锦鲤疱疹病毒病

<正>锦鲤疱疹病毒病(Koi herpes virus disease,KHVD)是20世纪末被确定的一种疾病,现已流行于世界各地,是严重威胁鲤和锦鲤养殖业安全的一种疾病,世界动物卫生组织(OIE)列为必须申报的疾病,我国将其列为二类疫病。一、病原学病原是锦鲤疱疹病毒(K o i herpes virus,KHV),暂列为疱疹病毒科(Herpesviridae),鲤疱疹病毒属(Cyprinid herpesvirus),又称鲤疱疹病毒Ⅲ型(CyHV-Ⅲ),与鲤痘疮病毒(鲤疱疹病毒Ⅰ型,CyHV-Ⅰ)和金鱼造血器官坏死病毒(鲤疱疹病毒Ⅱ型,CyHV-Ⅱ)同属。KHV和CyHV-Ⅰ存在交叉的抗原反应。KHV是球状病毒,成熟病毒颗粒有囊膜,直径约170nm～230nm。核衣壳为对称十面体,直径100nm～110nm,由31种病毒多肽组成,其中21种多肽分子量与鲤疱疹病毒相似,10种多肽与斑点叉尾鮰病毒(Channel catfish virus,CCV)相似。KHV含有双链DNA,基因组大小约为277kb,比疱疹病毒科的其他病毒基因组(250kb)大。二、流行病学目前该病流行范围已遍及欧亚美非各大洲的以色列、英国、德国、美国、南非、日本、韩国、中国、马来西亚、新加坡、印度尼西亚等国家,并在部分国家造成危害。1998年5月,该病在以色列首次发生,随后18个月内连续发生三次,造成以色列600吨食用普通鲤和400万美元出口锦鲤的损失。使以色列鲤和锦鲤养殖业遭受毁灭性打击。2002年4月印度尼西亚养殖锦鲤和鲤暴发该病,损失500万美元。2003年10月日本10余个县暴发该病,造成近千吨的鲤和锦鲤死亡。KHV的严重危害引起国际动物卫生组织(OIE)和世界粮农组织(FAO)的高度关注。KHV仅仅感染锦鲤、鲤和剃刀鱼(Solenostomus paradoxus)。其鱼苗、幼鱼、成鱼,均可感染。KHV发病最适温度是23℃～28℃(低于18℃,高于30℃不会引起死亡)。若鱼已感染KHV,水温18℃～27℃间持续时间越长,疾病暴发的可能性越大。该病多发于春、秋季,潜伏期14天,鱼发病并出现症状24小时～48小时后开始死亡,开始死亡至2天～4天内死亡率可迅速达80%～100%。KHV暴发后幸存的鱼成为疾病的传播者,可将病毒传染给其他健康的鱼。KHV主要通过水平传播,能否垂直传播目前尚未确定。三、临床症状病鱼停止游泳,鱼眼凹陷,皮肤上出现苍白的块斑与水泡,     

四川地区养殖鲫鲤疱疹病毒Ⅱ型的鉴定及病理学研究

为探究疾病病因和流行特点,对患病鲫进行剖检、病理学观察、分子生物学检测及人工感染实验。结果显示,患病鲫临床表现为离群独游、浮头、全身发黑及体表出血等。剖检发现鳃、肝脏、肾脏等器官出血、肿胀和坏死,鳔点状出血。组织病理观察显示,鳃呼吸上皮细胞肿胀、坏死及脱落;肾脏局灶性坏死;脾脏组织广泛变性坏死,造血系统崩解;肝脏发生空泡变性和坏死。透射电镜观察到脾脏和肾脏中存在4种大小不同的病毒颗粒,分别为DNA病毒内核、空衣壳、实心衣壳和有包膜的成熟病毒粒子。未成熟的病毒粒子存在于细胞核,成熟的病毒粒子存在于细胞质。电镜下可见鲤疱疹病毒Ⅱ型(CyHV-2)在细胞核内完成核酸复制和核衣壳装配,囊膜蛋白在出核后获得。细胞核萎缩、核染色质边缘化,线粒体肿胀、坏死、嵴断裂,细胞质空泡化。取患病鲫组织匀浆滤菌后接种到鲤上皮瘤细胞系(epithelioma papulosum cyprinid cell line,EPC)和胖头鲤肌肉细胞系(fathead minnow cell line, FHM),盲传3代后未见细胞病变。通过腹腔注射患病鲫组织匀浆进行人工感染实验,实验组鲫表现出与自然感染鲫相同的临床症状,对照组鲫正常,实验组累积死亡率为80%。用PCR方法检测CyHV-2的解旋酶基因片段,扩增出预期大小相符的目的条带。利用邻近法对该病毒的解旋酶基因进行系统发育分析,结果显示与CyHV-2(YZ-01)的同源性为99%。以上实验结果证实了CyHV-2是导致这次疾病暴发的原因。     

溶氧胁迫对异育银鲫感染鲫造血器官坏死症的影响

随着异育银鲫(Carassius auratus gibelio)养殖密度的增大,其主要养殖区爆发了以鳃部出血为典型特征的疾病,经确诊为鲫造血器官坏死症(Crucian Carp Hematopoietic Necrosis),引起该病的首要诱因为鲤疱疹病毒Ⅱ型（CyHV-2),给异育银鲫养殖造成了巨大的经济损失。溶氧是水生生物生存的必要条件之一,溶氧条件的改变直接影响水生生物的各项生理机能,在溶氧胁迫状态下,水生生物会表现出应激及免疫力下降,从而导致疾病的发生。为了探究溶氧浓度对异育银鲫感染CyHV-2及发生鲫造血器官坏死症的影响,将健康的异育银鲫与已感染CyHV-2的病鱼混合饲养,结合鲫的最低生存氧浓度,设定溶氧浓度分别为7、6、5、4、3 mg/L,不同溶氧浓度均设立对照组,在相同的饲养条件下,通过控制不同溶氧浓度,测定CyHV-2感染数及死亡数,分析不同溶氧浓度条件下鲫CyHV-2感染率及鲫造血器官坏死症的发病率。结果表明,随着溶氧浓度的降低,CyHV-2的感染率和死亡率都随之升高,当溶氧浓度为7、6、5、4 mg/L时,对照组均未出现感染及死亡状况;当溶氧浓度为 3 mg/L时,实验组感染率为100%并且对照组也出现了死亡,充分肯定了溶氧胁迫会增加异育银鲫对CyHV-2的感染率及死亡率。研究结果对异育银鲫的大规模养殖及造血器官坏死症防控具有重要的意义。     

冷水观赏鱼红鲫、锦鲤的亲本培育及病虫害防治试验

红鲫又名金鲫或草金鱼,属鲫鱼科,具有色彩艳丽、生长快、适应性强、性成熟早、繁殖力强等特点,因价格适中,易饲养,深得市场的青睐,是观赏鱼市场最基础,需求量最大的品种。锦鲤又名绯鲤,属鲤科,具有品系较多,色彩华丽,体形大,生长快,适应性强,繁殖能力强等特点,因色彩华丽,体格健美,花纹多变,泳姿优雅,被誉为水中的"活宝石"。近年来,云南省的红鲫、锦鲤的养殖取得了一定的发展,市场占有率也有一定提高,但产量和品质还远远不能满足云南市场的需求,大量的商品鱼,尤其中高端的商品鱼都要依赖省外提供。为了提高     

鲤疱疹病毒3型T分离株主要免疫原性蛋白的鉴定

为了鉴定鲤疱疹病毒3型(Cyprinid Herpesvirus 3,CyHV-3)主要免疫原性蛋白,研究采用CyHV-3-T分离株感染CCB细胞系,运用蔗糖密度梯度超速离心方法对CyHV-3进行纯化,纯化的病毒颗粒经SDSPAGE和考马斯亮蓝染色后,用锦鲤(Cyprinus carpio haematopterus)抗CyHV-3阳性血清进行Western blotting分析和液相色谱串联质谱鉴定。结果表明,透射电镜下可观察到大量完整囊膜包裹或只有裸露核衣壳的CyHV-3颗粒,Western blotting结果显示抗CyHV-3阳性血清与多种病毒蛋白具有明显特异性免疫反应,质谱鉴定表明其中4种免疫原性蛋白分别为ORF92、ORF66、ORF72和ORF81,其中ORF66和ORF72为首次鉴定的具有免疫原性衣壳蛋白。本研究将为CyHV-3血清学诊断方法的建立、亚单位疫苗或DNA疫苗的研制提供更多候选抗原。     

水晶彩鲫、红鲫、锦鲤、荷包红鲤杂交子代的生长和体色研究

利用人工授精的方法,进行水晶彩鲫、红鲫、锦鲤和荷包红鲤的相互杂交试验,测定各个杂交组合子代的成活率、生长速度和体色分离比例。结果表明：4种鱼能够相互杂交受精,孵出鱼苗。孵化率锦鲤自交最低为46．4％,其它组合为70％～80％;杂交鱼苗经28d的人工饲养,水晶彩鲫与荷包红鲤、锦鲤的正、反杂交,同其它杂交组合比较有明显的差别,其生长速度慢,个体之间差异大,成活率低;杂交子代的体型分为3类：鲫鱼型、鲤鱼型和鲤鲫型。鳞片反光组织(虹彩细胞或鸟粪素细胞)为2类：完全型、缺失型。体色分离复杂多样,水晶彩鲫与红鲫杂交是水晶彩鲫,红鲫与锦鲤、荷包红鲤杂交是青灰色鲤鲫杂种,水晶彩鲫与锦鲤、荷包红鲤杂交都会出现水晶彩色和青灰色鲤鲫杂种。     

三种杂交鲤对同种配合饲料的利用试验

<正> 本试验用含粗蛋白27.39％的配合饲料对三种杂交鲤子一代(散鳞镜鲤♀×鲤鲫移核鱼(?),鲤鲫移核鱼♀×散鳞镜鲤(?),荷元鲤♀×散鳞镜鲤(?);以下分别简称杂交鲤Ⅰ、Ⅱ、Ⅲ)夏花鱼种进行90天的喂养,结果报道如下。材料与方法一、试验池。为三个水泥池,编号1*、2*、3*,面积分别为50、50、25平方米,水深0.8～1米,分别放养杂交鲤Ⅰ、Ⅱ、     

鲤疱疹病毒Ⅱ型TaqMan real-time PCR 检测方法的建立及应用

针对鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2,CyHV-2)DNA解旋酶基因编码区序列设计特异性引物,利用PCR技术扩增出长度为1446 bp的基因编码区片段,克隆到pMD19T载体上,构建重组质粒.经PCR鉴定与测序分析确认正确后,以10倍梯度稀释重组质粒,作为标准模板进行TaqMan real-time PCR扩增,制作标准曲线,建立了鲤疱疹病毒Ⅱ型的荧光定量PCR检测方法.检测结果显示,标准曲线的相关系数(R2)达到0.9991,斜率为-3.412;对初始模板定量检测的范围为1×101～1×107 copies/μL;特异性试验结果表明,该方法可特异性地检测出鲤疱疹病毒Ⅱ型,而对大鲵虹彩病毒(GSIV)、锦鲤疱疹病毒(KHV)以及空白对照无检测信号.取江苏射阳和宝应两地疑似患病鲫组织核酸作为模板进行荧光定量PCR,结果表明反应体系中的病毒量分别为6.89×104 copies/μL和3.02×102 copies/μL.本研究建立的鲤疱疹病毒Ⅱ型TaqMan实时荧光定量PCR方法灵敏度高、特异性强,对因鲤疱疹病毒Ⅱ感染引起的养殖鲫造血器官坏死症的诊断与病毒病原定量检测有重要意义.     

鲤疱疹病毒3型ORF148 DNA疫苗对建鲤鱼苗的免疫保护

鲤疱疹病毒3型(Cyprinid herpesvirus 3, CyHV-3)又称为锦鲤疱疹病毒(Koi herpesvirus, KHV),是一种高传染性、致死性病毒。为开发新型CyHV-3 DNA疫苗,前期研究中将CyHV-3 ORF148基因插入pEGFP-N1构建重组质粒,在此基础上,本研究通过转染试验、间接免疫荧光试验证实pORF148-EGFP融合蛋白可以在CCB-J细胞系和建鲤体内表达;将重组质粒作为DNA疫苗,肌肉注射免疫建鲤鱼苗,ELISA检测表明免疫pEGFP-ORF148重组质粒可以显著提高建鲤血清特异性抗体水平; RT-qPCR检测显示,免疫pEGFP-ORF148重组质粒后,建鲤脾脏和头肾中的免疫相关基因如IFN-a1、Mx-1、CXCa、CXCR1、TNF-α、IL-1β及IgM基因表达量均显著提高。攻毒实验显示, CyHV-3攻毒21 d后, PBS组、pEGFP-N1组和pEGFP-ORF148组的建鲤存活率分别为30%、35%和85%,免疫pEGFP-ORF148可以显著提高建鲤的存活率(P0.01)。本研究旨在为CyHV-3 DNA疫苗的应用提供理论依据。     

鲤疱疹病毒Ⅱ型ORF66基因DNA疫苗载体的构建及其免疫效果

鲤疱疹病毒Ⅱ型(CyHV-2)能够引起鲫大量死亡,严重威胁我国水产养殖业的健康发展,目前,针对CyHV-2尚无有效的商业化疫苗或治疗措施。为构建CyHV-2DNA疫苗,本研究将其衣壳蛋白ORF66编码基因克隆至pVAX1真核表达载体上,构建重组质粒pVAX-ORF66。此外,在BL21(DE3)pLysS中对ORF66蛋白进行了原核表达,表达蛋白经纯化后免疫新西兰白兔制备了ORF66特异性抗体。酶联免疫吸附(ELISA)检测显示,制备抗体效价达1∶20000以上。将pVAX-ORF66质粒转染金鱼脑细胞系(GFB),利用制备的ORF66抗体进行间接免疫荧光(IFA)检测,结果显示,ORF66蛋白可以在细胞中大量表达,且主要定位于细胞质中。将pVAX-ORF66质粒肌肉注射鲫后进行CyHV-2免疫保护实验,结果表明,其相对免疫保护率达55.6%。本研究针对CyHV-2构建了一种制备简单、成本低廉的DNA疫苗,为鲫造血器官坏死病的免疫预防及感染分子机制研究奠定了前期实验基础。     

Anguilla huangi Teng, Lin, and Tzeng, 2009, is a junior synonym of Anguilla luzonensis Watanabe, Aoyama, and Tsukamoto, 2009

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.     

Atlantic Fisheries Technological Conference,Moncton, NB,August 21, 1995

No abstract available for this article.     

Age,growth, maturity and the overfishing of the iconic sciaenid,Argyrosomus japonicus,in south-eastern,Australia

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.     

Effects of Mosquitofish,Gambusia affinis,on Channel Catfish,Ictalurus puncatatus,Production Ponds

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.     

Surface disinfection of Pacific threadfin, Polydactylus sexfilis, and amberjack, Seriola rivoliana, eggs

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).     

Compositional differences,antioxidant activity,angiotensin-converting enzyme inhibition,and sensory characteristics of kamaboko fermented in wheat,rice, brown rice,and soybean koji

Fisheries Science - Fermentation of kamaboko in koji with wheat (KW), rice (KR), brown rice (KB), or soybean (KS) was performed by simulation of tofuyo processing. Moisture and protein content...     

Sperm Motilities in Waigieu Seaperch,Psammoperca waigiensis: Effects of Various Dilutions,pH, Temperature,Osmolality, and Cations

The objectives of this study were to evaluate the effects of varying dilutions, pH, temperature, osmolality, and cations on sperm motility parameters in waigieu seaperch, Psammoperca waigiensis. The maximum velocity of average in path (VAP), percentage of motile cells (MOT), and duration of sperm motility (DSM) were observed when semen was diluted in artificial seawater (ASW) at a ratio of 1:100 (144.9 ± 0.6 µm/sec, 95.6 ± 0.4%, and 230.3 ± 2.3 sec, respectively), at 30 C (142.0 ± 0.6 µm/sec, 93.6 ± 0.4%, and 238.3 ± 0.9 sec, respectively), and pH 8 (144.8 ± 0.6 µm/sec, 93.3 ± 0.4%, and 234.0 ± 1.5 sec, respectively). Maximum VAP, MOT, and DSM were obtained in each solution containing 0.6 M NaCl (143.8 ± 1.0 µm/sec, 91.3 ± 2.0%, and 230.6 ± 4.2 sec, respectively), 0.6 M KCl (135.1 ± 3.1 µm/sec, 91.1 ± 3.1%, and 230.3 ± 3.7sec, respectively), 0.2M CaCl₂ (105.3 ± 4.7μm/sec, 47.9 ± 2.7%, and 120.7 ± 1.3 sec, respectively), 0.2 M MgCl₂ (107.3 ± 3.0 m/s, 42.1 ± 3.3%, and 120.3 ± 4.8 sec, respectively), and osmolality of 400 mOsm/kg (145.1 ± 2.5 µm/sec, 93.0 ± 2.1%, and 346.5 ± 4.4s, respectively). We used these mediums as artificial insemination media for fertilizing matured eggs. The results showed that the fertilization and hatching rates in 0.6 M NaCl (75.3 ± 0.6% and 57.0 ± 2.4%, respectively), ASW (70.8 ± 1.2% and 51.2 ± 1.8%, respectively), or 400 mOsm/kg (72.9 ± 1.8% and 55.3 ± 1.6%, respectively) were higher than that in seawater (63.9 ± 1.2% and 39.2 ± 3.9%, respectively). In conclusion, using 0.6 M NaCl, ASW, or 400 mOsm/kg as an artificial insemination medium is effective for fertilizing of waigieu seaperch.     

Operating parameters affecting biofloc technology: carbon source,carbon/nitrogen ratio,feeding regime,stocking density,salinity, aeration,and microbial community manipulation

Aquaculture International - The world population continues to increase day by day. Therefore, aquaculture needs to grow in order to response the need for protein foods and to cope with the problem...