition of biochar may also have altered the microbial community structures within the compost and accelerated microbial degradation of the antibiotics.     

Antibiotic resistance patterns of Salmonella and Escherichia coli in the groundwater of Cyprus

In addition to diet-based vectors of disease, the contribution of water-borne zoonotic agents to gastrointestinal illnesses may be significant, but this has yet to be investigated for Cyprus. Our main objective was to evaluate antibiotic resistance patterns of Salmonella and Escherichia coli in groundwater samples collected at confined animal feeding operations. This is the first report on the occurrence of antibiotic-resistant Salmonella and E. coli strains in the groundwater of Cyprus. Most of Salmonella isolates belonged to the subgroup enterica, whereas none of the E. coli isolates expressed the verotoxin-encoding gene. Out of 27 isolated Salmonella strains, nearly half of them were resistant to at least one or more antibiotic, whereas the highest resistance was exhibited by sulphamethoxazole (85%), followed by streptomycin (39%), and tetracycline (31%). For the E. coli isolates, nearly a third of them showed resistance to at least one antibiotic, whereas the selection of antibiotic resistance was equal among sulphamethoxazole, tetracycline and streptomycin (20%). This study demonstrated that Salmonella and E. coli in groundwater could pose a public health risk via oral ingestion of contaminated water. Best management practices are needed for overexploited groundwater supplies of rural areas, minimizing human exposure to antibiotic-resistant pathogens.     

The economic dynamics of antibiotic efficacy under open access

We analyze the exploitation of an antibiotic in a market subject to open access on the part of antibiotic producers to the common pool of antibiotic efficacy. While the market equilibrium depends only on current levels of antibiotic efficacy and infection of the epidemiological system, the social optimum accounts for the dynamic externalities which relate those levels to the intertemporal use being made of the antibiotic. We show that depending on the parameters of the model, in particular the cost of production and the improvement in the recovery rate that results from antibiotic treatment, the positive steady-state level of antibiotic efficacy to which the system tends under open access can be lower or higher than the level which should prevail in the socially optimal steady state. In fact there are parameter configurations for which the steady states can be exactly the same. However, the paths leading to the steady state always differ.     

Pharmaceutical compounds in aquatic environment in China: locally screening and environmental risk assessment

An environmental risk assessment was performed for pharmaceutical compounds present in the aquatic environment of China. Predicted environmental concentration (PEC) of the compounds were calculated according to European Medicines Evaluation Agency (EMEA) guidelines. Available ecotoxicological data compromised by applying a very conservative assessment factor (AF) were employed to calculate the predicted no-effect concentration (PNEC). The screening principle and the risk assessment were based on risk quotient (RQ), which derived from the PEC and related PNEC values. PEC results indicated that all the compounds except sulfadimethoxine and levocarnitine, should carry out phase II risk assessment in EMEA guideline. RQ values suggested that more than 36 pharmaceuticals may be imposed health threats to the aquatic environment; especially the antibiotic therapeutic class including amoxicillin, sulfasalazine, trimethoprim, oxytetracycline and erythromycin showed high RQ values. These substances with high RQ value (RQ≥1) were regarded as top-priority pharmaceuticals for control in the aquatic environment of China. However, the antibiotic substances which had low risk quotient (RQ <1), should be reassessed by its potentially induced resistance under low concentration in future.     

Advances in the antimicrobial and therapeutic potential of siderophores

Environmental Chemistry Letters - The increasing bacterial resistance from antibiotic overuse has fostered the search for novel antimicrobial strategies. In particular, bacterial systems involving...     

Presence,dissemination and removal of antibiotic resistant bacteria and antibiotic resistance genes in urban drinking water system: A review

Reviewed the change of ARGs and ARB in full-scale urban drinking water systems. Conventional processes are more promising than BAC process in ARGs removal. Mechanisms of ARGs enrichment and spread in BAC filter and DWDSs are discussed. Raise the need of future research on ARGs and ARB change in building plumbing systems. Antibiotic resistance in aquatic environment has become an important pollution problem worldwide. In recent years, much attention was paid to antibiotic resistance in urban drinking water systems due to its close relationship with the biosafety of drinking water. This review was focused on the mechanisms of antibiotic resistance, as well as the presence, dissemination and removal of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the urban drinking water system. First, the presence of ARB and ARGs in the drinking water source was discussed. The variation of concentration of ARGs and ARB during coagulation, sedimentation and filtration process were provided subsequently, in which filtration was proved to be a promising technology to remove ARGs. However, biological activated carbon (BAC) process and drinking water distribution systems (DWDSs) could be incubators which promote the antibiotic resistance, due to the enrichment of ARGs and ARB in the biofilms attached to the active carbon and pipe wall. Besides, as for disinfection process, mechanisms of the inactivation of ARB and the promotion of conjugative transfer of ARGs under chlorine, ozone and UV disinfection were described in detail. Here we provide some theoretical support for future researches which aim at antibiotic resistance controlling in drinking water.     

Antimicrobial resistance among enteric bacteria, isolated from runoff of the Gangotri glacier, western Himalaya India

An attempt has been made to study antimicrobial resistance among the coliforms, faecal coliforms and faecal streptococci isolated from the runoff of the Gangotri glacier. The number of multiple antibiotic resistances (MAR) coliform isolates increases from upper stretch (33.33%) to lower stretch (83.33%). All faecal coliforms (100%) and faecal streptococci (100%) of lower stretch of study area showed multiple antibiotic resistances. Among coliforms, the value of Antibiotic resistance index (ARI) was found maximum in lower stretch (0.34) followed by middle stretch (0.29) and upper stretch (0.25).     

A review on anammox process for the treatment of antibiotic-containing wastewater: Linking effects with corresponding mechanisms

• Anammox is promising for nitrogen removal from antibiotic-containing wastewater. • Most antibiotics could inhibit the anammox performance and activity. • Antibiotic pressure promoted the increase in antibiotic resistance genes (ARGs). • Antibiotic-resistance mechanisms of anammox bacteria are speculated. Antibiotic is widely present in the effluent from livestock husbandry and the pharmaceutical industry. Antibiotics in wastewater usually have high biological toxicity and even promote the occurrence and transmission of antibiotic resistant bacteria and antibiotic resistance genes. Moreover, most antibiotic-containing wastewater contains high concentration of ammonia nitrogen. Improper treatment will lead to high risk to the surrounding environment and even human health. The anaerobic ammonium oxidation (anammox) with great economic benefit and good treatment effect is a promising process to remove nitrogen from antibiotic-containing wastewater. However, antibiotic inhibition has been observed in anammox applications. Therefore, a comprehensive overview of the single and combined effects of various antibiotics on the anammox system is conducted in this review with a focus on nitrogen removal performance, sludge properties, microbial community, antibiotic resistance genes and anammox-involved functional genes. Additionally, the influencing mechanism of antibiotics on anammox consortia is summarized. Remaining problems and future research needs are also proposed based on the presented summary. This review provides a better understanding of the influences of antibiotics on anammox and offers a direction to remove nitrogen from antibiotic-containing wastewater by the anammox process.     

畜禽养殖过程抗生素使用与耐药病原菌及其抗性基因赋存的研究进展

兽用抗生素在提高畜禽生产性能、防治疾病方面发挥着重要作用,目前全球超过一半以上抗生素用于畜禽养殖,畜禽养殖源耐药病原菌、抗性基因及其传播风险愈益得到人们的重视。我国是畜禽养殖和抗生素使用大国,但兽用抗生素使用、病原菌耐药水平及其抗性基因类型等数据却较为缺乏,不利于今后畜禽养殖源耐药病原菌及其传播风险的控制。因此,本文通过文献调研,对我国和主要发达国家的兽用抗生素使用情况、畜禽养殖源耐药病原菌及其携带的抗性基因、基因移动元件以及向环境传播的途径进行分析、总结,以期为规范合理用药、降低耐药病原菌及其抗性基因传播风险,建立从畜禽养殖场至公共环境全过程的抗性污染控制链条提供借鉴。     

昆虫抗菌肽及其基因工程研究进展

昆虫抗菌肽是一类具有免疫效应的小分子多肽,它具有热稳定、抗菌谱广、无免疫原性、作用机制独特等特性,是昆虫能够抵御自然界中许多有害微生物侵染的重要因素.近年来,随着有关抗菌肽作用机理及功能研究的深入,其在医药领域的应用越来越引起人们的关注.本文就昆虫抗菌肽的分类、生物功能、基因工程研究进展等作一综述.参33     

Antibiotika als Umweltkontaminanten — Effekte auf Bodenbakterien

Goal and scope

Among the human and veterinary pharmaceuticals, antibiotics form an important group. Recent research addressed the environmental ‘side effects’ of antibiotics. Evidence for environmental effects of antibiotics has for example been found for the respiratory activity of soil microorganisms. In the present contribution, results of studies on the ecotoxicology of antibiotics that are based on the utilization of carbon substrates in so-called Biolog plates are summarized.

Methods, Results and Conclusions

The method of pollution-induced community tolerance (PICT) takes centre stage. PICT is based on the changes in community composition of environmental communities brought about by a toxicant, which lead to an overall increase in community tolerance to this toxicant. The suitability of such an increase in community tolerance as ecotoxicological endpoint had been the subject of several experiments. It was shown that effect testing of antibiotics requires supplementation of the communities with nutrients, and that the PICT method reveals antibiotic effects with a high specificity. In an application of PICT, effects of three classes of antibacterial compounds were investigated. Dose-response relationships were obtained for all substances, and effect concentrations were partly in the range of expected environmental concentrations. The possible risks of antibiotic use for an increase in the occurrence of antibiotic resistance genes in the environment are also touched upon, including the extent of natural resistance.

Outlook

Due to its specificity, the PICT method has shown to be a suitable ecotoxicological assay. Due to possible effects of tetracyclines on the structure of the soil microbial community, it is advised to use veterinary antibiotics with caution. The risks of an increase in the occurrence of antibiotic resistance in the environment should be investigated in more detail.     

Effects of different concentrations of tetracycline and oxytetracycline on the growth and ecotoxicity in lettuce北大核心CSCD

四环素类抗生素(TCs)是目前我国应用广泛、用量最大的一类抗生素,畜禽粪便和土壤中存在TCs残留,影响蔬菜作物的生长发育. TCs因水溶性较高更容易被植物转运和积累,植物对TCs耐受性机理研究仍不足.为更全面探究土壤TCs对蔬菜的毒性作用,研究不同浓度四环素(TC)和土霉素(OTC)分别对生菜的抗生素残留、生长特征及抗氧化酶系统的影响.结果显示,在0(对照)、2、10、50、250 mg/kg 5个施用水平下,生菜叶片抗生素含量逐渐增加,且土霉素含量始终大于四环素含量.与对照相比,抗生素浓度在50 mg/kg以上时对生菜生长具有显著抑制作用,其中,株高、根长、地上部和地下部鲜重与叶片TC残留量具显著负相关.生菜叶片的脯氨酸含量随浓度增加呈先增加后降低的趋势,在10 mg/kg时达到最大.并且低浓度(2 mg/kg)促进抗氧化酶基因SOD、POD21和CAT的表达,高浓度抗生素(50、250 mg/kg)对其产生抑制作用,10 mg/kg的抗生素处理对SOD、POD21和CAT基因表达的影响在抗生素种类上存在差异.本研究表明抗生素浓度超过50 mg/kg对生菜生长产生抑制作用,脯氨酸和抗氧化酶SOD、POD、CAT的转录水平及其酶活性能快速响应抗生素胁迫,可作为生菜对抗生素抗性的辅助评价指标.(图8表3参19)