新疆罗布泊外围地区极端环境嗜碱放线菌生物学特性及活性物质筛选

2006～2007年从新疆罗布泊外嗣地区的泥土样品中分离了117株放线菌,初步筛选后对其中47株嗜碱放线菌进行了形态学鉴定,不同生长条件(pH值,盐NaCl、KCl、MgCl_2,碱性物质Na_2 CO_3、K_2CO_3、Mg_2CO_3、NaOH、KOH,温度)对菌株生长的影响,拮抗性、产酶活性检测及菌株LA4的分子进化鉴定等研究.结果发现,47株放线菌的15%属于诺卡氏菌属,70%属于链霉菌属,10%属于游动放线菌属,其中嗜碱链霉菌属占优势.在pH 7～11范围内,47株嗜碱放线菌均能生长.生长率为92%～95%.NaCl、KCl、MgCl,对嗜碱放线菌均具有抑制作用,NaCl、KCl的抑制作用高于MgCl_2,NaCl、KCl浓度超过10%时而MgCl_2 浓度超过15%时嗜碱放线菌完全停止生长.47株嗜碱放线菌对Na_2CO_3、K_2CO_3、MgCO_3、KOH、NaOH等碱性物质没有选择性,生长良好.47株嗜碱放线菌的最适生长温度为28～30℃.超过55℃后均不能生长.从47株嗜碱放线菌中筛选出了30株具有抗菌活性的菌株,其中对大肠杆菌有抑制作用的有11株,占36.7%,对金黄色葡萄球菌有抑制作用的有20株,占66.7%,对枯草杆菌有抑制作用的有16株,占53.3%;其中4株嗜碱放线菌LA4、LA24、LA33和LA19对3种指示菌均表现出抑制作用.在47株嗜碱放线菌中,74%具有淀粉酶活性,81%具有纤维素酶活性,43%具有蛋白酶活性,49%具有脂肪酶活性.通过分子鉴定,发现LA4为链霉菌中尚未报道的一个新亚种,其16S序列GenBank登录号为FJ182229.本研究结果表明新疆罗布泊外围地区存在大量的产纤维素酶、淀粉酶的嗜碱放线菌,并且潜藏着新的放线菌资源,为该地区嗜碱放线菌的种质资源开发和新药筛选有效菌源提供了参考.图5表4参20     

九孔鲍肠道及其养殖水体中异养细菌抗药性研究

为了解鲍鱼养殖环境中异养细菌的抗药性,对自广东汕尾粤顺鲍鱼养殖场九孔鲍养殖环境及其肠道中的异养细菌进行了分离鉴定,并对所分离鉴定的菌株进行了四环素等16种抗生素的抗药性试验、结果表明,鲍鱼养殖水体的异养细菌主要由鞘氨醇单胞Sphingomonas paucimobilis、巴斯德氏菌属Pasteurella sp、莫拉氏菌属Moraxella sp、假单胞菌属Pseudomonas sp、弧菌属Vibrio minicus等9种组成;鲍鱼消化道异养细菌类群菌株则由鞘氨醇单胞菌属Sphingomonas sp、气单胞菌属Aeromonas sp、黄杆菌属Flavobdcterium sp等7种组成。抗药性试验揭示,大多数异养菌菌株对四环素、青霉素G、卡那霉素、丁胺卡那霉素和新生霉素均产生了抗药性,而对氟哌酸、红霉素、氯霉素以及环丙沙星等的抗药性则相对较弱。     

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.     

发酵类抗生素废水处理工艺及运行效果

介绍了内蒙古某抗生素废水处理厂的处理工艺流程、处理单元类型及其详细设计参数,通过对该污水处理厂半年内运行效果的调研,分析评价该污水处理系统的处理效果及出水指标;该水厂工艺流程对COD、TOC、氨氮、总氮、总磷、BOD和急性毒性的平均去除率分别为96.52%、95.83%、50.24%、55.20%、62.05%、98.52%和99.14%,COD、氨氮、总氮、总磷出水水质分别为477、438、556.7和7.69 mg/L,达到排放到城区下水道要求,园区内所有废水汇集到园区污水处理厂进行集中深度处理达标后排放水体。通过对该水厂工艺的介绍和水质分析,为我国抗生素生产企业面临的废水处理工艺类型选择及工程改造升级提供一定参考。     

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。     

Fenton氧化法深度处理抗生素废水二级出水

采用Fenton氧化法对青霉素和土霉素混合废水二级处理出水进行深度处理,通过正交和单因素实验研究了废水初始反应pH值、H2O2投加量、Fe2+/H2O2摩尔比及反应时间等因素对废水处理效果的影响。实验结果表明,Fenton氧化法处理的最佳反应条件为:初始pH值4、H2O2(30%)投加量50 mL/L、Fe2+/H2O2摩尔比1/20和反应时间60 min,处理后出水COD小于120 mg/L,COD去除率在75%以上,急性毒性(HgCl2毒性当量)小于0.07 mg/L,满足《发酵类制药工业水污染物排放标准》(GB21903-2008)表2标准要求。     

The characteristics of nitrate removal by the psychrotolerant denitrifying bacterium Acinetobacter johnonii DBP-3, isolated from a low-temperature eutrophic body of water

A psychrotolerant denitrifying bacterial strain, DBP-3, was isolated from a eutrophic body of water by low-temperature-oriented acclimation culture. Based on its morphologicalandbiochemicalcharacteristics and 16S rDNA gene sequence, the bacterium was identified as belonging to the genus Acinetobacter and closely related to A. johnonii. The satisfactory growth of DBP-3 was observed at 10–30°C and pH 7–9. Strain DBP-3 was able to utilize three types of carbon sources (sodium acetate > sodium citrate > glucose) to support growth and denitrification. DBP-3 grew faster, but with lower nitrate removal efficiency and higher nitrite accumulation, under aerobic conditions than under anoxic conditions. DBP-3 was extremely susceptible to tetracycline and rifampicine and less sensitive to ampicillin and penicillin. The growth of DBP-3 was significantly affected by Hg (II), Cr (VI), Pb (II), Cd (II), and As (III) at 0.32, 0.63, 1.25, 2.5, and 25.0 mg L^?1, respectively. Interestingly, chromium (VI) significantly promoted DBP-3 growth at concentrations lower than 0.32 mg L^?1. These data may be helpful to support the use of strain DBP-3 in the purification of eutrophic water bodies at low temperatures.     

Speciation and persistence of doxycycline in the aquatic environment: Characterization in terms of steady state kinetics

The aim of the present work was to establish the kinetics for the degradation of doxycycline in the aquatic environment with a view to arriving at a kinetic model that can be used to predict the persistence of antibiotic with confidence. The degradation of doxycycline in both water and sediment phases of aquatic microcosm experiments, as well as in distilled water control experiments, was studied over a period of 90 days. An initial 21% loss due to adsorption by the sediment was observed in the microcosm experiment soon after charging. Biphasic zero-order linear rates of degradation, attributed to microbial degradation of the free and sediment or colloidal particle-adsorbed antibiotic, were observed for both water phase (2.3 × 10^?2 and 4.5 × 10^?3 μgg^?1 day^?1) and sediment phase (7.9 × 10^?3 and 1.5 × 10^?3 μgg^?1 day^?1) of the microcosm experiment. The covered distilled water control experiment exhibited a monophasic zero-order linear rate (1.9 × 10^?3 μgg^?1 day^?1) attributed to hydrolysis, while the distilled water experiment exposed to natural light exhibited biphasic liner rates attributed to a combination of hydrolysis and photolysis (2.9 × 10^?3 μgg^?1 day^?1) and to microbial degradation (9.8 × 10^?3 μgg^?1 day^?1). A kinetic model that takes into account hydrolysis, photolysis, microbial degradation as well as sorption/desorption by colloidal and sediment particles is presented to account for the observed zero-order kinetics. The implications of the observed kinetics on the persistence of doxycycline in the aquatic environment are discussed.     

山东省主要河流中抗生素污染组成及空间分布特征

采用固相萃取-液相色谱/串联质谱法研究了山东省境内四大流域主要河流中抗生素污染组成及空间分布特征,涉及我国用量最大的6类共20种抗生素。结果表明:20种抗生素均有检出,且大环内酯类、喹诺酮类和四环素类抗生素整体检出浓度较高。就流域而言,半岛诸河流域抗生素污染较小,平均总质量浓度61.4 ng/L;海河流域、小清河流域和淮河流域相对较为严重,平均总质量浓度分别为232、175、118 ng/L。抗生素空间分布呈现一定的规律,检出浓度较高的点位主要集中在人口密集区下游,抗生素污染与周边生活污水、养殖企业废水和城市污水处理厂排水密切相关,而且抗生素组成可从一定程度上反映出污染来源。     

Fecal source tracking by antibiotic resistance analysis on a watershed exhibiting low resistance

The ongoing development of microbial source tracking has made it possible to identify contamination sources with varying accuracy, depending on the method used. The purpose of this study was to test the efficiency of the antibiotic resistance analysis (ARA) method under low resistance by tracking the fecal sources at Turkey Creek, Oklahoma exhibiting this condition. The resistance patterns of 772 water-isolates, tested with nine antibiotics, were analyzed by discriminant analysis (DA) utilizing a five-source library containing 2250 isolates. The library passed various representativeness tests; however, two of the pulled-sample tests suggested insufficient sampling. The resubstitution test of the library individual sources showed significant isolate misclassification with an average rate of correct classification (ARCC) of 58%. These misclassifications were explained by low antibiotic resistance (Wilcoxon test P < 0.0001). Seasonal DA of stream E. coli isolates for the pooled sources human/livestock/deer indicated that in fall, the human source dominated (P < 0.0001) at a rate of 56%, and that human and livestock respective contributions in winter (35 and 39%), spring (43 and 40%), and summer (37 and 35%) were similar. Deer scored lower (17–28%) than human and livestock at every season. The DA was revised using results from a misclassification analysis to provide a perspective of the effect caused by low antibiotic resistance and a more realistic determination of the fecal source rates at Turkey Creek. The revision increased livestock rates by 13–14% (0.04 ≤ P ≤ 0.06), and decreased human and deer by 6–7%. Negative misclassification into livestock was significant (0.04 ≤ P ≤ 0.06). Low antibiotic resistance showed the greatest effect in this category.