凡纳滨对虾不同养殖密度高位池水体细菌群落动态

通过对养殖水体环境基因组DNA中细菌16S rRNA基因V4–V5区的高通量测序和生物信息学分析,研究了两种养殖密度的凡纳滨对虾(Litopenaeus vannamei)高位池水体中细菌群落在养殖过程中的动态。结果显示,养殖过程中各菌群相对丰度变化明显,细菌多样性随时间逐渐提高,优势菌群为变形菌门(Proteobacteria)、蓝藻门(Cyanobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)和浮霉菌门(Planctomycetes)。随着养殖时间增长,蓝藻丰度所占比例逐渐减少,而变形菌、拟杆菌和浮霉菌丰度逐渐增大,同时养殖前期高密度池浮霉菌丰度显著高于低密度池(P0.01),而其他菌群无显著差异。结果表明,养殖期前50 d不同养殖密度水体细菌群落差异较大,而后30 d内细菌群落的时间异质性大于空间异质性,这意味着高位池水体菌相被划分为两类,到养殖后期菌相快速转变,养殖密度所带来的影响被减弱。     

3种磺胺类药物在中国对虾(Fenneropenaeus chinensis)体内的药代动力学特征

采用高效液相色谱法研究了3种磺胺类药物在中国对虾体内的药物代谢动力学特征,这3种磺胺类药物包括磺胺二甲嘧啶(SM2)、磺胺嘧啶(SD)及磺胺对甲氧嘧啶(SMD)。实验期间,中国对虾的养殖水温为(24.6±2.4)℃,单次口服3种磺胺类药物的剂量均为100 mg/kg。结果显示,3种磺胺类药物在中国对虾体内的血药经时过程均符合一级吸收二室开放模型,SM2的主要药动学参数T1/2β、AUC、Vd、CL、Tmax、Cmax分别为25.812 h、34.066 mg/L·h、94.553 L/kg、2.608 L/h·kg、2 h、1.07 mg/L;SD的主要药动学参数T1/2β、AUC、Vd、CL、Tmax、Cmax分别为46.446 h、45.39 mg/L·h、97.207 L/kg、1.504 L/h·kg、1 h、1.17 mg/L;SMD的主要药动学参数T1/2β、AUC、Vd、CL、Tmax、Cmax分别为66.296 h、65.917 mg/L·h、40.015 L/kg、0.763 L/h·kg、2 h、2.00 mg/L。结果表明,SMD在中国对虾体内分布比SM2、SD更广泛;中国对虾体内SM2的消除相半衰期最短,SD次之,SMD消除相半衰期最长;3种磺胺类药物在中国对虾体内72 h药物吸收量SMD最高,SD次之,SM2最低;且SMD药物清除率最低,SD次之,SM2药物清除率最高,所以口服3种磺胺类药物72 h中国对虾体内SMD残留最多,SD次之,SM2残留最少。SMD在中国对虾体内药效更加持久,故在不考虑使用成本及毒副作用等其他因素的前提下,比较这3种磺胺类药物的药物代谢动力学特征,更加推荐使用SMD。     

基于线粒体基因组COI和12S rRNA序列的魁蚶中国群体分类学地位研究

东北亚地区不同地理种群魁蚶(Scapharca brouhtonii)的分类学地位仍存在较大争议。为探讨魁蚶中国群体的分类地位,测定了魁蚶中国群体线粒体基因组COI和12S rRNA序列,并从GenBank中下载了蚶科26种贝类的同源序列进行分析。以贻贝科的贻贝(Mytilus edulis)为外群,用邻近法和最大简约法构建了蚶科贝类的分子系统进化树,分析其分子系统进化关系和分类等级。结果显示,魁蚶中国群体的COI和12S rRNA基因的碱基组成、密码子使用率及变异位点等与魁蚶日本群体和粗饰蚶属的Anadara sativa相似,且遗传距离很小,进化树中聚为一支,亲缘关系很近,初步确定魁蚶中国群体的分类地位和魁蚶日本群体一致。本研究结果初步阐明了魁蚶中国群体的分子系统进化地位,有利于掌握魁蚶的遗传背景和资源状况,以应用于自然资源的保护和推动养殖业的发展。     

脱二氧化碳装置对循环水养殖系统pH的影响

为了调节循环水系统中养殖水体的pH,根据气体交换原理,设计一种脱二氧化碳(CO_2)装置。采用该装置去除养殖水体中的CO_2,并对由于CO_2含量累积造成的pH下降进行调节,使养殖鱼类处在一个适宜的pH环境中。试验时水温控制在(25±0.5)℃,每1 h取水样测1次pH,每4 h测1次碱度。水样取自养鱼桶内的水,检测前先对水样用40μm孔径针头过滤器进行过滤处理,实验周期24 h。结果显示,循环水系统加装脱二氧化碳装置能有效去除CO_2,使水体稳定在一个适宜的pH范围(7.39~7.42);CO_2质量浓度呈降低趋势,24 h后由开始的13.16 mg/L降低到7~8 mg/L,降低近50%,而不加装脱二氧化碳装置的循环水系统CO_2质量浓度持续上升,24 h后增加到37 mg/L左右,pH持续降低,最终降低到6.72~6.81。研究表明,脱二氧化碳装置能够有效去除水体中的CO_2,使水体pH维持在一个适宜鱼类生长的范围。     

Determination of primary structure of amberjack myosin heavy chain and its relationship with structural stability of various fish myosin rods

The structural stability of fish myosin depends upon species and temperatures of water in which fish live. Primary, secondary, and quaternary structures of myosin heavy chain (MyHC) from three species of fish living at different temperature ranges have been compared with those of rabbit MyHC in order to investigate the differences in stability. Primary structure of MyHC, although being accessible for warm-water and cold-water fish (carp and walleye pollack), was not available in previous for tropical-water fish literature; so in this study primary structure of MyHC of the tropical-water fish amberjack has been determined by cloning and sequencing its cDNA. The MyHC has 1938 amino acid residues (AA), which are almost as much as as those of carp and walleye pollack. The amberjack MyHC is 91–95% homologous with other fish and rabbit MyHCs. There is a discernible difference between animal species with stable myosin rod (amberjack, carp, and rabbit) and walleye pollack with unstable rod. Stable rod species have a high probability of forming coiled-coil around the COOH-terminal end of the rod, while the pollack has a low coiled-coil formation probability. In addition, the average scores of the coiled-coil for myosin rod were rabbit (1.738) > amberjack (1.691) > carp (1.680) > walleye pollack (1.674) which correlated exactly with the observed stability. The results suggest that coiled-coil forming ability, particularly around the COOH-terminal end, directs structural stability of fish myosin rod.     

First report of Shewanella sp. and Listonella sp. infection in freshwater cultured loach,Misgurnus anguillicaudatus

A novel disease outbreak occurred among cultured loach, Misgurnus anguillicaudatus in China. The diseased fish displayed lethargy, loss of appetite, black discolouration, anasarca and ventral skin hyperaemia. Clinical signs were observed in most of the fish examined, which consisted of discoloured liver, swollen spleen and petechial haemorrhage in the intestine and peritoneal membranes. The causative agent was found to be two species of bacteria by experimental infection, identified as Shewanella sp. and Listonella sp. The result showed that these were likely to be Shewanella putrefaciens and Listonella anguillarum respectively. Bacterial identification consisted of physiological and biochemical tests as well as 16S rRNA sequence analysis. In addition, antibiotic susceptibility testing was performed, revealing that Shewanella putrefaciens and Listonella anguillarum were susceptible to enrofloxacin, ciprofloxacin, levofloxacin, pipemidicacid and norfloxacin. This is the first report of S. putrefaciens and L. anguillarum from cultured loach.     

Effects of Cu2+-exchanged montmorillonite on intestinal microflora, digestibility and digestive enzyme activities of Nile tilapia

A total of 360 Nile tilapia at an average initial body weight of 3.9 g were randomly allocated to four treatments, each of which had three replicates of 30 fish per tank and used to investigate the effects of Cu²⁺‐exchanged montmorillonite (Cu‐MMT) on intestinal microflora, digestibility and digestive enzyme activities. The dietary treatments were: (i) basal diet, (ii) basal diet + 1.5 g kg^?1 MMT; (iii) basal diet + 30 mg kg^?1 copper as CuSO₄ (equivalent to the copper in the Cu‐MMT treatment group), or (iv) basal diet + 1.5 g kg^?1 Cu‐MMT. The results showed that supplementation with Cu‐MMT significantly improved growth performance as compared with the control and fish fed with Cu‐MMT had higher growth performance than those fed with MMT or CuSO₄. Supplementation with Cu‐MMT reduced (P < 0.05) the total intestinal aerobic bacterial counts and affected the composition of intestinal microflora with Aeromonas, Vibrio, Pseudomonas, Flavobacterium, Acinetobacter, Alcaligence, Enterobacteriaceae decreasing as compared with the control. Addition of MMT improved (P < 0.05) the apparent digestibility of dry matter and crude protein, and the activities of intestinal amylase, lipase, and alkaline phosphatase as compared with the control. Addition of Cu‐MMT improved (P < 0.05) the apparent digestibilities of dry matter, crude protein, ether extract, and crude ash, and the activities of total protease, amylase, lipase, and alkaline phosphatase as compared with the control. While supplementation with CuSO₄ had no (P > 0.05) effect on intestinal microflora, digestibility, and digestive enzyme activities the results showed that Cu‐MMT exhibited antibacterial activity in vivo resulting in a positive effect on digestive enzyme activities, and then promoted the digestion and absorption of dietary nutrition.     

光棘球海胆(♀)×中间球海胆(♂)杂交育苗试验

以光棘球海胆为母本,中间球海胆为父本进行杂交试验及苗种培育.试验结果表明,通过筛选同时得到自然成熟的父母本精、卵,在比正常海胆自交精子量高40倍以上的精子作用下,杂交受精率为2.5%.在适宜的水温、光照及饵料等的培养条件下,杂交海胆能够生长附着变态,变态时间与母本相近.培育出的杂交海胆外部颜色及棘色、棘长介于两亲本之间.杂交海胆经过6～7个月的中间暂养,出苗量为0.6×10~4个/m~3,个体壳径3～30 mm,平均壳径8.8 mm.     

罗氏沼虾育苗水质变动因子及处理技术的研究

针对目前罗氏沼虾育苗生产操作流程,作者研究了育苗池中三态氮、ＣＯＤ 及ｐＨ 等水质变化状况,其特点是这些化学指标无明显的昼夜变化,但ＮＨ３－Ｎ、ＮＯ２－ －Ｎ 含量较高,并呈现波动式上升的趋势,ＣＯＤ 含量较为稳定。从育苗安全性需要出发,还研究了育苗水的预处理模式以及投饵、换水、施药对育苗池水质的影响,结果表明,对河口天然水进行预处理是必要的,作为常规在操作管理上坚持每天彻底排污、适时适量换水、合理用药能有效控制育苗水的水质状况,即使采取高密度（ Ｚ１：２２万尾／ｍ ２）育苗也能获得较高的出苗率     

厚壳贻贝5-羟色胺2A受体(5-HT2AR)基因克隆和时空表达

为探究5-羟色胺2A受体（5-HT 2A receptor, 5-HT2AR）基因在海水贝类生长发育中的作用,本研究通过RACE技术和实时荧光定量PCR（qRT-PCR）克隆了厚壳贻贝5-HT2AR 基因的cDNA全长,并分析该基因的时空表达。结果显示,5-HT2AR基因全长2 636 bp,开放阅读框（ORF）2 124 bp,共编码707个氨基酸。序列分析结果显示该序列与人、小鼠、斑马鱼、长牡蛎和虾夷扇贝等物种分别具有45%、45%、48%、49%和67%的同源性。厚壳贻贝雌雄成体各组织和器官中均有5-HT2AR基因表达,雄性中的鳃表达量最高,而在雌性鳃、外套膜和性腺中的表达稍高于其他组织和器官;推测该基因可能与厚壳贻贝的摄食、对外界环境的感知及促进卵母细胞成熟有关。5-HT2AR基因在厚壳贻贝各发育阶段均有表达,且稚贝的表达量为眼点幼虫的1.4倍,推测5-HT2AR基因可能参与了调控厚壳贻贝幼虫的生长发育过程。本研究为进一步了解5-HT基因家族在双壳贝类中的功能奠定了基础。