Characterization and catabolic genes detection of three oil-degrading Rhodococcus spp.北大核心CSCD

Oil pollution is one of the major factors causing environmental deterioration. Bioremediation of oil contaminated environments by microorganisms attracts much research attention. This study aimed to screen efficient oil-degrading bacteria from oil contaminated soil and analyze their characteristics and catabolic genes. Oil-degrading bacteria were screened from oil contaminated soil in minimal medium containing crude oil and identified by morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis. Their growth and degradation characteristics were studied with ultraviolet spectroscopy and GC-MS analysis. The surfactant production was studied by adopting culture method. The major oil-degrading related genes were detected by t he PCR a mplification. As a result, t hree oil-degrading bacteria strains named KB1, 2182 and JC3-47 were isolated from the oil contaminated soil samples. The strains could use crude oil as the sole carbon source to degrade oil with a degrading rate of 41.02%, 32.26% and 55.90%, respectively, when cultured in minimal medium containing crude oil for 3 days. The bacteria were identified to belong to genus Rhodococcus. With 100% similarity of 16S rDNA sequences of the three strains with known ones of Rhodococcus, KB1 was preliminarily identified as Rhodococcus erythropolis, 2182 as Rhodococcus equi, and JC3-47 as Rhodococcus qingshengii. They grew well at 10-50 °C, with the initial pH of 3-9 and the NaCl concentration of 0-5%. The optimal temperature for bacterial growth was 35 °C, 35 °C and 30 °C respectively. KB1 and 2182 could grow at pH 2 and 9.0% of NaCl. The bacteria grew well in broth containing different organic substrates as sole carbon source, such as n-dodecane, n-octadecane, benzene, methylbenzene, xylene and naphthaline. KB1 and JC3-47 could grow well in broth containing pyrene. GC-MS analysis revealed that the bacteria could effectively degrade medium- and long-chain alkane components in crude oil. The bacteria produced biosurfactants and decreased the surface tension of the culture broth. They also showed adhesion activities to n-hexadecane. The oil-degrading related genes such as alkane monooxygenase, aromatic-ring-hydroxylating dioxygenase and catechol dioxygenase genes were detected in all the three strains. Besides, biphenyl dioxygenase genes were detected in KB1 and 2182. The isolated Rhodococcus spp. strains could effectively degrade petroleum hydrocarbons with high adaptabilities to extreme environments such as high salt and low temperature. They are supposed to be applied broadly in the bioremediation of oil contaminated soil in such environments.     

多溴联苯醚微生物降解过程与机理的研究进展

多溴联苯醚(PBDEs)属溴代阻燃剂,曾因其优良的阻燃性能而广泛应用于电子电器、石油化工和建材纺织等工业产品中.然而,随着大量生产和使用,PBDEs已成为大气、水体、土壤和生物体等多环境介质中普遍检出且极具生态风险的有机污染物.因此,开展微生物降解研究对于典型环境中PBDEs污染风险消除和污染修复,具有重要的科学意义.本文从PBDEs环境归趋行为及其暴露风险出发,综述了PBDEs微生物厌氧降解和好氧降解的最新研究动态,比较分析了两种降解过程的降解特性与影响因素,并针对微生物,尤其是好氧微生物降解机理,阐述了bph A或etb A功能基因及其编码酶对PBDEs好氧降解过程的调控作用,同时就PBDEs微生物高效降解菌种选育、降解机理等方面的研究趋势进行了展望.     

长期施用猪粪水稻土抗生素抗性基因污染研究

为了全面研究猪粪有机肥对水稻土抗生素抗性及分布格局的影响,选择295对抗生素抗性基因引物,采用高通量荧光定量PCR对水稻土中的抗生素抗性基因污染情况进行了研究.结果表明,未施用猪粪水稻土检测出66种抗性基因,而施用猪粪水稻土则检测出107种抗生素抗性基因,施用猪粪后水稻土中抗生素抗性基因种类显著增加(P<0.05).相对于未施用猪粪水稻土,施用猪粪水稻土有49种抗生素抗性基因丰度显著增加(P<0.05),其中施用猪粪水稻土中喹诺酮类/氯霉素类抗性基因的mexF的丰度相对于未施用猪粪水稻土增加1791倍.高通量定量结果表明,施用猪粪的水稻土抗生素抗性基因分布格局发生显著变化,高通量定量技术是研究环境抗生素抗性基因的一个非常实用的工具.     

The exposure risks associated with pathogens and antibiotic resistance genes in bioaerosol from municipal landfill and surrounding area

Pathogenic microbes with antibiotic resistance can thrive on municipal solid waste as nutrients and be aerosolized and transported to vicinities during waste disposal processes. However, the characterization of pathogenic bioaerosols and assessment of their exposure risks are lacking. Herein, particle size, concentration, activity, antibiotic resistance, and pathogenicity of airborne microorganisms were assessed in different sectors of a typical landfill. Results showed that active sector in downwind direction has the highest bioaerosol level (1234 CFU/m³), while residential area has the highest activity (14.82 mg/L). Botanical deodorizer from mist cannon can effectively remove bioaerosol. Most bioaerosols can be inhaled into respiratory system till bronchi with sizes ranging from 2.1−3.3 and 3.3−4.7 µm. Pathogenic bacteria (Bacilli, Bacillus, and Burkholderia-Paraburkholderia) and allergenic fungi (Aspergillus, Cladosporium, and Curvularia) prevailed in landfill. Although high abundance of microbial volatile organic compounds (mVOCs) producing bioaerosols were detected, these mVOCs contributed little to odor issues in landfill. Notably, surrounding areas have higher levels of antibiotic-resistance genes (ARGs) than inner landfill with tetC, acrB, acrF, mdtF, and bacA as dominant ones. Most ARGs were significantly correlated with bacterial community, while environmental parameters mainly influenced fungal prevalence. These findings can assist in reducing and preventing respiratory allergy or infection risks in occupational environments relating to waste management.     

人工湿地中抗生素抗性大肠杆菌和抗性基因的去除与分布

抗生素的滥用导致抗生素抗性菌和抗性基因随生活污水和养殖废水的排放在环境中肆意散播,其去除及环境行为越来越受到关注。采用K-B纸片法测定了9套不同工艺构型模拟人工湿地中大肠杆菌对7种抗生素的抗性率,并应用多重PCR检测磺胺类sul1、2、3与四环素tetA、B、C、D抗性基因,探究人工湿地对抗性菌的去除效率及抗性菌、抗性基因的分布规律。结果显示,人工湿地能有效去除污水中70%左右的抗性大肠杆菌,有效降低了细菌抗性的传播风险;共计分离出535株大肠肝菌中有378株对一种以上抗生素有抗性性,以四环素、磺胺类和氨苄西林抗性率最高,达到25%以上,其他4种抗性率较低,不足20%;2种抗性基因的检出率都在70%以上;对不同采样点大肠杆菌的抗性性及抗性基因的比较发现,各部分大肠杆菌的抗性水平、多重抗性指数(MRI)以及抗性基因(sul、tet)检出率和组合数表现出:基质≥出水>进水,推测抗性菌被湿地基质截留,在基质生物膜上发生抗性基因的重组,并释放抗性菌,提高了出水中抗性水平和抗性基因检出率。     

微塑料及金属在黄浦江地表水环境的赋存特征及与金属抗性基因的相关性分析

城市河流是微塑料(MPs)及重金属污染的“热点”区域,在污染迁移中起到重要作用.然而同样作为抗性基因赋存及释放的重要媒介,鲜有研究考察了流域MPs、金属与金属抗性基因(MRGs)的关联.为此,调研了10个黄浦江站点水样的金属及MPs的污染特征,并基于宏基因组学对水样及提取的MPs上的MRGs及可移动遗传元件(MGEs)进行分析.结果显示,表层水中ρ[锑(Sb)]平均值最高,为(3.16±0.419)μg·L^-1. MPs的丰度平均值为(1.78±0.84)n·L^-1,位于工业区和人口密集区的MPs丰度显著高于农业区和低人口密度的区域.纤维状、“小粒径”(<0.5 mm)的聚对苯二甲酸乙二醇酯(PET)是最主要的MPs类型.所有样本中共检出18类MRGs,其中水样中MRGs的相对丰度为1.68±0.21.最主要的MRGs亚型是merR和ruvB,分别为汞和多重金属抗性基因的亚型.相关性分析表明,水中的铬和镍与MRG-Cr、MRG-Ni和多重金属抗性基因之间存在显著正相关.MPs上MRGs的相对丰度为1.63±0.53,最主要的MRGs亚型...     

荒漠绿洲土壤抗生素抗性基因分布特征及驱动机制

荒漠绿洲农田生态系统是干旱区环境下人类活动显著的复合生态系统.土壤微生物抗生素抗性与人类健康和生态平衡关系密切.研究荒漠绿洲环境不同土地利用类型模式下土壤抗生素抗性基因的多样性、分布特征和影响因素,对于评估干旱区土壤环境健康风险,促进绿洲农业生态的发展具有重要意义.采用高通量测序和高通量定量PCR技术对荒漠绿洲土壤微生物的群落结构和抗生素抗性基因多样性开展了研究,旨在探究干旱区土壤抗性基因的分布特征及其驱动机制.结果表明,从沙漠边缘到绿洲,荒漠沙生植物土壤、棉花地土壤、玉米地土壤、芦苇地土壤和湖泊沉积物中抗生素抗性的种类和丰度显著增加,与土地利用变化关系密切,农田土壤是抗性基因的重要存储库;荒漠绿洲土壤微生物群落与抗生素抗性基因显著相关,硫杆菌属(Thiobacillus)、沙漠细菌属(Pontibacter)、诺卡氏菌属(Nocardioides)、耐盐微杆菌属(Salinimicrobium)、土壤红杆菌属(Solirubrobacter)和链霉菌属(Streptomyces)等是各类抗性基因重要的潜在携带者;干旱区土壤中重(类)金属元素和可移动基因元件,与微生物群落共同塑造了抗生...     

Function of arsATorf7orf8 of Bacillus sp. CDB3 in arsenic resistance

Bacillus sp. CDB3 isolated from an arsenic contaminated cattledip site possesses an uncommon arsenic resistance (ars) operon bearing eight genes in the order of arsRYCDATorf7orf8. We investigated the functions of arsA, arsT, orf7 and orf8 in arsenic resistance using a plasmid-based gene knockout approach in the ars genedeficient Escherichia coli strain AW3110. The CDB3 arsA genewas shown to play a significant role in resistance, suggesting that the encoded ArsA may couplewith the arsenite transporter, forming an ArsAY complex that can enhance arsenite extrusion efficiency. Thedisruption of either arsT or orf7was not observed to affect arsenic resistance in the heterologous E. coli host, but their involvement in arsenic resistance can not be excluded. The orf8 gene is predicted to encode a putativedual-specificity protein phosphatasewhich also shares certain homology to arsenate reductases. The function loss of orf8 resulted in a remarkabledecrease in resistance to arsenate, though not arsenite. To examine if this effectwasdue to the reduction of arsenate by orf8, the arsC genewithin the 8-gene operonwasdisrupted. The resulting abolishment of arsenate resistance suggests that the involvement of orf8 in arsenic resistance is not via reductase activity.     

九龙江下游水源水中新发病原微生物和抗生素抗性基因的定量PCR检测

新发水传病原微生物和抗生素抗性基因对于饮用水的生物安全性有重要意义.以7种典型的水传病原微生物和4种抗生素抗性基因作为检测指标,对闽西南地区的重要水源九龙江下游水源水进行了检测.从九龙江下游4个采样点和附近某水厂进出水采集水样,用0.22μm的滤膜过滤后,进行DNA提取及荧光定量PCR检测.结果表明,九龙江下游水源水中Salmonella enterica(Salmonella spp.)、Legionella pneumophila(L.pneumophila)和Pseudomonas aeruginosa(P.aeruginosa)等个别指标可达到103、104、105copies.mL-1的水平.有机物浓度可能对病原微生物和抗生素抗性基因的水平有较大影响.水厂常规工艺对于大多数病原微生物和抗生素抗性基因可以实现有效去除,但对Salmonella spp.的去除效果不理想.因此为保证饮用水安全,需要在清理污染源、提高处理工艺水平以及末端控制方面加强管理.     

华北平原典型城市土壤微生物群落时空变化规律及其驱动因子

微生物群落结构和功能受多种环境因素的共同影响,为阐明典型城市土壤中微生物群落的时空变化规律及其主要影响因素,亟需开展典型城市土壤中微生物群落时空动态变化特征研究.鉴于此,选取华北平原典型城市——石家庄市为研究区,共设置12个采样点,分别于2021年6月（夏季）和9月（秋季）采集表层土壤,并利用16S rRNA高通量测序技术对土壤中微生物群落结构及功能进行研究,探究其时空变化规律;同时运用Pearson相关性分析建立微生物群落与环境因子间的相关性,识别微生物群落时空变化的主要驱动因子.结果表明：①石家庄市表层土壤中主要优势菌门为放线菌（Actinobacteriota）和变形菌（Proteobacteria）;就季节变化而言,在门水平上,Actinobacteriota和Proteobacteria相对丰度均呈降低趋势;在属水平上,夏季主要优势菌属为节杆菌属（Arthrobacter）和未知分类菌属,秋季主要优势菌属为Arthrobacter和Candidatus_Nitrocosmicus,且优势菌属呈显著季节差异（P<0.05）;②就季节变化而言,Simpson、Ace和Chao指数均值呈升高趋势,而OTU均值呈降低趋势;就空间差异而言,Shannon指数和Simpson指数呈显著空间差异（P<0.01和P<0.05）;③各类功能基因无显著季节和空间差异;其中能源生产和转换类功能基因相对丰度最高,在夏季和秋季的相对丰度分别为24.06%~24.84%和24.63%~25.98%;④微生物群落结构组成、多样性指数和功能基因与喹诺酮类抗生素（QNs）、总磷（TP）和硝氮（NO₃^--N）呈显著相关,且与QNs显著相关性最强（|r| >0.900）;这表明在石家庄市表层土壤中抗生素成为影响微生物群落变化的主要驱动因素.因此,为保障城市土壤中微生物群落结构和功能稳定,应进一步加强土壤中抗生素污染的综合管控.