973 resultados para Prawn Penaeus-japonicus
Resumo:
Juvenile tiger prawns (Penaeus semisulcatus De Haan and P. esculentus Haswell) show a strong association with vegetated habitats and are rarely caught on non-vegetated areas. This pattern of distribution may be caused by postlarvae selecting vegetation when they settle, or to differences in post-settlement mortality in different habitats. In this study, we examined whether the postlarvae and early juvenile stages of P. semisulcatus would distinguish between seagrass (Zostera capricorni Aschers) without epiphytes, artificial seagrass and bare substratum in the laboratory. The responses of prawns reared from the egg to different stages of postlarval and juvenile development were tested to determine whether, and when, each size class showed a response to a particular habitat. Five size classes of postlarvae (average carapace lengths [CL] of 1.2, 1.4, 1.6, 1.7 and 2.0 mm) were offered a choice between Z. capricorni and bare sand. Small size classes of postlarvae either did not respond to Z. capricorni (1.2 and 1.6 mm CL), or were more abundant on bare substratum than Z. capricorni. In contrast, the largest size classes of postlarvae (1.7 and 2.0 mm CL) were more abundant on Z. capricorni during the day but not at night. The behaviour of postlarvae changed markedly at a size of 1.7 mm CL (22 days from the first nauplius): smaller postlarvae frequently swam in the water column; 1.7 and 2.0 mm CL postlarvae spent much more. time resting on the substrate and perched on seagrass leaves. This size at which postlarvae first respond to seagrass during the day, and show mainly benthic behaviour, is similar to the size at which they are found on shallow seagrass beds in northern Australia. Large postlarvae (2.7 mm CL) and juveniles (4.1 mm CL) both were more abundant on artificial seagrass than bare sand during the day but not at night, indicating that they respond to structured habitats. When large postlarvae (2.4 mm CL) and juveniles (3.5 mm CL) were offered a choice between Z. capricorni without epiphytes and artificial seagrass, they were more abundant on the Z. capricorni, which suggests that chemical cues from seagrass may explain some of the responses of P. semisulcatus to seagrass. (C) 1997 Elsevier Science B.V.
Structural analysis of SNARE motifs from sea perch, Lateolabrax japonicus by computerized approaches
Resumo:
Three cDNA sequences encoding four SNARE (N-ethylmaleimide-sensitive fusion protein attachment protein receptors) motifs were cloned from sea perch, and the deduced peptide sequences were analyzed for structural prediction by using 14 different web servers and softwares. The "ionic layer" structure, the three dimensional extension and conformational characters of the SNARE 7S core complex by using bioinformatics approaches were compared respectively with those from mammalian X-ray crystallographic investigations. The result suggested that the formation and stabilization of fish SNARE core complex might be driven by hydrophobic association, hydrogen bond among R group of core amino acids and electrostatic attraction at molecular level. This revealed that the SNARE proteins interaction of the fish may share the same molecular mechanism with that of mammal, indicating the universality and solidity of SNARE core complex theory. This work is also an attempt to get the protein 3D structural information which appears to be similar to that obtained through X-ray crystallography, only by using computerized approaches. (C) 2007 Elsevier Ltd. All rights reserved.
Resumo:
Genetic differentiation of the shrimp Penaeus chinensis in the Yellow Sea and Bohai Sea was investigated using the mitochondrial control region (CR). RFLP of a partial CR segment (613 bp) shows that 106 out of 122 (86.9%) individuals from six sampling localities along the coast of northern China and the west coast of the Korean Peninsula share the same haplotype, and the haplotype frequencies among localities are not significantly different. The findings are further confirmed by sequencing the complete CR. Divergence of the complete CR (992 bp) is less than 1.6% in 14 individuals from the six localities. F-statistics based on RFLP data and the TCS network of sequencing data suggest little genetic differentiation of P. chinensis in the Yellow Sea and Bohai Sea. Mismatch analysis suggests a rapid expansion of P. chinensis population to the Yellow Sea and the Bohai Sea, which probably occurred with the rapid rise in sea level after the last glacial maximum. Despite the lack of genetic heterogeneity, we propose that P. chinensis populations in this region should be treated as separate management units, as fishery management programs have to be applied on a local basis by different governments.
Resumo:
RNA interference (RNAi) is an evolutionarily conserved mechanism by which double-stranded RNA (dsRNA) initiates post-transcriptional silencing of homologous genes. Here we report the amplification and characterisation of a full length cDNA from black tiger shrimp (Penaeus monodon) that encodes the bidentate RNAase III Dicer, a key component of the RNAi pathway. The full length of the shrimp Dicer (Pm Dcr1) cDNA is 7629 bp in length, including a 51 untranslated region (UTR) of 130 bp, a 3' UTR of 77 bp, and an open reading frame of 7422 bp encoding a polypeptide of 2473 amino acids with an estimated molecular mass of 277.895 kDa and a predicted isoelectric point of 4.86. Analysis of the deduced amino acid sequence indicated that the mature peptide contains all the seven recognised functional domains and is most similar to the mosquito (Aedes aegypti) Dicer-1 sequence with a similarity of 34.6%. Quantitative RT-PCR analysis showed that Pm Dcr1 mRNA is most highly expressed in haemolymph and lymphoid organ tissues (P 0.05). However, there was no correlation between Pm Dcr1 mRNA levels in lymphoid organ and the viral genetic loads in shrimp naturally infected with gill-associated virus (GAV) and Mourilyan virus (P > 0.05). Treatment with synthetic dsRNA corresponding to Pm Dcr1 sequence resulted in knock-down of Pm Dcr1 mRNA expression in both uninfected shrimp and shrimp infected experimentally with GAV. Knock-down of Pm Dcr1 expression resulted in more rapid mortalities and higher viral loads. These data demonstrated that Dicer is involved in antiviral defence in shrimp. (c) 2007 Elsevier Ltd. All rights reserved.
Resumo:
An electrochemical technique for the real-time detection of hydrogen peroxide (H2O2) was employed to describe respiratory burst activity (RBA) of phagocytes in plasma which can be used to evaluate the ability of immune system and disease resistance. The method is based upon the electric current changes, by redox reaction on platinum electrode of extracellular hydrogen peroxide (H2O2) released from phagocytes stimulated by the zymosan at 680 mV direct current (d.c.). Compared with the control, activation of respiratory burst by zymosan particles results in a high amperometric response, and a current peak was obtained during the whole monitoring process. The peak current was proved by addition Of Cu2+ and other controls, to be the result of intense release of H2O2 from phagocytes. The peak area was calculated and used to evaluate the quantity of effective H2O2, which represents the quantity of H2O2 beyond the clearance of related enzymes in plasma. According to Faraday's law, the phagocytes' ability of prawns to generate effective H2O2 was evaluated from 1.253 x 10(-14) mol/cell to 6.146 x 10(-14) mol/cell, and carp from 1.689 x 10(-15) Mol/Cell to 7.873 x 10(-1)5 mol/cell. This method is an acute and quick detection of extracellular effective H2O2 in plasma and reflects the capacity of phagocytes under natural conditions, which could be applied for selecting species and parents with high immunity for breeding in aquaculture. (c) 2007 Elsevier Ltd. All rights reserved.
Resumo:
To develop genetic and physical maps for shrimp, accurate information on the actual number of chromosomes and a large number of genetic markers is needed. Previous reports have shown two different chromosome numbers for the Pacific whiteleg shrimp, Penaeus vannamei, the most important penaeid shrimp species cultured in the Western hemisphere. Preliminary results obtained by direct sequencing of clones from a Sau3A-digested genomic library of P. vannamei ovary identified a large number of (TAACC/GGTTA)-containing SSRs. The objectives of this study were to (1) examine the frequency of (TAACC)(n) repeats in 662 P. vannamei genomic clones that were directly sequenced, and perform homology searches of these clones, (2) confirm the number of chromosomes in testis of P. vannamei, and (3) localize the TAACC repeats in P. vannamei chromosome spreads using fluorescence in situ hybridization (FISH). Results for objective I showed that 395 out of the 662 clones sequenced contained single or multiple SSRs with three or more repeat motifs, 199 of which contained variable tandem repeats of the pentanucleotide (TAACC/GGTTA),, with 3 to 14 copies per sequence. The frequency of (TAACC)n repeats in P. vannamei is 4.68 kb for SSRs with five or more repeat motifs. Sequence comparisons using the BLASTN nonredundant and expressed sequence tag (EST) databases indicated that most of the TAACC-containing clones were similar to either the core pentanucleotide repeat in PVPENTREP locus (GenBank accession no. X82619) or portions of 28S rRNA. Transposable elements (transposase for Tn1000 and reverse transcriptase family members), hypothetical or unnamed protein products, and genes of known function such as 18S and 28S rRNAs, heat shock protein 70, and thrombospondin were identified in non-TAACC-containing clones. For objective 2, the meiotic chromosome number of P. vannamei was confirmed as N = 44. For objective 3, four FISH probes (P1 to P4) containing different numbers of TAACC repeats produced positive signals on telomeres of P. vannamei chromosomes. A few chromosomes had positive signals interstitially. Probe signal strength and chromosome coverage differed in the general order of P1 > P2 > P3 > P4, which correlated with the length of TAACC repeats within the probes: 83, 66, 35, and 30 bp, respectively, suggesting that the TAACC repeats, and not the flanking sequences, produced the TAACC signals at chromosome ends and TAACC is likely the telomere sequence for P. vannamei.
Resumo:
Peritrophin, one of the components of the peritrophic matrix, was first isolated from the intestine of insects. It is thought to protect insects from invasion of microorganisms and to stimulate digestion of food. Peritrophin-like proteins have also been found in crustaceans, as a component of the egg layer. In this study, one fragment of the peritrophin-like gene was obtained from fleshy prawn (Chinese shrimp) (Fenneropenaeus chinensis) by panning the T7 phage display library constructed with the shrimp hemocyte cDNA. The total sequence of the peritrophin cDNA was cloned by modified SMART cDNA and LD-PCR methods. The full cDNA is 1048 bp and the deduced protein is composed of 274 amino acids, including 21 amino acid signal peptide, and four peritrophin A domains and the latter three forming three chitin-binding domains. Similarity analysis results showed that the peritrophin-like protein from F chinensis has significant similarities with peritrophin-like and cortical rod proteins from other shrimp. It was inducing expression in hemocytes, heart, stomach, gut, and gills of the infected shrimp, and constitutive expression in the ovaries. No expression signal was detected in the hepatopancreas of either infected or noninfected shrimp. The recombinant peritrophin-like protein has the activity of binding Gram-negative bacteria and strong binding activity to chitin. Therefore, the bacteria and chitin binding activities of the peritrophin-like protein suggest that it may plays a role in immune defense and other physiological resposes. (c) 2005 Elsevier Ltd. All rights reserved.
Resumo:
Fully grown oocytes of Apostichopus japonicus have a cytoplasmic protuberance where the oocyte attaches to the follicle. The protuberance and the oolamina located on the opposite side of the oocyte indicate the animal-vegetal axis. Two pre-meiotic centrosomes are anchored to the protuberance by microtubules between centrosomes and protuberance. After meiosis reinitiation induced by DTT solution, the germinal vesicle (GV) migrates towards the protuberance. The GV breaks down after it migrates to the oocyte membrane on the protuberance side. The protuberance then contracts back into the oocyte and the first polar body extrudes from the site of the former protuberance. The second polar body forms beneath the first. Thus the oocyte protuberance indicates the presumptive animal pole well before maturation of the oocyte.
Resumo:
Triploid Penaeus (Fenneropenaeus) chinensis was successfully produced by heat shock. Their metamorphosis, the relationship between body weight and length and difference in appearance between triploids and their diploid siblings under laboratory culture were studied. Hematological studies showed a smaller number of haemocytes, but larger cell volume, in triploids than in diploids. Triploid shrimp did not show higher growth during the immature stage, but exhibited superior growth during the maturation stage. Characteristics of reproductive organs indicated that triploid shrimp may be sterile and sex ratio can be changed through triploidization of shrimp. This paper summarizes the progress made in triploid shrimp research which would be helpful in understanding more about triploids of crustaceans. (c) 2006 Elsevier B.V. All rights reserved.
Resumo:
本文描述了中国对虾Panaeus chinensis (Osbeck)卵母细胞的发育及其内部结构变化和卵巢的周年变化。根据卵母细胞的大小,核及核仁的形态,以及卵黄粒的有无,皮质棒的出现及卵母细胞与滤泡细胞的关系,将中国对虾卵母细胞的发育分为七个主要时期:(1)中央核仁期,(2)核仁染色质期,(3)周边核仁期,(4)滤泡期,(5)卵黄粒期,(6)皮质特化期和(7)长成期。卵原细胞含有酸性粘多糖。从滤泡期始,卵母细胞核外周出现中性粘多糖,并逐渐充满细胞质。根据卵母细胞的发育,内部结构及卵巢的周年变化特征,将卵巢发育分为五期:卵黄未发生期,卵黄发生早期,卵黄发生后期,皮质棒期和产后期。本文还讨论了卵黄的形成,卵巢发育的分期及亲虾产卵少,即孵化率低等问题。
Resumo:
针对目前刺参养殖过程中亟需解决的种质、环境和产品品质之间日益突出的矛盾,本论文从刺参的自身防御和内分泌机制入手,比较系统地研究了环境胁迫与细胞免疫和应激激素之间的作用机理;并从生理和分子的角度对刺参的夏眠机理进行了初步研究,主要研究成果如下: 1. 查阅国内外大量文献资料,较为系统地综述了海参的免疫体系、生态免疫学研究进展、当前夏眠机理的研究状况和刺参夏眠的基础研究。对刺参生态免疫及其夏眠机理研究提出了新的见解和思路。 2. 研究了温度(0、8、16、24、32℃)和盐度(20、25、35)的急性胁迫(72h)作用对刺参免疫指标的影响。刺参体腔液内几种免疫酶类活性受升温胁迫影响显著;而降温胁迫的影响并不显著。高盐胁迫可使刺参体腔液细胞吞噬活性、超氧化物歧化酶和过氧化氢酶活性显著变化,而对髓过氧化物酶和溶菌酶活性都没有显著影响。从这几种免疫指标变化情况来看,温度胁迫对刺参的影响要强于盐度胁迫,且高温胁迫的影响要强于低温胁迫。 3. 研究了温度(8、16、24℃)、盐度(20、25、35)和露空等急性胁迫(24h)作用对刺参体腔液儿茶酚胺类激素含量的影响。急性温度胁迫能够显著影响刺参体腔液内肾上腺素、去甲肾上腺素和多巴胺的含量;而盐度和露空胁迫对刺参体腔液肾上腺素含量、去甲肾上腺素含量以及多巴胺含量都没有产生显著影响。 4. 完成了刺参体腔液几种酶类变化的周年测定(2006年7月到2007年6月)。一年内刺参体腔液内几种酶类变化的转折点为9月份,10月份,1月份,2月份,4月份和5月份。刺参体腔液内几种酶类活性变化与温度和盐度等环境因素并不存在简单的相关关系,因此推测这些显著变化并不是单一温度、盐度等因素作用的结果,还可能与刺参生长、繁殖及其它环境因素的作用有关。 5. 开展了夏眠期间刺参免疫指标的现场研究(2006年7月到11月)。在此期间刺参体腔液细胞总数显著下降;刺参体腔液内免疫相关酶类均在8月份、9月份达到最高值;肾上腺素和去甲肾上腺素的含量均在8月下旬和11月下旬显著升高,而多巴胺含量则没有显著变化。刺参完全进入夏眠的8、9月份,体腔液内各项免疫指标也都具有显著变化,说明夏眠对刺参机体的免疫状态产生影响。 6. 探讨了刺参夏眠前后肠道基因表达的差异(2007年7月至12月)。抑制消减杂交实验结果表明接头连接效率大于50%,说明连接效率较高。消减效率分析结果表明,消减后进行cDNA保守序列扩增比未消减落后约10个循环才能明显看到扩增产物,说明对共同序列的消减效率比较高。利用抑制消减杂交技术成功构建了符合技术要求的夏眠刺参与未夏眠刺参肠道抑制消减杂交基因文库。
Resumo:
无脊椎动物免疫是一个新兴的研究领域。本文研究了中国对虾的血细胞形态,亚显微结构以及定量测定细胞吞噬活性的方法。中国对虾血细胞分三种:透明细胞,呈现球形或梭形,内部不含颗粒,半颗粒细胞及颗粒细胞卵球形椭球形,内部含有电子致密颗粒,颗粒又分均匀和非均匀颗粒。半颗粒细胞富含线粒体,可能与它对异物敏感易发生胞吐作用有关,半颗粒细胞是识别外源异物的关键细胞。可以观察到高尔基器,它可能与酶原颗粒的形成有关。姬姆萨染色细胞为紫红色,细胞核与颗粒染色较深,呈现蓝紫色与人白细胞中的粒细胞染色结果相近,这或许与两者功能相似有关。应用医学上常用的化学发光法(CL)定量测定中国对虾血细胞的吞噬活性,加入脂多糖及弧菌悬液的血细胞的发光值较只另菌液未加脂多糖的血细胞和对照组的发光值高。实验表明,脂多糖(<10μg/ml)对中国对虾血细胞的吞噬活性具有激活加强作用,而酵母聚糖不能引发血细胞的吞噬作用。上述测定方法可为监测对虾免疫机能的强弱提供定量指标,在对虾养殖生产实战中具有潜在的应用前景。
Resumo:
本论文选择在我国分离得到的一株有毒赤潮甲藻-塔玛亚历山大藻(Alexandrium tamarense,ATHK株),研究了其对一种我国沿海常见和典型养殖鱼类鲈鱼(Lateolabrax japonicus)的危害机制。首先研究了塔玛亚历山大藻(ATHK)对鲈鱼鳃结构的影响及其溶血毒性; 然后采用腹腔注射的方法,研究了高剂量塔玛亚历山大藻毒素(ATHK毒素:约为1.6×105 细胞,相应PSP为0.886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal)和低剂量塔玛亚历山大藻毒素(ATHK毒素: 约为0.16×105 细胞,相应PSP为0.0886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.00118µg STX Equal)在鲈鱼体内代谢过程中对鲈鱼肝脏、肾脏和鳃组织的超微结构、Na+K+-ATPase活性、肝脏功能和肾脏功能的影响、以及对抗氧化系统酶活性和异生物质代谢酶的影响,以期从不同方面了解塔玛亚历山大藻及其所产水溶性毒素(ATHK毒素)对鲈鱼的毒害效应及机制,为有毒赤潮的有效管理提供一定的科学依据。 塔玛亚历山大藻(ATHK)对鲈鱼鳃组织影响的实验结果表明,该藻使鲈鱼鳃组织出现水肿现象,细胞间隙变大;粘液细胞颗粒不规则,颜色加深,颗粒发生凝集,有板结状;氯细胞线粒体内部基质凝集。不产PSP的一种亚历山大藻(AT-6)也使鲈鱼鳃出现水肿,且使细胞出现一定的固缩现象。显微镜观察发现鲈鱼鳃丝间存在有这两种亚历山大藻细胞。由此推测塔玛亚历山大藻ATHK和AT-6的表面结构可能具有能导致鲈鱼鳃组织水肿的机械作用。对人血细胞溶血实验结果表明塔玛亚历山大藻(ATHK)具有较强的溶血毒性,大小与藻的生长阶段和细胞密度都有一定的关系:指数期的溶血毒性最大,随细胞数目的增多,活性逐渐加大;藻细胞、细胞碎片、细胞内容物都有一定的溶血毒性,其中细胞碎片的活性最大。通过11种(株)产PSP的亚历山大藻、不产PSP的亚历山大藻以及标准PSP的实验结果表明这种溶血毒性是由藻细胞的其它非PSP物质造成的,且这种溶血毒性在产PSP的亚历山大藻中具有一定的普遍性。 塔玛亚历山大藻(ATHK)对鲈鱼组织超微结构实验结果表明:ATHK毒素(约为1.6×105 细胞,相应PSP为0.886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal)能导致鲈鱼组织细胞超微结构发生剧烈的变化,主要表现在:肝细胞细胞膜有肿胀现象,部分膜边缘溶解;细胞质糖原颗粒化;核糖体脱落,仅见滑面内质网;细胞质和线粒体内都出现空泡,且线粒体的嵴状结构也发生变化;核膜溶解比较严重,核质外溢,且异染色质边际化。前肾细胞超微结构的变化主要是淋巴细胞核质出现空泡,核膜有溶解迹象;Ⅰ型粒细胞颗粒膨大,伪足增多且变长;Ⅱ型颗粒细胞颗粒增多,内部出现空腔,细胞膜和核膜溶解,胞质、细胞器和核质外溢。鳃组织中氯细胞的核膜局部溶解,核仁弥散,线粒体膜溶解,微细小管膨大;粘液颗粒膜溶解,内部结构遭受破坏;扁平细胞核膜及线粒体膜几乎全部溶解。因此,我们的结果表明,ATHK毒素能作用于鲈鱼细胞的内膜和外膜系统,使膜发生溶解、脱落等变化;比较注射同样剂量大小ATHK毒素120h和240h时鲈鱼组织超微结构发现,细胞超微结构在一定程度上能够恢复。 ATHK毒素对Na+K+-ATPase活性、肝脏功能以及肾脏功能的影响结果表明,0.16×105―1.6×105细胞范围内的ATHK毒素可以显著影响肝脏和鳃组织中的Na+K+-ATPase,使这两种组织中的Na+K+-ATPase活性出现不同程度的下降; 而且还能够显著抑制肝脏中谷丙转氨酶的活性,最大抑制率为95%。但此范围内的ATHK毒素不能显著影响肾脏中的Na+K+-ATPase活性以及尿素氮含量。因此,ATHK毒素对Na+K+-ATPase活性的抑制则会导致细胞能量的缺失,使细胞进一步发生其它变化,而ATHK毒素对肝脏功能完整性的影响则可能会抑制对蛋白质的分解代谢。 ATHK毒素对鲈鱼肝脏、肾脏和鳃组织中的、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱肝肽过氧化物酶(GSH-Px)以及谷胱肝肽转硫酶(GST)活性变化影响的结果表明:高剂量(约为1.6×105 细胞,相应PSP为0.886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.0118µg STX Equal)ATHK毒素能显著诱导鲈鱼肝脏和鳃组织中SOD、GSH-Px以及GST酶活性,最大变化范围为正常状态下的3-4倍,对肝脏中CAT酶活性具有一定的抑制作用,对鳃中的CAT抑制效应则不显著;但此剂量的ATHK毒素仅对肾脏鳃中的GSH-Px活性有一定的诱导作用,对SOD、CAT以及GST的活性没有显著影响。低剂量(约为0.16×105 细胞,相应PSP为0.0886µg STX Equal,相当于每克湿重的鲈鱼PSP注射量为0.00118µg STX Equal)ATHK毒素也能诱导鲈鱼肝脏和鳃组织中SOD、GSH-Px以及GST酶活性,其在第一个24h内的诱导效果与临界致死毒素剂量相似; 且连续注射低剂量ATHK毒素则对肝脏和鳃中这三种酶活性具有累加的诱导作用,使这三种酶活性的变化范围为正常的5倍;低剂量ATHK毒素对肝脏中CAT酶活性也具有抑制作用,但对鳃中CAT酶活性的抑制作用并不显著。 同样,低剂量ATHK毒素除对肾脏中GSH-Px活性具有一定诱导效应外,对SOD、CAT以及GST都没有显著影响。 SOD、GSH-Px以及GST酶活性的显著升高表明ATHK毒素在鲈鱼体内代谢过程中能诱导鲈鱼产生活性氧自由基,且GST活性的升高则说明作为细胞色素P450依赖的异生物质代谢酶,GST在ATHK毒素代谢过程中可能可以加速ATHK毒素的代谢。推测鲈鱼可以通过这三种酶降低ATHK毒素以及次生毒物活性氧自由基对鲈鱼的危害。
Resumo:
1)本文描述了南美白对虾正常的卵子形态及发生特点,以及受精卵和非受精卵的形态学鉴别方法。2)从生产的角度出发提出了评估卵子质量的五项指标:同批卵子规格的一致程度,卵子的沉降速度,孵化膜的抗变形能力,4-8细胞期的分裂特点,扩膜卵在同批卵中所占的比例。除此外,对于亲虾持续产卵与所产卵子质量的关系等也作了相应的探讨。