In vitro gossypol induced spermatozoa motility alterations in rabbits

The objectives of the present study were to: (i) examine the in vitro dose response of rabbit spermatozoa motility to the antifertility agent gossypol (GOS) and (ii) determine whether filtered (FIL) and unfiltered (UNFIL) GOS differ in their magnitude of effect. Rabbit semen belonging to adult males (n = 5; 12–14 months) were cultured with UNFIL GOS and FIL GOS (5% solution) and subsequently diluted (1:1–7) for analysis using a Computer Assisted Semen Analyzer (CASA) system in 5 time periods (0, 60, 120, 180 and 360 minutes). At Time 0, no significant change in rabbit spermatozoa motility (MOT) and progressive motility (PROG) with GOS FIL was noted, while increases were observed with GOS UNFIL. At Time 60, weak changes were noted for MOT and PROG. After 120 minutes of culture with both GOS FIL and GOS UNFIL, MOT and PROG decreased significantly in some experimental groups. However, no differences were recorded for both the parameters at Times 180 and 360, with the exception of PROG in the GOS UNFIL category (groups A, B, E, F and G), where a significant decrease was noticed. Detailed evaluation of the distance and velocity parameters revealed reduction in all these studied markers after 60 and 120 minutes of in vitro culture with both GOS FIL and GOS UNFIL, indirectly confirming the PROG decrease. Straightness (STR), linearity (LIN), wobble (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) mostly remained unaltered at all time periods for GOS FIL, where as some minor alterations were noticed in GOS UNFIL category for STR, LIN, WOB, ALH and BCF parameters at Time 0, 60 and 120. The present study confirms the dose and time dependent alterations of rabbit spermatozoa motility parameters by GOS. The GOS dynamics in our experiment shows that rabbit spermatozoa as a biological material can indicate a GOS inhibition of motility. Obtained data for the first time indicates a higher immobilizing potential of unfiltered GOS in comparison to filtered GOS in its inhibitory action of spermatozoa motility parameters in rabbits.     

Field Detector Evaluation of Organic Clay Soils Contaminated with Diesel Fuel

Due to numerous types of uncontrolled petroleum releases into the environment such as leaking storage tanks, spills, and improper disposal of petroleum wastes, there is a need for quicker, more efficient methods to determine the levels of soil contamination for site remediation. The portable field detectors used most often in preliminary site evaluations are the flame ionization detector (FID) and the photoionization detector (PID). This research explored the relationship between these two instruments in analysis of two clay soil sites contaminated with diesel fuel. As in previous research, a log-log linear correlation was found between the PID and FID instruments for diesel fuel-contaminated soil at each site (R 2 > 0.91). Also, the correlation factors (0.64 and 0.60) between the field instruments at each site were found to be similar. It was asserted that either field instrument can be used to delineate the diesel fuel contamination at that site based upon a previously calculated correlation between the two instruments, and an overall numerical correlation between the field instruments can be used at various sites of similar soil and contamination characteristics. The implementation of the relationships between these two instruments could facilitate and accelerate site characterization in the future.     

堆肥对土壤中石油烃的去除及微生物群落的影响

为研究堆肥对石油污染土壤中不同组分烃的去除作用及土壤微生物群落结构变化的影响,利用重量法和GC-MS测定土壤中总石油烃、烷烃和多环芳烃的含量,采用高通量测序技术研究了堆肥对土壤微生物群落结构和多样性的影响作用.结果表明,向石油污染土壤中施加堆肥进行42d的修复处理,土壤中石油烃、烷烃、多环芳烃去除率分别为（12.4±0.01）%、（10.2±0.01）%、（9.38±0.02）%;自然放置的土壤中3种烃去除率分别为（3.21±0.02）%、（-3.00±0.01）%、（-6.59±0.02）%.自然放置的土壤香农指数、ACE指数和Chao1指数分别为4.30、3489.3和2691.0,加入堆肥进行修复处理后,土壤香农指数、ACE指数和Chao1指数分别增加为5.80、4684.7和3851.8.油污土壤中放线菌门（Actinobacteria）所占丰度由47.3%降低为28.2%,拟杆菌门（Bacteroidetes）丰度由0.78%增加至16.2%.变形菌门（Proteobacteria）丰度为37.4%,修复结束后几乎不变.属水平上,油污土壤中的优势菌属包括原小单孢菌属（promicromonospora）、微小杆菌属（Exiguobacterium）、诺卡氏菌属（Nocardioides）、分支杆菌属（Mycobacterium）、柠檬酸细菌属（Citrobacter）.施入堆肥使土壤中的这些优势菌属丰度降低,土壤中出现氮单胞菌属（Azomonas）、藤黄单胞菌属（Luteimonas）、假鞘氨醇杆菌属（Pseudosphingo bacterium）、紫单胞菌属（Parapedobacter）等新菌属.研究结果表明,与自然放置的土壤相比,向石油污染土壤中施入有机堆肥可有效去除土壤中的石油烃、烷烃和多环芳烃.并使土壤微生物群落结构发生明显变化.     

Morphological and molecular diversity of arbuscular mycorrhizal fungi in revegetated iron-mining site has the same magnitude of adjacent pristine ecosystems

Arbuscular mycorrhizal fungi(AMF) are important during revegetation of mining sites, but few studies compared AMF community in revegetated sites with pristine adjacent ecosystems. The aim of this study was to assess AMF species richness in a revegetated iron-mining site and adjacent ecosystems and to relate AMF occurrence to soil chemical parameters. Soil samples were collected in dry and rainy seasons in a revegetated iron-mining site(RA) and compared with pristine ecosystems of forest(FL), canga(NG),and Cerrado(CE). AMF species were identified by spore morphology from field and trap cultures and by LSU r DNA sequencing using Illumina. A total of 62 AMF species were recovered, pertaining to 18 genera and nine families of Glomeromycota. The largest number of species and families were detected in RA, and Acaulospora mellea and Glomus sp1 were the most frequent species. Species belonging to Glomeraceae and Acaulosporaceae accounted for 42%–48% of total species richness. Total number of spores and mycorrhizal inoculum potential tended to be higher in the dry than in the rainy season, except in RA. Sequences of uncultured Glomerales were dominant in all sites and seasons and five species were detected exclusively by DNA-based identification. Redundancy analysis evidenced soil p H,organic matter, aluminum, and iron as main factors influencing AMF presence. In conclusion, revegetation of the iron-mining site seems to be effective in maintaining a diverse AMF community and different approaches are complementary to reveal AMF species, despite the larger number of species being identified by traditional identification of field spores.     

典型生活垃圾填埋场覆盖土微生物群落分析

采用第二代高通量测序技术Illumina MiSeq对典型生活垃圾填埋场覆盖土样(山东莱芜,SD;广东深圳,GD;上海老港,SH;重庆长生桥,CQ)进行16S rDNA V3~V4区高通量测序,并分析了Alpha多样性、物种组成和丰度、菌群结构及环境因子对群落结构的影响.结果表明:取自垃圾填埋场GD土样的物种种类多于其他土样,GD、SD、SH、CQ土样的Shannon指数分别为5.52±0.026、4.76±0.030、4.89±0.037、3.43±0.027;所有覆盖土样的优势菌为Alphaproteobacteria(α-变形杆菌纲)和Betaproteobacteria(β-变形杆菌纲),所占比例范围分别为12.67%~25.54%,14.35%~18.88%;SD、GD和SH三种覆盖土样的优势菌为Sphingomonas(鞘氨醇单胞菌属),分别占7.25%、10.67%、11.30%; Deltaproteobacteria(德耳塔变形杆菌纲)和Gammaproteobacteria(γ-变形菌杆纲)的相对丰度分别与TN(r=1.00,P<0.001)和TP(r=1.00,P<0.001)呈正相关关系,且结合RDA图,TN、TP和OM含量可能是SD土样区别于其他土样群落组成的主要因素.     

不同材质水窖沉积物中真菌群落差异及其与环境因子的相关性

集雨窖水沉积物中蕴含丰富多样的真菌群落,这些真菌作为连接窖水生态系统与沉积物之间物质交换的桥梁和纽带,在窖水生态系统中发挥着重要作用;真菌群落结构特征的变化也通常与环境因子的变化有着紧密的联系.运用16S rRNA基因-Illumina MiSeq高通量测序技术,对两种不同集流面环境下的水窖沉积物中真菌群落多样性及差异进行研究.结果表明,混凝土集流环境下的水窖沉积物较黄土地集流环境下的水窖沉积物具有更高的真菌群落多样性与丰富度,两种集流环境下的真菌群落优势菌门相同,分别为子囊菌门（Ascomycota）、担子菌门（Basidiomycota）和接合菌门（Zygomycota）,它们构成了菌门丰度的90%以上,但前者具有更好的均一性和稳定性.基于LEfSe分析得到的标志物种显示,在黄土地集流环境中对差异性贡献最大的菌群是Basidiobolales,贡献最小的是Mycosphaerella;在混凝土集流环境中对差异性贡献最大的菌群是Saccharomycetales,贡献最小的是Periconia.微生物群落-环境因子共现性网络显示真菌和真菌之间,真菌和环境因子之间均为正向关系大于负向关系.研究结果加深了对水窖沉积物中真菌群落多样性的认识,可为保障集雨窖水人畜饮用安全和改善窖水水质提供了借鉴.     

沼液施用对麦稻茬口期土壤微生物群落结构特征及功能的影响

为揭示麦稻茬口期沼液施用对土壤微生物群落及功能的影响,通过土柱试验,设置麦秆还田后3种处理［无氮肥（CM）、常规施肥（SN）和添加沼液（SZ）］,采用Illumina高通量测序技术分析了土壤淹水后1 d和21 d土壤细菌及真菌群落的组成、多样性和结构差异,并对其功能进行预测.微生物多样性分析表明,1 d时各处理的真菌α多样性指数均显著高于21 d处理且各处理间无显著差异;21 d时各处理细菌Simpson指数开始出现差异,SZ-21处理的细菌Simpson指数高于SN-21处理,而细菌Chao1指数显著低于SN-21处理;细菌群落结构分析表明,1 d时,SN处理的厚壁菌门、绿弯菌门和放线菌门与其它处理差异较大,而21d时,SZ和SN处理的各细菌菌门相对丰度相似;真菌群落结构分析表明,1 d时,SZ处理中的子囊菌门和接合菌门相对丰度高于SN和CM处理;21 d时,SN和SZ处理的子囊菌门相对丰度均低于CM处理,而接合菌门的相对丰度均高于CM处理.NMDS分析表明,21 d时,SN和SZ处理的细菌和真菌群落组成均有逐渐相似的趋势.PICRUSt功能分析表明,不同时期和不同处理的土壤细菌群落在功能层上表现相似,FUNGuild功能预测反映SZ-21和SN-21处理的真菌功能主要差异体现在腐生营养型和病理营养型上.综上说明茬口期施沼液在一定程度上可以替代化肥平衡土壤养分和维持土壤生态功能,但仍可能存在真菌致病风险.     

利用高通量测序对封存CO2泄漏情景下土壤细菌的研究

全球CO2浓度不断升高的背景下,地质封存CO2为控制大气CO2浓度上升提供了全新的思路,但封存CO2潜在的泄漏风险也对生态系统产生重大威胁.土壤微生物与土壤健康密切联系.鉴于目前地质封存CO2泄漏对土壤微生物影响的研究鲜有报道,本研究通过模拟地质封存CO2泄漏情景,利用Miseq平台Illumina第二代高通量测序土壤细菌16S rRNA基因,结合相关生物信息学分析,初步探讨了封存CO2泄漏情景下农田土壤细菌群落丰富度、多样性和群落结构的变化.试验共获得15个土壤样本的43 017个OTU共486 645条读数,生物学分析结果表明,封存CO2泄漏通量和时间不同的情景下,农田土壤细菌丰富度和细菌群落多样性的变化存在差异,初步推测CO2泄漏量增大和泄漏时间持续更长的情景下,土壤细菌丰富度和多样性下降幅度较大,土壤细菌群落中优势菌群向若干细菌集中,其中土壤酸杆菌门细菌相对增多可能作为地质封存CO2泄漏对土壤生态系统影响的生物监测指标.     

菲反硝化降解菌群的富集及其群落结构解析

从潜在多环芳烃(Polycyclic Aromatic Hydrocarbons,PAHs)污染的油田区域采集土壤样品,以菲为唯一碳源且添加硝酸根的培养基来富集土壤中的菲反硝化降解菌群.随后,通过定量PCR(Polymerase Chain Reaction)测定了获取的富集菌群中反硝化相关功能基因(硝酸还原酶基因nar G、亚硝酸还原酶基因nir S)的丰度,并通过Illumina Mi Seq测序对其中的细菌群落结构进行解析.结果表明,获取到的3个菌群(PDN-1、PDN-2和PDN-3)12 d内对菲的降解率分别为45.18%、34.04%和25.92%.各富集培养菌群中nar G的丰度均高于nir S,且菲降解率最高的PDN-1中的反硝化相关基因丰度较低.Illumina Mi Seq测序结果表明,菲降解率最高的富集菌群PDN-1同时也具有较高的细菌多样性指数,变形菌门(Proteobacteria)、疣微菌门(Verrucomicrobia)和拟杆菌门(Bacteroidetes)是各富集菌群中的优势菌门,且Proteobacteria在3个富集菌群PDN-1(97.78%)、PDN-2(96.57%)、PDN-3(93.90%)中的比例均最高.变形菌门的Pseudomonas(γ-Proteobacteria)和Methylophilus(β-Proteobacteria)则是各富集菌群中最大的优势菌属,前者为公认的PAHs降解菌,而后者则为能够利用还原型"一碳化合物"的特殊菌属.细菌多样性与菲的降解率呈正相关,表明菲的反硝化降解可能是多种细菌参与的共同结果.上述结果可为揭示典型PAHs反硝化降解的微生物机制提供理论依据,同时为深入研究反硝化与菲代谢的偶联机理打下基础.     

李家河水库春季分层期nirS型反硝化菌群特征分析

为探究水源水库春季分层期（3～5月）nirS型反硝化细菌群落结构组成及其与水质的关系,以李家河水库为例,采用原位监测耦合Illumina高通量测序技术,分析了水体水质及反硝化细菌群落相对丰度及结构特征.结果表明:（1）通过高通量测序,鉴定为4门13属,优势菌门为变形菌门（Proteobacteria）,相对丰度在52.5%～70.6%,时间尺度总体呈降低趋势（P<0.05）,空间尺度上表层和中层高于底层（P<0.05）,表层与中层无差异（P>0.05）;识别出具有反硝化功能细菌8属,其中优势菌属（相对丰度>1%）为脱氯单胞菌属（Dechloromonas）和假单胞菌属（Pseudomonas）,脱氯单胞菌属相对丰度在时间上呈先降低后升高的趋势,假单胞菌属在时间上呈先升高后降低的趋势,此2属在空间上无差异（P>0.05）;细菌多样性及丰富度变化基本一致,时间上呈先升高后减低的趋势,空间上随深度逐渐升高;（2）本研究期间水库水体ρ（总氮）为2.35～2.91 mg·L^-1,氮素污染较为严重,3月和4月垂向水体总氮基本一致且呈降低趋势,5月...