Ecology of Vibrios in the Oliveri-Tindari Lagoon (Messina), Two-Year Study

Abstract. A two-year ecological study in the Oliveri-Tindari (Messina) brackish water ponds evaluated the quantitative and qualitative distribution of Vibrio species in relation to the physicochemical conditions of the ponds.
The evaluation of 204 samples shows a general relationship between the halophilic vibrios and water temperature. In particular this relationship is stronger for the Vibrio fraction able to grow at 37°C, i. e. , those species related to human infections. On the other hand, the halophilic vibrios show no relation with the index of fecal contamination.
The ecology of V. parahaemolyticus is discussed in relation to the salinity of Marinello pond.
The distribution of V. cholerae non O1, isolated from all ponds, shows that this microorganism can be considered as autochthonous in coastal environments.     

皱纹盘鲍脓疱病病原菌──河流弧菌-Ⅱ的抗药机制的初步研究

1993年9月至1995年末，大连地区的一龄鲍至成鲍都出现了脓疱病，用常规方法从3个养殖厂的病鲍中分离到了3个菌株，分别为D株、N株和T株。经鉴定这3个菌株为同一种──河流弧菌-Ⅱ。这3个菌株来自3个不同的海区、不同的单位，并且这3个单位使用的抗生素种类和浓度也木同，所以这3个菌株对18种抗生素的敏感度也木同。D株对环丙沙星（抑菌坏直径30mm）、复方新诺明（抑菌坏直径27mm）等敏感，而对青霉素、青霉素Ⅱ、氨苄青霉素（抑菌环直径0mm）等耐药。N株除对青霉素Ⅱ、白霉素、吡哌酸等耐药外，对其它12种抗生素都敏感或中度敏感（抑菌环直径13．5－30mm）。T株对氟派酸（抑菌环直径22mm）、环丙沙星（抑菌环直径20．3mm）等敏感，而对氯霉素、复方新诺明等耐药。研究发现，经常使用同一种抗生素很容易使病原菌产生耐药性．连续3d使用单一的抗生素（青霉素）就会产生耐药的菌株。为证明上述3个菌株的抗药性是属于哪一类，以T株为例，提取总DNA，并对总DNA进行EcoRⅠ酶切图谱分析，图谱有明显的异同，说明3个菌株的抗药性不同可能与基因突变有关。不管是非遗传型还是遗传型，都是由于抗生素为细菌提供了耐药突变株的选择环境，从而使耐药菌株得以大量繁殖。     

一种新的中国对虾弧菌病原菌──产气弧菌

于１９９３年８月，从山东省昌邑县下营镇患败血病的中国对虾血淋巴内分离到多株细菌，其中２株人工感染证实为病原菌；经５０多项形态，生理，生化特性测试鉴定为产气弧菌Ｖｉｂｉｏｇａｚｏｇｅｎｅｓ。其主要特性为：革兰氏阴性，弧状，极生单鞭毛，产生红色素，氧化酶呈阳性，精氨酸双水解酶，赖氨酸脱羧酶，鸟氨酸脱羧酶均为阴性，淀粉酶，明胶酶为阳性，硝酸盐还原阴性，利用木糖，水杨苷，山梨醇发酵葡萄糖产气，利用柠檬酸盐     

养殖牙鲆鳗弧菌疫苗的研究

以牙鲆病原性鳗弧菌M3为抗原,制备了细胞（CV）、细胞－胞外产物（CEV）、脂多糖（LPS）三种疫苗,通过浸泡和注射两种途径对牙鲆进行两次免疫,检测疫苗的免疫保护效果。免疫后4周可检测到免疫组牙鲆具有明显的凝集抗体效价,其中注射组效价显著高于浸泡组,而对照组变化不明显。与对照组相比,免疫组牙鲆获得良好的免疫保护力,其中注射免疫组高于浸泡组。在注射免疫途径中,以CEV的免疫保护效果最好;在浸泡免疫途径中,以CEV和LPS的免疫保护效果最好。结果表明鳗弧菌M3的细胞－胞外产物苗有显著的免疫保护作用。     

Ecotoxicological evaluation of sewage sludge using bioluminescent marine bacteria and rotifer

Bioassay using the marine bacteria,Vibrio fischeri and rotifer,Brachionus plicatilis, and chemical analyses were conducted to assess the toxicity of the various sewage sludges, one of the major ocean dumped materials in the Yellow Sea of Korea. Sludge elutriates extracted by filtered seawater were used to estimate the ecotoxicity of the sludge. Chemical characterization included the analyses of organic contents, heavy metals, and persistent organic pollutants in sludge. Bacterial bioluminescent inhibition (15 min), rotifer mortality (24 hr) and rotifer population growth inhibition (48 hr) assay were conducted to estimate the sludge toxicity. EC50 15 min (inhibition concentration of bioluminescence after 15 minutes exposed) values by Microtox® bioassay clearly revealed different toxicity levels depending on the sludge sources. Highest toxicity for the bacteria was found with the sludge extract from dyeing waste and followed by industrial waste, livestock waste, and leather processing waste. Clear toxic effects on the bacteria were not found in the sludge extract from filtration bed sludge and rural sewage sludge. Consistent with Microtox® results, rotifer neonate mortality and population growth inhibition test also showed highest toxicity in dyeing waste and low in filtration bed and rural sewage sludge. High concentrations of persistent organic pollutants (POPs) and heavy metals were measured in the samples from the industrial wastes, leather processing plant waste sludge, and urban sewage sludge. However, there was no significant correlation between pollutant concentration levels and the toxicity values of the sludge. This suggests that the ecotoxicity in addition to the chemical analyses of various sludge samples must be estimated before release of potential harmful waste in the natural environment as part of an ecological risk assessment.     

嵊泗列岛海域三种贻贝贝体框架性状对壳重的影响效应

以嵊泗列岛海域2+龄厚壳贻贝同生群养殖个体、同域生长的1+龄紫贻贝同生群养殖个体以及混于厚壳贻贝和紫贻贝养殖筏架中的"杂交贻贝"为实验对象,以壳宽SW、壳长SL、壳高SH(BD)、OA(壳顶至韧带末端的直线距离)、OB(壳顶至壳背面最高点的直线距离)、OC(壳顶至壳后端最远点的直线距离)、OD(壳顶至壳高性状在腹缘的落点的直线距离)、AB(韧带末端至壳背缘最高点的直线距离)、BC(壳背缘最高点至壳后端最远点的直线距离)、CD(壳后端最远点至壳高性状在腹缘的落点的直线距离)为贝体框架变量,采用多元分析方法系统比较了三者贝体框架性状对壳重的影响效应。结果表明:(1)3种实验贝壳重的变异系数均大于各贝体框架性状,"杂交贻贝"各测定性状的变异系数均为最大,而厚壳贻贝和紫贻贝间则均接近;(2)3种实验贝各贝体框架性状与壳重的相关系数均达到极显著水平(P0.01),三者中仅"杂交贻贝"各贝体框架性状间的相关系数也均达到极显著水平(P0.01);(3)经通径分析,厚壳贻贝被保留的3个贝体框架性状与壳重的复相关指数为0.868,它们对壳重的直接作用呈OCSHSW,紫贻贝被保留的2个贝体框架性状与壳重的复相关指数为0.700,它们对壳重的直接作用呈OCSW,"杂交贻贝"被保留的3个贝体框架性状与壳重的复相关指数为0.931,它们对壳重的直接作用呈SLSHOA;(4)经多元回归分析,得到了用于估算厚壳贻贝、紫贻贝和"杂交贻贝"壳重的回归方程。     

鳗弧菌(Vibrio anguillarum)侵染对青蛤(Cyclina sinensis)髓样分化因子88基因表达的影响

经转录组测序后筛选并克隆得到青蛤(Cyclina sinensis)髓样分化因子88(myeloid differenttiation factor 88,My D88)的c DNA序列。在鳗弧菌(Vibrio anguillarum)胁迫下,采用荧光定量PCR法分析了My D88基因在青蛤体内的表达过程。结果显示,青蛤My D88基因的开放阅读框为1521bp,编码506个氨基酸,分子量约为57.14k Da,氨基酸N段存在DEATH结构域,C段存在TIR结构域(Toll/Interleukin-1 receptor domain)。My D88基因在青蛤血淋巴、肝脏、外套膜、鳃和闭壳肌等组织中普遍表达,但在血淋巴中表达量最高,与其它组织出现显著性差异(P0.05)。通过检测鳗弧菌刺激下My D88基因在青蛤血淋巴中的表达值,发现My D88基因在24h开始升高,48h达到最大值,约为对照组的10倍,实验组与对照组及空白组均出现了极显著性差异(P0.01);研究结果表明,该基因在软体动物的免疫应答反应中对革兰氏阴性菌有识别作用。     

Multiplex PCR allows simultaneous detection of pathogens in ships' ballast water

There is enormous potential for global transfer of microorganisms, including pathogens, in ships' ballast water. We contend that a major advancement in the study of ballast-water microorganisms in particular, and of aquatic pathogens in general, will be expedited sample analysis, such as provided by the elegant technology of DNA microarrays. In order to use DNA microarrays, however, one must establish the appropriate conditions to bind target sequences in samples to multiple probes on the microarrays. We conducted proof-of-concept experiments to optimize simultaneous detection of multiple microorganisms using polymerase chain reaction (PCR) and Southern hybridization. We chose three target organisms, all potentially found in ballast water: a calicivirus, the bacterium Vibrio cholerae, and the photosynthetic protist Aureococcus anophagefferens. Here, we show simultaneous detection of multiple pathogens is possible, a result supporting the promising future use of microarrays for simultaneous detection of pathogens in ballast water.     

海水网箱养殖大黄鱼病原菌研究

从福建宁德地区患病大黄鱼肝、肾、脾等组织分离出３株病原菌,进行纯化培养、人工感染、药敏试验及菌种鉴定等研究,根据形态及生理生化特征,其中２株致病菌归为哈维氏弧菌,另一株为腐败假单胞菌。药敏实验的结果表明,氟哌酸、环丙氟哌酸、氯霉素、庆大霉素和复方新诺明等对致病菌有较强的抑制作用。     

光合细菌对致病弧菌的抑制作用

本文主要介绍5株光合细菌对3株致病弧菌的抑制作用研究,结果表明,在不同的培养基中其抑菌效果不同,在普通培养基中No.3PSB对副溶血弧菌和河弧菌有明显的抑制作用,但在TCBS培养基中,则无抑菌效果,在培养基I中,No.1和No.2 PSB对3株致病弧菌的抑制作用皆取得良好的效果。