Antibiotic resistant bacteria/genes dissemination in lacustrine sediments highly increased following cultural eutrophication of Lake Geneva (Switzerland)

This study investigates faecal indicator bacteria (FIB), multiple antibiotic resistant (MAR), and antibiotic resistance genes (ARGs), of sediment profiles from different parts of Lake Geneva (Switzerland) over the last decades. MARs consist to expose culturable Escherichia coli (EC) and Enterococcus (ENT) to mixed five antibiotics including Ampicillin, Tetracycline, Amoxicillin, Chloramphenicol and Erythromycin. Culture-independent is performed to assess the distribution of ARGs responsible for, β-lactams (blaTEM; Amoxicillin/Ampicillin), Streptomycin/Spectinomycin (aadA), Tetracycline (tet) Chloramphenicol (cmlA) and Vancomycin (van). Bacterial cultures reveal that in the sediments deposited following eutrophication of Lake Geneva in the 1970s, the percentage of MARs to five antibiotics varied from 0.12% to 4.6% and 0.016% to 11.6% of total culturable EC and ENT, respectively. In these organic-rich bacteria-contaminated sediments, the blaTEM resistant of FIB varied from 22% to 48% and 16% to 37% for EC and ENT respectively, whereas the positive PCR assays responsible for tested ARGs were observed for EC, ENT, and total DNA from all samples. The aadA resistance gene was amplified for all the sediment samples, including those not influenced by WWTP effluent water. Our results demonstrate that bacteria MARs and ARGs highly increased in the sediments contaminated with WWTP effluent following the cultural eutrophication of Lake Geneva. Hence, the human-induced changing limnological conditions highly enhanced the sediment microbial activity, and therein the spreading of antibiotic resistant bacteria and genes in this aquatic environment used to supply drinking water in a highly populated area. Furthermore, the presence of the antibiotic resistance gene aadA in all the studied samples points out a regional dissemination of this emerging contaminant in freshwater sediments since at least the late nineteenth century.     

Transcriptome analysis provides insights for understanding the adverse effects of endosulfan in Drosophila melanogaster

Indiscriminate use of agrochemicals worldwide, particularly, persistent organic pollutants (POPs), is of concern. Endosulfan, a POP, is used by various developing/developed nations and is known to adversely affect the development and the hormonal profiles of humans and animals. However, little is known about the molecular players/pathways underlying the adverse effects of endosulfan. We therefore analyzed the global gene expression changes and subsequent adverse effects of endosulfan using Drosophila. We used Drosophila melanogaster keeping in view of its well annotated genome and the wealth of genetic/molecular reagents available for this model organism. We exposed third instar larvae of D. melanogaster to endosulfan (2.0 μg mL⁻¹) for 24 h and using microarray, we identified differential expression of 256 genes in exposed organisms compared to controls. These genes are associated with cellular processes such as development, stress and immune response and metabolism. Microarray results were validated through quantitative PCR and biochemical assay on a subset of genes/proteins. Taking cues from microarray data, we analyzed the effect of endosulfan on development, emergence and survival of the organism. In exposed organisms, we observed deformities in hind-legs, reminiscent of those observed in higher organisms exposed to endosulfan. In addition, we observed delayed and/or reduced emergence in exposed organisms when compared to their respective controls. Together, our studies not only highlight the adverse effects of endosulfan on the organism but also provide an insight into the possible genetic perturbations underlying these effects, which might have potential implications to higher organisms.     

Hydrocarbon degradation, plant colonization and gene expression of alkane degradation genes by endophytic Enterobacter ludwigii strains

The genus Enterobacter comprises a range of beneficial plant-associated bacteria showing plant growth promotion. Enterobacter ludwigii belongs to the Enterobacter cloacae complex and has been reported to include human pathogens but also plant-associated strains with plant beneficial capacities. To assess the role of Enterobacter endophytes in hydrocarbon degradation, plant colonization, abundance and expression of CYP153 genes in different plant compartments, three plant species (Italian ryegrass, birdsfoot trefoil and alfalfa) were grown in sterile soil spiked with 1% diesel and inoculated with three endophytic E. ludwigii strains. Results showed that all strains were capable of hydrocarbon degradation and efficiently colonized the rhizosphere and plant interior. Two strains, ISI10-3 and BRI10-9, showed highest degradation rates of diesel fuel up to 68% and performed best in combination with Italian ryegrass and alfalfa. All strains expressed the CYP153 gene in all plant compartments, indicating an active role in degradation of diesel in association with plants.     

Expression of alkane monooxygenase (alkB) genes by plant-associated bacteria in the rhizosphere and endosphere of Italian ryegrass (Lolium multiflorum L.) grown in diesel contaminated soil

For phytoremediation of organic contaminants, plants have to host an efficiently degrading microflora. To assess the role of endophytes in alkane degradation, Italian ryegrass was grown in sterile soil with 0, 1 or 2% diesel and inoculated either with an alkane degrading bacterial strain originally derived from the rhizosphere of Italian ryegrass or with an endophyte. We studied plant colonization of these strains as well as the abundance and expression of alkane monooxygenase (alkB) genes in the rhizosphere, shoot and root interior. Results showed that the endophyte strain better colonized the plant, particularly the plant interior, and also showed higher expression of alkB genes suggesting a more efficient degradation of the pollutant. Furthermore, plants inoculated with the endophyte were better able to grow in the presence of diesel. The rhizosphere strain colonized primarily the rhizosphere and showed low alkB gene expression in the plant interior.     

上海沙田湖养殖区及周边水体中氟喹诺酮类抗性基因的分布特征及其与环境因子关系

集约化水产养殖区常被认为是水体抗生素和抗性基因的主要潜在来源,然而对其该类水体中抗生素和抗性基因污染情况的研究仍相对匮乏.本文以上海市沙田湖养殖区及周边水体为调查对象,利用宏基因组学高通量测序技术,于2020年9月对水体中的氟喹诺酮类抗生素(fluoroquinolones,FQs)和氟喹诺酮类抗性基因(fluoroq...     

Short-term inhalation exposure evaluations of airborne antibiotic resistance genes in environments

Antibiotic resistance is a sword of Damocles that hangs over humans. In regards to airborne antibiotic resistance genes (AARGs), critical knowledge gaps still exist in the identification of hotspots and quantification of exposure levels in different environments. Here, we have studied the profiles of AARGs, mobile genetic elements (MGEs) and bacterial communities in various atmospheric environments by high throughput qPCR and 16S rRNA gene sequencing. We propose a new AARGs exposure dose calculation that uses short-term inhalation (STI). Swine farms and hospitals were high-risk areas where AARGs standardised abundance was more abundant than suburbs and urban areas. Additionally, resistance gene abundance in swine farm worker sputum was higher than that in healthy individuals in other environments. The correlation between AARGs with MGEs and bacteria was strong in suburbs but weak in livestock farms and hospitals. STI exposure analysis revealed that occupational intake of AARGs (via PM₁₀) in swine farms and hospitals were 110 and 29 times higher than in suburbs, were 1.5 × 10⁴, 5.6 × 10⁴ and 5.1 × 10² copies, i.e., 61.9%, 75.1% and 10.7% of the overall daily inhalation intake, respectively. Our study comprehensively compares environmental differences in AARGs to identify high-risk areas, and forwardly proposes the STI exposure dose of AARGs to guide risk assessment.     

鲤鱼对2,6-二硝基甲苯的生物浓缩、消除与代谢研究

 以鲤鱼为实验生物，应用流动式摄取及释放装置，进行了2，6－二硝基甲苯（2，6－DNT）的生物浓缩与释放研究。取得了2，6－DNT在鲤鱼（全鱼）、肝及肠部的生物浓缩曲线。全鱼的浓缩曲线可以稳态（平台式）曲线描述，但是肝、肠部浓缩曲线均呈峰形。肝部峰快速下降后渐趋稳态，肠部峰下降未得稳态。峰形，肝在先，肠在后。对比2，6－DNT在全鱼及肝、肠的生物浓缩曲线，并参照报道_（1，2）指出，全鱼摄取曲线上各点所表现的浓度值为各时刻鱼体内各组织摄取化学品总重对全鱼重量的比值。肝、肠部峰形曲线预示了在肝、肠内生物浓缩过程发生了生物转化作用。对全鱼的生物浓缩过程应用单区一级动力学模型，求得BCF值与消除速率常数K₁与K₂，由K₁/K₂计算得到的BCF值与实测值相符。释放较慢，释放曲线呈双曲线形，半减期为4.8h。     

Amoxicillin effects on functional microbial community and spread of antibiotic resistance genes in amoxicillin manufacture wastewater treatment system

This study aimed to reveal how amoxicillin(AMX) affected the microbial community and the spread mechanism of antibiotic resistance genes(ARGs) in the AMX manufacture wastewater treatment system. For this purpose, a 1.47 L expanded granular sludge bed(EGSB) reactor was designed and run for 241 days treating artificial AMX manufacture wastewater. 454 pyrosequencing was applied to analyze functional microorganisms in the system. The antibiotic genes OXA_(-1), OXA_(-2), OXA_(-10), TEM_(-1), CTX-M_(-1), class I integrons(intI1) and 16 SrRNA genes were also examined in sludge samples. The results showed that the genera Ignavibacterium, Phocoenobacter,Spirochaeta, Aminobacterium and Cloacibacillus contributed to the degradation of different organic compounds(such as various sugars and amines). And the relative quantification of eachβ-lactam resistance gene in the study was changed with the increasing of AMX concentration.Furthermore the vertical gene transfer was the main driver for the spread of ARGs rather than horizontal transfer pathways in the system.     

嗜盐菌群对菲的降解及萘双加氧酶基因的表达规律

从胜利油田富集了一个能够降解多环芳烃的嗜盐菌群,通过克隆文库技术解析了菌群萘双加氧酶(ndo)基因的多样性.结果表明,该嗜盐菌群有6种ndo基因型,其中3种主要基因型(占总克隆子92.7%)与经典nah-like基因的相似度为89%.采用real-time RT-PCR等技术分析了不同盐度下菲降解过程中ndo基因的表达量、降解速率、生长曲线以及菲的生物可利用度,探索了菌群对菲的降解及ndo基因的表达规律.结果表明,当盐度由10%升高到20%,菌群完全降解100mg/L菲的时间从6d延长到9d,菌群生长的迟滞期由1d延长为3d.溶解菲的浓度在菌群生长过程中呈增加的趋势.Ndo基因的表达量在降解过程中呈先降低后随溶解菲浓度升高而升高的趋势,表明高浓度菲在初始阶段可能抑制ndo基因的表达.单因素方差分析(n=3, P>0.05)表明溶解菲浓度在10%和20%盐度下没有显著差异,表明一定范围的盐度不会影响菌群降解过程中菲的溶解度.菌群ndo基因的相对表达量在10%盐度下较在20%盐度下高,说明盐度对嗜盐菌群功能基因的表达具有抑制作用.