Chitosan extends the shelf-life of filleted tilapia (Oreochromis niloticus) during refrigerated storage

Shelf-life extension of aquatic products is of significant economical importance. To determine the potential effect of chitosan on the shelf-life of filleted tilapia, this study analyzed the bacterial community diversity in fresh and spoiled tilapia fillets stored at (4 ± 1)℃ and examined the antimicrobial activity of chitosan against relevant bacteria isolates. Results showed that Pseudomonas (20%) and Aeromonas (16%) were abundant in fresh tilapia fillets, whereas Pseudomonas (52%), Aeromonas (32%) and Staphylococcus (12%) were dominant in the spoiled samples. Chitosan showed wide-spectrum antibacterial activity against bacteria isolated from tilapia and 5.0 g L-1 chitosan was selected for application in preservation. We further determined the shelf-life of chitosan-treated, filleted tilapia stored at (4 ± 1)℃ based on microbiological, biochemical and sensory analyses. Results showed that the shelf-life of chitosan-treated, filleted tilapia was extended to 12 d, whereas that of untreated, control samples was 6 d. These indicate that chitosan, as a natural preservative, has great application potential in the shelf-life extension of tilapia fillets.     

Generation of reactive oxygen species in different fractions of the coelomocytes of holothurian Eupentacta fraudatrix in response to the thermostable toxin of Yersinia pseudotuberculosis in vitro

Pure fraction (92%–95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38%)-FPMC- of the holothurianEupentacta fraudatrix' (Holothuroidea, Dendrochirota) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin ofYersinia pseudotuberculosis (TST) at different concentrations (0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation. In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively. Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared to that under toxin treatment alone. TST stimulated superoxide dismutase activity in concentration-dependent manner (maximum at 0.5 μg/ml concentration in FP) after 24 h treatment, and this stimulation was prevented by a commercial catalase. Plant lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously, ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production by these cells may be one of the mechanisms of antibacterial protection of holothurians. This work was financially supported by the Russian Foundation for Fundamental Research Grant (No. 00-04-48949).     

Design ofVibrio 16S rRNA gene specific primers and their application in the analysis of seawaterVibrio community

The pathogenic species of genus Vibrio cause vibriosis, one of the most prevalent diseases of maricultured animals and seafood consumers. Monitoring their kinetics in the chain of seafood production, processing and consumption is of great importance for food and mariculture safety. In order to enrich Vibrio-representing 16S ribosomal RNA gene （rDNA） fragments and identify these bacteria further real-timely and synchronously among bacterial flora in the chain, a pair of primers that selectively amplify Vibrio 16S rDNA fragments were designed with their specificities and coverage testified in the analysis of seawater Vibrio community. The specificities and coverage of two primers, VF169 and VR744, were determined theoretically among bacterial 16S rDNAs available in GenBank by using BLAST program and practically by amplifying Vibrio 16S rDNA fragments from seawater DNA. More than 88.3% of sequences in GenBank, which showed identical matches with VR744, belong to Vibrio genus. A total of 33 clones were randomly selected and sequenced. All of the sequences showed their highest similarities to and clustered around those of diverse known Vibrio species. The primers designed are capable of retrieving a wide range of Vibrio 16S rDNA fragments specifically among bacterial flora in seawater, the most important natural environment of seafood cultivation.     

Comparison of intestinal bacterial communities in grass carp, Ctenopharyngodon idellus, from two different habitats

The intestinal bacteria of vertebrates form a close relationship with their host.External and internal conditions of the host,including its habitat,affect the intestinal bacterial community.Similarly,the intestinal bacterial community can,in turn,influence the host,particularly with respect to disease resistance.We compared the intestinal bacterial communities of grass carp that were collected from farm-ponds or a lake.We conducted denaturing gradient gel electrophoresis of amplified 16S rRNA genes,from which 66 different operational taxonomic units were identified.Using both the unweighted pair-group method with arithmetic means clustering and principal component analysis ordination,we found that the intestinal bacterial communities from the two groups of pond fish were clustered together and inset into the clusters of wild fish,except for DF-7,and there was no significant correlation between genetic diversity of grass carp and their intestinal bacterial communities(Mantel one-tailed test,R=0.157,P=0.175).Cetobacterium appeared more frequently in the intestine of grass carp collected from pond.A more thorough understanding of the role played by intestinal microbiota on fish health would be of considerable benefit to the aquaculture industry.     

新疆乌鲁木齐二叠系湖相微生物白云岩成因

新疆乌鲁木齐地区养牛场剖面中二叠统芦草沟组以发育浅湖至半深湖背景下的中层深灰色、灰色碳酸盐岩、粉砂岩、细砂岩与中、厚层灰黑色泥岩、油页岩的互层沉积为特点.湖相碳酸盐岩以微晶白云岩为主体,其次为微晶灰岩.微晶白云岩主要由白云石、铁白云石及少量方解石等组成,常混有泥质组分且富含有机质.镜下观察白云石主要为微晶(＜4μm)及微亮晶(4-10 μm),极少数为亮晶(＞10μm).扫描电镜分析发现微晶白云岩中存在微球状(直径约9μm)、微棒状(长度约0.3～1.2μm)及微米级它形(＜ 5μm)等3种微形貌的白云石,其中微米级它形白云石在白云岩中占绝大多数.在微晶灰岩中还发现了直径约70-150nm,形态与球菌相似的纳米微粒,具有微生物矿化的特征.研究区白云岩Sr丰度及Sr/Ca比总体持平或略高于微晶灰岩,Mn丰度远高于微晶灰岩,C、O同位素均高于微晶灰岩,暗示了白云岩可能形成于比微晶灰岩更深及盐度更大的水下还原环境,二者之间缺乏明显的交代关系.芦草沟组白云岩的δ13CPDB介于9.2 ‰～15.6‰,强烈正偏的δ13CPDB可能是产甲烷古菌的代谢活动引起有机质碳同位素分馏的结果.以上特征表明,研究区芦草沟组白云石的沉淀可能与产烷带厌氧微生物的代谢活动引起的甲烷生成作用有关.     

伊犁蒙其古尔铀矿床含矿层砂岩中黄铁矿形成机制及对铀成矿的指示意义

蒙其古尔铀矿床为伊犁盆地南缘大型层间氧化带砂岩型铀矿床,为查明该矿床含矿层中黄铁矿成因及其形成机制,探讨微生物参与铀成矿过程。文章对含矿层砂岩中黄铁矿与铀矿物矿物学特征、黄铁矿S同位素与碳酸盐胶结物的C-O同位素开展细致研究。研究表明:①蒙其古尔铀矿床中铀主要以铀矿物与吸附铀形式存在,吸附铀主要为有机质吸附铀,铀矿物以沥青铀矿为主,多与黄铁矿、炭屑共生;②蒙其古尔铀矿床含矿层砂岩中黄铁矿主要以自形晶、草莓状和不规则状集合体产出,多与沥青铀矿、碳酸盐胶结物共生,其中黄铁矿S同位素(δ~(34)S_(V-CDT)=-68.4‰～22.1‰)与碳酸盐胶结物的C-O同位素(δ~(13)C_(V-PDB)=-10.2‰～-7.4‰,δ~(18)O_(V-PDB)=-9.6‰～-5.8‰)分析表明黄铁矿具有细菌硫酸盐还原(BSR)与有机物热解2种成因,并探讨了这2种不同成因黄铁矿的形成机制。③结合前人研究成果,认为硫酸盐还原菌(SRB)参与蒙其古尔铀矿床铀成矿过程,以间接还原方式为主,在有机质、黏土矿物与颗粒表面吸附U(Ⅵ)的基础上,通过硫酸盐还原菌(SRB)还原SO_4~(2-)产生的H_2S将U(Ⅵ)被还原成U(Ⅳ),形成铀矿物。     

海水养殖鱼类细菌性疾病研究概况

本文概述性地介绍了发生在海水养殖中的常见细菌性疾病包括弧菌现、链球菌症、卡诺氏菌症、细菌性肾病、巴斯德氏杀菌症、爱德华氏症、滑走细菌症和杀鲑气单胞菌症等的主要症状、流行情况及病原菌特征，通过遗传育种筛选有抗病力的养殖品种是人们正在努力的方向，目前的研究重点主要在鱼类抗病能力方差分析研究及抗病育种的方法研究，免疫接种是提高养殖鱼类对某一特定性病原菌的最有效的方法，目前已有三十多种菌苗上市，通过使用免疫刺激剂提高鱼类的非特异性免疫力已显示一定的应用前景，虽然抗菌素由于存在抗药菌株和鱼体残留，但它仍是目前对细菌性疾病疾病的常规疗法，而海洋藻类提取物对细菌的抑制作用已被人们所认识，它将会成为一种有潜力的抗菌药物。     

海鞘血细胞培养和细菌污染控制

以海鞘(Ciona savignyi)为研究对象,考察了海鞘血细胞原代培养方法及培养中细菌污染的鉴定和控制.血细胞在培养1 h后贴壁,细胞基本以圆形细胞为主,变形细胞一般呈不规则形.变形细胞存活时间较短,而圆形细胞存活时间较长.对于培养过程中的细菌污染,通过细菌分离、培养和纯培养发现两类菌株检出率较高,均为革兰氏阴性菌.经PCR扩增16S rDNA基因序列片断,结果显示这两类菌株分别属于弧菌属(Vibrio)和施万氏菌属(Shewanella).药敏试验结果显示,弧菌属对氯霉素和环丙沙星等具有较强的敏感性;对施万氏菌属较敏感的药物依次为亚胺培南和氯霉素等.最后比较了几种抗生素组合控制血细胞培养中细菌污染的使用效果,其中氯霉素、亚胺培南加双抗的组合有较好的抑菌效果并对培养细胞的贴壁和生长没有影响.     

斑节对虾GPx3a基因的分子克隆及表达分析

谷胱甘肽过氧化物酶(Glutathione peroxidase,GPx)是生物体内重要的抗氧化酶,能防止过氧化氢对生物体的氧化应激。该研究利用RACE技术获得了斑节对虾(Penaeus monodon)GPx3a(PmGPx3a)的全长c DNA序列,进行了相关生物信息学分析。作者利用荧光定量PCR方法研究了PmGPx3a在斑节对虾不同组织的表达情况。探究了PmGPx3a在不同胁迫条件下(盐度、重金属和细菌)的表达情况。结果表明PmGPx3a c DNA全长1135 bp,其中开放阅读框(ORF)长651 bp,预测编码216个氨基酸。PmGPx3a推导的氨基酸序列与其他动物的GPx3a氨基酸序列具有高度一致性。实时定量PCR结果显示,在高低盐胁迫下,PmGPx3a在肝胰腺中相对表达量都为上升的(p0.05)。在铜、锌、铬胁迫中,鳃中的PmGPx3a的相对表达量总体呈现下降趋势,在肝胰腺中呈现上升趋势。在哈维弧菌(Vibrio harveyi)刺激下,PmGPx3a在血淋巴中的相对表达量总体呈现上升趋势,24 h后表达量最大,显著高于对照组2.2倍(P0.05)。以上研究结果表明,PmGPx3a基因参与了斑节对虾对环境胁迫和氧化应激的适应性反应。     

Contribution of the Bacterial and Microphytobenthic Microflora in the Energetic Demand of the Meiobenthos in an Intertidal Muddy Sediment (Kerguelen Archipelago)

Abstract. An intertidal mud sediment in the Kerguelen Archipelago was sampled weekly from January 1982 to March 1983. Measurements were taken for organic matter (carbon and nitrogen) and biological (bacteria, microphytobenthos, and mciofauna) parameters. The sediment is rich in organic matter, with a mean organic carbon content of 130gC- m-². Benthic living components yield low mean biomasscs in relation to the total organic pool. Besides some marine inputs, the main allochthonous source was identified as terrestrial inputs. Benthic activities seemed strongly governed by food availability and not by temperature. Obvious seasonal changes in bacterial abundances (direct counts), primary production, and mciofauna distributions were observed. Photosynthctic production reached high values (up to 2275 mg C-m^-2-d^-1), reflecting the presence of a dense microphytobenthos. Preliminary energy budgets for two seasons (spring and winter) have been constructed. In both periods, the measured primary production can support the high mciofauna production.