饲料添加β-1,3-葡聚糖对凡纳滨对虾生长性能、血清代谢、免疫相关基因表达和抗亚硝酸氮应激能力的影响

实验旨在研究β-1,3-葡聚糖的不同投喂方式对凡纳滨对虾(Litopenaeus vannamei)生长、血清代谢和抗亚硝酸氮应激能力的影响。选用480尾初体重(0.43±0.01) g的凡纳滨对虾, 随机分为4组, 即G0(全程投喂基础饲料)、G1组(全程投喂0.1%β-1,3-葡聚糖饲料)、G2组(0.1% β-1,3-葡聚糖饲料7d+基础饲料7d循环)和G3组(0.1% β-1,3-葡聚糖饲料14d+基础饲料14d循环)。在养殖84d后, 应用亚硝酸钠进行120h亚硝酸氮应激实验。结果显示, 各实验组凡纳滨对虾生长性能和全虾营养成分没有显著性差异。在养殖84d后, G2和G3组凡纳滨对虾肝胰腺脂肪酶活性显著高于G0和G1组(P<0.05), G1、G2和G3组凡纳滨对虾血清胆固醇和甘油三酯含量显著高于G0组(P<0.05), G3组凡纳滨对虾肌肉脂多糖/?-1,3-葡聚糖结合蛋白(LGBP)、酚氧化物酶原(proPO)和超氧化物歧化酶(SOD)mRNA表达显著高于G0和G1组(P<0.05)。亚硝酸氮应激120h, G1、G2和G3组凡纳滨对虾累计死亡率显著低于G0组(P<0.05), G3组凡纳滨对虾累计死亡率显著低于G0、G1和G2组(P<0.05)。在亚硝酸氮应激120h后, 与G0组相比, G1、G2和G3组凡纳滨对虾血清总蛋白含量显著升高(P<0.05), 葡萄糖含量、谷丙转氨酶和谷草转氨酶活性显著降低(P<0.05); G3组凡纳滨对虾肌肉LGBP、proPO和SOD mRNA表达显著高于G0 (P<0.05), G1组凡纳滨对虾肌肉丝氨酸蛋白酶(SP)mRNA表达显著高于G0、G2和G3组(P<0.05)。结果表明, 14d间隔投喂0.1% β-1,3-葡聚糖可能通过促进能量代谢和LGBP、proPO和SOD mRNA表达提高凡纳滨对虾抗亚硝酸氮应激能力。     

基于Miseq测序技术分析黄颡鱼不同养殖模式下池塘微生物群落结构多样性

利用Miseq测序技术分析了黄颡鱼Pelteobagrus fulvidraco (Richardson)三种养殖模式[(黄颡鱼单养、黄颡鱼混养草鱼(Ctenopharyngodon idellus, Valenciennes)和黄颡鱼混养草鱼、鳙(Aristichthys nobilis, Richardson)、鲢(Hypophthalmichthys molitrix, Valenciennes)]下养殖水体及底泥微生物群落结构多样性。结果显示, 12个样本共获得了655100条合格16S rDNA序列, 每个样本获得的有效序列数目为41256—77508, 可归纳为608—3686个分类操作单元。α多样性指数结果表明, 在三种养殖模式下末期水体的微生物群落丰度和多样性均高于养殖初期, 黄颡鱼混养草鱼模式下水体的群落丰度最高(Chao1指数: 2199.25, ACE指数: 2374.60), 黄颡鱼混养草鱼、鲢、鳙模式下水体微生物群落多样性最高(Shannon指数: 4.88, Simpson指数: 0.021)。样本聚类分析及PcoA主坐标分析结果表明, 黄颡鱼单养及黄颡鱼、草鱼混养模式下的水体和底泥的菌群组成及群落结构相似性较高, 黄颡鱼、草鱼、鲢、鳙混养模式下样本单独形成一类。从门水平的比较分析发现, 所有样本检测到细菌有8门, 其中变形菌门(Proteobacteria)的物种丰度在所有样本中占绝对优势地位。放线菌门(Actinobacteria)在养殖末期升高成为水体中占比第二的优势门, 尤其是黄颡鱼混养草鱼模式下(37.6%)。黄颡鱼在单养模式下, 养殖末期水体拟杆菌门(Bacteroidetes)细菌含量均明显增加(1.95%到10.98%)。从属水平的比较分析发现, 各样本之间的优势属组成有较大差异, 其中黄颡鱼在单养模式下水体样本中气单胞菌属(Aeromonas)在养殖末期丰度占优势(5.81%)。从微生物群落结构和多样性的角度考虑, 黄颡鱼、草鱼、鲢、鳙混养是一种值得推荐的养殖模式。     

凡纳滨对虾ANT2 基因的克隆及低温表达谱分析

为研究凡纳滨对虾适应低温的分子机理, 实验对从低温处理凡纳滨对虾抑制性消减文库中筛选出的一个360 bp EST 序列进行了研究。首先, 同源比对显示该EST 片段与其他物种的ANT2 基因高度同源, 命名为凡纳滨对虾ANT2 基因(LVANT2); 其次, 通过构建凡纳滨对虾肝胰腺全长cDNA 文库, PCR 扩增获得LVANT2 基因的全长cDNA1540 bp, 其中包括1011 bp 的完整开放阅读框, 编码336 个氨基酸残基。然后, 对基因进行了不同组织和低温处理的表达谱分析: (1)组织表达谱的结果显示, 该基因在凡纳滨对虾肌肉组织中表达量最高; (2)在不同低温处理下的表达结果显示, 该基因在15℃处理下基因表达量发生显著变化, 13℃开始呈下调表达, 11℃时表达量又升高; 13℃低温处理不同时间发现该基因在12h 内表达量发生显著变化,48h 后表达量最高。LVANT2 基因的低温诱导表达模式说明其可能在凡纳滨对虾低温适应中发挥作用       

维生素C对凡纳滨对虾生长及抗病力的影响

以不同水平维生素C 2 磷酸酯 (添加量分别为 0、75、15 0、30 0和 6 0 0mg/kg)的饲料喂养凡纳滨对虾 10周 ,研究维生素C 2 磷酸酯对凡纳滨对虾生长及抗病力的影响。结果显示 :在养殖前 4周 ,饲料中添加维生素C 2 磷酸酯显著促进凡纳滨对虾的生长 ,然而对对虾的成活以及饲料利用不产生影响 (P >0 0 5 ) ;而到实验后期添加维生素C 2 磷酸酯不能促进凡纳滨对虾的生长 ,却显著提高凡纳滨对虾的成活率 (P <0 0 5 )。维生素C 2 磷酸酯对对虾体水分、脂肪、蛋白质和维生素C在肝胰脏中的积累量的影响显著 (P <0 0 5 ) ,对对虾体灰分影响不显著 (P >0 0 5 )。维生素C 2 磷酸酯对对虾血清中超氧化物歧化酶活力无显著影响 ,饲料中未添加维生素C或过量添加 (超过 30 0mg/kg饲料 )均导致血清中酚氧化酶活力、血细胞总数和溶菌酶活力的显著下降。以生长、成活和酚氧化酶活力为指标 ,饲料中维生素C 2 磷酸酯的适宜添加量为 15 0mg/kg。     

一株凡纳滨对虾源维氏气单胞菌的分离鉴定及药敏特性

【目的】凡纳滨对虾(Litopenaeus vannamei)是世界范围内最主要的对虾养殖品种之一,2017年5-6月上海某凡纳滨对虾养殖场出现不明原因的死亡病例,发病急,死亡率高。从患病凡纳滨对虾体内分离到一株优势菌AVZ01,旨在确定病因并筛选出敏感药物,为今后凡纳滨对虾维氏气单胞菌(Aeromonas veronii)的防治提供参考。【方法】从患病凡纳滨对虾肝胰腺和肠道中分离致病菌,通过理化特性及16S r RNA基因序列分析进行鉴定,通过人工感染试验确定病原,使用Bliss法计算出半数致死剂量(LD50),并通过纸片扩散法进行药敏试验。【结果】从患病凡纳滨对虾体内分离到一株优势菌AVZ01,进行人工回归感染试验后,对虾发病症状与自然发病症状相似,凡纳滨对虾的LD50为8.7×105 CFU/m L。根据该菌株的形态特征、理化特性、16S r RNA基因序列分析,综合判断该病原菌为维氏气单胞菌。药敏试验结果显示,该菌株对米诺环素、诺氟沙星、庆大霉素等16种抗生素高度敏感,对青霉素、苯唑西林、头孢氨苄等9种抗生素耐药。【结论】分离菌株AVZ01对凡纳滨对虾有较强的致病性,养殖过程中可选用庆大霉素及新霉素等药物进行防控。     

两种微藻对凡纳滨对虾幼体成活率的影响

试验旨在研究钝顶螺旋藻(Sprinulian platensis)和Picochlorum sp.两种微藻培育凡纳滨对虾(Litopenaeus vannamei)幼体效果, 为开发凡纳滨对虾开口饵料提供参考依据。实验对照组D0 投喂商品螺旋藻粉, 实验组D1、D2 分别投喂钝顶螺旋藻和Picochlorum sp., 每组3 个重复,每个重复2 万尾凡纳滨对虾幼体, 在300 L 育苗桶中饲养七天。 结果表明, D0 和D1 组间凡纳滨对虾存活率无显著差异(P＞0.05), 但两组极显著高于D2 组(P＜0.01)。在凡纳滨对虾育苗过程中, 水体总氨氮浓度小于50 umol·L^-1、亚硝酸盐氮浓度小于2 umol·L^-1。钝顶螺旋藻可以保证较高的幼体变态存活率, 可以作为凡纳滨对虾开口饵料新资源开发利用。     

饲料中添加壳聚糖季铵盐对凡纳滨对虾生长及非特异免疫的影响

实验研究壳聚糖季铵盐对凡纳滨对虾生长性能及非特异性免疫能力的影响。在饲料中分别添加0、0.05%、0.10%、0.15%和0.20%的壳聚糖季铵盐, 制成5组等氮等能饲料。将900尾[初体质量(3.820.34) g]健康的凡纳滨对虾随机分成5组(45尾4平行), 养殖时间56d。结果表明:饲料中添加壳聚糖季铵盐显著影响凡纳滨对虾的生长, 0.15%实验组凡纳滨对虾的增重率和特定生长率最佳(P0.05)。饲料中添加壳聚糖季铵盐0.1%、0.15%和0.20%能显著提高凡纳滨对虾血清溶菌酶和碱性磷酸酶及酚氧化酶的活性(P0.05)。饲料中添加壳聚糖季铵盐可显著提高凡纳滨对虾抗副溶血弧菌感染的能力(P0.05), 0.15%组的保护效果最好, 其相对免疫保护率为33.24%。壳聚糖季铵盐能显著提高凡纳滨对虾的生长性能和抗病能力, 本实验条件下适宜的添加量为0.15%。     

海水池塘中国对虾与罗非鱼施肥混养的实验研究

利用 8个海水池塘陆基围隔 ,进行了封闭式施肥混养中国对虾与罗非鱼的实验研究 .结果表明 ,在对虾放养密度为 0 .92～ 2 .36尾·m- 2 的条件下 ,对虾成活率为 75.72～89.98% ,平均为 82 .40 % .产量在 1 1 .4～ 2 4 .1 g·m- 2 之间 (养殖期为 91d) .生产力为0 .2 39g·m- 2 .d- 1 .在罗非鱼放养量为 0 .2 4尾·m- 2 (体重 1 0 0 g左右 )的条件下 ,鱼虾的数量最适配比为 1 :1 0 ,中国对虾 (体长 3.0 0cm左右 )的最适放养量约为 2 .40尾·m- 2 .     

夏季混养鳙和银鲫对凡纳滨对虾低盐养殖池塘浮游藻类群落结构的影响

基于2011年6-8月在浙江省慈溪市庵东镇凡纳滨对虾低盐(2-3.2)养殖塘调查取得的数据, 分析了混养鳙和银鲫对凡纳滨对虾低盐养殖塘浮游藻类群落结构的影响。结果显示: (1) 调查期间共检出浮游藻类5门32属48种, 其中蓝藻门16种, 绿藻门20种, 硅藻门5种, 裸藻门5种, 隐藻门2种, 主要优势种为螺旋藻Spirulinasp.)、湖泊鞘丝藻(Planktolyngbya limnetica)和湖生卵囊藻(Oocystis lacustris)等。(2) 混养鳙和银鲫的虾塘浮游藻类平均生物量(22.7 mg·L−1)要低于仅混养银鲫的(32.6 mg·L−1)和单养对虾的(34.0 mg·L−1), 其中混养鳙和银鲫与仅混养银鲫的蓝藻生物量所占比例分别为46.4%与45.1%, 要低于对虾单养池塘的68.1%。(3) 混养鳙和银鲫的虾塘浮游藻类群落相似度在中、后期较高,而仅混养银鲫的虾塘在后期相似度变化较大。因此, 混养适当密度的鳙和鲫能够改善虾塘藻相, 防止藻过度增长, 促进养殖期间生态系统的稳定性。     

凡纳滨对虾各月龄性状的主成分与判别分析

为了研究凡纳滨对虾各性状增长规律和判定最佳生长季节凡纳滨对虾的体格与月龄的关系,选择1～6月龄凡纳滨对虾各1000只,选择全长、体长、第一腹节背高、第三腹节背高、第一腹节背宽、头胸甲长和体重共7个性状,进行主成分与判别分析.结果表明:各月龄凡纳滨对虾性状之间均呈现显著的正相关(P<0.01), 其中以全长与体长的相关性最为明显,1月龄凡纳滨对虾体重与形态性状的相关系数较小.各月龄凡纳滨对虾的主成分有所不同,1～2月龄凡纳滨对虾的第一主成分为长度因子,第二主成分为宽度因子,第三主成分为高度因子;3月龄凡纳滨对虾的第一主成分与1～2月龄凡纳滨对虾一致,但第二主成分为高度因子,第三主成分为体重因子;4～6月龄凡纳滨对虾的第一主成分为体重因子,第二主成分为高度因子,第三主成分为宽度因子.1～3月龄凡纳滨对虾形态性状的增长优先于体重, 4～6月龄凡纳滨对虾体重优先于形态性状的增长.错过最佳生长季节的凡纳滨对虾的与体格大小相符的月龄可通过建立的判别式来判断,总的判别准确率为98.98%,其中2～4月龄凡纳滨对虾的判别准确率为100%.     

长江中下游四个湖泊鱼类与渔业研究

选择长江中下游的牛山湖、东汤逊湖、龙感湖和黄湖作为研究地点,调查了这些湖泊鱼类区系组成、鱼种放养量、渔获物组成、渔产量和渔产值,现已查明牛山湖鱼类有64种,东汤逊湖53种,黄湖76种,龙感湖80种。这些湖泊的鱼类区系组成相似,鲤科鱼类最多,占总种数的60.4%-65.6%。牛山湖主要放养草鱼,东汤逊湖主要放养鲢和鳙,黄湖除鱼类外重点放养河蟹。这三个湖泊年均鱼种放养量分别为9.5kg/hm2、36.8kg/hm2、8.1kg/hm2;年均鱼产量分别为128kg/hm2、126kg/hm2、88kg/hm2;年均每公顷渔产值分别为623元、680元、1202元。龙感湖为非放养型湖泊,年均鱼产量为64kg/hm2,其中鲫、红鳍原泊和小杂鱼占有较高的比例;年均每公顷渔产值183元。文中分析了各湖泊鱼产量构成特征,讨论了未来湖泊渔业可持续发展的对策,为合理利用鱼类资源,发展可持续渔业提供重要依据。       

Evaluation of production performance in carp polyculture with different stocking densities and species combinations

Experiments were conducted in eight ponds of 0.08 ha each for a period of 6 months to evaluate the production performance combinations of Indian major carps and exotic carps at varied stocking densities and species. The four different treatments evaluated were stocking densities of 5000 and 10 000 fingerlings/ha with three Indian major carps (catla, rohu, mrigal) and combination of three Indian major carps with three exotic carp species (silver carp, grass carp and common carp) in replicate ponds. The mean net production levels recorded in different treatments were 1791 kg/ha, 2385 kg/ha, 2532 kg/ha and 3472 kg/ha in 6 months in treatments with three species combinations at 5000/ha, six species at 5000/ha, three species at 10 000/ha and six species at 10 000/ha, respectively. Correspondingly, the mean growth, survival and feed conversion ratios recorded in four treatments were 432 g, 84.7% and 1.49; 567 g, 86.2% and 1.47; 318 g, 82.1% and 1.69; and 444 g, 80.4% and 1.67, respectively.     

Structural conservation and food habit-related liver expression of uncoupling protein 2 gene in five major Chinese carps

The full-length cDNA of grass carp (Ctenopharyngodon idellus) and silver carp (Hypophthalmichthys molitrix) uncoupling protein 2 (UCP2) was obtained from liver. The grass carp UCP2 cDNA was determined to be 1152 bp in length with an open reading frame that encodes 310 amino acids. Five introns (Intron 3, 4, 5, 6 and 7) in the translated region, and partial sequence of Intron 2 in the untranslated region of grass carp UCP2 gene were also obtained. Gene structure comparison between grass carp and mammalian (human and mouse) UCP2 gene shows that, the UCP2 gene structure of grass carp is much similar to that of human and mouse. Partial UCP2 cDNA sequences of bighead carp (Aristichthys nobilis) and mud carp (Cirrhinus molitorella), were further determined. Together with the common carp (Cyprinus carpio) UCP2 sequence from GenBank (AJ243486), multiple alignment result shows that the nucleotide and amino acid sequences of the UCP2 gene, were highly conserved among the five major Chinese carps that belong to four subfamilies. Using beta-actin as control, the ratio UCP2/beta-actin mRNA (%) was determined to be 149.4 +/- 15.6 (common carp), 127.4 +/- 22.1(mud carp), 96.7 +/- 12.7 (silver carp), 94.1 +/- 26.8 (bighead carp) and 63.7 +/- 16.2 (grass carp). The relative liver UCP2 expression of the five major Chinese carps, shows a close relationship with their food habit: benthos and detritus-eating fish (common carp and mud carp) > planktivorious fish (silver carp and bighead carp) > herbivorous fish (grass carp). We suggest that liver UCP2 might be important for Chinese carps to detoxify cyanotoxins and bacteria in debris and plankton food.     

池塘混养模式下生态基对鲤生长及水质的影响

为探索池塘混养模式下生态基对鱼类生长的影响, 在6个土池中进行了一个饲养试验, 将土池分为2组, 一组为不放生态基的对照组, 另一组为放置生态基的生态基组, 每组3个重复。将尾均重为(310±11) g鲤3867尾、尾均重(810±15) g鲢及鳙370尾平均分别分为2组, 平均放养于6个土池中。对池塘鲤每天饲喂颗粒饲料3次, 饲养周期62d。在饲养期内, 每隔10天左右采集水样与底泥样品, 检测其中的浮游生物与微生物群落。在饲养结束后, 将试验鱼捕出并称重, 计算鲤的饲料效率。结果表明, 生态基组鲤鱼的增重率与饲料效率显著高于对照组; 生态基组的鲢、鳙末重显著低于对照组; 生态基组水体透明度与微生物群落多样性显著高于对照组; 生态基组浮游生物浓度低于对照组。结果表明, 在混养模式下土池中生态基的应用有利用促进鲤的生长, 然而池塘生态基的应用对于滤食性鲢、鳙的生长并无促进作用。     

Cloning of the Candida albicans homolog of Saccharomyces cerevisiae GSC1/FKS1 and its involvement in beta-1,3-glucan synthesis.

Saccharomyces cerevisiae GSC1 (also called FKS1) and GSC2 (also called FKS2) have been identified as the genes for putative catalytic subunits of beta-1,3-glucan synthase. We have cloned three Candida albicans genes, GSC1, GSL1, and GSL2, that have significant sequence homologies with S. cerevisiae GSC1/FKS1, GSC2/FKS2, and the recently identified FKSA of Aspergillus nidulans at both nucleotide and amino acid levels. Like S. cerevisiae Gsc/Fks proteins, none of the predicted products of C. albicans GSC1, GSL1, or GSL2 displayed obvious signal sequences at their N-terminal ends, but each product possessed 10 to 16 potential transmembrane helices with a relatively long cytoplasmic domain in the middle of the protein. Northern blotting demonstrated that C. albicans GSC1 and GSL1 but not GSL2 mRNAs were expressed in the growing yeast-phase cells. Three copies of GSC1 were found in the diploid genome of C. albicans CAI4. Although we could not establish the null mutation of C. albicans GSC1, disruption of two of the three GSC1 alleles decreased both GSC1 mRNA and cell wall beta-glucan levels by about 50%. The purified C. albicans beta-1,3-glucan synthase was a 210-kDa protein as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and all sequences determined with peptides obtained by lysyl endopeptidase digestion of the 210-kDa protein were found in the deduced amino acid sequence of C. albicans Gsc1p. Furthermore, the monoclonal antibody raised against the purified beta-1,3-glucan synthase specifically reacted with the 210-kDa protein and could immunoprecipitate beta-1,3-glucan synthase activity. These results demonstrate that C. albicans GSC1 is the gene for a subunit of beta-1,3-glucan synthase.     

葛洲坝水利枢纽兴建后对青、草、鲢、鳙繁殖生态效应的研究

本文分析了葛洲坝枢纽截流后,大坝上、下游草、青、鲢、鳙的繁殖条件,产卵场位置,产卵规模,鱼卵、鱼苗成色以及4种鱼的群体组成等。指出除原宜昌产卵场发生变化外,其余产卵场基本存在,并在上游新发现了几处产卵场。讨论了大坝对4种鱼的影响程度和上游存在草鱼地方性群体,论证了4种鱼不必过坝产卵的理由。       

The contribution of silver carp (Hypophthalmichthys molitrix) to the biological control of Netofa reservoirs

When silver carp were introduced into the Netofa reservoirs at an initial density of 300–4500 fish per hectare in order to control phytoplankton and zooplankton, there was a significant reduction of algae, zooplankton, and suspended organic matter; the silver carp prevents the growth of blue-green algae.Annual yield ranged from 600 to 1500 kg per hectare. The growth of individual fish after 6 to 8 years was 6 to 15 kg per fish.Introducing silver carp to reservoirs as a means of biological control creates a balanced ecological system in which the interspecific competition is minimal and the environmental improvements are considerable.Silver carp and bottom-feeding fish create a positive synergism in the water-body by filtering phytoplankton and zooplankton from the water, excreting a major part of it to the bottom and enriching it with organic matter suitable for zoobenthos.     

Performance of different fish species for controlling weeds in rainfed lowland rice field

A field experiment was conducted during wet season to evaluate the performance of different fish species for biocontrol of weeds in rainfed waterlogged rice fields with fingerlings of three exotic carps – grass carp, Ctenopharyngodon idella (Val.) (Cyprinidae, Cypriniformes), silver barb, Puntius gonionotus (Bleeker) (Cyprinidae, Cypriniformes) and common carp, Cyprinus carpio (L.) (Cyprinidae, Cypriniformes) – and three Indian major carps – rohu, Labeo rohita (Ham.) (Cyprinidae, Cypriniformes), catla, Catla catla (Ham.) (Cyprinidae, Cypriniformes), mrigal, Cirrhinus mrigala (Ham.) (Cyprinidae, Cypriniformes). A total of 13 major weeds under the categories of grassy, sedges, broadleaf and aquatic weeds were observed in the rice fields. Grass carp reduced maximum weed biomass (weed control efficiency [WCE] 63% at 60 days after transplanting [DAT] and 62% at 100 DAT) followed by silver barb and common carp. Among the Indian carps, only rohu was effective in control of weeds (WCE, 23% at 60 DAT). The grain yield of rice (variety Varshadhan) slightly increased (4.2–4.5 t/ha), but straw yield was significantly higher (10.2–10.7 t/ha) under rice-fish farming. Fish yield was significantly higher in exotic carps (270–288 kg/ha/90days) due to higher specific growth rates (1.8–2% body weight/day). The study indicated that exotic carps (grass carp, silver barb and common carp in order) were more effective than Indian carps for control of weed in rainfed lowland rice fields and among the Indian carps, rohu showed potential for weed control.