凡纳滨对虾海水养殖系统内细菌群落的PCR-DGGE分析

采用PCR-DGGE(PCR-denaturing gradient gel electrophoresis)技术对一个典型的凡纳滨对虾(Litopeneausvannamei)海水养殖系统细菌群落进行分子分析.结果表明,沿岸水、蓄水池、养殖池水具有较高的细菌种类多样性,而蓄水池进水、对虾粪样、肠壁定植细菌样以及排水渠污水的细菌多样性程度低.每种环境群落的优势种明显.3个养殖池水样(Y1、Y2、Y3)、2个沿岸水样(W1、W2)、2个粪样(F1、F2)、蓄水池水样(B1、B2)及2个肠壁定植细菌样(G1、G2)各自具有高度群落相似性.BLAST结果表明,12个条带克隆序列所代表优势种很可能来源于以下几个属:柔发菌属(Flexithrix)、黏纤维菌属(Cytophaga)、Dyella属、聚球菌属(Synechococcus)、Chlorarachnion属、支原菌属(Mycoplasma)、草螺菌属(Herbaspirillum)、河氏菌属(Hahella)、Ruegeria属.本研究表明,PCR-DGGE技术可以用于海水对虾养殖系统的细菌群落结构分析.对于海水对虾养殖系统来说,一些序列所代表的主要细菌种类极有可能是很少被注意到或研究过的,具有潜在的研究价值.[中国水产科学,2009,16(1):31-38]     

养殖长毛对虾虾体长体重关系式研究

长毛对虾(Penaeus Penicillatus Aicock)是福建省近海大型虾类优势种群,也是我国南方对虾养殖优良的种质资源之一。福建省于1979年开展生产性养殖试验获得成功,尔后逐步推广发展到目前20多万亩的养殖规模。虽然近几年开展多种虾和多季虾养殖,但是长毛对虾养殖面积仍占重要地位。     

长毛对虾北移养殖初试

从浙江温州购进０．７ｃｍ长毛对虾苗１４万尾,暂养和盐度驯化后在５亩池塘养殖６３天,平均体长由１．２ｃｍ,增至８．４ｃｍ,旬增长１．１４ｃｍ。至９月底收获时,平均体长１０．５ｃｍ,亩产６２ｋｇ,亩效益６４０元。     

长毛对虾生物学及其养殖

长毛对虾(Fenneropenaeus penicillatus Alcock)又名大虾、白虾、红尾虾、红虾、大明虾,隶属对虾科对虾属,是对虾类中的大型虾之一,广泛分布在印度洋、西太平洋的巴基斯坦到印度尼西亚沿海一带,我国福建、台湾及广东东部沿海最为常见,是南方省区重要的海洋捕捞和养殖对象;养殖120～150天,体长可达120mm 以上,每667m~2产量100kg 左右。 1 形态特征长毛对虾体淡棕黄色,额角上缘7～8齿,下缘4～ 6齿,额角基部侧视比中国对虾的高,而比墨吉对虾的低,额角后脊伸至头胸甲后缘附近,无中央沟,第1触角鞭比头胸甲稍长,雄虾第3颚足末节有毛笔状长毛,其长度为末第2节的1.2～2.7倍,额角脊上有断续的凹点, 雌交接器前片顶端的疣突比墨吉对虾的小。 2 生态习性长毛对虾存活的最高温度为40%,最低10℃C,生长     

循环海水养殖系统硝化滤器中氨氧化微生物分析

研究循环水养殖硝化滤器载体上附着生物膜的微生物群落结构可以为提高其处理速率和效率,并为特异性工程菌构建提供依据。采用改良的AFLP方法分析了循环水养殖硝化滤器载体上附着的氨氧化细菌16S rRNA基因和氨单加氧酶amoA基因片段及其系统发育情况。结果表明:分析16S rRNA基因得到的序列片段比分析amoA基因片段得到了更多信息,准确度较高,可作为分析循环水养殖硝化滤器氨氧化菌群组成的有效方法。克隆测序所得序列与网上公布数据比对,可见存在于循环水养殖硝化滤器载体上的氨氧化细菌与Nitrosomonas cryotolerans、Nitrosomonas oligotropha、Nitrosospira tenuis、Nitrosomonas marina相似度达100%,与Nitrosomonas aestuarii相似度为87%。大部分属于亚硝化单胞菌属(Nitrosomonas),仅少数序列属于亚硝化螺菌属(Nitrosospira)。采用16S rRNA基因和amoA片段分析方法得到的附着于封闭循环海水养殖硝化滤器载体上的氨氧化细菌主要为变形菌(Proteobacteria)的β-亚类的亚硝化单胞菌属(Nitrosomonas)和少量的亚硝化螺菌属(Nitrosospira)氨氧化细菌,以及一定数量的γ-亚类氨氧化细菌。     

凡纳滨对虾养殖环境及肠道微生物群落特征分析

为探究凡纳滨对虾养殖过程中其肠道和养殖环境微生物群落的结构及变化,本实验采用Illumina MiSeq测序平台,基于16S rRNA基因的测序结果,对46和86 d虾龄的凡纳滨对虾肠道以及养殖水样、底泥中的微生物群落进行分析。结果显示,86 d时微生物Shannon-Wiener多样性指数显著高于46 d,水样中的Shannon-Wiener指数显著低于底泥和凡纳滨对虾肠道;有35个门、70个纲、152个目、274个科以及420个属在水样、底泥和虾肠道中均能检测到,其中虾肠道和底泥间共有菌类较水样中更多。微生物群落随养殖时间增加发生变化,46与86 d的差异性门类为放线菌门、绿菌门、丝状杆菌门、浮霉状菌门和TM6,相对丰度随时间增加而增高。水样、底泥和虾肠道中有相对固定的优势菌群,在水样、底泥和虾肠道中主要的门类均为变形菌门、拟杆菌门和放线菌门,优势纲类为α变形菌纲、β变形菌纲以及放线菌纲,除此以外,虾肠道与底泥共有的优势纲为δ变形菌纲、γ变形菌纲以及芽单胞菌纲;从目、科、属的分类水平上看,水样、底泥和虾肠道中菌类相对丰度各异,鲜有明显重叠的优势菌类,在水样、底泥和虾肠道中均为一种未...     

海水对虾工厂化循环水养殖系统模式分析

对虾养殖由于受到水资源和虾病的困扰,工厂化循环水养殖已经成为今后对虾养殖的一个重要方向。对虾工厂化循环水养殖系统的结构包括了养虾池、水处理技术、消毒杀菌、增氧技术、水温调节装置等。目前,典型的养殖模式有美国德州跑道式对虾养殖系统、台南室内自动化循环水养虾系统、美国佛罗里达三阶段养殖系统和美国夏威夷基于微藻的循环水对虾养殖系统。文中对这4种典型的对虾工厂化循环水养殖系统的养殖试验情况进行分析比较。     

石斑鱼循环水养殖系统微生物群落结构

通过16S rRNA基因片段高通量测序研究了褐点石斑鱼(Epinephelus fuscoguttatus)和东星斑(Plectropomus leopardus)循环水养殖系统水体细菌种群结构。结果显示石斑鱼循环水养殖系统中优势细菌类群为变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、蓝藻门(Cyanobacteria)、梭杆菌门(Fusobacteria)、厚壁菌门(Firmicutes)和硝化螺旋菌门(Nitrospira)。其中养殖塘、固液分离池、沉淀池和蛋白分离池中主要优势细菌为γ-变形菌纲(Gammaproteobacteria),紫外消毒池和补氧池中则以a-变形菌纲(Alphaproteobacteria)为主,而生物滤池中两者比例接近,均为优势种群。从养殖塘到补氧池细菌多样性指数先降低后升高,生物滤池中达到最高值,之后再次下降至紫外消毒池中达到最低值,补氧池中有所回升。PCoA分析和聚类分析结果表明,养殖水在紫外消毒前后其细菌群落组成有着明显的差异。水质理化指标检测表明,经过净化处理后,循环系统的养殖水溶解氧含量上升,氨氮和亚硝酸盐含量下降并维持在较低的浓度。环境因子与细菌群落结构相关性分析结果表明,磷酸盐、pH、溶解氧和温度等指标可能对细菌群落结构影响相对较大,但未发现两者间有很显著的相关性,这可能与实验样本较少有关。     

长毛对虾仔虾对不饱和脂肪酸需求量的初步研究

用正交法对不饱和脂肪酸进行６因素２水平的饲料含量设计，并在对虾营养试验系统中进行饲养实验。结果表明；长毛对虾仔虾的生长率，存活率受不２饱和脂肪酸种类与含量的影响。在本实验条件下，长毛对虾仔虾期Ｃ１６：ｎ，Ｃ１８：ｎ，Ｃ１８；２ｎ，Ｃ１８；３ｎ，Ｃ２０：１－５ｎ和Ｃ２２－２４的最适宜含量分别上饲料的０．２４％，１．４０％，１．８５％，０．４３％，０．７６％和２．２５％。     

长毛对虾配合饵料试验研究

<正> 各种不同品种的对虾,由于分布的地理位置、栖息环境条件及生物饵料品种构成的不同,形成了它们对饵料中主要营养成份要求的不同。作为对虾饵料中主要组成成份的蛋白质各种对虾要求上存在着差异。过去,人们对日本对虾、斑节对虾、墨吉对虾、龙虾及中国对虾等饵料中各种主要营养成份研究得     

Serotonin stimulates ovarian maturation and spawning in the black tiger shrimp Penaeus monodon

Serotonin (5-hydroxytryptamine, 5HT) has been reported to induce ovarian maturation and spawning in the crayfish Procambarus clarkii and white Pacific shrimp Litopenaeus vannamei. The aim of this study was to explore the role of exogenous 5HT on the reproductive performance of the black tiger shrimp Penaeus monodon. 5HT solution was injected into domesticated P. monodon broodstock at 50 μg/g body weight and ovarian maturation and spawning were recorded. The presence of 5HT in the ovary and oviduct of P. monodon was also studied by immunohistochemistry and its levels in the ovary by enzyme link immunoabsorbance assay (ELISA). The 5HT-injected P. monodon developed ovarian maturation and spawning rate at the level comparable to that of unilateral eyestalk-ablated shrimp. Hatching rate and the amount of nauplii produced per spawner were also significantly higher in the 5HT-injected shrimp, compared to the eyestalk-ablated shrimp. 5HT-positive reactions were found in the follicular cells of pre-vitellogenic oocytes, in the cytoplasm of early vitellogenic oocytes and on the cell membrane and cytoplasm of late vitellogenic oocytes. 5HT in the ovary was present at 3.53 ± 0.26 ng/mg protein level in previtellogenic stage and increased to 17.03 ± 0.57 ng/mg protein level in the mature stage of the ovary. The results suggest a significant role of 5HT, possibly directly on the ovary and oviduct, on the reproductive function of female P. monodon.     

副溶血弧菌对斑节对虾非特异性免疫酶活性的影响

为了研究副溶血弧菌对斑节对虾非特异性免疫酶活性的影响,分别对斑节对虾注射生理盐水和副溶血弧菌,于不同时间点测定肝胰腺和鳃中的总抗氧化能力(T-AOC)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)和溶菌酶(LSZ)的活性变化。结果显示,与对照组相比,感染副溶血弧菌后,肝胰腺和鳃中T-AOC活性分别于12和6 h达到最大值(P0.05),随后逐渐降低;肝胰腺中ACP活性于3 h显著升高,随后逐渐降低,并于24 h达到最小值(P0.05),而鳃中ACP活性于12 h达到最大值,随后逐渐降低,并于48 h显著低于对照组(P0.05);肝胰腺和鳃中ALP活性整体均呈现出先升高后下降的趋势,但未出现显著低于对照组的现象;肝胰腺和鳃中LSZ活性均于3 h显著升高至最大值(P0.05),随后逐渐恢复至初始水平。研究表明,副溶血弧菌感染对斑节对虾非特异性免疫酶活有显著影响,对其机体免疫防御系统有明显的破坏作用。     

Denaturing Gradient Gel Electrophoresis (DGGE) as a tool for the characterisation of Brachionus sp. strains

Many zooplanktonic organisms, like the cyclic parthenogenetic rotifer Brachionus plicatilis (Rotifera: Monogononta), are actually a complex of species and biotypes with a high degree of morphological similarity (i.e. cryptic species). Various phylogenetic studies with molecular markers (e.g. ITS1 and COI) on wild Brachionus populations described the presence of at least nine genetically divergent Brachionus species and biotypes. Because different studies found evidence that these cryptic species and biotypes differ significantly in ecological preferences and thus probably behave differently in response to rearing conditions in the hatchery, questions rise on the actual identity of the rotifer strains used in aquaculture, where Brachionus discrimination is still based on morphology. This study is a part of an investigation of the genetic make-up of strains used in hatcheries, aquaculture research institutes and laboratories, and describes the rapid and sensitive PCR-DGGE method for the detection of Brachionus species and biotypes based on nucleotide sequence variation within the mitochondrial 16S rRNA gene. Considerable genetic diversity was found, albeit smaller within hatcheries than within laboratories and aquaculture research institutes. All 16S haplotypes produced an unambiguous DGGE fingerprint out of which a database was constructed.     

Effects of biological water purification grid on microbial community of culture environment and intestine of the shrimp Litopenaeus vannamei

Biological water purification grid (BWPG), a type of biofilm, could increase aquaculture production by improving the growth of shrimp and culture environment. In this study, the microbial community composition and diversity of water, sediment and shrimp intestine were analysed using the Illumina Miseq high‐throughput sequencing technique. Installation of BWPG did not only increase the dissolved oxygen, pH and decreased inorganic nutrients, but also increased shrimp body length by 5.39% and weight by 16.35%. Furthermore, it was found that the microbial community diversity of water and shrimp intestine in test pond with BWPG was higher than that of control pond without BWPG. Cyanobacteria, Bacillus and Lactococcus were enriched in the test pond with BWPG. However, Rhodobacter was mainly identified in the test pond. It thus seems to suggest that the application of BWPG in shrimp culture pond enhanced the microbial species richness, types of species and proportion of beneficial bacteria in culture environment as well as shrimp intestine. The results from this study will therefore provide some scientific basis for the improvement and development of shrimp aquaculture.     

AFLP-based genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis

This paper presents the genetic linkage map of the Chinese shrimp Penaeus (Fenneropenaeus) chinensis constructed with 472 AFLP markers. A hundred F₁ progeny from an intercross between a female from the new variety “Yellow Sea No. 1” and wild caught male used for the mapping study. Two separate maps were constructed for each parent. The female linkage map consisted of 197 marker loci forming 35 linkage groups and spanned a total length of 2191.1 cM, with an average marker space of 13.5 cM. The male map consisted of 194 marker loci mapped to 36 linkage groups and spanned a total length of 1737.3 cM, with an average marker spacing of 11.0 cM. The level of segregation distortion observed in this study was 12.2%. The estimated genome length of P. chinensis was 3150.3 cM for the female and 2549.3 cM for the male, respectively. The observed genome coverage was 69.6% for the female and 68.1% for the male map. The linkage maps constructed in this study provide basic information for further linkage studies on Chinese shrimp, and more importantly, the construction of the maps are part of the work of the genetic breeding programs which will be used for growth discovered in the QTL analysis of P. chinensis.     

Comparison of conventional and molecular techniques to investigate the intestinal microflora of rainbow trout (Oncorhynchus mykiss)

The concept of the use of probiotic organisms or prebiotic compounds to modify the fish gut microflora is becoming a popular topic for investigation. A major flaw in many such studies is a failure to consider fully the nature of the established microflora, which is to be modified pre-, or probiotically. Since it is widely accepted that a large proportion of bacteria are non-culturable, the use of conventional bacteriological (culture) techniques alone to investigate fish intestinal microflora may be expected to bias results. We report a study designed to investigate the normal intestinal microflora of rainbow trout (Oncorhynchus mykiss) using both conventional bacteriological and molecular methods.Over an 18 month period, the intestinal microflora of a single population of laboratory-raised rainbow trout was investigated. Bacteria isolated using bacteriological techniques were identified using the BiOLOG system and 16S rRNA gene sequencing. Dominant bacteria consistently were Aeromonas sp. and Carnobacterium piscicola, demonstrating that the microflora is stable in fish kept in defined conditions. Restriction Fragment Length Polymorphism (RFLP) and 16S rRNA gene sequence analysis was used to investigate anaerobic and non-culturable bacteria. An obligate anaerobe, Clostridium gasigenes, was shown to be among the dominating intestinal microflora.     

拟穴青蟹丝裂原活化蛋白激酶激酶（MAPKK）基因的克隆与表达分析

采用qRT-PCR、RACE等方法,获得了拟穴青蟹丝裂原活化蛋白激酶激酶(MAPKK)基因cDNA全长序列。该基因全长1 558 bp,开放阅读框长度为1 224 bp,编码407个氨基酸残基。同源分析显示,该基因编码的蛋白与昆虫的相似性高达70%,推测MAPKK基因在节肢动物具有较高的保守性。经荧光定量PCR检测,MAPKK基因在拟穴青蟹多个组织中有表达,且在脑神经节和卵巢中表达量较高。在拟穴青蟹卵巢发育过程中,MAPKK基因在卵巢发育期(Ⅲ期)表达量最高,发育期为卵母细胞快速生长期,推测MAPKK具有促进卵母细胞快速生长的作用。     

不同施肥方法对鱼蚌综合养殖池塘浮游植物群落结构的影响

通过155 d围隔实验检验了不同施肥条件下鱼蚌综合养殖水体中的浮游植物群落结构。实验设3个处理:施鸭粪、施化肥、兼施鸭粪和化肥。放养种类为三角帆蚌、草鱼、鲫、鲢和鳙,放养量分别为每围隔20、15、5、5和5个。结果发现,围隔内浮游植物生物量平均值为(2.1~6.0)×108个/L。不同施肥方法对浮游植物种类组成和优势种、叶绿素a(Chl.a)、生物量以及蓝藻在浮游植物生物量中的比例无显著影响。浮游植物群落变化表现出较明显的季节性特点,影响围隔浮游植物群落的理化因子为TN、NH3-N和DO。研究表明,采用不同施肥方法的围隔内浮游植物群落结构未表现出显著差异,难以从浮游植物角度解释兼施鸭粪和化肥的围隔珍珠产量高于施鸭粪或施化肥的围隔的事实,也难以确定珍珠产量与浮游植物群落结构之间存在必然的联系。