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1.
云斑尖塘鳢肿大细胞病毒属虹彩病毒的分离与鉴定   总被引:2,自引:0,他引:2  
2009年10月,广东顺德地区一云斑尖塘鳢养殖场暴发不明病因疾病,发病尖塘鳢体长15-18 cm不等,死亡率约85%,濒死尖塘鳢从池塘底层游至水面,呈现游动失衡状态直至死亡.死亡尖塘鳢腹部膨大,剖检可见肝脏、脾脏、肾脏肿大,有出血斑点,从内脏器官肝脏、脾脏和肾脏未分离到致病菌.病鱼内脏组织研磨过滤除菌后,腹腔注射20尾尖塘鳢,7d后开始出现死亡,10d后全部死亡,对照组无死亡.自然发病鱼和人工感染鱼的病理切片显示肝脏、脾脏和肾脏出现大量肿大细胞,超薄切片经电子显微镜观察,肝脏、脾脏和肾脏观察到大量病毒颗粒.电镜下病毒呈六边形,直径约135 nm,形态与虹彩病毒相似.针对虹彩病毒主衣壳蛋白(Major capsid protein,MCP)序列设计引物,提取自然发病鱼和人工感染鱼的DNA作为模板,均能扩增出预期大小的特异性产物.利用NCBI的Blast搜索,结果显示扩增序列与肿大细胞病毒属的传染性脾肾坏死病毒(ISKNV)、闪电丽鱼虹彩病毒(DGIV)和条石鲷虹彩病毒(RBIV)MCP核苷酸序列同源性分别为98.8%、98.1%和94.7%.利用MCP序列构建的系统发育树显示,导致云斑尖塘鳢发病死亡的病毒为肿大细胞病毒属虹彩病毒,暂命名云斑尖塘鳢虹彩病毒(MSGIV).  相似文献   

2.
眼斑拟石首鱼虹彩病毒病的诊断   总被引:1,自引:0,他引:1  
对广东省珠海市每年6—10月池塘养殖的眼斑拟石首鱼(Sciaenop socellatus)暴发不明原因疾病进行了研究。通过对该病的流行病学调查,结合症状观察、病原分离鉴定、病理学变化,结果显示:该病具有明显的传染性,解剖病鱼可见鳃丝贫血、脾脏和肾脏肿大、出血,通过镜检未检测到大量的寄生虫;从发病鱼的肝、脾、肾脏和肠中未分离到致病菌;在病鱼肝、脾和肾组织病理切片中观察到有大量肿大细胞;通过使用OIE 推荐的虹彩病毒特异性引物进行PCR扩增出570 bp左右的目的片段,MCP基因序列比对显示扩增获得的基因序列与真鲷虹彩病毒(RSIV)的基因序列同源性高达99.2%。综合以上结果表明导致眼斑拟石首鱼发病死亡的原因是感染了细胞肿大属虹彩病毒而引起的虹彩病毒病。  相似文献   

3.
根据已知的海水鱼类C型凝集素基因的核苷酸保守区序列设计一对简并引物,先后通过RT-PCR和RACE PCR法从红笛鲷脾脏中首次克隆获得红笛鲷C型凝集素(CTL)基因的c DNA全长(登录号:AGT37609)。该序列长828 bp,开放阅读框663 bp,编码220个氨基酸。氨基酸序列分析显示,红笛鲷CTL基因氨基酸序列与其他物种CTL的相似性在30%-68%之间。系统进化分析表明,红笛鲷C型凝集素与鳉鱼、条石鲷、斜带石斑鱼CTL蛋白亲缘关系最近,聚成一支。通过荧光定量PCR分析红笛鲷基因的组织差异表达,红笛鲷CTL基因在肝、脾以及皮肤中表达水平较高,其次是头肾、肾、胃、肠及肌肉,在心脏与脑中表达量较低。  相似文献   

4.
鳜传染性脾肾坏死病毒主衣壳蛋白基因结构及序列分析   总被引:4,自引:1,他引:3  
分析了鳜传染性脾肾坏死病毒(infectious spleen and kidney necrosis virus,ISKNV)的主衣壳蛋白(MCP)基因结构及其序列。对ISKNV DNA Kpn I L酶切片段的序列分析结果发现,该序列中含有完整的MCP基因。ISKNV MCP基因完整读码框为1362bp,比含量为56.24%,编码一个长为453aa、分子量为49.61kD、等电点为6.25的推定蛋白。结构分析发现,该基因具有启动子元件TATA框和CAAT基序。根据对虹彩病毒MCP系统进化树和脊椎动物虹彩病毒的生物学特性的分析比较发现,ISKNV、RSIV、SBIV、GIV和ALIV等在养殖海、淡水鱼类中引起其脾、肾、固有层和表皮细胞肿大的虹彩病毒,是独立于蛙病毒属和淋巴囊肿病毒属的又一新脊椎动物虹彩病毒类群。  相似文献   

5.
新疆分离的0507JS60鉴定为辽宁病毒   总被引:1,自引:0,他引:1  
0507JS60是从中国新疆喀什地区采集的库蚊标本分离的病毒株,可以在C6/36细胞上稳定传代.电镜观察显示完整病毒颗粒呈球形,直径55nm(n=10),无包膜,衣壳表面壳粒结构非常清晰.基因组核酸电泳显示基因组包括12条双链RNA(Double stranded RNA,dsRNA)片段.病毒第12基因片段序列测定结果显示该片段全长760bp(GenBank ID:FJ157354),具有长度为525bp的单一开放读码框(Single open reading frame,ORF),编码长度为174个氨基酸的蛋白,分子量约18.9kD.病毒第12基因片段序列比对显示与辽宁病毒(Liaoning virus,LNV)核酸序列同源性大于89%,氨基酸序列同源性大于91%.病毒第12基因片段系统进化分析显示该病毒与LNV病毒位于同一进化分枝内,与LNV-NE9712病毒株进化关系更接近.0507JS60经系统鉴定为LNV,这是首次在东北以外的地区分离到该病毒.  相似文献   

6.
0507BS3是从中国新疆喀什地区采集的库蚊和按蚊混合蚊标本分离的病毒株,对C6/36细胞致病变而对Ve-ro和BHK-21细胞不致病变。电镜观察显示病毒颗粒呈圆球形,直径约60nm(n=20),无包膜,单层衣壳,衣壳内有中央核。基因组核酸电泳显示基因组包括10条双链RNA(double stranded RNA,dsRNA)片段。病毒第10基因片段核酸序列测定显示该片段全长964bp(GenBankID:FJ150869),具有单一开放读码框,编码长度为275个氨基酸的蛋白,分子量约30.8kD。病毒第10基因片段核酸序列比对未发现相似的病毒核酸序列,氨基酸序列与胞质多形体病毒(Cytoplasmic polyhedrosis virus,CPV)第10基因片段编码的多形体蛋白部分区段匹配。病毒第10基因片段和已知各型CPV第10基因片段核酸序列共同进行系统进化分析显示该病毒位于独立的进化分枝,提示0507BS3病毒可能是一种新型CPV病毒。  相似文献   

7.
患病中国大鲵中分离到一株虹彩病毒及其特性的研究   总被引:4,自引:0,他引:4  
从陕西某大鲵养殖场患病的大鲵体内分离到一株病毒。患病大鲵以体表溃疡,特别是肢体远端溃烂为主要临床特征。该病毒于10℃~30℃能在BF-2(Caudal trunk cells of blue-gillfry)、CO(Gorad cells of grass carp)、CHSE(Embryo cells of Chinook salmon)、FHM(cells of fathed minnow)等细胞中较好地增殖,最适生长温度为25℃~30℃。病毒对氯仿、热、pH3、pH10敏感,DNA抑制剂5-氟-2′-脱氧尿苷(5-fluro-2-′deoxyuridine,FUDR)能抑制病毒在细胞中的增殖,提示该病毒是有囊膜的DNA病毒。经电镜观察,在感染了病毒的细胞切片中可见到大量直径约130~150 nm有囊膜的六角形病毒颗粒成晶格排列在细胞质里,病毒呈典型的虹彩病毒形态。抽提病毒核酸后进行PCR扩增,用已知蛙病毒主要衣壳蛋白(MCP)基因的保守序列设计的引物能扩增出431bp的片段。扩增的片段测序后,和已知的几种蛙病毒属成员的主要衣壳蛋白基因中的相应片段进行比对,相似性在96%以上。血清学试验结果显示该病毒和IPNV(Infectious pancreatic necrosis virus,IPNV)、GCRV(Grass carp reovirus,GCRV)、SVCV(Spring viraemia of carp virus,SVCV)I、HNV(Infectious hematopoietic necrosis virus,IHNV)在血清学上没有相关性。以上结果提示该病毒可能是虹彩病毒科蛙病毒属的成员,暂时命名为大鲵虹彩病毒(Andrias davidianus iridovirus,ADIV)。该病毒与大鲵发病的关系有待进一步研究。  相似文献   

8.
用电子显微镜观察了稀有(鱼句)鲫出血病(Hemorrhage of Gobiocypris rarus)鱼的病理切片及对照鱼的超薄切片。在对照鱼的鳃、肠道、肾脏、脾脏、肌肉和肝脏中没有见到任何病毒颗粒。在病鱼鳃血管内皮细胞质中观察到聚集的直径70nm左右的GCHV颗粒,说明鳃是GCHV侵袭的主要器官之一,并讨论了鳃对GCHV敏感在GCHV传播中的意义。还在病鱼肠道,肾脏中观察到成片的病毒颗粒,它们也是GCHV侵袭的主要组织。在病鱼脾脏的电子密度低的细胞质中观察到散在的病毒颗粒。很少见到病毒聚集体,脾脏中的病毒可能是脾脏的吞噬细胞吞噬而来。作者认为在病鱼组织中观察到的大小不同的病毒颗粒是处于不同成熟阶段的GCHV。  相似文献   

9.
应用RT—PCR技术,从兔出血症病毒中国分离株WX84中成功扩增出预期大小为1.7kb的特异性条带,将扩增产物提纯后克隆入pGEM^R—T载体,经转化、筛选及酶切鉴定后,获得了该株病毒衣壳蛋白基因的克隆,序列分析表明扩增的中国株BHD衣壳蛋白基因片段长度为1740bp,共编码580个氨基酸。该核酸序列与其它国家报道的多株BHDV序列相互间同源性高达98.2%一99.0%,其推导的氨基酸序列同源性也达98.3%--99.1%,为极度保守片段。  相似文献   

10.
针对近年来由虹彩病毒所引起的海水养殖鱼类疾病呈日趋严重的态势,该文在证实了引发我国大黄鱼大规模流行病的病原为一种虹彩病毒及测定病毒全基因组序列(111,760bp;GenBank accession numberl.AY779031)的基础上,通过与已报道虹彩病毒核酸序列进行分析比较,结合生物信息学手段,确定了以虹彩病毒ATPase基因保守区序列(295bp)作为扩增靶序列,设计合成了一对特异性引物,通过改进PCR模板的制备方法和优化扩增条件,建立了大黄鱼虹彩病毒PCR快速检测技术,并开发成简便、快速、实用的检测试剂盒,该试剂盒的检测灵敏度相当于30个病毒粒子,模板制备时间约30min、回收率为52%、半个工作日即可得到准确的结果,无非特异性扩增带,适用于大黄鱼虹彩病毒病的早期快速诊断、苗种的检疫及水质环境的监测,目前正在推广应用。  相似文献   

11.
Interleukin-1 receptor activated kinases (IRAKs) play crucial roles in the Toll-like receptor (TLR) mediated signal transduction pathways that control host innate immune responses. Here we report the cloning of an IRAK1 cDNA (named ScIRAK1) from the mandarin fish. The predicted ScIRAK1 peptide contains a death domain and a serine/threonine-specific kinase domain. Quantitative RT-PCR showed that ScIRAK1 mRNA was primarily expressed in blood cells and posterior kidney. Seven days following infection with infectious spleen and kidney necrosis virus (ISKNV), the ScIRAK1 mRNA level was significantly higher in the blood cells of clinically symptomatic fish than in the blood cells of asymptomatic fish or control fish injected with phosphate-buffered saline. Additional experiments showed that overexpression of ScIRAK1 in the 293T cells could induce NF-κB activation. These results suggest that ScIRAK1 may play a role in the pathology of ISKNV infection in the mandarin fish.  相似文献   

12.
A better understanding of the immune responses in fish elicited by oral immunisation is of importance for the development of new and effective oral vaccines for cultured fish. In the present study, we characterized specific cell-mediated cytotoxic responses in isogeneic ginbuna crucian carp (Carassius auratus langsdorfii) following oral immunisation with cellular antigens. Trinitrophenyl- (TNP) or dinitrophenyl- (DNP) modified syngeneic and allogeneic cells were used for studying the fine specificity and genetic restriction of orally-induced cytotoxic cells. Hapten-specific cytotoxic responses were detected in peripheral blood leukocytes (PBLs) of fish orally immunised with haptenated syngeneic cells. PBLs from orally immunised fish had cytolytic activity for haptenated syngeneic cells, but they showed little reactivity against both haptenated and unmodified allogeneic targets. Similarly, oral immunisation of fish with hapten-modified allogeneic cells did not induce hapten-specific cytotoxic cells which can lyse haptenated syngeneic targets. Although ginbuna crucian carp possess spontaneous cytotoxic cells that are capable of killing mammalian tumour cells, cold target inhibition studies suggested that such spontaneous cytotoxic cells were not involved in the killing of haptenated syngeneic targets. Oral immunisation of fish with haptenated syngeneic cells also induced hapten-specific cytotoxic memory responses. Oral administration of haptenated fixed cells also effectively induced hapten-specific cytotoxic cells in the treated fish. These findings suggest that oral immunisation with antigens can elicit antigen-specific cytotoxic cells that are capable of recognizing antigens in an MHC-restricted manner. In addition, our results provide indirect evidence that fish possess a mechanism for taking up exogenous non-replicating antigens from the alimentary tract and generating antigen-specific cytotoxic cells.  相似文献   

13.
Immunocytochemical, light microscopy and ultrastructural studies were conducted on gill of sea bream, Sparus aurata L., naturally parasitized with the important parasitic copepod Ergasilus sp. to assess pathology and cellular responses. Thirty-seven S. aurata were examined from a fish farm; 26 (70%) were parasitized, with infection intensity ranging from 3 to 55 parasites per fish. Hosts were divided into two groups, lightly infected fish (<15 parasites per fish) and heavily infected fish (>15 parasites per fish). In histological sections, the copepod encircled gill lamellae with its second antennae, compressed the epithelium, provoked hyperplasia and hemorrhage, occluded arteries and often caused lamellar disruption. Fusion of the secondary lamellae due to epithelial hyperplasia was common in all infected fish; heavily infected fish showed more intense branchial inflammation. In both healthy and infected fish, mast cells (MCs) were free within the connective tissue inside and outside the blood vessels of the primary lamellae and made close contact with vascular endothelial cells, mucous cells and rodlet cells (RCs). MCs were irregular in shape with a cytoplasm filled by numerous electron-dense, membrane-bound granules. Immunostaining of primary and secondary gill filaments with an antibody against the antimicrobial peptide (AMP) piscidin 3 (anti-piscidin 3 antibody, anti-HAGR) revealed a subpopulation of MCs that were positive. These MCs were more abundant in gills of heavily infected fish than in either lightly infected or uninfected fish (ANOVA, P<0.05). Our report documents the response of gill to ectoparasite infection and provides further evidence that mast cells and their AMPs may play a role in responding to branchial ectoparasite infections.  相似文献   

14.
采用常规石蜡组织切片的方法对野生和养殖黄鳍鲷(Sparus latus)消化道的形态组织结构进行了比较观察。结果表明,野生和养殖黄鳍鲷的消化道存在一定差异。(1)形态学研究表明,食道粗而短,胃呈V形,分为贲门部、胃体部和幽门部,胃与肠的连接处有4条幽门盲囊,肠道在体腔内迂回两个回折。野生黄鳍鲷牙齿更为坚硬锋利,体腔中脂肪较少,消化道更为粗短。野生和养殖黄鳍鲷的肠道系数分别为0.71±0.03和0.94±0.12。(2)组织学研究表明,食道黏膜上皮由扁平细胞层和杯状细胞层组成,杯状细胞发达。胃黏膜由单层柱状上皮组成,无杯状细胞,贲门部和胃体部胃腺发达。幽门盲囊组织学特征与肠相似,上皮为柱状上皮,其中的杯状细胞少于肠。肠中,前肠杯状细胞最多,中肠次之,后肠最少。直肠杯状细胞多于肠。野生与养殖黄鳍鲷组织学的区别在于,消化道相同部位养殖鱼的杯状细胞多于野生鱼,野生鱼的肌层厚度大于养殖鱼。黄鳍鲷消化道的形态组织结构与其生活环境和食物是相关的。  相似文献   

15.
This study evaluated the influence of temperature on the immune responses and hematological parameters in channel catfish Ictalurus punctatus immunized via intraperitoneal injection with live theronts of Ichthyophthirius multifiliis. Fish were distributed in 18 aquaria and received 9 treatments: 4 groups of fish were vaccinated with live theronts and maintained at constant temperature 15 °C, 20 °C, 25 °C and 30 °C; 3 groups of fish vaccinated and subjected to cycling temperature regime from 15-25 °C, 20-25 °C and 20-30 °C, changed 5 °C each day; 2 groups of fish were not vaccinated and served as controls at 25 °C, one with Ich challenge and the other without challenge. Non vaccinated fish and those vaccinated at 15 °C or 15-25 °C did not show anti-Ich antibodies in the serum 14 and 21 days post-immunization. The antibody levels were significantly higher from fish vaccinated at 25 °C, 30 °C, 20-25 °C and 20-30 °C compared to fish at 15 °C, 20 °C and 15-25 °C both 14 and 21 days post-immunization. At constant water temperature, fish vaccinated at 15 °C showed significantly higher mortality rate (67.8%, P < 0.05) than those vaccinated at 20 °C, 25 °C, and 30 °C (0-10.7% mortalities). At cycling water temperature, fish vaccinated at 15-25 °C showed significantly higher mortality rate (67.8%) than those vaccinated at 20-25 °C and 20-30 °C (P < 0.05). Twenty days after immunization fish vaccinated at 30 °C and 20-30 °C showed significant increase in the red blood cells, white blood cells, thrombocytes and monocytes. Six days after challenge with I. multifiliis theronts the fish showed decreased white blood cells, thrombocytes and monocytes. This study suggests that vaccinated catfish were severely impacted by low temperature, either at 15 °C constant temperature or at 15-25 °C cycling temperature. The fish showed no anti-Ich antibodies and suffered high mortality similar to non vaccinated control fish.  相似文献   

16.
Two groups of transgenic rainbow trout (Oncorhynchus mykiss, Walbaum) have been produced and compared. One group harbored the reporter gene of chloramphenicol acetyltransferase (CAT) associated with mouse immunoglobulin (Ig) promoter/enhancer (pUCL-CAT-E). The other group carried the same reporter gene under the control of the cytomegalovirus promoter/enhancer (pCMV-CAT). Slot blot analysis of DNA from blood cells and other tissues from pUCL-CAT-E fish showed variation of copy number between the major tissues but not between red and white blood cells. Southern blot analysis indicated that multiple copies organized in concatemers were incorporated into the genome. The pCMV-CAT fish had a pronounced expression of CAT in both white and red blood cells. In contrast, activity of CAT was found in the white blood cells of all pUCL-CAT-E fish but not in their red blood cells. Expression in white blood cells was found preferentially in sIg+ cells, indicating that B cells are the major expressors. High expression was also found in spleen and kidney, but the activity found in thymocytes was equal to the background level. Analysis of some major tissues showed high white blood cell expression associated with low tissue expression, except that liver (known to contain lymphoid tissue in fish) was higher. Thus the regulatory elements of the Ig gene from mouse induce a tissue-specific expression in fish.  相似文献   

17.
Ultrastructural changes of the pineal organ were investigated in the blind cave fish, Astyanax mexicanus, kept under continous artificial light (5000 lux), in continuous darkness, and under natural light conditions. The pineal end-vesicle of the fish kept under natural photoperiod consisted of photoreceptor cells and supporting cells mixed with a few ganglion cells. The photoreceptor cells possessed well-developed outer segments with regularly arranged lamellar membranes. The supporting cells contained a number of lipid droplets and large globular cisternae filled with fine granules. In the fish kept under continuous light or in darkness, the pineal end-vesicle displayed a dilated lumen, and the outer segments of the receptors showed signs of degeneration. Furthermore, alterations of cell organelles were observed in the photoreceptor and supporting cells.  相似文献   

18.
实验模拟自然氧化条件制备氧化鱼油, 替代普通鱼油加入饲料中, 研究其对黄颡鱼(Pelteobagrus fulvidraco)表皮黑色素、酪氨酸酶活力、血清与脑内分泌激素含量的影响。处理组分别为未替代组(C组)、50%氧化鱼油替代组(Y50组)和100%氧化鱼油替代组(Y100组), 结果表明: 随着氧化鱼油替代普通鱼油的比例提高, 黄颡鱼5级(黑色素占细胞面积80%以上)黑色素细胞比例上升; 酪氨酸酶活力上升, 但各组差异不显著(P>0.05); 皮质酮(CORT)在血清和脑组织中各组含量差异均不显著(P>0.05); 肾上腺皮质激素(ACTH)在脑组织中含量增加 (P<0.05), 而血清中Y50组含量高于C组及Y100组(P<0.05); 血清中促肾上腺皮质激素释放激素(CRH)含量降低, Y50与Y100组之间差异不显著(P>0.05), 而脑组织中各组差异不显著(P>0.05); 血清中α-黑素细胞刺激素(α-MSH)含量, Y100组高于其他组(P<0.05), 而脑组织中各组差异不显著(P>0.05)。上述结果表明, 随着氧化鱼油替代普通鱼油的比例提高, 5级黑色素细胞比例上升, 黄颡鱼血清和脑组织中能互相转换的ACTH与α-MSH总量提高, 变化趋势与表皮黑色素含量和酪氨酸酶活性相一致。  相似文献   

19.
The histopathology and ultrastructure of a skin neoplasm in rudd Scardinius erythrophthalmus (L.) are described and the neoplasm is diagnosed as squamous cell carcinoma. In the early stage, tumour cells appear singly or in clusters close to the epidermis; then tumour cells are seen in the dermis as epithelial pearls/nests, cords or sheets with central keratin; and with further development of the neoplasm, tumour cells appear as large sheets and masses, enclosing extensive keratinous formations and foci of necrotic cells, infiltrating deeper into the dermis and penetrating the muscle layers. Increasing vascularity and inflammation are associated with all stages of progression. In some cases metastases are observed in the viscera. Electron microscopic examination shows that the neoplastic cells are joined by desmosomes; the cytoplasm contains abundant mitochondria and rough endoplasmic reticulum. The neoplasm was successfully transmitted experimentally to healthy rudd when tumour cells were inoculated subcutaneously. Eight of 19 surviving test fish developed tumour growth at the site of tumour cell injection and/or in a corresponding site on the opposite of the body. One of these 8 fish, which developed a neoplasm, also showed a microscopic internal tumour in the viscera. One of the 19 test fish showed a microscopic tumour in the spleen, even though no skin tumour was visible in this fish. No tumours were found in control fish. In contrast, intraperitoneal injection of tumour cells into healthy rudd did not result in transmission of the neoplasm.  相似文献   

20.
几种激素在鳜胃肠道内分泌细胞中存在的免疫细胞化学证据   总被引:24,自引:2,他引:22  
用链酶亲和素-生物素过氧化物酶(Strept Avidin-Biotin-Complex,SABC)免疫细胞化学方法,使用4种兔抗消化 道激血清对鳜胃肠道中的内分泌细胞进行鉴别和定位。结果在鳜鱼胃的贲门、幽门及肠道中均发现不同程度地存在着生长抑素和五羟色胺免疫活性内分泌细胞。在鳜的贲门上皮和贲门腺之间,幽门上皮和幽门腺之间均分布有生长抑素免疫活性阳性细胞,在贲门腺和幽门腺处的分布较密。这些含五羟色胺的肽能神经元具有典型的APUD(Amine Precursor Uptake and Decarboxylation)细胞特征,提示鱼类的胃肠道同哺乳动物的胃肠道一样富含肽能神经元;为神经-内分泌系统的研究提供有力的形态学依据,另外两种抗血清-高血糖素和胃泌素的免疫细胞化学染色在鳜的胃肠道的各部位均未发现阳性反应。  相似文献   

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