养殖鱼塘底泥微生物抗生素耐药基因分布分析

针对水产养殖中抗生素滥用引起微生物耐药基因污染的现象,本实验以杭州市某水产养殖区底泥样品为研究对象,测定了底泥样品中抗生素含量,并从其中分离出74株可培养细菌.采用PCR方法对分离菌株中磺胺类、四环素类、喹诺酮类、氯霉素类抗性基因(antibiotic resistance genes,ARGs)和整合子基因进行了检测.结果表明,除tet A(四环素类)耐药基因未被检出外,其余3种耐药基因及整合子基因均被检出,其中qnr S(喹诺酮类)耐药基因的检出率最高,为50.00%.另外,利用16S r DNA序列分析技术将分离菌株鉴定为12个属、19个种,包括环境中多种常见土著细菌及部分致病菌,其中气单胞菌(Aeromonas spp.)数量最多(29株),占分离菌株的39.19%.掌握水产养殖区中ARGs的污染现状,对控制其传播、保障食品安全和保护微生态环境具有重要指导意义.     

Field released transgenic papaya effect on soil microbial communities and enzyme activities

Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase （RP）-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, significant differences （P〈0.05） were observed in total nitrogen in soils grown with transgenic papaya. There were also significant differences （P〈0.05） in the total number of colony forming units （CFUs） of bacteria, actinomycetes and fungi between soils amended with RP-transgenic plants and non-transgenic plants. Compared with non-transgenic papaya, the total CFUs of bacteria, actinomycetes and fungi in soil with transgenic papaya increased by 0.43-1.1, 0.21-0.80 and 0.46-0.73 times respectively. Significantly higher （P〈0.05） CFUs of bacteria, actinomycetes and fungi resistant to kanamycin （Km） were obtained in soils with RP-transgenic papaya than those with non-transgenic papaya in all concentrations of Km. Higher resistance quotients for Km＇ （kanamycin resistant） bacteria, actinomycetes and fungi were found in soil planted with RP-transgenic papaya, and the resistance quotients for Km＇ bacteria, actinomycetes and fungi in soils with transgenic papaya increased 1.6-4.46, 0.63-2.5 and 0.75-2.30 times. RP-transgenic papaya and non-transgenic papaya produced significantly different enzyme activities in arylsulfatase （5.4-5.9x）, polyphenol oxidase （0.7-1.4x）, invertase （0.5-0.79x）, cellulase （0.23-0.35x） and phosphodiesterase （0.16-0.2x）. The former three soil enzymes appeared to be more sensitive to the transgenic papaya than the others, and could be useful parameters in assessing the effects of transgenic papaya. Transgenic papaya could alter soil chemical properties, enzyme activities and microbial communities.     

不同喷雾方式下低阻文丘里振弦栅除尘配置优化实验研究

为了对比径向外喷和轴向喷雾下的低阻文丘里振弦栅耦合除尘效果,以除尘效率和系统阻力为参照,选定喉管气速、振弦栅数、纤维丝间隙和喉管液气比进行单因素实验。结果表明2种喷雾方式的除尘效率和系统阻力的变化趋势相同。利用SPSS软件进行4因素3水平正交实验设计,在特定参数条件下,2种喷雾方式的因素影响排序相同,除尘效率为纤维栅数＞纤维丝间隙＞喉管气速＞喷雾量;系统阻力为喉管气速＞纤维栅数＞纤维丝间隙＞喷雾量;径向外喷最佳配置为喉管气速24 m/s、喷雾量2.3 L/min、2块间隙为0.8 mm纤维栅板;轴向喷雾为喉管气速28 m/s、喷雾量2.4 L/min、2块间隙为0.8 mm纤维栅板;在最优配置下的除尘性能为径向外喷＞轴向喷雾。     

通风机站风流间相互影响阻力特性研究*

为研究双风机并联机站进口、出口风流相互影响的机站局部阻力特性,构建机站通风局部阻力计算的数学模型,同时运用相似原理建立双风机并联通风机站实验模型,并佐以数值模拟对比验证,分析计算不同风机间距的无量纲数A、不同扩散器结构参数条件下的综合影响系数Kc。结果表明:A与Kc具有二次函数关系,且存在较优的A,使得Kc值最小;Kc值随着L/Df增加而线性减小。研究结果有利于Kc的选取,为高效率通风机站结构设计提供技术及数据支撑。     

屏蔽服冷却系统对人体表面温度影响的研究*

为科学有效地论证多种方式组合的冷却屏蔽服在不同环境条件下对人体表面温度控制的效果,需要对冷却系统及人体敏感部位发热量进行客观评估。通过对5名健康男性的高温测试,探究人体在不同环境温度下体表温度的变化,得出胸部、背部及额头为热量最高部位,并构建以“人体-降温屏蔽服-外界环境”为主体的冷却系统数值模型,对不同环境中的屏蔽服冷却效果展开研究,分析穿戴冷却屏蔽服时人体躯干部分的温度分布及影响。结果表明:在屏蔽服中靠近胸部、背部部位引入相变材料和风扇,均可帮助人体降低体温,提高舒适度。     

TSLCDM长袋低压脉冲布袋除尘器及其应用

为了使电炉烟气治理过程中污染源的排放达到国家排放标准,提出一种TSLCDM长袋低压脉冲除尘器(简称TSLCDM除尘器)。通过对该除尘器结构、除尘原理以及不同工况下除尘效率和过滤阻力的分析,得到该除尘器的除尘效率>99.9%,压力损失<1200 Pa,最终排出的烟气含尘质量浓度约5.54 mg/m3,满足了国家的超低排放要求,以泉州某公司电炉项目除尘系统为研究对象,证明了该新型脉冲布袋除尘器在电炉烟气治理除尘系统的应用效果,具有很好的经济和环境效益。     

浮游动物DNA宏条形码标志基因比较研究

DNA宏条形码技术作为一种新型生物监测方法,在未来生态环境监测中有巨大的应用潜力。目前,浮游动物DNA宏条形码监测仍在发展阶段,需要首先对其(采样方法、引物选择和数据分析等)进行标准化和调整,然后才能用于常规流域生态监测。其中,如何选择合适的PCR扩增引物是DNA宏条形码生物监测标准化的关键问题之一。本研究比较了COI、18SV9和16S通用引物在浮游动物DNA宏条形码监测中的差异,为初步建立规范化的浮游动物DNA宏条形码监测方法提供技术支撑。结果表明,16S引物对浮游动物具有更好的特异性,其产生的操作分类单元(operational taxonomic unit, OTU)有88.1%属于浮游动物。虽然18SV9引物具有更高的物种覆盖度,不仅能扩增出浮游动物,还能扩增出大量藻类和真菌,但其物种识别敏感性较差,不适合浮游动物物种水平多样性监测。COI引物的浮游动物物种特异性、物种覆盖度和物种识别敏感性都适中,检出的浮游动物物种数量高于18SV9引物和16S引物,更加适合浮游动物DNA宏条形码监测。     

全氟辛烷磺酸盐(PFOS)对半滑舌鳎(Cynoglossus semilaevis)免疫相关基因表达的影响

为了解全氟辛烷磺酸盐(perfluorooctane sulfonate, PFOS)暴露对半滑舌鳎(Cynoglossus semilaevis)免疫功能的影响,在实验室条件下,运用RT-PCR方法分析了PFOS暴露对半滑舌鳎热休克蛋白hsp70、hsp90、C型凝集素(c-type lectin)和细胞色素c氧化酶(cytochrome c oxidase, cox)等4种免疫相关基因表达水平的影响。实验测定了上述4种基因在半滑舌鳎肝、鳃、肠及肌肉4种不同组织中随时间(0、24 h、48 h、96 h和7 d)的表达变化情况。结果表明,在4种组织中,hsp70基因的表达与对照相比为上调,其中,肝组织hsp70基因的表达量显著高于其他各组织,且表达高峰值的出现也早于其他各组织;hsp90基因在肝和鳃组织中表达量随时间不同而波动,在肠组织中表达上调,在肌肉中表达显著下调;c-type lectin基因表达量与对照组相比表达显著下调或无明显差异;cox基因在肝组织和肠组织中表达下调,在鳃和肌肉中表达上调。上述研究结果表明,PFOS能引起免疫相关基因的表达变化,对半滑舌鳎具有潜在的免疫毒性。肝组织中各免疫基因对PFOS胁迫的响应高于其他组织。本研究可为阐明全氟辛烷磺酸盐对半滑舌鳎的免疫毒性提供基础数据。     

Validation of Bacteroidales-based microbial source tracking markers for pig fecal pollution and their application in two rivers of North China

• Pig feces is the predominant excrement produced by animal husbandry in China. • The PF, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan MST assays showed better performance. • The pig-specific MST assays can contribute to managing the pig fecal pollution. In China, pig feces is the predominant source of excrement produced by animal husbandry. Improper use or direct discharge of pig feces can result in contamination of natural water systems. Microbial source tracking (MST) technology can identify the sources of fecal pollution in environmental water, and contribute to the management of pig fecal pollution by local environmental protection agencies. However, the accuracy of such assays can be context-dependent, and they have not been comprehensively evaluated under Chinese conditions. We aimed to compare the performance of five previously reported pig-specific MST assays (PF, Pig-Bac1^SYBR, Pig-Bac2^SYBR, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan, which are based on Bacteroidales 16S rRNA gene markers) and apply them in two rivers of North China. We collected a total of 173 fecal samples from pigs, cows, goats, chickens, humans, and horses across China. The PF assay optimized in this study showed outstanding qualitative performance and achieved 100% specificity and sensitivity. However, the two SYBR green qPCR assays (Pig-Bac1^SYBR and Pig-Bac2^SYBR) cross-reacted with most non-pig fecal samples. In contrast, both the Pig-1-Bac^TaqMan and Pig-2-Bac^TaqMan assays gave 100% specificity and sensitivity. Of these, the Pig-2-Bac^TaqMan assay showed higher reproducibility. Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand, Japan, and America. Using the PF and Pig-2-Bac^TaqMan assays, a field test comparing the levels of pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that pig fecal pollution was effectively controlled at this location.     

Biofiltration and disinfection codetermine the bacterial antibiotic resistome in drinking water: A review and meta-analysis

• Published data was used to analyze the fate of ARGs in water treatment. • Biomass removal leads to the reduction in absolute abundance of ARGs. • Mechanism that filter biofilm maintain ARB/ARGs was summarized. • Potential BAR risks caused by biofiltration and chlorination were proposed. The bacterial antibiotic resistome (BAR) is one of the most serious contemporary medical challenges. The BAR problem in drinking water is receiving growing attention. In this study, we focused on the distribution, changes, and health risks of the BAR throughout the drinking water treatment system. We extracted the antibiotic resistance gene (ARG) data from recent publications and analyzed ARG profiles based on diversity, absolute abundance, and relative abundance. The absolute abundance of ARG was found to decrease with water treatment processes and was positively correlated with the abundance of 16S rRNA (r² = 0.963, p<0.001), indicating that the reduction of ARG concentration was accompanied by decreasing biomass. Among treatment processes, biofiltration and chlorination were discovered to play important roles in shaping the bacterial antibiotic resistome. Chlorination exhibited positive effects in controlling the diversity of ARG, while biofiltration, especially granular activated carbon filtration, increased the diversity of ARG. Both biofiltration and chlorination altered the structure of the resistome by affecting relative ARG abundance. In addition, we analyzed the mechanism behind the impact of biofiltration and chlorination on the bacterial antibiotic resistome. By intercepting influent ARG-carrying bacteria, biofilters can enrich various ARGs and maintain ARGs in biofilm. Chlorination further selects bacteria co-resistant to chlorine and antibiotics. Finally, we proposed the BAR health risks caused by biofiltration and chlorination in water treatment. To reduce potential BAR risk in drinking water, membrane filtration technology and water boiling are recommended at the point of use.