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1.
根据斜带石斑鱼(Epinephelus coioids)神经坏死病毒(orang-spotted nervous necrosis virus,0GNNV)的主衣壳蛋白(major capsid protein,MCP)基因的保守序列,设计一对引物,从感染0GNNV的斜带石斑鱼组织匀浆液提取RNA为模板进行RT-PCR扩增,得到426bp的cDNA片断。用得到的RT-PCR产物加上地高辛(DIG)标记作为核酸探针。通过注射病毒提取液人工感染一组斜带石斑鱼,解剖感染病毒的斜带石斑鱼,从中分离出脑和眼睛,运用原位杂交技术检测组织中的OGNNV。实验表明,原位杂交具有较高的灵敏性和特异性,可以用原位杂交的办法来检测养殖的石斑鱼是否有携带该病毒,达到监控和预防神经坏死病爆发的目的。本实验还采用了H&E染色方法检测了感染NNV的石斑鱼脑部和眼部组织,观察细胞内的坏死部分。与原位杂交做对比,提高了检测的可靠性和准确性。本实验建立的石斑鱼神经坏死病毒的ISH检测方法有着较高的特异性和敏感性,易于操作,有助于石斑鱼神经坏死病毒的组织定位、发病机理的研究。  相似文献   

2.
网箱养殖青石斑鱼河流弧菌病研究   总被引:9,自引:1,他引:9  
随着养殖规模的扩大 ,病害问题日益严重 ,严重地制约着石斑鱼养殖业健康发展。刘秀珍等1994年报道创伤弧菌是石斑鱼的病害弧菌病的病原。本文作者对1999年夏季厦门市同安区琼头村网箱养殖石斑鱼的严重病害进行了研究 ,试图能为养殖青石斑鱼(Epinephelusawoara)的病害防治提供理论依据。1材料与方法1.1试验用鱼患病石斑鱼取自同安区琼头村养殖网箱 (体重97~245g) ;健康石斑鱼购自厦门火烧屿养殖网箱 (体重81~114g) ,暂养备用。1.2病原菌的分离将病鱼体表清洗消毒后 ,用无菌操作方法从肝脏、…  相似文献   

3.
云纹石斑针淋巴囊肿病病变过程的超微研究   总被引:3,自引:1,他引:3  
张永嘉  郭青 《海洋与湖沼》1997,28(4):406-410
于19891-990年,对海水网箱养殖的云纹石斑鱼自然感染和人工感染的淋巴囊肿病进行电镜观察。观察结果表明,自然感染和人工感染的淋巴囊肿细胞之间并无区别,经们均含有许多包涵体和病毒颗粒;主要描述用患淋巴囊肿病鱼的皮肤结节匀浆对实验鱼 下注射而感染的淋巴囊肿病的超微病理过程,包括病毒颗粒在成纤维细胞胞膜上的附着和成纤维细胞及其内含物的形态变化等过程;此外,病鱼心脏、肝脏、脾脏、肾脏和肠上皮也有不同程  相似文献   

4.
云纹石斑鱼淋巴囊肿病病变过程的超微研究   总被引:9,自引:1,他引:9  
于1989—1990年,对海水网箱养殖的云纹石斑鱼自然感染和人工感染的淋巴囊肿病进行电镜观察。观察结果表明,自然感染和人工感染的淋巴囊肿细胞之间并无区别,它们均含有许多包涵体和病毒颗粒;主要描述用患淋巴囊肿病病鱼的皮肤结节匀浆对实验鱼作皮下注射而感染的淋巴囊肿病的超微病理过程,包括病毒颗粒在成纤维细胞胞膜上的附着和成纤维细胞及其内含物的形态变化等过程;此外,病鱼心脏、肝脏、脾脏、肾脏和肠上皮也有不同程度的病理变化。  相似文献   

5.
溶藻弧菌脂多糖在两种鱼类体内的组织分布   总被引:3,自引:0,他引:3  
用溶藻弧菌Vibrio alginolyticus的VAL02和VAL00尾静脉和腹腔注射点带石斑鱼Einephelus malabaricus,收集感染后1,4,24,48和144h鱼的各器官标本,用斑点免疫ELISA(dot Enzyme-linked immunosorbent Assay)法检测其中的脂多糖(lipopolysaccharide,LPS),发现肠,肝,脾,肾前部,肾后部均可吸收溶藻弧菌的LPS,而心脏吸收LPS的能力比其它器官差;检测结果还表明肝,肾组织中的LPS比脾组织中的LPS清除快。用斑点免疫ELISA法检测VAL02号菌感染的野生五带豆娘鱼Abudefduf vaigiensis器官组织中溶藻弧菌的LPS,结果在肠,肾,脾和肝脏中检测出LPS,在心脏组织中没有检测出LPS。实验结果表明溶藻弧菌LPS不易被点带石斑鱼和五带豆娘鱼的心脏组织吸收,但可被它们的肠,肾脏,肝脏和脾脏吸收。  相似文献   

6.
记述华南地区网箱养殖石斑鱼鱼虱病病原──南海鱼虱,新种Caligusnanhaiensisnsp.形态的光镜和电镜观察结果,并对南海鱼虱与相似种C.seriolaeYamaguti,1936和C.tanagoYamaguti,1939的形态特征进行详细比较。  相似文献   

7.
记述华南地区网箱养殖石斑鱼鱼虱病病原-南海鱼虱,新种Caligus nanhaiensis n.sp.形态的光镜和电镜观察结果,并对南海鱼虱与相似种C.seriolae yamaguti,1936和C.tanago Yamaguti,1939的形态特征进行详细比较。  相似文献   

8.
为了确定鱼类神经坏死病毒对军曹鱼(Rachycentron canadum)仔鱼的致病性,本研究通过肌肉注射、浸泡和共浴3种途径分别感染军曹鱼仔鱼,观察了由其导致的发病死亡情况和病理学变化。结果表明,3种感染途径都能造成军曹鱼仔鱼出现厌食、体色发黑和螺旋游泳等典型的病毒性神经坏死症症状,累计死亡率分别达到78%、44%和34%。肌肉注射18h后,利用RT-PCR方法在鱼的脑、眼、脊髓中均可检测到该病毒。患病鱼的脑、视网膜、脊髓等组织发生空泡化,与其他鱼类感染该病毒发生的变化一致。因此,军曹鱼仔鱼是鱼类诺达病毒的易感种群之一,病毒能通过水平传播感染军曹鱼仔鱼。  相似文献   

9.
建立了一种同时检测对虾白斑综合征病毒(WSSV)、传染性皮下和造血器官坏死病毒(IHHNV)的二温式多重PCR技术。根据基因库中WSSV(AF369029)和IHHNV(NC002190)基因序列,设计了两对分别与WSSV和IHHNV某段保守基因序列互补的引物,用这两对引物对同一样品中的WSSV和IHHNVDNA模板进行多重PCR扩增,结果均同时得到了两条大小与实验设计相符的593bp(WSSV)和356bp(1HHNV)特异性多重PCR扩增条带,而对其他对虾疾病病原的PCR扩增结果均为阴性;敏感性测定结果表明,该多重PCR技术最低能检测到WSSV和IHHNVDNA各100pg。用该多重PCR对400份临床病料进行检测,结果230份检出WSSV,阳性率57.5%;96份检出IHHNV,阳性率24%;56份同时检出WSSV和IHHNV,阳性率14%。18份为阴性,阴性率为4.5%。结果提示了WSSV和IHHNV广泛存在于中国南方的养殖对虾中,作者建立的二温式多重PCR可以用于这两种病毒的临床快速检测和鉴别诊断。  相似文献   

10.
血液病理学测定表明,患病赤点石斑鱼Epinephelusakaara组列的血红蛋白平均值约5.49g.(100ml)-1,颗粒白细胞平均值为48.09%,单核细胞平均值为2.82%,健康鱼组列的血红蛋白平均值为7.26g·(100ml)-1,颗粒白细胞平均值为31.33%,单核细胞平均值为0.11%,两组列间有明显差异(p<0.05)。患病石斑鱼出现较多星个体小、膜薄易破、难着色的异形红细胞;颗粒白细胞的核分叶较多,病鱼表现出严重贫血症状。  相似文献   

11.
The number of harbour seals in Japan has been rebounding since protection began in the mid‐1980s. With the increase in seal numbers, increased conflict with fisheries has occurred through depredation and the belief that seals compete with fisheries for prey. However, competition can only be determined if the prey species and quantities seals consume over time are known. We studied the diet of harbour seals in Erimo, site of the largest population of harbour seals in Japan, from 2011 to 2012 and assessed the degree of prey overlap with local fisheries. We used both hard parts and DNA techniques to identify prey items in seal scats, and compared these results to local fisheries data. A total of 46 prey occurrences was detected by both methods, of which 17 matched between techniques at least to the family level. Hard parts methods identified five incidences of prey undetected by DNA methods in five scats (one incidence per scat). DNA methods identified 24 additional prey occurrences in 13 scats, for which no hard part evidence for that prey had been found. This more than doubled the total number of prey occurrences across the 15 scats compared. Overall, the most frequently occurring harbour seal prey were walleye pollock (Theragra chalcogramma), sculpins and snailfishes. In contrast, the top three groups targeted by fisheries were codfishes, salmon and invertebrates. Many species common in the harbour seal's diet such as snailfishes and blennies were not targeted by fisheries. Fishes such as greenlings, sculpins, rockfishes and Japanese anchovy (Engraulis japonicus) were common in the diet of harbour seals, but made up a very small proportion of fisheries catches in Erimo. The importance of other prey species varied between seasons. Sculpins, greenlings and forage fishes were the top three prey groups for harbour seals in spring by percent modified frequency of occurrence, whereas the most important target groups by percentage mass caught by fisheries were codfishes, cephalopods and other invertebrates. Sculpins and rockfishes remained as important prey items for harbour seals in summer and autumn, in addition to codfishes, snailfishes and blennies. In contrast, the main groups caught by fisheries in summer were other fishes, particularly Japanese amberjack (Seriola quinqueradiata) and various species of sharks and eels, and invertebrates. By autumn, Erimo fisheries had focussed on catching salmon. Salmon were taken by harbour seals as well during this period, but at a relatively lower frequency compared to other prey groups. The results of our study show that although harbour seals consumed several of the prey species targeted by fisheries, the relative importance of these species to seals and fisheries and the seasons in which they were targeted were different.  相似文献   

12.
Porcelain crabs possess a ‘hair‐trigger’ propensity to autotomize their chelipeds (claws), and laboratory studies have demonstrated that this ability is highly effective in avoiding predation from other crabs. However, porcelain crabs are also subject to predation from fishes, which use a very different means of capture. In this study, we investigated whether autotomy in porcelain crabs is also effective against predation by fishes. To do this, we examined stomach‐contents data from four common species of kelp‐forest rockfishes and determined the frequency of disassociated chelipeds (those with no associated bodies) in porcelain crabs and in brachyuran crabs, which do not readily autotomize their chelipeds. We found that disassociated chelipeds of porcelain crabs were six times as common as those of brachyuran crabs (35% of the remains of all porcelain crabs versus 6% of the remains of all brachyuran crabs). We interpret this difference to be evidence that, through autotomy, porcelain crabs escaped ingestion of their entire bodies, and thus certain mortality, at a higher rate than did brachyuran crabs. We conclude that autotomy constituted an effective escape strategy from rockfish predation. This evidence, in conjunction with previous studies, suggests that autotomy of chelipeds in porcelain crabs is an effective anti‐predatory strategy against phylogenetically disparate predatory groups.  相似文献   

13.
Mytilus edulis digestive gland microsomes were prepared from indigenous populations sampled from a clean reference site (Port Quin) and an urban-industrial contaminated site (Blackpool) in the UK. Samples were collected in March/April, May, August and December 1998. Western blot analysis was performed using polyclonal antibodies to fish CYP1A and rat CYP2E using partially purified M. edulis CYP as a positive control, to aid identification. CYP1A- and CYP2E-immunopositive protein levels showed different site-specific seasonal variation with higher levels of CYP2E determined in May (P < 0.05). At both sites, lower levels of CYP1A-immunopositive protein but not CYP2E-immunopositive protein were observed in the samples collected in December (P < 0.05). This correlated with lower levels of nuclear DNA damage (Comet assay expressed as per cent tail DNA) observed in December compared to August (P < 0.05).  相似文献   

14.
Recruitment of pelagic larval fishes to the nearshore environment is dependent on a suite of biological and physical processes operating at many spatial and temporal scales. Nearshore circulation processes associated with coastal upwelling are widely upheld as major determinants of year class strength for many rockfishes (Sebastes spp.), but the mechanism by which these processes drive recruitment is largely unknown. We used Standard Monitoring Units for the Recruitment of Fishes (SMURFs) to monitor recruitment of two rockfish complexes (Sebastes spp.) and cabezon (Scorpaenichthys marmoratus) from March to September of 2004 and 2005 at 3 sites along the central California coast. We examined the relationship between recruitment of these fishes and measurements of oceanographic variability associated with upwelling dynamics, including in situ water temperature, AVHRR sea surface temperature, the Bakun upwelling index, and an index of alongshore surface water transport. We found that rockfish comprising the KCGB complex (Sebastes atrovirens, Sebastes caurinus, Sebastes carnatus, Sebastes chrysomelas) recruit during early summer, while fishes of the BYO complex (Sebastes melanops, Sebastes flavidus, Sebastes serranoides), as well as cabezon recruit during late summer. Our results provide limited support for an association between the arrival of juvenile pelagic rockfish and cabezon to the nearshore environment and physical processes related to upwelling and relaxation. Beyond the limitations of our bimonthly sampling scheme, the lack of a clear pattern may be related to the near absence of upwelling–relaxation cycles along this stretch of coast during these two study periods. Moreover, the settlement and recruitment of nearshore fishes may be closely tied to processes occurring earlier in the larval stage.  相似文献   

15.
Edwardsiella tarda is one of the most important emerging pathogens in the global aquaculture industries. As such, an accurate diagnosis and quantitative analytical methods are urgently needed for this bacterium. In this study, primers and a TaqMan probe specific to the conservative sequences of the 16S rRNA gene of E. tarda were designed. The concentration of primers and TaqMan probe were optimized to 200 nmol/L and 120 nmol/L, respectively. The detection sensitivity of the FQ-PCR assay was determined to be as low as five copies of the target sequence per reaction using the pGEM-16S rDNA recombinant plasmid as a template, which was 100 times more sensitive than conventional PCR. A standard curve by plotting the threshold cycle values (y) against the common logarithmic copies (log10 nc as x; nc is copy number) of pGEM-16S rDNA was generated. The results of intra-and inter-assay variability tests demonstrate that the established FQ-PCR method was highly reproducible. The assay was specific for E. tarda as it showed that there was no cross-reactivity to eight additional bacterial pathogen strains in aquaculture. Thus, the FQ-PCR assay has the potential for diagnostic purposes and for other applications, especially for the rapid detection and quantification of low-grade E. tarda infections.  相似文献   

16.
When cultured in Dulbecco's minimal essential medium the established epithelioma papulosum cyprini cell line from carp was found to be vitamin E-deficient due to the very low level of vitamin E in the medium and the foetal calf serum used as supplement. The toxicity of oxidative stressors to this cell line was evaluated by means of the neutral red cytotoxicity assay and it was found that an organic hydroperoxide, t-butylhydroperoxide was extremely cytotoxic and that the redox-cycling agents diquat and menadione were less toxic. When grown under vitamin E supplementation (25 microM), the toxicity of these chemicals was reduced by at least an order of magnitude in concentration demonstrating the protective effect of vitamin E. These data show the importance of vitamin E status for interpretation of in vitro and in vivo data and that this in vitro system is useful for mechanistic studies.  相似文献   

17.
吴翠琴 《台湾海峡》2009,28(1):60-64
采用半静态暴露的方式研究己烯雌酚(DES)和辛基酚(OP)对真鲷幼鱼血浆性激素含量的影响.将真鲷幼鱼暴露于一定浓度的DES和OP时间为6周,用放射免疫法测定对照组和染毒组血浆中的17β-雌二醇(E2)和睾酮(T)浓度.结果表明,DES和OP严重干扰了真鲷幼鱼的内分泌系统,特别是对17β-雌二醇含量的影响非常显著.  相似文献   

18.
采用酶联免疫吸附法测定了野生中华鸟塘鳢(Bostrichthyssinensis)不同季节血清E2和1l—KT含量的变化,并比较了生殖季节养殖雌雄同体与野生雌雄异体之间E,、11一KT和17仪,2013一P三种性类固醇激素含量的差异.结果表明,野生雌鱼和雄鱼血清E:和11一KT的含量均具有明显的季节变化,其高低均依次为5〉8〉2〉11月.在相同月份,雌鱼的E,含量显著高于雄鱼,而雄鱼的ll—KT含量显著高于雌鱼.生殖高峰期,雌雄同体E:含量高于雌雄异体雄鱼,但低于雌雄异体雌鱼;雌雄同体l1一KT和17仅,20]3一P含量均高于雌雄异体雌鱼和雄鱼;野生雌鱼和雄鱼血清170L,2013一P含量没有显著差异.性类固醇激素在中华鸟塘鳢的生殖过程中起着重要作用,E:是野生中华乌塘鳢雌鱼的主要雌激素,而11一KT则是野生雄鱼的主要雄激素.  相似文献   

19.
Benzothiazole, a common chemical associated with tire manufacturing and industrial wastewater, is a principal component of both fresh water and estuarine tire leachate, a neurotoxicant to larval sheepshead minnows (Cyprinodon variegatus) in in vivo estuarine studies. The neurotoxic potential of benzothiazole was investigated following in vivo and in vitro exposure of sheepshead minnows to 3.75, 7.5, 15, 30 and 60 mg/l benzothiazole. Following benzothiazole exposure, fish were evaluated for survival, growth and histological alterations. Fish mortality occurred after 5 days of exposure to 60 mg/l (LC50 = 41.9). Significant decreases in larval growth were noted at all concentrations. Histologically, gills had cellular alterations but the central nervous system lacked the severe cellular damage seen in previous tire leachate exposure studies. Benzothiazole cytotoxicity to primary cultures of brain cells from sheepshead minnow and tilapia (Oreochromis niloticus) and two epithelial cell lines was evaluated using a tetrazolium salt assay (MTT) at 1 and 4 days. In vitro results indicate primary cultures of brain cells are less sensitive to benzothiazole than epithelial cell lines. Significant cytotoxicity to the epithelial cell lines was noted at 30 and 60 mg/l concentrations. Histologically and cytotoxicologically, the present study indicates that benzothiazole is a gill toxicant and not a neurotoxicant.  相似文献   

20.
In the present study, we report N_2 fixation rate(~(15)N isotope tracer assay) and the diazotroph community structure(using the molecular method) in the western tropical North Pacific Ocean(WTNP)(13°–20°N, 120°–160°E). Our independent evidence on the basis of both in situ N_2 fixation activity and diazotroph community structure showed the dominance of unicellular N_2 fixation over majority of the WTNP surface waters during the sampling periods.Moreover, a shift in the diazotrophic composition from unicellular cyanobacteria group B-dominated to Trichodesmium spp.-dominated toward the western boundary current(Kuroshio) was also observed in 2013. We hypothesize that nutrient availability may have played a major role in regulating the biogeography of N_2 fixation.In surface waters, volumetric N_2 fixation rate(calculated by nitrogen) ranged between 0.6 and 2.6 nmol/(L·d) and averaged(1.2±0.5) nmol/(L·d), with 10 μm size fraction contributed predominantly(88%±6%) to the total rate between 135°E and 160°E. Depth-integrated N_2 fixation rate over the upper 200 m ranged between 150 μmol/(m~2·d)and 480 μmol/(m~2·d)(average(225±105) μmol/(m~2·d). N_2 fixation can account for 6.2%±3.7% of the depthintegrated primary production, suggesting that N_2 fixation is a significant N source sustaining new and export production in the WTNP. The role of N_2 fixation in biogeochemical cycling in this climate change-vulnerable region calls for further investigations.  相似文献   

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