养鸡场空气微生物污染及工人呼吸暴露风险

动物养殖场是空气环境微生物污染的重要来源,然而目前关于养殖场空气中微生物污染特征的时间规律少有报道.针对以上情况,以蛋鸡场为例,采用16S rRNA基因扩增子测序分别对养殖场空气和粪便环境中细菌分布特点及呼吸暴露展开为期80余周的研究.结果表明,空气和粪便样本中16S rRNA含量范围分别在6.08×10⁵ ~4.90×10⁶ copies·m^-3和4.27×10⁸ ~1.15×10¹⁰ copies·g^-1之间.空气中细菌浓度的平均值在冬季显著高于夏季,而生物多样性则呈现相反趋势.蛋鸡场空气与粪便中的优势细菌门均为厚壁菌门（Firmicutes）.在所调查时间内,空气中前3个优势菌属的种类较为稳定,依次为乳杆菌属（Lactobacillus）、拟杆菌属（Bacteroides）和栖粪杆菌属（Faecalibacterium）,而粪便中优势菌属则随养殖时间的增加波动较大.空气和动物粪便中细菌和致病菌群落结构的相关性均不显著,但不同介质中两种目标微生物的含量均显著相关.粪便中细菌的气溶胶化指数随养殖时间的增加而呈上升趋势,而致病菌趋势相反.其中,瘤胃菌科torques属（[Ruminococcus]_torques_group）、拟杆菌属（Bacteroides）和栖粪杆菌属（Faecalibacterium）为最易发生气溶胶化的前3个致病菌属.养鸡场工人的细菌呼吸暴露具有季节性差异,其中细菌和致病菌摄入量的平均值分别为2.54×10⁷ copies·d^-1和2.87×10⁵ copies·d^-1.研究结果将为系统评估养殖场空气微生物的污染特征和潜在健康风险,对以及制定相应的职业暴露行业标准和防控措施提供科学依据.     

Impacts of cadmium addition on the alteration of microbial community and transport of antibiotic resistance genes in oxytetracycline contaminated soil

The large-scale development in livestock feed industry has increased the chances of antibiotics and heavy metals contamination in the soil. The fate of antibiotic resistance genes (ARGs) and microbial community in heavy metals and antibiotic contaminated soil is still unclear. In this study, we investigated the effect of cadmium (Cd) addition on the transport of ARGs, microbial community and human pathogenic bacteria in oxytetracycline (OTC) contaminated soil. Results showed that the addition of OTC significantly increased the abundance of ARGs and intI1 in the soil and lettuce tissues. The addition of Cd to OTC treated soil further increased the abundance and translocation of ARGs and intI1. Moreover, Cd promoted the transfer of potential human pathogenic bacteria (HPB) into lettuce tissues. Compared with O10 treatment, the addition of Cd decreased the concentration of OTC in soil and lettuce tissue, but slightly increased the fresh weight of lettuce tissues. Redundancy analysis indicated that bacterial community succession is a major factor in ARGs variation. Network analysis indicated that the main host bacteria of ARGs were mainly derived from Proteobacteria. Correlation analysis showed that intI1 was significantly correlated with tetG, tetC, sul1, sul2, ermX, and ermQ. Meanwhile, potential HPB (Clostridium, and Burkholderia) was significantly correlated with intI1 and eight ARGs (tetG, tetC, tetW, tetX, sul1, sul2, ermX, and ermQ.). The findings of this study suggest that the addition of heavy metals to agricultural fields must be considered in order to reduce the transfer of ARGs in the soil and crops.     

城市污水处理工艺对宿主特异性标记物和肠道病原菌的去除效果

采用自制小型反应器模拟实验和实时荧光定量PCR检测方法,针对指示人类、犬类粪便污染的宿主特异性标记物,以及病原性大肠埃希氏菌、沙门氏菌和志贺氏菌等典型的肠道病原菌,分别考察了序批式活性污泥法(SBR)、混凝、过滤工艺对它们的去除效果。研究结果表明,SBR工艺的去除效果优于混凝和过滤工艺,对标记物和病原菌毒力基因的去除率都在93.5%以上。SBR工艺中初期短时间的搅拌即可去除大部分标记物和病原菌。在这3种处理工艺中,宿主特异性标记物与肠道病原菌毒力基因的去除率具有显著相关性,这说明宿主特异性标记物能够在一定程度上反映污水处理过程中肠道病原菌的含量。     

畜禽养殖场排放物病原微生物危险性调查

以粪大肠菌群和沙门菌为指标,采集江苏省内10家不同养殖类型畜禽养殖场的排放物及其周边水、土样共37个进行观察与计数,同时分析排放污水中大肠杆菌的耐药性以及禽畜排泄物对土壤和水体耐药细菌数的影响.结果表明:调查的10家养殖场中有9家粪便未经无害化处理直接排放到水体或施于农田,且排放物的粪大肠菌群数全部严重超标,沙门菌检出率达19%,施新鲜粪肥的土壤中粪大肠菌群数(MPN,最大可能数)在105 g-1以上,水体中分离出的大肠杆菌表现出多重耐药性,抗生素抗性细菌总数也远高于未施新鲜粪肥的土壤及水体.     

九龙江流域潜在病原菌污染分析

近年来,由于流域经济快速发展和城镇化进程加快等原因,九龙江水污染问题日趋严重.为全面认识九龙江流域病原菌的分布状况,应用16S rRNA基因-454焦磷酸测序技术测定九龙江支流西溪、北溪水体和沉积物中细菌16S rRNA基因V1~V3高变区,共获得204 216条高质量序列.通过与病原菌参考数据库对比分析发现,九龙江分布有68个潜在病原菌属,占序列总量的6.1%.其中梭菌属(Clostridium)、分支杆菌属(Mycobacterium)和鞘氨醇单胞菌属(Sphingomonas)在所有样品中都有检出,且丰度最高,分别占病原菌属序列总量的54.5%、5.9%和5.6%.在种水平上,九龙江分布有48种病原菌,占序列总量的0.76%.其中Afipia felis、Mycobacterium asiaticum、Clostridium baratii、Brucella melitensis和Delftia tsuruhatensis是丰度最高的5种病原菌,分别占病原菌种序列总量的48.9%、20.3%、8%、2.7%和1.7%.统计分析表明,九龙江水体中分布的病原菌种类数(属或种)显著高于沉积物中,且西溪水体检出的病原菌种类(属或种)和丰度最高,说明九龙江水体,尤其是西溪水体受病原菌污染的风险较高.此外,相关分析表明,九龙江水体中病原菌的种类数(属或种)和丰度与营养盐(氮、磷)有着显著的正相关关系,说明九龙江水体中分布的病原菌与沿岸的人类活动,如养殖业、污水排放等密切相关.因此,为保证流域的公共卫生安全,需进一步加强污染源清理整顿工作,开展水环境病原菌的实时监测.     

农村污水膜生物反应器系统中微生物群落解析

农村污水的随意排放和未加限制的灌溉回用可能会导致水源地环境污染加剧,并且其中潜在的病原微生物对周边淡水资源安全和农村居民的身心健康造成潜在的威胁.为了解农村污水的微生物群落组成,为后续污水灌溉的病原微生物风险评价提供理论依据,采用末端限制性片段长度多态性(T-RFLP)和16S r DNA基因克隆文库技术研究农村污水膜生物反应器(MBR)系统中的微生物群落多样性,并结合实时荧光定量PCR方法监测处理前后典型病原菌——弓形菌(Arcobacter spp.)以及总细菌数量的变化.从未处理的农村污水中获取的73个阳性克隆测序结果表明,其主要细菌类群为变形菌门(91.8%)、厚壁菌门(2.70%)、拟杆菌门(1.40%),以及部分不可培养细菌(4.10%).其中弓形菌属是ε-变形菌门的优势菌,也是农村污水中的优势菌株,占克隆总数的68.5%.TRFLP的结果表明,不同处理工艺阶段的主要细菌类群及丰度明显不同,调节池的物种丰度(S)、Shannon-Wiener(H)和物种均匀度(E)最高,分别为43.0、3.56和0.95.定量PCR结果显示未处理的农村污水中弓形菌数量高达(1.09±0.064 0)×1011copies·L-1,该结果与克隆文库结果均表明弓形菌在污水中确实占较高比例.与未处理的农村污水相比,处理后的出水中所监测的弓形菌及细菌总拷贝数分别减少了2~3个数量级,说明MBR处理系统在一定程度上能够去除微生物.处理后的再生水理化指标及指示菌卫生指标均符合农田灌溉用水标准,但其中残留病原微生物引发的健康风险仍需进一步评价.     

Antibacterial activity of different natural honeys from Transylvania,Romania

Honey is used in food industry and medicine due to its nutritive, therapeutic and dietetic qualities. The microbiological characteristics of 10 unpasteurized honey samples of known origin, collected from Transylvania beekeepers (Romania) were determined. The antibacterial activity of these types of honey against Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Salmonella enteritidis, Salmonella anatum, Salmonella choleraesuis, Bacillus cereus, Bacillus subtilis subsp. spizizenii and Listeria monocytogenes strains was studied. The most sensitive to the antibacterial activity were the two staphylococus strains (the largest diameter of inhibition zone was 18 mm) and B. subtilis strains (13.5 mm). The strains of B. cereus, E. coli, L. monocytogenes and Salmonella spp. were found to present resistance to some of the honey samples. Manna, sunflower and polyfloral honeys presented high antibacterial activity while acacia and linden honeys had a lower activity in terms of the number of sensible strains. Statistical analysis shows that the type of strains and the type of honey have influence on the diameter of inhibition.     

浅论微生物技术控藻

富营养化水体会引起藻类增殖，影响人类的生存环境。阐述了利用病毒、原生动物、细菌等微生物技术控制富营养化藻类的研究进展，并对病毒控藻技术应用时的两个问题进行了讨论，认为利用微生物技术控藻需要对生态系统进行更加深入的认识和掌握。     

Quantification of multi-antibiotic resistant opportunistic pathogenic bacteria in bioaerosols in and around a pharmaceutical wastewater treatment plant

Pharmaceutical wastewater treatment plants(WWTPs) are thought to be a "seedbed" and reservoirs for multi-antibiotic resistant pathogenic bacteria which can be transmitted to the air environment through aeration. We quantified airborne multi-antibiotic resistance in a full-scale plant to treat antibiotics-producing wastewater by collecting bioaerosol samples from December2014 to July 2015. Gram-negative opportunistic pathogenic bacteria(GNOPB) were isolated, and antibiotic susceptibility tests against 18 commonly used antibiotics, including 11 β-lactam antibiotics, 3 aminoglycosides, 2 fluoroquinolones, 1 furan and 1 sulfonamide, were conducted.More than 45% of airborne bacteria isolated from the pharmaceutical WWTP were resistant to three or more antibiotics, and some opportunistic pathogenic strains were resistant to 16 antibiotics, whereas 45.3% and 50.3% of the strains isolated from residential community and municipal WWTP showed resistance to three or more antibiotics. The calculation of the multiple antibiotic resistance(MAR) index demonstrated that the air environment in the pharmaceutical WWTP was highly impacted by antibiotic resistance, while the residential community and municipal WWTP was less impacted by antibiotic resistance. In addition, we determined that the dominant genera of opportunistic pathogenic bacteria isolated from all bioaerosol samples were Acinetobacter, Alcaligenes, Citrobacter, Enterobacter, Escherichia, Klebsiella, Pantoea, Pseudomonas and Sphingomonas. Collectively, these results indicate the proliferations and spread of antibiotic resistance through bioaerosols in WWTP treating cephalosporin-producing wastewater, which imposed a potential health risk for the staff and residents in the neighborhood, calling for administrative measures to minimize the air-transmission hazard.     

城市环境生物安全研究的进展与挑战

近年来世界范围内新发和再发传染病的暴发严重威胁生物安全,危害人类健康.人类活动包括快速城市化、集约化养殖、全球贸易和旅行等加速了传染性疾病的传播.人群健康、动物健康和环境健康紧密相关,国际上健康一体化（One Health）的理念已被广泛接受用以管理和预防人畜共患病的暴发.城市环境是人群-微生物组-环境要素相互作用最为强烈和最为复杂的场所,使得城市环境生物安全显得尤为重要.本文系统分析了城市环境生物安全领域的研究进展,厘清了该领域研究的主要挑战,提出了建立城市环境生物安全大数据平台、评估微生物污染健康风险和构建城市微生物安全风险防控及治理体系的建议.