邻苯二甲酸二甲酯及其异构体的好氧微生物降解

3种苯二甲酸二甲酯异构体(邻、间和对苯二甲酸二甲酯)主要应用于化学工业,作为增塑剂和生产聚酯的原料。用邻苯二甲酸二丁酯为惟一碳源,从红树林底泥中驯化、富集、培养、分离得到的微生物对邻苯二甲酸二甲酯(Dimethylphthalate,DMP)及其异构体对苯二甲酸二甲酯(Terephthalate,DMT)和间苯二甲酸二甲酯(Isophthalate,DMI)具有较强的降解作用。此菌株16SrDNA分子生物学的鉴定为Rhodococcusruber1k。实验得出该菌能够在苯二甲酸二甲酯作为惟一碳源和能源的培养基中生长。浓度为50mg·L-1的DMP、DMI和DMT分别在6、10、11d内可以完全被降解;DMP能够在好氧条件下被该菌快速降解,生成邻苯二甲酸一甲酯(monomethylphthalate,MMP)和邻苯二甲酸(phthalicacid,PA)2种主要中间产物,最终可以完全矿化成CO2和H2O;该菌对DMI和DMT的降解速度则比DMP慢。两者的降解中间产物间苯二甲酸一甲酯(MMI)和对苯二甲酸一甲酯(MMT)却不能被Rhodococcusruber1k继续降解而在培养基中积累。结果表明苯二甲酸二甲基酯的3种异构体能够被红树林底泥中的土著微生物降解。降解速度及降解途径与底物的化学结构有密切关系。     

中国南海Bukholderia sp.DA2菌株降解邻苯二甲酸二甲酯研究

以邻苯二甲酸二甲酯(dimethyl phthalate,DMP)为惟一碳源,从中国南海底泥中分离筛选到一株高效降解菌株,即伯克霍尔德氏菌Bukholderia.sp.DA2.该菌株降解DMP及其邻苯二甲酸二甲酯(Dimethyl phthalate,DMT)和间苯二甲酸二甲酯(Dimethyl dimethyl isophthalate,DMI)2种异构体的效果显著.B.sp.DA2菌株能够在8d内好氧降解矿化100mg·L-1的DMT,15d内好氧降解矿化400mg·L-1的DMP.但对于200mg·L-1的DMI,则不能彻底降解矿化,反应终止在生成间苯二甲酸一甲酯中间产物的阶段.研究表明,B.sp.DA2菌株降解DMT及DMP的途径相似,首先脱酯生成单酯和甲醇,继而水解酸化,直至彻底矿化.降解速率及其途径,与化学结构密切相关.     

冬季刺参养殖环境与肠道内细菌菌群的研究

运用传统细菌分离培养与分子生物学技术相结合的方法,对2008年11月至2009年1月冬季刺参(Apostichopus japonicus Liao)养殖池塘环境(养殖水、底泥、附着基)及刺参肠道内的细菌菌群进行了分析。应用平板稀释涂布培养计数法测得刺参养殖池塘水体、底泥、附着基和肠道细菌数量分别为0.75×102～1.4×104cfu/mL、8.7×104～8.1×105cfu/g、3.8×105～2.8×106cfu/g、7.1×105～1.5×107cfu/g;根据形态学差异从培养所得的细菌中筛选得到22株菌,用限制性内切酶Rsa I和Msp I对所分离菌株进行ARDRA(Amplified rDNA Restriction Analysis)分析,这22株菌被分为8种不同的分类单元(Operational Taxonomic Unit,OTU),其中OTU2与OTU3所包含的菌株分别占分离菌株种数的30%和20%;此外,作者对不同环境培养所得的优势度最高的细菌进行分子鉴定分析。结果表明:水环境中优势菌为施氏假单胞菌(Pseudomonas stutzeri)、门多萨假单胞菌(Pseudomonas mendocina)及巨大芽孢杆菌(Bacillus megaterium),沉积物中优势菌为巨大芽孢杆菌(B.megaterium)、施氏假单胞菌(P.stutzeri)、苏云金芽孢杆菌(Bacillus thuringiensis),附着基中优势菌为巨大芽孢杆菌(B.megaterium)、苏云金芽孢杆菌(B.thuringiensis),灿烂弧菌(Vibrio splendidus),肠道中优势菌为巨大芽孢杆菌(B.megaterium)、苏云金芽孢杆菌(B.thuringiensis),灿烂弧菌(V.splendidus)、施氏假单胞菌(P.stutzeri)。通过对冬季刺参池塘细菌菌群多样性分析和优势菌鉴定,为筛选低温益生菌和防治刺参疾病提供了有益参考。     

褐藻酸降解酶特性的初步研究

对从海带(Laminaria japonica)病烂处分离出的2株别单胞菌属的褐藻酸降解菌(简称菌株A1,A2)进行了褐藻酸钠降解能力及胞外产物中褐藻酸酶特性的初步分析。结果表明,2株褐藻酸降解菌中,菌株A2表现出较强的对褐藻酸钠的降解能力;同时菌株A2所分泌的褐藻酸酶在pH 6.0~8.0相对稳定,30℃以下酶不易失活。在较低离子浓度时,Mn2 ,Ba2 对反应有较强的促进作用,Ag 和Pb2 则有明显的抑制作用。     

文蛤鳃组织内共生菌分离鉴定和多样性分析

为探究文蛤（Meretrix petechialis）鳃组织内共生菌群的群落组成及结构，采用纯培养方法，对文蛤鳃组织的内共生菌进行了分离、纯化和16S rRNA鉴定，共筛选到215株细菌，分属于变形菌门、厚壁菌门、拟杆菌门和放线菌门4个门级类群中，其中变形菌门是实现纯培养菌株数量最多的类群。基于16S rRNA序列信息，进一步对变形菌门的系统演化关系进行了分析，结果显示不同纲级类群的菌株分别聚集在了相应的分支内。另外，对获得的具有潜在抑菌活性或致病性的共生细菌进行了探讨，为后续对文蛤鳃内生菌的深入研究奠定了基础。     

菲降解菌的分离鉴定及其降解性研究

采用微生物富集培养分离法对南洋油田不同油样中的菲降解菌进行了分离鉴定。分离得到2株以菲作为唯一碳源和能源的细菌菌株S17和S28,经革兰氏染色及显微镜形态观察,发现两者都是具有极生单鞭毛的革兰氏阴性、无芽孢杆菌。根据生理生化特性分析,以及16S rDNA序列同源性分析,两者属于假单胞菌属的不同种。菌株S17与食树脂假单孢菌(Pseudomonas resinovorans)序列同源性为97%;菌株S28则为高温假单胞菌(Pseudomonas thermaerum),其序列同源性达到100%。在以菲为唯一底物的条件下,菌株S28的生长速度是S17的2倍多,第4天即达到最大生长量,但是对于菲的降解而言,两者都在第10天达到最大,降解率分别为88.86%和82.02%,但是二者在起始的两天内对菲的降解效率最高,分别达到70.21%和72.74%,因此可用于菲污染的快速治理领域。     

养殖鲟鱼出血症病原鲁氏耶尔森菌的分离鉴定和致病性研究

鲁氏耶尔森菌(Yersinia ruckeri)是鲑鳟鱼类和温水性鱼类肠炎红嘴病的主要病原,可引起病鱼体表出血、肠道肿胀发炎等临床症状。本研究从患出血症施氏鲟(Acipenser schrenckii)中分离到一株致病性菌株(YR-H01),生理生化鉴定和16S rRNA基因序列分析表明YR-H01株为鲁氏耶尔森菌。人工感染该菌后发病鱼表现为肛门红肿、外突明显,部分鱼鳍基部和下颌处有出血现象,内脏器官不同程度出血,且从组织中再分离的细菌特性与原感染菌相同。腹腔注射后该菌株对施氏鲟的半致死浓度为7.2×106CFU,且攻毒剂量越大,临床病症出现越快。病理组织切片显示,该菌感染鲟鱼后肝细胞发生明显病变,淋巴细胞侵润,肝索结构消失,细胞肿胀,核空泡变性或核仁边缘化。     

深海沉积物中龙须菜降解菌群的筛选与鉴定及多样性分析

以龙须菜为唯一的营养源对深海沉积物进行富集和筛选,获得一个可降解龙须菜(Gracilaria lemaneiformis)、紫菜(Porphyra umbilicalis)和海带(Laminaria japonica)产生还原性寡糖的菌群.通过16S rRNA序列及16S rRNA-RFLP分析对该菌群中的细菌多样性进行了研究.结果表明,降解菌群中的细菌组成主要为弧菌属(Vibrio,8株)、火色杆菌属(Flammeovirga,7株)和希瓦氏菌属(Shewanella,5株),其中希瓦氏菌属和火色杆菌属的细菌在菌群中的丰度较高.采用龙须菜为唯一营养源的筛选培养基对菌群中的细菌进行分离,获得4株具有琼胶酶活力的细菌,包括2株火色杆菌属和2株希瓦氏菌属细菌.培养和未培养的结果均表明火色杆菌和希瓦氏菌这2个属为所研究的深海沉积物主要的龙须菜降解菌.对原始降解菌群和所分离的关键菌株进行了龙须菜酶解产糖能力的比较,结果发现菌群中的弧菌属菌株虽然自身没有酶解龙须菜的能力,但可能可以协助关键菌株,提高菌群对龙须菜的降解效率.因此本研究中所获得的菌群和菌株有望在琼胶寡糖的绿色生产中得到广泛应用.     

工厂化养殖杂色鲍致病菌副溶血弧菌的分离鉴定及其生理特性

从广东汕尾地区工厂化养殖的患病和死亡的杂色鲍(Haliotis diversicolor)及养殖海水中分离到3株优势菌,经回归感染试验,证实所分离的细菌为杂色鲍的致病菌。经形态、生理、生化等多项指标鉴定,该菌株为副溶血弧菌(Vibrio parahaemolyticus),并经ATB微生物自动鉴定系统证实鉴定结果。在中国大陆,由副溶血弧菌使杂色鲍致病并大量死亡的事件,本文属首次报道。     

一种AHL降解菌株的高效筛选方法

酰化高丝氨酸内酯化合物(acyl-homoserine lactone,AHL)是革兰氏阴性细菌(G-)群体感应的信号分子。本文建立1种从环境微生物中高效地筛选AHL降解菌株的方法。首先利用AHL作为唯一能源对环境微生物进行初筛,再利用顶层琼脂法和报告菌株Chromobacterium violaceumATCC 12472、Agrobacterium tumefaciensA136等进行复筛,共筛选得到21株具有AHL降解活性的菌株。对其中活性最高的菌株A11进行初步的菌种鉴定,通过形态观察、16S ribosomal RNA(16S rDNA)基因分析最终确定为假单胞菌属细菌Pseudomonassp.A11。     

VE对河蟹血清和组织中超氧化物歧化酶及磷酸酶活性的影响

通过投喂添加VE 量分别为 0、0 .1× 1 0 - 3 、0 .2× 1 0 - 3 、0 .4× 1 0 - 3 、0 .6× 1 0 - 3(m m)的 5种实验饲料饲养河蟹 6 0d ,以探讨VE 对其体内SOD、ALP和ACP等酶活性的影响 .结果表明 ,饲料中未添加VE 时 ,河蟹血清、肝胰腺、卵巢和肌肉各组织中SOD、ALP和ACP活性各异 ,其中SOD的活性从高到低依次为肝胰腺 >卵巢 >血清 >肌肉 ,ALP活性的为肝胰腺 >血清 >卵巢 >肌肉 ,而ACP活性则是肝胰腺 >血清 >肌肉 >卵巢 .饲料中添加VE 时 ,与对照组相比 ,各组织中SOD活性随着VE添加量的升高而显著降低 ( p <0 .0 5或 p <0 .0 1 ) ;ALP和ACP活性则随着VE 添加量的增加而显著升高 ( p <0 .0 5或 p <0 .0 1 ) .这表明VE 作为天然抗氧化剂在河蟹体内发挥了很大的作用 ,并促进机体的新陈代谢 ,从而增强机体的非特异性免疫力 .VE 作为免疫刺激剂能有效地增强河蟹的非特异性免疫能力 ,能有效发挥其增强河蟹免疫功能的VE 适宜添加量为 ( 0 .2～ 0 .4)× 1 0 - 3(m m) .     

Proposing a coordinated environmental effects monitoring (EEM) program structure for the offshore petroleum industry,Nova Scotia,Canada

Environmental effects monitoring (EEM) identifies, quantifies, and compares predicted environmental impacts of a proposed industrial project to those observed after the project has been developed. Several EEM program structures exist, while the structure used by Environment Canada to monitor impacts of the pulp and paper industry has proven to be effective. The aim of this study is to provoke discussion on the development of an effective and coordinated petroleum EEM program structure for the offshore of Nova Scotia, Canada. A structure that supports development of a national EEM program for the petroleum industry is then proposed. Advantages of a coordinated EEM program structure include: (1) reduced cost of the EEM; (2) fewer gaps and less overlapping of responsibilities; (3) fewer calculated risks to the marine environment; and (4) improved environmental safeguards.     

Sources and implications of particulate organic matter from a small tropical river——Zuari River,India

Transitional ecosystems,estuaries and the coastal seas,are distinctively affected by natural and anthropogenic factors.Organic matter(OM)originating from terrestrial sources is exported by rivers and forms a key component of the global biogeochemical cycles.Most previous studies focused on the bulk biochemical and anthropogenic aspects affecting these ecosystems.In the present study,we examined the sources and fate of OM entrained within suspended particulate matter(SPM)of the Zuari River and its estuary,west coast of India.Besides using amino acid(AA)enantiomers(L-and D-forms)as biomarkers,other bulk biochemical parameters viz.particulate organic carbon(POC),δ13C,particulate nitrogen(PN),δ15N and chlorophyll a were analyzed.Surprisingly no significant temporal variations were observed in the parameters analyzed;nonetheless,salinity,POC,δ13C,PN,δ15N,glutamic acid,serine,alanine,tyrosine,leucine and D-aspartic acid exhibited significant spatial variability suggesting source differentiation.The POC content displayed weak temporal variability with low values observed during the post-monsoon season attributed to inputs from mixed sources.Estuarine samples were less depleted than the riverine samples suggesting contributions from marine plankton in addition to contributions from river plankton and terrestrial C3 plants detritus.Labile OM was observed during the monsoon and post-monsoon seasons in the estuarine region.More degraded OM was noticed during the pre-monsoon season.Principal component analysis was used to ascertain the sources and factors influencing OM.Principally five factors were extracted explaining 84.52%of the total variance.The first component accounted for 27.10%of the variance suggesting the dominance of tidal influence whereas,the second component accounted for heterotrophic bacteria and their remnants associated with the particulate matter,contributing primarily to the AA pool.Based on this study we ascertained the role of the estuarine turbidity maximum(ETM)controlling the sources of POM and its implications to small tropical rivers.Thus,changes in temporal and regional settings are more likely to affect the natural biogeochemical cycles of small tropical rivers.     

高压液相色谱分析外源类固醇在真鲷血液中的代谢

本研究利用高压液相色谱法（ＨＰＬＣ）法测定：外源睾酮（Ｔ）和甲基睾酮（ＭＴ）（１０μｇ／ｇ）腹腔注射给入真鲷（Ｐａｇｒｏｓｏｍｕｓｍａｊｏｒ）鱼体后，在血液中的代谢状况．证明，Ｔ和ＭＴ的吸收和清除很快。ＭＴ在注射１ｈ达最大吸收峰（２８３ｎｇ／ｍｌ），２４ｈ内很快降低。Ｔ的吸收和清除状况与ＭＴ相似，不同的是其吸收值（最大值为６４ｎｇ／ｍｌ）比ＭＴ低许多。随着鱼类养殖至商品规格，类固醇激素的含量将降至很低，因此，ＭＴ可做为一种生长促进因子用于水产养殖生产。     

A Late Winter Hydrography in Hidaka Bay, South of Hokkaido, Japan

Hydrographic observations in Hidaka Bay, south of Hokkaido, Japan were carried out in late winter 1996 and 1997 to examine the spatial distributions and circulation features of two different water masses, i.e., Coastal Oyashio Water (COW) and Tsugaru Warm Water (TWW), and their modifications. It is known that COW is mostly composed of cold and low-salinity water of the melted drift ice coming from the Okhotsk Sea and flows into Hidaka Bay from winter to spring and TWW with high-salinity continuously supplies from the Tsugaru Strait to the North Pacific. Cold surface mixed layers (<26.2σ_θ, 0–100 m depth) were found mainly over the shelf slope, confirming that anti-clockwise flow of COW was formed. TWW was relatively high in salinity and low in potential vorticity, and had some patch-like water masses with a temperature and salinity maximum in the limited area in the further offshore at the deeper density levels of 26.6–26.8σ_θ. The fine structure of vertical temperature and salinity profiles appeared between TWW and COW is an indication of enhanced vertical mixing (double-diffusive mixing), as inferred from the estimated Turner angles. At a mouth of the Tsugaru Strait in late winter 1997, a significant thermohaline front between TWW and the modified COW was formed and a main path of TWW spreaded south along the Sanriku coast, probably as the bottom controlled flow. Hence, the patch-like TWW observed in late winter is isolated from the Tsugaru Warm Current and then rapidly modified due to a diapycnal mixing. This revised version was published online in August 2006 with corrections to the Cover Date.     

固相吸附毒素跟踪技术(SPATT)在浙江南麂海域的应用

浙江南麂岛海域是我国近岸贝类的重点养殖区,近年来一直遭受脂溶性贝类毒素(lipophilic shellfish toxins,LSTs)的污染,威胁着人类健康。为了建立有效的食品安全预警方法,本研究利用固相吸附毒素跟踪技术(solid phase adsorption toxin tracking,SPATT)在该海域进行了为期一年的野外监测和LSTs毒素分析。结果表明:7种LSTs毒素在SPATT(DIAION?HP20)中被检出,分别是大田软海绵酸(okadaic acid,OA)、鳍藻毒素-1(dinophysistoxin-1,DTX-1)、虾夷扇贝毒素(yessotoxin,YTX)及其衍生物homo YTX、扇贝毒素-2(pectenotoxin-2,PTX-2)及其衍生物7-epi-PTX2sa和环亚胺毒素(gymnodimine,GYM);有8种毒素在厚壳贻贝(Mytilus galloprovincialis)中被检出,分别是OA、DTX1、homo YTX、PTX2、7-epi-PTX2sa、GYM、原多甲藻酸贝类毒素-3(azaspiracid-3,AZA-3)和虾夷扇贝毒素衍生物45-OH-homo YTX。整体而言,SPATT(HP20)中吸附的毒素种类与贝肉中监测出的毒素种类大体上一致,且两者间OA、DTX1和PTX2的浓度在时间上具有较好的相关性,可见SPATT(HP20)对这些LSTs毒素具有较高的灵敏度,有望作为水体中LSTs毒素的预警监测材料。南麂岛海域LSTs分布特征明显,夏季最高,其次是春秋季,冬季最低,其中2014年夏季贝肉中OA毒素含量高达77.19ng/g贝肉,超出国家限量标准(45ng/g贝肉)比例达11.76%,为浙江南麂海域贻贝的食用安全带来隐患。     

栉孔扇贝不同组织SOD、CAT和MPO活力的比较分析

对2龄和3龄栉孔扇贝外套膜、鳃、消化盲囊、肾、闭壳肌、性腺组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和髓过氧化物酶(MPO)活力分析表明,检测组织均存在这三种酶,且在不同组织中活力表现存在明显差别,其中肾组织SOD、CAT活力最高,而MPO在消化盲囊中表现最高活力,与其它组织相比均呈显著性差异(P＜0.05).不同贝龄的栉孔扇贝酶活力比较表明,三龄贝各组织SOD、CAT和MPO活力均高于二龄贝.     

Multivariate analysis of fluorescence and source identification of dissolved organic matter in Jiaozhou Bay, China

Hierarchical clustering analysis and principal component analysis (PCA) methods were used to assess the similarities and dissimilarities of the entire Excitation-emission matrix spectroscopy (EEMs) data sets of samples collected from Jiaozhou Bay, China. The results demonstrate that multivariate analysis facilitates the complex data treatment and spectral sorting processes, and also enhances the probability to reveal otherwise hidden information concerning the chemical characteristics of the dissolved organic matter (DOM). The distribution of different water samples as revealed by multivariate results has been used to track the movement of DOM material in the study area, and the interpretation is supported by the results obtained from the numerical simulation model of substance tracing technique, which show that the substance discharged by Haibo River can be distributed in Jiaozhou Bay.     

VC对河蟹血清和组织中超氧化物歧化酶及磷酸酶活性的影响

研究了饲料中添加不同含量VC 对河蟹体内超氧化物歧化酶 (SOD)、碱性磷酸酶 (ALP)和酸性磷酸酶 (ACP)活性的影响 .结果表明 ,饲料中不添加VC 时 ,河蟹肝胰腺、卵巢、血清和肌肉各组织中SOD、ALP和ACP活性各异 ,其中SOD活性大小顺序为肝胰腺 >卵巢 >血清 >肌肉 ,ALP活性顺序为肝胰腺 >血清 >卵巢 >肌肉 ,而ACP活性则为肝胰腺 >血清 >肌肉 >卵巢 .饲料中VC 添加量分别 2 .5× 1 0 - 3 、5.0×1 0 - 3 、1 0 .0× 1 0 - 3 和 1 5.0× 1 0 - 3 (m m)时 ,与对照组相比 ,河蟹不同组织中SOD活性显著下降 (p <0 .0 5或 p <0 .0 1 ) ,而ALP、ACP这两种酶的活性却显著升高 (p <0 .0 5或 p <0 .0 1 ) .但它们的变化幅度不同 ,其中就肝胰腺、卵巢、血清和肌肉中SOD活性而言 ,VC 添加量为 1 5.0× 1 0 - 3 (m m )饲料组分别比对照组的低1 41 .3 4 %、1 2 5.94%、1 1 6.67%和 1 1 5.79%;ALP活性 ,VC 添加量为 1 0 .0× 1 0 - 3(m m)饲料组分别比对照组高 93 .0 %、77.5%、1 66.3 %和 2 66.4%;ACP活性 ,VC的添加量为 1 5.0× 1 0 - 3 (m m)饲料组分别比对照组高 1 3 0 .7%、2 3 7.4%、43 7.6%和 2 65.5%.饲料中VC 添加量为 (5.0～ 1 0 .0 )× 1 0 - 3 (m m )时 ,可有效地增强河蟹非特异性免疫能力 .此外 ,本实验还就VC