Cloning and expression of a surface immunogenic protein in Streptococcus dysgalactiae isolated from fish and its application in enzyme‐linked immunosorbent assays to diagnose S. dysgalactiae infections in fish

Lancefield group C Streptococcus dysgalactiae (GCSD) causes severe necrotic lesions in the caudal peduncle in the genus Seriola farmed in Japan. To develop a sero‐diagnostic method for GCSD infection in farmed fish, we attempted to identify a surface immunogenic protein that induces an antibody after infection with GCSD by immunoblot analysis using sera collected from infected fish. A protein obtained from sodium dodecyl sulfate (SDS) extracts of GCSD was identified as S. dysgalactiae surface immunogenic protein (Sd‐Sip). Sd‐Sip exhibited more than 94% homology with a surface antigen or a hypothetical protein from S. dysgalactiae mammalian isolates at the nucleotide sequence level. Expression of the recombinant Sd‐Sip (rSd‐Sip) was confirmed by immunoblot analysis, that is, its reactivity to GCSD‐infected sera. Antibody detection ELISA using rSd‐Sip and their usefulness for diagnosis of GCSD infection were examined. GCSD‐infected sera collected from farmed amberjack, Seriola dumerili (Risso), showed strong reaction with immobilized rSd‐Sip. Meanwhile, sera immunized by other pathogenic bacteria of fish were showed ELISA values similar to those of non‐infected sera. These results of this study suggest that the antibody detection ELISA using rSd‐Sip is an effective diagnostic method for GCSD infection in fish.     

Combined effect of temperature,salinity and density on the growth and feed utilization of Nile tilapia juveniles (Oreochromis niloticus)

Based on Box–Behnken experimental design and response surface method, the joint effect of temperature (16–36°C), salinity (0–22 ppt) and rearing density (200–1000 fish.m^?3) on the specific growth rate (SGR) and feed conversion rate (FCR) of Nile tilapia juveniles were studied under laboratory conditions. The entire experiment lasted for 1 month (30 days). Results showed that the linear and quadratic effects of temperature, salinity on both growth and feed utilization were highly statistically significant (P < 0.01). The linear and quadratic effect of rearing density on the growth was highly significant (P < 0.01); the linear effect of rearing density on feed utilization was significant (P < 0.05), but the quadratic effect nonsignificant (P > 0.05). Interactions between temperature and salinity, and between salinity and rearing density on the growth statistically differed from zero (P < 0.05). Interactions between temperature and salinity, between temperature and density on feed utilization was significant (P < 0.05). Model equations of the growth and feed utilization on temperature, salinity and density were established, with the coefficient of determination being 98.34% for growth and 98.11% for feed utilization, and could be applied to projection. The optimal temperature/salinity/density combination was obtained utilizing statistical optimization approach: 29°C/6 ppt/500 fish.m^?3, at which the maximal specific growth and feed utilization reached 4.228%.d^?1 and 0.520 respectively, with the desirability being 0.989.     

2013年威海南部近岸海域表层海水重金属分布特征及污染评价

于2013年春季、夏季及秋季对威海南部近岸海域表层海水重金属进行调查,分析了重金属含量分布特征并利用重金属综合污染指数(WQI)进行污染评价。采用多元线性回归方法分析了重金属含量与表层海水盐度、pH及化学需要量之间的相关性。结果表明,威海南部近岸海域表层海水水质良好,符合二类海水水质标准,受外源输入的影响较大;综合污染指数均远低于1,呈现从海岸线向外海逐渐降低的趋势;重金属元素含量与环境要素测值(pH、盐度及COD)之间相关性顺序为:As>Hg>Cu>Cd>Pb>Zn。     

2015年靖海湾及五垒岛湾海域表层海水化学要素时空分布特征及潜在富营养化评价

于2015年春季、夏季及秋季3个航次对威海南部靖海湾及五垒岛湾海域进行表层海水水质调查并进行了富营养化评价。结果表明:调查海域表层海水水质符合二类水质标准,受磷限制影响;叶绿素-a春季航次(0.660μg/L)含量偏低,夏季航次(0.808mg/L)达到最高值;溶解氧春季航次(10.711mg/L)高于夏季航次(8.940mg/L)及秋季航次(8.678mg/L),近岸区域出现低值区;无机氮秋季航次(0.146mg/L)最高,活性磷酸盐(0.013mg/L)及化学需氧量(1.316mg/L)夏季航次最高,三者受外源河流输入的影响呈现从近岸区域向外海逐渐降低的趋势;活性硅酸盐主要受海洋本底的影响;夏秋航次在靖海湾河流入海口区域出现富营养化状态。     

通过小型探鱼无人船探测大型围网养殖区大黄鱼的分布特性

为了明确大黄鱼在大型围网养殖区内的分布规律,实验通过集成、构建具备大黄鱼探测以及水温、盐度、光照、pH、溶解氧等海洋环境探测功能的小型探鱼无人船,并在浙江省舟山市桃花岛大黄鱼围网养殖基地内开展了6次现场监测。结果发现,(1)在水平范围,大黄鱼主要分布在大型围网养殖区内礁石丰富且水流相对较缓的区域;(2)在垂直范围,大黄鱼主要分布在加权相对深度0.6～0.9的养殖海区中下层;(3)围网养殖海区内温度、盐度、pH、溶解氧等海洋环境条件整体变化较小,大黄鱼往往分布在光照强度分别为(5 921±2 702)、(50 799±50 988)、(5 990±542)、(3 494±695)、(6 836±4 761)及(15 395±5 531)lx等相对较弱的区域。本实验首次通过小型探鱼无人船系统研究了围网养殖区内大黄鱼的分布特性及相应的海洋环境因子对其分布的影响,结果为大黄鱼围网养殖区的操作平台选址、投喂管理、起网设计等提供了依据。     

Surface-exposed expression of Edwardsiella tarda EseB in live attenuated Vibrio anguillarum based on novel surface display systems

Live, attenuated Vibrio anguillarum strains can serve as vectors for the delivery of heterologous antigens for development of multivalent recombinant vaccines. Based on the outer membrane anchoring elements of V. anguillarum , we have previously constructed several efficient surface display systems Lpp-Omporf1, Lpp-OmpU, Lpp-Omp26La, Wza-Omporf1, Wza-OmpU and Wza-Omp26La. In this study, with these constructed surface display systems, a putative antigen protein EseB from pathogenic Edwardsiella tarda was successfully expressed on the surface of an attenuated V. anguillarum strain to get multivalent vaccine candidates. Further immune protection evaluation in zebra fish ( Danio rerio ) demonstrated that the V. anguillarum EseB-display strain AV/pW-26La-B could trigger full protection against V. anguillarum infection and early protection against E. tarda infection in the immunized fish. These results suggest that surface display of heterologous protective antigens in attenuated V. anguillarum could be used as a tool to develop potential V. anguillarum vector vaccine.     

鳖源嗜水气单胞菌M-96株表面抗原分析

用ＳＤＳ－ＰＡＧＥ、免疫印迹技术分析嗜水气单胞菌Ｍ－９６株表面抗原,表明该菌体表面抗原是由分子量为３０ＫＤ、３３ＫＤ、３６Ｋ和３９ＫＤ的蛋白质和脂多糖组成,其中部分对胃蛋白酶敏感。     

洞庭湖表层沉积物营养盐和重金属污染特征及生态风险评价

为探究洞庭湖表层沉积物中营养盐和重金属的空间分布特征与生态风险,2012年2月和2013年4月在东、南、西洞庭湖各取3个共计9个具有代表性样点的表层沉积物,测定其有机质（OM）、总氮（TN）、总磷（TP）、镉（Cd）、汞（Hg）、砷（As）、铅（Pb）和铬（Cr）的含量。结果表明,洞庭湖表层沉积物中 OM、TN 和 TP 的含量分别为1．48％～4．22％、382～2217 mg／kg 和142～716 mg／kg,与国内其他湖泊（水库）相比,OM和 TN 含量处于中等水平;OM和 TN 的空间分布相似,表现为南洞庭湖区＞东洞庭湖区＞西洞庭湖区,TP 表现为东洞庭湖区＞西洞庭湖区＞南洞庭湖区。洞庭湖表层沉积物中,Cd、Hg、As、Pb 和 Cr 的含量分别为0.60～20．70 mg／kg、0.090～0．640 mg／kg、10．4～83．7 mg／kg、16．9～95．8 mg／kg 和59．0～199．0 mg／kg,其中 Cd 和 As 超过土壤环境质量三级标准,是主要的重金属污染物;Cd 和 Hg 的空间分布相似,表现为南洞庭湖区＞西洞庭湖区＞东洞庭湖区;As、Pb 和 Cr 的空间分布相似,表现为南洞庭湖区＞东洞庭湖区＞西洞庭湖区。初步评价结果显示,除 TP 外,其余营养盐和重金属均具有引起较低生态风险的可能性,横岭湖和虞公庙的 Cd、As 和 Cr 以及洞庭湖出口的 Cr 具有引起较高生态风险的可能性;受 Cd 和 As 含量较高的影响,南洞庭湖区具有较高的生态风险。     

氨氮与拥挤胁迫对吉富品系尼罗罗非鱼幼鱼生长和肝脏抗氧化指标的联合影响

采用中心复合试验设计(CCD)和响应曲面方法(response surface methodology,RSM),探讨了氨氮(0.02 ～ 2.00 mg/L)和养殖密度(1～5尾/10 L)对吉富罗非鱼幼鱼生长和肝脏抗氧化指标的联合影响.结果表明,本试验条件下,氨氮和养殖密度的一次与二次效应对特定生长率有显著影响(P＜0.05),随着氨氮或养殖密度的上升,特定生长率呈先上升后下降的变化.氨氮与养殖密度之间存在互作效应(P＜0.05),氨氮浓度为0.02 ～0.20 mg/L,养殖密度在1～2尾/10 L时,幼鱼特定生长率较高;而氨氮浓度高于0.20 mg/L,养殖密度在3尾/10 L左右时,生长速度较快.肝脏丙二醛(MDA)含量随氨氮浓度和养殖密度的上升而上升,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活力呈先上升后下降的变化.氨氮与养殖密度的一次效应对MDA含量和两种酶活力均有显著影响(P＜0.05),二次效应对两种酶活力的表达有极显著影响(P＜0.01);氨氮与养殖密度对CAT活力有互作效应,高浓度氨氮与高养殖密度环境会抑制SOD和CAT活力的表达.因子与响应值间二次多项回归方程的决定系数分别达到0.972 4、0.913 2、0.938 9和0.969 2(P＜0.01),可用于预测;氨氮效应对生长和抗氧化酶活力的影响较养殖密度明显.建议在罗非鱼的养殖过程中合理安排好养殖密度,保持溶氧充足,降低氨氮胁迫,提高罗非鱼的生长与抗病力.     

Tomato yield, biomass accumulation, root distribution and irrigation water use efficiency on a sandy soil, as affected by nitrogen rate and irrigation scheduling

Florida is the largest producer of fresh-market tomatoes in the United States. Production areas are typically intensively managed with high inputs of fertilizer and irrigation. The objectives of this 3-year field study were to evaluate the interaction between N-fertilizer rates and irrigation scheduling on yield, irrigation water use efficiency (iWUE) and root distribution of tomato cultivated in a plastic mulched/drip irrigated production systems. Experimental treatments included three irrigation scheduling regimes and three N-rates (176, 220 and 230 kg ha⁻¹). Irrigation treatments included were: (1) SUR (surface drip irrigation) both irrigation and fertigation line placed right underneath the plastic mulch; (2) SDI (subsurface drip irrigation) where the irrigation line was placed 0.15 m below the fertigation line which was located on top of the bed; and (3) TIME (conventional control) with irrigation and fertigation lines placed as in SUR and irrigation being applied once a day. Except for the “TIME” treatment all irrigation treatments were controlled by soil moisture sensor (SMS)-based irrigation set at 10% volumetric water content which was allotted five irrigation windows daily and bypassed events if the soil water content exceeded the established threshold. Average marketable fruit yields were 28, 56 and 79 Mg ha⁻¹ for years 1-3, respectively. The SUR treatment required 15-51% less irrigation water when compared to TIME treatments, while the reductions in irrigation water use for SDI were 7-29%. Tomato yield was 11-80% higher for the SUR and SDI treatments than TIME where as N-rate did not affect yield. Root concentration was greatest in the vicinity of the irrigation and fertigation drip lines for all irrigation treatments. At the beginning of reproductive phase about 70-75% of the total root length density (RLD) was concentrated in the 0-15 cm soil layer while 15-20% of the roots were found in the 15-30 cm layer. Corresponding RLD distribution values during the reproductive phase were 68% and 22%, respectively. Root distribution in the soil profile thus appears to be mainly driven by development stage, soil moisture and nutrient availability. It is concluded that use of SDI and SMS-based systems consistently increased tomato yields while greatly improving irrigation water use efficiency and thereby reduced both irrigation water use and potential N leaching.