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1.
为比较凡纳滨对虾(Litopenaeus vannamei)的氧化反应和信号转导相关基因在白斑综合征病毒(WSSV)感染中的变化情况,采用实时荧光定量PCR,分析WSSV感染凡纳滨对虾6、12、24、48、72 h后,鳃和类淋巴组织中硫氧还原蛋白(TRx)、p38信号通路(Lv P38)、过氧化氢酶(CAT)及过氧化物酶(POD)在m RNA转录水平的变化。结果显示,感染对虾的类淋巴组织中,TRx、Lv P38、CAT、POD在72 h时表达量最高,与对照组差异显著(P0.05);而在鳃组织中,该4种基因在12 h时表达量最高,呈先升高后下降的趋势。推断TRx、Lv P38、CAT、POD与WSSV感染密切相关。  相似文献   

2.
采用实时荧光定量PCR技术分析白斑综合征病毒(WSSV)感染凡纳滨对虾不同时段鳃、肠、肝胰腺、肌肉、类淋巴和血细胞组织中热休克蛋白(HSP)60和90基因mRNA转录水平的变化。结果显示,HSP60和HSP90在这6个组织中都有表达,WSSV感染影响各组织中HSP60和HSP90 mRNA转录水平的表达,WSSV感染抑制HSP60在鳃、肠、肌肉和血细胞的表达,促进HSP90在鳃、肠、肝胰腺、肌肉、血细胞和类淋巴的表达,表明HSP60和HSP90参与WSSV感染免疫反应。  相似文献   

3.
为了探讨凡纳滨对虾转录因子AP-1在病毒引发的免疫应答过程中的潜在作用,实验根据前期的转录组和表达谱结果提示信息,首次克隆了凡纳滨对虾AP-1基因(LvAP-1,GenBank注册号:KF999956),利用在线软件进行了生物信息学分析,运用半定量的方法进行了组织表达分析,并利用实时荧光定量PCR(qPCR)技术分析了该基因在白斑杆状病毒(WSSV)侵染过程中的表达变化特征。结果显示,AP-1基因ORF区全长882 bp,编码293个氨基酸。预测分析显示该基因编码的蛋白质含有1个Jun结构域和1个高度保守的亮氨酸拉链结构域(bZIP),其中Jun结构域在非脊椎动物中保守性不高。组织表达分析表明,该基因在凡纳滨对虾各组织中广泛表达,其中在血细胞中表达量最高。在WSSV感染早期(0.5 hpi),该基因表达没有显著改变,感染后5 h(5 hpi),AP-1基因开始显著上调表达,在人工感染后24 h,该基因的表达量达到最高(P0.01)。研究表明,该基因在一定程度上参与了凡纳滨对虾体内由WSSV引发的先天免疫应答过程,为进一步研究LvAP-1在对虾应答病毒侵染过程中的功能和作用机制奠定了基础。  相似文献   

4.
刘强  周俊芳  房文红  王元  李新苍  周帅 《水产学报》2013,37(7):1106-1112
为了评价养殖水环境中毒死蜱对凡纳滨对虾生存的危害性,开展了毒死蜱胁迫下白斑综合征病毒(WSSV)对凡纳滨对虾致死实验,分析了毒死蜱胁迫下凡纳滨对虾鳃组织WSSV含量和肌肉组织乙酰胆碱酯酶活性变化。通过急性毒性实验测定了毒死蜱对凡纳滨对虾的半致死浓度(LC50),随着暴露时间的延长,LC50值显著下降,存在着浓度-反应的正向关系,96 h LC50为0.758 μg/L(0.521~0.987 μg/L)。在此基础上,确定了毒死蜱胁迫实验浓度为0.2 μg/L,此浓度下药浴4 d后对凡纳滨对虾注射WSSV,结果显示:毒死蜱胁迫下注射WSSV组的对虾死亡率(83.33?Ee4.7%)极显著高于乙醇-WSSV组(40.00?Ee0.9%);对虾鳃组织WSSV荧光定量PCR检测结果显示:感染72 h后,毒死蜱-WSSV组WSSV含量约是乙醇-WSSV组的4倍;感染96 h后,毒死蜱-WSSV组WSSV含量显著增加,约是72 h毒死蜱-WSSV组的4.9倍,是96 h乙醇-WSSV组的5.9倍。毒死蜱胁迫下,对虾肌肉组织乙酰胆碱酯酶(AchE)活性低于对照组20%左右。由此可见,毒死蜱胁迫下,WSSV增殖速率加快,导致对虾死亡率升高。  相似文献   

5.
为了分析凡纳滨对虾JAK(Lv-JAK)和STAT(Lv-STAT)基因应答病原菌侵染的表达变化特征,本实验利用半定量PCR技术分析了Lv-JAK和Lv-STAT基因的组织表达特征,并利用实时荧光定量PCR技术分析了其在苏云金芽孢杆菌侵染过程中的表达变化特征。结果显示,Lv-JAK和Lv-STAT基因在凡纳滨对虾的9种组织中有不同程度的表达,均在鳃、肠道和心脏中表达量较高。苏云金芽孢杆菌感染后,在鳃组织中,Lv-JAK和Lv-STAT基因的相对表达量在感染的中晚期(24~72 h)显著上调表达;在肠道组织中,Lv-JAK基因的相对表达量在感染后的6和24 h显著上调,Lv-STAT基因的相对表达量在感染后的24和72 h显著上调。综上表明,JAK和STAT基因在一定程度上参与了凡纳滨对虾体内由苏云金芽孢杆菌引发的天然免疫应答过程,探讨凡纳滨对虾JAK和STAT基因应答苏云金芽孢杆菌感染的表达变化特征,有助于研究对虾JAK/STAT通路在应答病原菌侵染过程中的功能和作用机制。  相似文献   

6.
为了研究含Armadillo重复蛋白8在凡纳滨对虾抗病感染过程中所起的免疫作用,本实验采用RACE-PCR技术首次克隆得到凡纳滨对虾ARMC8基因(LvARMC8,Gen Bank注册号:KX058562)的c DNA序列全长,并利用在线软件进行生物信息学分析;采用实时荧光定量PCR(Rt-PCR)技术对LvARMC8基因在凡纳滨对虾不同组织及感染白斑综合征病毒和副溶血弧菌过程中的表达变化特征进行分析。结果显示,LvARMC8基因全长为2917bp,其中开放阅读框长2046 bp,编码681个氨基酸,5′非编码区长49 bp,3′非编码区长822 bp。预测分析显示LvARMC8编码的蛋白质含有6个ARM结构域。同源性分析发现,LvARMC8基因与内华达古白蚁ARMC8基因的相似度最高,为71%。系统进化分析结果显示,LvARMC8和多种无脊椎动物ARMC8聚为一支,其中与昆虫类动物赤拟谷盗、致倦库蚊和柑橘凤蝶的亲缘关系最近。Rt-PCR分析发现,LvARMC8基因在凡纳滨对虾多个组织中均能检测出,在表皮中表达量最高,眼柄中表达量最低。WSSV感染后12 h,LvARMC8基因在凡纳滨对虾血液中的表达量显著降低,但48 h后显著升高,72 h达到最高水平。除副溶血弧菌感染后24 h外的时间点,LvARMC8基因在凡纳滨对虾血液中的表达量均显著升高。研究表明,LvARMC8基因可能参与凡纳滨对虾抗病免疫应答途径。  相似文献   

7.
氨氮和亚硝基氮共同胁迫对凡纳滨对虾感染WSSV的影响   总被引:1,自引:0,他引:1  
为了评价养殖水环境中氨氮和亚硝基氮对凡纳滨对虾(Litopenaeus vannamei)的危害性,开展了氨氮和亚硝基氮共同胁迫对凡纳滨对虾感染WSSV后的死亡率、WSSV在患病对虾体内增殖速率和对虾主要免疫相关酶活性影响的研究。实验设置氨氮(NH+4)和亚硝基氮(NO-2)的共同胁迫浓度均为20 mg·L-1,分别注射10-4和10-5稀释度的WSSV提取液。结果显示,胁迫下感染10-4WSSV的凡纳滨对虾144 h死亡率达到100%,显著高于无胁迫组(76.67%),相同实验条件下高浓度病毒感染组死亡率高于低浓度组。对虾鳃组织WSSV荧光定量PCR检测结果显示,氨氮和亚硝基氮共同胁迫下凡纳滨对虾体内WSSV的增殖加快,感染48 h后胁迫组病毒量是无胁迫组的1.6倍,72 h时病毒量达到无胁迫组的2.0~3.7倍。此外,免疫相关酶活性结果显示,氨氮和亚硝基氮浓度突变会促使对虾血清中酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)活性先短暂升高然后降低。由此可见,氨氮和亚硝基氮共同胁迫会加快WSSV在患病凡纳滨对虾体内的增殖,导致更高死亡率,这可能是因为胁迫造成了对虾免疫相关酶活性的降低和抗病原感染能力下降所致。  相似文献   

8.
氨氮胁迫下白斑综合征病毒对凡纳滨对虾的致病性   总被引:1,自引:0,他引:1  
为了评价养殖水环境中氨氮(NH_4-N)对凡纳滨对虾(Litopenaeus vannamei)的危害性,开展了NH_4-N胁迫对凡纳滨对虾感染白斑综合征病毒(WSSV)后的死亡率、WSSV增殖速率和对虾主要免疫相关酶活性影响的实验。在NH_4-N胁迫质量浓度为15.6 mg·L-1,分别注射2×105和2×106个WSSV粒子,结果显示,NH_4-N胁迫下注射2×105个WSSV粒子的凡纳滨对虾第144小时死亡率达到53.3%,显著高于无胁迫组(40.0%)。对虾鳃组织WSSV荧光定量PCR检测结果显示,NH_4-N胁迫下凡纳滨对虾鳃组织内WSSV的增殖加快。此外,免疫相关酶活性结果显示,NH_4-N浓度突变会促使对虾血清中酚氧化酶(PO)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性短暂升高后持续降低。由此可见,NH_4-N胁迫会加快WSSV在患病凡纳滨对虾体内的增殖,导致更高死亡率,这可能是因为胁迫造成了对虾免疫相关酶活性降低和抗病原感染能力下降。  相似文献   

9.
将VP110基因的部分序列克隆到pET-28a载体中构建pET28a-vp110b重组质粒并进行原核表达,获得重组表达的蛋白rVP110-B;用rVP110-B注射凡纳滨对虾Litopenaeus vannamei后,经WSSV感染,实验表明,该蛋白注射使凡纳滨对虾感染WSSV的半数死亡时间比对照组延长了20%。用表达纯化的该重组蛋白制备了兔抗rVP110-B多克隆抗体,该抗体用于凡纳滨对虾鳃细胞膜蛋白与rVP110-B的Far-western分析显示,凡纳滨对虾鳃细胞膜蛋白中除90 kDa左右的血蓝蛋白外,在41.7 kDa存在结合条带,经质谱分析表明这条鳃细胞膜蛋白是肌动蛋白。  相似文献   

10.
为了评估饲料中添加酵母提取物对凡纳滨对虾免疫灵敏性和平衡性的相关基因表达的影响,以鱼粉含量为24.5%的基础饲料(对照组)及在基础饲料中添加2.5%酵母提取物的实验饲料,分别饲喂凡纳滨对虾56 d,检测两种饲料投喂的对虾在急性感染溶藻弧菌前后鳃组织Toll受体、IM D和溶菌酶mRNA表达量变化及感染后的对虾死亡情况。结果表明:摄食添加酵母提取物饲料的凡纳滨对虾鳃组织中Toll受体mRNA和溶菌酶mRNA表达量均显著高于对照组对虾(P0.05),IMD mRNA表达量与对照组无显著性差异(P0.05)。急性感染溶藻弧菌后,两组对虾鳃组织Toll受体、IMD和溶菌酶mRNA表达量随感染进程均出现显著变化,Toll受体、IMD和溶菌酶mRNA表达量峰值出现的时间分别为24、42和36 h,且摄食添加酵母提取物组对虾各基因的表达量峰值分别为对照组峰值的201.06%、481.46%和276.77%,均显著高于对照组对虾(P0.05)。摄食添加酵母提取物饲料组对虾鳃组织中Toll受体和IM D mRNA表达量在感染溶藻弧菌后12和24 h分别出现显著上调,而对照组对虾鳃组织中Toll受体和IMD mRNA表达量分别在感染弧菌后24和42 h才出现显著上调。两组对虾经溶藻弧菌人工急性感染后72 h内的累积死亡率无显著差异(P0.05)。实验表明,饲料中添加酵母提取物上调了溶藻弧菌感染前凡纳滨对虾Toll受体和溶菌酶mRNA的表达量,提早了溶藻弧菌感染后对虾Toll受体和IMD mRNA表达量上调时间,且增加了对虾Toll受体、IM D和溶菌酶mRNA表达量的峰值,在一定程度上提高了凡纳滨对虾免疫相关基因表达的灵敏性。  相似文献   

11.
Anguilla luzonensis and A. huangi were each described in 2009 using eels obtained from northern Luzon Island. We examined the taxonomic status of these two groups of eels using morphological and molecular genetic characters. There were no significant differences in two vertebrae counts between eels of A. luzonensis and A. huangi. Mitochondrial 16S ribosomal RNA and cytochrome b genes sequences were obtained and compared among 28 specimens of A. luzonensis, the holotypes of A. luzonensis and A. huangi, and one specimen of the other 15 anguillid species. The specimens of A. luzonensis exhibited almost identical sequences, including the holotype, with only a few site differences, and the genetic difference between the holotypes of A. luzonensis and A. huangi was within the range of differences of specimens of A. luzonensis. The other anguillid species were genetically very different from A. luzonensis and A. huangi, although A. interioris is a closely related species. It is clear that A. luzonensis and A. huangi are the same species, and according to the principle of priority in zoological nomenclature, A. luzonensis Watanabe, Aoyama, and Tsukamoto, 2009 is the valid species name, and A. huangi Teng, Lin, and Tzeng, 2009 is a junior synonym of A. luzonensis.  相似文献   

12.
The effects of exposing the eggs of Pacific threadfin and amberjack eggs (AEs) to different concentrations of hydrogen peroxide for 5 min on hatch rate and survival were assessed in a series of experiments using a petri dish model rearing system. Despite significant inter‐batch variation in hatch rate, it was shown that eggs of both species could be safely exposed to up to 11 340 mg L−1 H2O2 for 5 min. Exposure to 34 230 mg L−1 H2O2 for 5 min was shown to be lethal to AEs at a late stage of development. In two further experiments, it was demonstrated that Pacific threadfin eggs were resistant to all tested concentrations of a range of polyvinylpyrrolidone iodine (PVP‐I) concentrations and contact times (up to 1000 mg L−1 PVP‐I for 10 min). The level of bacteria adhering to the eggs of both species was highly variable. Where eggs were heavily colonized (>104 cfu egg−1), hydrogen peroxide concentrations of at least 11 340 mg L−1, or PVP‐I concentrations higher than 500 mg L−1 for 10 min, were required for effective sterilization. In less colonized batches, rinsing in sterile seawater or exposure to lower (550 mg L−1) concentrations of H2O2 was sufficient to result in high apparent levels of surface sterility (<1 cfu egg−1).  相似文献   

13.
The purpose of this study was to evaluate the acute toxicity of lead for different life stages of shrimp, Litopenaeus vannamei, and to determine the effect of its sublethal concentrations on osmoregulatory capacity (OC) as well as the possible histological alteration in the gills of juvenile shrimp. The 24‐, 48‐, and 96‐h LC50 values for lead to L. vannamei increased progressively with increasing life stage, from nauplii < zoeae < mysis < postlarvae < juvenile. After 15‐d exposure to 3.25, 6.50, and 9.75 mg Pb/L, the OC values of exposed shrimp were reduced by 39, 73, and 157%, respectively compared to control animals. Compared to controls, the lead concentrations in gill tissues increased significantly by 127,500, 137,500, and 141,000% in shrimps exposed to 3.25, 6.50, and 9.75 mg Pb/L, respectively. After lead exposure, hemocytic congestion in efferent vessels and multiple hyperplasia were observed in gill filaments, resulting in narrowed hemolymphatic lacuna. The histopathological effects increased with increasing Pb concentration.  相似文献   

14.
对驼背鲈(Chromileptes altivelis)的胚胎发育及仔、稚、幼鱼的形态特征进行了详细的观察与研究,描述了从受精卵到仔、稚、幼鱼各发育期的时间和形态特征变化。结果表明,在水温25-26℃、盐度30的海水中,受精卵历时27 h 25 min完成整个胚胎发育过程,经历从卵裂、囊胚、原肠、神经胚到肌节形成、各器官的逐渐形成、变化、完善等一系列的胚胎发育和变化过程;根据其卵黄囊消长情况、鳞片的覆盖状态、体色发生的不同变化以及第二背鳍和腹鳍的消长,将胚后发育分为仔、稚、幼鱼3个阶段。在水温22-26℃、盐度29-31、DO≥5 mg/L的条件下,2-3 d仔鱼卵黄囊消失,开口摄食;生长发育至31 d,仔鱼已变态进入稚鱼期;培育至57 d,稚鱼完成变态,成为幼鱼。第二背鳍棘和腹鳍棘的生长与收缩等石斑鱼类早期发育的共性生长特征及其体表特性体色变化特征为驼背鲈胚后发育过程中最明显的特征。  相似文献   

15.
As an intertidal species, Apostichopus japonicus is subject to diel and seasonal temperature fluctuations. Color variation is a distinct characteristic of A. japonicus, and a new color morph, purple A. japonicus, was recently found on the coast of Rushan, Weihai City. This study was conducted to compare the performance of green, white, and purple color morphs of A. japonicus at different water temperatures to help improving aquaculture technology and management. In this study, green, white, and purple color morphs of juvenile A. japonicus (1.5 ± 0.1 g) were cultured at 10, 14, 18, 22, and 26 C for 60 d, according to the suitable temperature range for sea cucumber growth. Temperature was shown to have a significant effect on growth, energy budget, and physiology of sea cucumbers (P < 0.05). Specific growth rates, feed conversion efficiencies, and the proportion of energy allocated to growth of all three color morphs were optimal at 18 C. Activities of the enzymes related with oxidative stress, immune response, and digestion were determined. Superoxide dismutase (SOD) and trypsin activities generally reached a maximum in all three color morphs at 18 C. The purple color morph of A. japonicus was higher in growth rate, food conversion efficiency, and proportion of energy allocated to growth than green and white color morphs at 18 and 22 C, consistent with its higher trypsin and SOD activity, indicating the mutual effect of growth and physiology. At 18, 22, and 26 C, the green color morph had the highest level of lysozyme activity and purple color morph had the lowest, indicating possible infection by external pathogens of green color morph, which might be caused by high temperature. These results suggested that the optimum temperature for culture of the three color morphs was approximately 18 C, the purple had a better adaption to high temperature, and the green and white color morphs may be more susceptible to thermal stress than the purple color morph.  相似文献   

16.
Mosquitofish, Gambusia sp., have been spread throughout the world to biologically control mosquitoes. However, the fish has gained a reputation as an invasive species and has been implicated in displacing native aquatic species. Gambusia affinis are native to the southeastern United States and commonly occur in commercial channel catfish, Ictalurus punctatus, production ponds. We investigated effects of mosquitofish presence on zooplankton populations, water quality, disease occurrence, and fish production in experimental ponds. There were no differences between ponds with or without mosquitofish in numbers of calanoid copepods, cyclopoid copepods, total copepods, Bosmina sp., Ceriodaphnia sp., Moina sp., Daphnia sp., or total cladocerans. There were also no differences in copepod and cladoceran sizes. Copepod nauplii were more numerous during the summer months in ponds with mosquitofish. There were no differences in water quality variables (soluble reactive phosphorus, nitrate, nitrite, ammonia, total nitrogen, total phosphorus, pH) or phytoplankton density between ponds stocked with and without mosquitofish. Catfish production and disease occurrence were also similar between ponds with and without mosquitofish. Although mosquitofish may cause problems when stocked outside their native range, there does not appear to be any adverse effects of mosquitofish presence in catfish production ponds.  相似文献   

17.
Mulloway (Argyrosomus japonicus) occur in estuarine and coastal waters surrounding Australia, Africa, India, Pakistan, China, Korea and Japan, where they are important in fisheries. This study identified that mulloway in south-eastern Australia had similar growth rates, but matured at smaller lengths and younger ages, to those in South Africa and Western Australia. Growth of both sexes was similar to about 5 years, after which females grew faster and attained a greater maximum length than males. Female mulloway matured at 4–5 years of age with a L50 of 68 cm, whereas males matured at 2–3 years of age with a L50 of 51 cm. The commercial fishery in New South Wales was characterised by declining catches and a reduction in the proportion of mulloway of mature lengths in landings. During 2002–2005 commercial landings were dominated (83%) by fish within 15 cm of the current minimum legal total length of 45 cm and aged 2 and 3 years (>80%), even though mulloway can attain lengths of 200 cm and live >30 years. Estimates of the rates of instantaneous total mortality ranged between 0.34 and 0.45, whilst the rate of instantaneous natural mortality (M) was estimated to be approximately 0.12. Yield-per-recruit analyses indicated that mulloway in New South Wales are being growth overfished and substantial increases in yield could be achieved by increasing the length at first harvest. Values of the spawning potential ratio were below 0.2 under a range of mortality estimates, suggesting that mulloway are at risk of recruitment overfishing. These results suggest that the spawning stock of mulloway in south-eastern Australia has been depleted and that remedial management action is required to protect this iconic species.  相似文献   

18.
The recruitment strategy of swimming crabs, Portunus trituberculatus, was investigated by field observations and numerical experiments in Hiuchi-Nada, Japan. Calculated dispersal patterns of zoeal and megalopal stages in a three-dimensional numerical model reproduce observed patterns accurately. The temporal change of vertical migration patterns between the 1st and 2nd zoeas and the 3rd and 4th zoeas plays an important role in the process of recruitment of P. trituberculatus from spawning ground to nursery ground.  相似文献   

19.
Abstract

Effects of salinity on embryonic development and growth of African catfish, Clarias gariepinus, eggs and larvae were studied. Eggs were incubated at 27-29°C in 2,4,6,8, and 10 ppt sodium chloride. Rate of embryonic development was delayed in all salt solutions by 15, 15,28 and 30 minutes, in 2,4,6, and 8 ppt sodium chloride, respectively, when compared with the control group (0% salt); total mortality occurred at 12 hours after gastrula stage in the 10 ppt concentration. Percentage hatching was 45.1,47.7, 59.5,49.2, and 26.6% while percentage deformity was 10.4, 16.1, 52.0, 28.6, and 71.6% in 0, 2, 4, 6, 8, and 10 ppt salt treatments, respectively. There were significant differences (P <0.05) in the hatching percentage and in deformity percentage between 4, 6, and 8 ppt. Rate of yolk absorption was significantly faster in the control and 2 ppt salt treatments, but slower in 4, 6, and 8 ppt. Rate of increase in length was slower with increasing salinity. The optimum salinity for African catfish eggs and was between 0-2 ppt and acceptable up to 6 ppt. The results suggest that increasing salinity delayed hatching and development of African catfish eggs and larvae, respectively, as well as increased the deformity of the larvae.  相似文献   

20.
Abstract

Rainbow trout, Oncorhynchus mykiss, were raised in culture cages (1 m3) to determine the effect of stocking density on growth, survival, and percentage of market-size fish. Large fingerling rainbow trout (20-25 cm, 232 g average weight) were stocked into six cages located in a 0.4-ha pond. Two stocking densities (100 or 200 fish/cage) were used, and fish were grown for 140 days (2000-April 2001). Average total harvest weight (35.0 kg) in the low-density cages was approximately one-half the average total harvest weight (61.2 kg) in the high-density cages. Average weight gain (11.7 kg to 15.1 kg) and feed conversion (1.2 to 1.5) were also smaller for the low-density cages. Average survival was 96.7% for the low-density cages and 94.2% for the high-density cages, with the percentage of market-size fish (< 29 cm) averaging 50.3% and 52.0%, respectively. Production costs for the actual experiment and the revenues from fish sold at the end of the study were collected. An enterprise budget based on the experimental results for the two densities was developed to determine if a culture operation of this size would produce a net return. Production costs and revenues from the experiment resulted in a large negative return (-$3,124) and high breakeven price ($13.53/kg).  相似文献   

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