桑沟湾贝类筏式养殖与环境相互作用研究

2003年6~10月和2004年3月利用YSI6600多参数水质监测仪和SD6000全自动海流计以及石膏块对桑沟湾扇贝筏式养殖区养殖设施内外、扇贝养殖笼内外部分环境参数(pH、DO、叶绿素a、海流)的动态变化进行了系统研究。实验结果表明:(1)扇贝筏式养殖区内外流速及叶绿素a浓度衰减显著:经过20排筏架(约105m)后,养殖区内的海水平均流速由区外的7·50±5·29降到5·28±3·23cm·s~(-1),降低了29·6%,区内外海流的主流向没有显著改变;进入养殖系统的叶绿素a浓度由2·48降低到了1·19μg·L~(-1),减少了52%;养殖区内的pH和溶解氧与区外相比变化不大;(2)扇贝养殖笼内外环境因子发生显著的变化:2003年7月1日养殖笼内外水交换率可达80·5%,1个月后下降到只有23·6%;2004年10月份养殖笼内叶绿素a浓度降低到仅有1·57μg·L~(-1),较之笼外的4·20μg·L~(-1)来说,减少了62·6%;养殖笼外pH基本保持在8·0左右,笼内的pH却一直趋于下降趋势,到9月份降到了最低值7·46;在附着生物附着盛期的8、9月份,笼内溶解氧最低值降低到了5·13mg·L~(-1),比起笼外的7·04mg·L~(-1)来说,减少了27·1%。     

胶州湾栉孔扇贝大规模死亡的流行病学调查

2 0 0 0年 1月 - 12月 ,跟踪调查了胶州湾养殖栉孔扇贝大规模死亡情况 ,对养殖海域环境因子进行了测定 ,并对大规模死亡期间发病栉孔扇贝进行了病理学观察。调查结果显示 ,胶州湾养殖栉孔扇贝大规模死亡呈暴发性并集中在 7月下旬的高水温期。发病高峰期在 7月 2 1日至 7月 2 7日 ,最高时点患病率达 14 .5 % ,7月份的月死亡率为 5 8.0 %。至 8月 3日累积死亡率达 90 .0 % ,患病贝死亡率接近 10 0 %。发病期扇贝壳高平均为 5 .0± 0 .9cm。,养殖栉孔扇贝半数死亡时间在 7月 2 3日。通过对水环境因子、病原性生物因子的调查分析及病理学观察表明 ,栉孔扇贝的大规模死亡可能是由生物性因子 (病毒 )所致 ,而与环境中理化因子可能无直接的相关关系     

汞及甲基汞在栉孔扇贝全组织内的积累与净化

以栉孔扇贝Chlamys farreri为研究对象,暴露于不同浓度HgCl2溶液35d后,转入自然海水中净化30d,检测不同时间点栉孔扇贝全组织中总汞与甲基汞含量,研究二者在栉孔扇贝组织中积累与净化情况.实验采用Kahle的双箱模型,数据处理采用两组不同动力学参数的双曲线毒性动力学模型.拟合发现,总汞和甲基汞在栉孔扇贝组织内的积累与净化规律符合双箱动力学模型;浓度低于2μg/L的汞溶液对栉孔扇贝的慢性毒性较小;各实验组暴露35d后栉孔扇贝组织中的总汞含量达到最高,暴露浓度与扇贝组织内的总汞积累量呈正比例关系.总汞在栉孔扇贝组织内的富集高峰出现在第35天,与理论点一致,而甲基汞富集高峰出现在40d后,有延迟现象;对比总汞和甲基汞在栉孔扇贝组织内的积累与净化动力学参数发现,总汞在栉孔扇贝组织内的积累因子（BCF）总体上大于甲基汞在栉孔扇贝组织内的BCF;汞与甲基汞在扇贝组织内的BCF和生物学半衰期（B1/2）与暴露液浓度均呈正相关关系;海水环境中总汞浓度低于0.064 8μg/L时,对栉孔扇贝的食品安全性无影响（以甲基汞为限量）.     

栉孔扇贝病原感染与病害发生关系探讨

利用电镜负染技术检测自然海区养殖和人工感染试验的栉孔扇贝组织提取液,分别发现病毒和类立克次氏体(RLO)两种病原体。动态检测两类病原体感染状况与整个养殖期间栉孔扇贝发病死亡情况,结果表明,养殖期间病毒感染率在80％—100％之间,最大感染强度出现在栉孔扇贝大规模死亡的7、8月份。人工感染病毒的栉孔扇贝,其病毒感染率和感染强度与养殖栉孔扇贝大规模死亡时期相当。而两种情况下栉孔扇贝类立克次氏体的感染率和感染强度均未见明显变化。综合分析表明,病毒可能是造成栉孔扇贝大规模死亡的直接病原体。     

栉孔扇贝夏季大规模死亡的神经网络预测模型

根据2002年和2003年对山东荣成桑沟湾栉孔扇贝养殖海区的水温、盐度、pH、氨氮浓度、亚硝氮浓度等环境因子和扇贝血清中的蛋白浓度、酸性磷酸酶活力、碱性磷酸酶活力、超氧化物歧化酶活力和过氧化氢酶活力等免疫学指标及栉孔扇贝养殖密度和死亡率的监测数据,运用人工神经网络(artificial neurd network,ANN)的原理和误差反相传播(back propagefion,BP)网络的方法,利用MATLAB软件初步建立养殖栉孔扇贝夏季大规模死亡的BP人工神经网络预测模型.预测模型经过300次的学习训练,误差平方和由67.46下降至0.009 1.该预测模型对未参与模型构建的样本预测的结果与实际监测结果的符合率达到87.5%.首次将人工神经网络与水产动物病害死亡的预测相结合,建立的预测模型具有对数据适应能力强,可适时学习,预测结果准确等突出优点,为水产养殖动物病害死亡程度的预测提供了一个新的研究方法.     

栉孔扇贝烟台养殖区海水水质及扇贝体内重金属含量评价

为了解栉孔扇贝集中上市季节扇贝养殖区海水水质状况及扇贝体内重金属蓄积情况,本研究在烟台选取长岛、开发区和牟平区3处具有代表性的栉孔扇贝养殖区,于2021年4月分别采集表层海水及栉孔扇贝样品,对海水常规指标及海水和扇贝中的重金属(Pb、Cd、Cu、Hg、Zn和Cr)含量进行测定和分析。结果表明：在栉孔扇贝集中上市时间,3个栉孔扇贝主要养殖海域海水中溶氧(DO)、化学需氧量(COD)和营养盐均符合国家一类海水水质标准,重金属含量均未超出二类海水水质标准值,其中,从Cu含量来看,长岛养殖区海水水质明显优于另外2个养殖区海水。栉孔扇贝对重金属Zn和Cu的富集有明显的选择性,富集系数超过1 000。本研究中3个养殖海域栉孔扇贝整体软体组织及闭壳肌中Pb、Cd、Hg和Cr 4种重金属含量均符合中国食品安全国家标准,Cu和Zn含量均符合《海洋生物质量》限量要求,且为一类标准。[中国渔业质量与标准,2022,12(4):36-41]     

两种扇贝病害的ELISA分析

运用ELISA分析技术,对烟台海区不同时期的栉孔扇贝和海湾扇贝体内感染的病毒粒子的季节性变化进行了研究。结果表明一龄栉孔扇贝体内病毒粒子的含量在8月份最高,其P/N值和OD值分别为0.8305和2.23,与栉孔扇贝大规模死亡时期相一致。一龄海湾扇贝的OD值在8月份的检测中大于1,即表示海湾扇贝中可能存在栉孔扇贝病毒粒子。其他月份的检测结果均为完全阴性,即没有检测到栉孔扇贝病毒粒子。     

栉孔扇贝夏季大规模死亡的神经网络（ANN）预测模型

根据2002和2003年对山东荣成桑沟湾栉孔扇贝养殖海区的水温、盐度、pH、氨氮浓度、亚硝氮浓度等环境因子和扇贝血清中的蛋白浓度、酸性磷酸酶活力、碱性磷酸酶活力、超氧化物歧化酶活力和过氧化氢酶活力等免疫学指标及栉孔扇贝养殖密度和死亡率的监测数据,运用人工神经网络（ANN）的原理和误差反相传播（BP）网络的方法,利用MATLAB软件初步建立养殖栉孔扇贝夏季大规模死亡的BP人工神经网络预测模型。预测模型经过300次的学习训练,误差平方和由67.46下降至0.0091。该预测模型对未参与模型构建的样本预测的结果与实际监测结果的符合率达到87.5％。本研究首次将人工神经网络与水产动物病害死亡的预测相结合,建立的预测模型具有对数据适应能力强,可适时学习,预测结果准确等突出优点,为水产养殖动物病害死亡程度的预测提供了一个新的研究方法。     

养殖密度对循环水系统中大菱鲆(Scophthalmus maximus)生长的影响

将初始体重为(580.9±44.65)g的大菱鲆成鱼按照低密度A组14.30 kg/m2、中密度B组20.49 kg/m2、高密度C组31.32 kg/m2的标准分为3个不同养殖密度组,并放养于循环水养殖系统中120 d,同时对大菱鲆成活率、体重差异、饵料系数、溶菌酶水平及养殖水体中总氨氮(TAN)、亚硝酸氮(NO2--N)、COD浓度的变化进行测定。研究表明,实验结束时A、B、C三组大菱鲆养殖密度分别达到30.09、41.30、60.07 kg/m2,各实验组成活率都在95%以上。大菱鲆养殖密度对增重率的影响主要体现在研究前期,并且随着养殖密度的增加,各实验组体重差异度出现显著变化(P0.01)。大菱鲆A、B、C组的饵料系数分别为0.73、0.75、0.82,与养殖密度呈正相关。研究开始第5天,高密度组大菱鲆溶菌酶水平升高,20 d后血液溶菌酶水平逐渐降低,40 d之后显著低于低密度组。研究期间系统运行稳定,循环水养殖大菱鲆的不同密度对系统各项水质指标总氨氮(TAN)、亚硝酸氮(NO2--N)、COD浓度的变化有显著影响(P0.05)。研究结果显示,随着养殖密度的升高,各项水质指标显著升高,但高密度组各项水质指标均未超过渔业水质标准所规定的浓度。     

5-羟色胺对栉孔扇贝血淋巴中髓过氧化物酶活性和一氧化氮含量的影响

用质量浓度为0.05、0.5、5μg/ml的5-羟色胺与栉孔扇贝血淋巴孵育,采用生化方法,测定栉孔扇贝血清和血细胞中髓过氧化物酶活性和一氧化氮含量。结果表明,3个5-羟色胺质量浓度下各试验组的血清中髓过氧化物酶活性均低于对照组,差异显著;各试验组的血细胞中髓过氧化物酶活性均低于对照组,差异显著。5-羟色胺质量浓度为0.05μg/ml时,各试验组的血清一氧化氮含量低于对照组,差异不显著;5-羟色胺质量浓度为0.5、5μg/ml时,各试验组的血清一氧化氮含量高于对照组,差异显著。3个5-羟色胺质量浓度下,各试验组的血细胞一氧化氮含量均低于对照组,差异显著。说明5-羟色胺对栉孔扇贝血淋巴中的髓过氧化物酶活性和一氧化氮含量起到了调节作用。5-羟色胺可能在栉孔扇贝的免疫调节中发挥重要作用。     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.