Herpesvirus cyprini: lethality and oncogenicity

Abstract. Cyprinid herpesvirus 1 (CHV) or Herpesvirus cyprini was virulent for carp, Cyprinus carpio L., fry following 1 h immersion in water at 20 °C. Cumulative mortality for carp fry was 86–97% in 2-week-old common carp, 20% in 4-week-old fancy carp, and 0% in both 8-week-old common and fancy carp. The virus did not produce mortality in fry of crucian carp, grass carp or other cyprinids. It was also oncogenic in carp, inducing papillomas to the extent of 55% among both common and fancy carp fry. The neoplasms appeared 5–6 months after carp had been exposed to the virus by immersion and recurred at an incidence of 83% in carp 7·5 months post-desquamation of the tumour. The CHV was reisolated from all moribund fish and from all survivors. It also induced papillomas at an incidence of 13% in adult mirror carp and at 10% in adult fancy carp 5 months after intraperitoneal inoculation of 10⁵ TCID₅₀ ml^-1 fish. The virus was rcisolated only from the ncoplastic tissue and not from internal organs. The neoplasms were normally located on fin, skin or mandible, at the intraperitoneal inoculation site. Specific fluorescence for CHV antigen was frequently detected in the gills, liver, kidneys and intestine of 2-week-old fry from 3 to 21 days following challenge with CHV. It was found in greater concentrations in experimentally induced papillomata on 2-week-old carp fry survivors examined 24 weeks after challenge than in naturally occurring neoplasms.     

一例冰下低温爆发的锦鲤疱疹病毒病的鉴定

2013年4月初低温下,吉林省白山地区某网箱养殖场鲤（Cyprinus carpioL．）大量死亡,病死鲤无明显临床病变。采用PCR扩增技术检测了这些病鱼,结果表明,该低温爆发的疾病为锦鲤疱疹病毒病（KHVD）。这种病常爆发在水温为18-28℃的春秋季,冰下爆发还未有报道。本次锦鲤疱疹病毒病的鉴定,对于锦鲤疱疹病毒感染鲤温度条件的变化和疾病的预防提供了参考。     

Dynamics of experimental production of Thelohanellus hovorkai (Myxozoa: Myxosporea) in fish and oligochaete alternate hosts

The dynamics of development and production of Thelohanellus hovorkai (Myxozoa) were examined to investigate factors inducing haemorrhagic thelohanellosis in carp, Cyprinus carpio L. Fresh actinospores of T. hovorkai were harvested from the oligochaete alternate host, Branchiura sowerbyi, and used for infection experiments with myxosporean-free carp. Visualization of actinospores by fluorescent labelling revealed that sporoplasms penetrated the gill filaments of carp immersed in an actinospore suspension as early as 30 min post-exposure (PE). Plasmodia of T. hovorkai developed in the connective tissues of various organs and matured 3-5 weeks PE; dispersion of myxospores from degenerate plasmodia occurred 5-7 weeks PE. Challenges with a high dose of actinospores (4.5 x 10(6) spores per fish) resulted in the onset of disease, which was more easily achieved by the oral intubation of actinospores than by immersion in an actinospore suspension. Actinosporean-free B. sowerbyi were exposed to different densities of myxospores (10(4)-10(6) spores per oligochaete) and subsequently reared at different temperatures (15, 20, 25 degrees C). At 20 and 25 degrees C, actinospore releases were first detected 40-43 days PE, with multiple peaks of release (max. 7 x 10(5) actinospores day(-1)) during the next 60 days. We concluded that the developmental cycle of T. hovorkai was completed within 3-5 months at 20-25 degrees C, and that the ingestion of large numbers of actinospores orally, possibly by feeding on infected oligochaetes, resulted in a disease condition in carp.     

Fatty acid composition of the ingested food only slightly affects physicochemical properties of liver total phospholipids and plasma membranes in cold-adapted freshwater fish

The fatty acid content, fatty acid molecular species composition, and fluidity of phospholipid vesicles and plasma membranes of livers of fish feeding on phytoplankton (silver carp, Hypophthalmichtys molitrix), zooplankton, (big head carp, Hypophtalmitrix nobilis), higher plants, (grass carp, Ctenopharyngodon idella), and omnivorous fish, (common carp, Cyprinus carpio), adapted to reduced temperatures, was determined. Levels of long chain polyunsaturated fatty acids (PUFAs) were highest in silver and big head carp and lowest in common carp. Differences in long chain PUFAs were poorly reflected in fluidity, determined either by fluorescence anizotropy of 1,6-diphenyl 1,3,5-hexatriene or rotational correlation time of 16-doxyl- stearic acid, of phospholipid vesicles. Phospholipid vesicles of C. caripo and C. idella proved to be slightly more rigid than of H. molitrix and H. nobilis below ca. 15 °C. These differences were not seen with isolated plasma membranes assessed with 1,6-diphenyl 1,3,5-hexatriene. Using electron spin resonance spectroscopy plasma membranes from H. molitrix proved to be more fluid than those from C. carpio below ca. 15 °C. Phosphatidylethanolamines were abundant in 1-monounsaturated, 2-polyunsaturated (18:1/22:6, 18:1/20:4) species with H. molitrix and H. nobilis more rich in these species than C. carpio and C. idella. It is suggested that differences in fluidities of phospholipid vesicles below 15 °C is due to differences in levels of the above molecular species. In native membranes these differences may be masked by proteins and other membrane components. It is also suggested that it is not the fatty acid composition of the ingested food but the specific response of the fish to temperature that is the most important factor controlling the physicochemical properties of membranes during thermal adaptation.     

Distribution of the introduced cyprinid herpesvirus 3 in a wild population of common carp, Cyprinus carpio L.

Cyprinid herpesvirus 3 (CyHV-3), which causes a lethal disease in common carp, Cyprinus carpio L., and koi, C. carpio koi , first occurred in Lake Biwa, Japan in 2004. To elucidate distribution of CyHV-3 in a wild common carp population, we conducted a PCR survey of CyHV-3 among such fish in Lake Biwa in 2006. Only 6% (1/18) of the common carp smaller than 300 mm were positive with PCR, whereas 31% (18/58) of fish larger than 300 mm were positive. To evaluate their past exposure to CyHV-3 infection based on the presence of antibodies, we also measured the levels of serum anti-CyHV-3 antibodies in the carp, using an enzyme-linked immunosorbent assay. None (0/26) of the fish smaller than 300 mm was positive for the antibodies, whereas 54% (33/61) of fish larger than 300 mm were positive. Of the antibody-positive individuals, 44% (14/32) were also positive by PCR strongly suggesting that wild common carp that survived infection become CyHV-3 carriers. Five individuals were positive by PCR but negative for antibodies indicating that their infection with CyHV-3 had occurred recently. These results suggest that transmission of CyHV-3 from carriers to naïve common carp is still occurring in Lake Biwa.     

Antibody response of two populations of common carp, Cyprinus carpio L., exposed to koi herpesvirus

Common carp, Cyprinus carpio L., exposed to koi herpesvirus (KHV) may become persistently infected and populations containing such virus-infected individuals may transmit the virus to other fish when co-habited. Detection of virus-infected fish in a population is thus critical to surveillance and control programmes for KHV. A study was therefore designed to detect anti-KHV serum antibodies, with an enzyme-linked immunosorbent assay, in common carp following experimental exposures to KHV under varying environmental conditions. The study determined that a proportion of fish within a population experimentally exposed to KHV (at least 10–25%) develop high antibody titres (1/1600 or greater) to the virus, and this immunological response was detectable for several months (observed at the termination of the experiments at 65, 46 and 27 weeks post-exposure). Furthermore, this response was detected in one population of fish that did not succumb to a high level of mortality when maintained at water temperatures that were non-permissive for KHV. Elevating the water temperatures to permissive conditions for KHV resulted in recurrence of disease despite the presence of anti-virus antibodies, suggesting that serum antibodies alone are not protective under the conditions of our trials.     

Movement, social cohesion and site fidelity in adult koi carp, Cyprinus carpio

Abstract  Koi carp is an ornamental variant of the common carp, Cyprinus carpio L., that was introduced to New Zealand in the 1960s and has since become a major aquatic pest. A total of 1265 wild adult koi carp were caught by boat electric fishing, dart tagged and released at multiple sites in the lower Waikato River and associated lakes and wetlands between September 2002 and February 2005. Subsequent recaptures by boat electric fishing and recreational fishing returned 76 koi carp (6% of all tagged fish). Of these, 85% were recaptured less than 5 km from their release site; only one fish moved more than 50 km. On seven occasions, pairs or small groups of koi carp (20% of all tag returns) that had previously been tagged and released at the same locations and times were subsequently recaptured together after considerable periods of time at liberty (mean 551 days ± 419 SD). Adult koi carp in the Waikato River showed a high degree of site fidelity, exhibited prolonged social groupings and females moved downstream more often than males.     

鲤病毒病原的感染性测定

为查明养殖鲤（Cyprinus carpio）突然大批发病死亡的原因，对鲤病样品进行了细胞攻毒、空斑测定、电镜观察，以及鱼体感染等实验。先以患病鲤的组织匀浆液，经过滤后，分别接种到草鱼鳍条细胞（GCF）、鲤上皮瘤细胞（EPC）等14种培养细胞中。利用倒置显微镜观察显示，在1-2d内，该病鱼组织匀浆液可使其中9种细胞出现典型的细胞病变。收集出现病变的细胞液（即病毒悬液），进一步进行病毒滴度检测、空斑测定和鱼体感染实验。结果显示，在GCF细胞上的病毒滴度为10^7.3TCID50／mL；在FHM，TSB和GCO等细胞中可产生直径1～4mm的圆形空斑，空斑的大小与宿主细胞的种类和接种的病毒浓度有关。通过对感染细胞制备的超薄切片和病毒负染样品进行电镜观察，显示这是一类呈典型子弹头样的弹状病毒颗粒。感染了病毒悬液的鲫和鲤先后在第2天和第3天开始出现病症，间隔1～2d后发病的鱼开始死亡，至第14天，两种感染鱼的死亡率均达到83．3％。收集人工感染后濒死的鲫和鲤，分别制备组织匀浆液，回接感染鱼类培养细胞，24h内能使其出现与原发病鲤组织匀浆液所引起的类似的细胞病变。因此证实患病鲤是由病毒病原感染所致。[中国水产科学，2006，13（4）：617—623]     

Growth promoting effects of carp serum components on goldfish culture cells

ABSTRACT:   The growth promoting effect of carp Cyprinus carpio serum was investigated in a medium for goldfish Carassius auratus cell culture. The cell growth rate was decreased in the L-15 medium containing carp serum free from low molecular weight (LMW) fraction of less than 10 000. The addition of this fraction to the medium recovered the original growth rate, whereas the fraction itself did not enhance cell growth. Similar experiments were carried out with carp serum lipoprotein and albumin-like protein fractions. The high density lipoprotein (HDL) fraction containing apolipoprotein (apo) A-I and apoA-II enhanced cell growth at 20°C and 35°C when added with the above-mentioned LMW fraction. A good proliferation was observed at both 20°C and 35°C with the HDL fraction at 0.58 mg/mL. Carp albumin-like protein fraction, mainly containing a 71 kDa-component, also enhanced goldfish cell growth with the LMW fraction at the two temperatures. These results suggest that HDL and albumin-like proteins play an important role in the goldfish cell growth cooperatively with LMW substances contained in carp serum.     

Response of Israeli carp, Cyprinus carpio L., to lysine supplementation of a practical ration at varying conditions of fish size, temperature, density and ration size

Abstract. Four feeding trials with Israeli carp, Cyprinus carpio L., were performed in cages or in ponds in order to investigate responses to lysine supplementation of low protein carp feeds under different practical conditions. In each of the trials one of the basic conditions was different: fish size—small (80–125g) or large (560g); temperature—22°C or 26°C; density—cages or ponds. In each trial 2–3 ration sizes × 2–3 lysine levels were tested. A basal diet of 25% protein and 1·35% lysine served for all the trials.
At low feeding levels, no responses to lysine supplements were found either at 22 ± 2°C or at 26 ± 2°C (0600 h) for all fish sizes. At higher feeding levels, at both temperatures, 0·5% lysine-HCl supplements to 1·7% total lysine yielded significant growth increases. Further supplements to 2·1% lysine (=8·4% of the protein) brought an additional response only from small fish of 90g in cages at the highest ration and temperature. In ponds at 26°C, the effects on small fish of 80g were smaller, probably due to available natural food. The reduction in lysine retention suggested that 1·7% lysine might have been adequate even at optimal conditions in ponds.     

Inactivation of koi‐herpesvirus in water using bacteria isolated from carp intestines and carp habitats

Since its first outbreak in Japan in 2003, koi‐herpesvirus (KHV) remains a challenge to the carp Cyprinus carpio L. breeding industry. In this study, inactivation of KHV in water from carp habitats (carp habitat water) was investigated with the aim of developing a model for rapidly inactivating the pathogen in aquaculture effluent. Experiments with live fish showed that, in carp habitat water, KHV lost its infectivity within 3 days. Indications were that inactivation of KHV was caused by the antagonistic activity of bacteria (anti‐KHV bacteria) in the water from carp habitats. Carp habitat water and the intestinal contents of carp were therefore screened for anti‐KHV bacteria. Of 581 bacterial isolates, 23 showed anti‐KHV activity. An effluent treatment model for the disinfection of KHV in aquaculture effluent water using anti‐KHV bacteria was developed and evaluated. The model showed a decrease in cumulative mortality and in the number of KHV genome copies in kidney tissue of fish injected with treated effluent compared with a positive control. It is thought that anti‐KHV bacteria isolated from the intestinal contents of carp and from carp habitat water can be used to control KHV outbreaks.     

利用OneMap软件构建鲤遗传连锁图谱

首次使用R环境中的OneMap软件包,以荷包红鲤抗寒品系(♂)和云南大头鲤(♀)为祖父母本所培育的110个F2个体为作图群体,以荧光扩增片段长度多态性(fluorescent amplification fragment length polymorphism,fAFLP)为主要分子标记,采用远交全同胞家系模型构建鲤的遗传连锁图谱。结果显示,110个F2个体中共产生1513个清晰的fAFLP标记,其中多态性标记911个;另开发多态性的EST标记12个,最后总计923个标记用于构建遗传连锁图谱;采用OneMap软件包构建的遗传图谱含有238个fAFLP标记和8个EST标记分布在50个连锁群上,总图距为2876.64cM,标记间平均间距为14.68cM,图谱覆盖率为66.56%。     

Responses of fingerling carp, Cyprinus carpio L., to dietary lysine

Abstract. The lysine requirements of fingerling carp, Cyprinus carpio L., for optimum growth and efficiency of food utilization were determined in two experiments by the addition of graded supplements of lysine to a basal diet deficient in this amino acid.
The lysine requirement was estimated to be around 14.5 g/kg dry matter at an ambient temperature of 20°C. At 25°C continuing growth responses occurred up to a dietary lysine concentration of 22.4 g/kg DM. However, when these responses are considered in relation to daily lysine intake, the gross efficiency of utilization of dietary lysine for growth appeared to be unaffected by environmental temperature.
Dietary lysine concentrations exerted no consistent effect upon carcass composition of carp. However, fat deposition tended to increase as food intakes increased in response to lysine additions or to elevated ambient temperature.     

辛基酚对鲤的雌激素效应

研究了辛基酚对鲤的雌激素效应。经10、50、100、300和500μg/L辛基酚暴露32d后,鲤存活率和肥满度与对照组无差异;性腺指数变化明显,雌鱼性腺指数随暴露剂量的增大而增大,雄鱼性腺指数随暴露剂量的增大而减小;50μg/L及以上暴露组与对照组差异显著(P<0.05)。辛基酚能诱导鲤雄鱼产生卵黄蛋白原,暴露剂量为10μg/L组有部分雄鱼肝脏匀浆和血液中检出卵黄蛋白原,50、100和300μg/L组卵黄蛋白原含量随辛基酚浓度的增大而显著增大,500μg/L组卵黄蛋白原含量低于300μg/L组、高于100μg/L组,均为极显著差异(P<0.01)。     

Challenge studies of European stocks of redfin perch, Perca fluviatilis L., and rainbow trout, Oncorhynchus mykiss (Walbaum), with epizootic haematopoietic necrosis virus

A challenge model for comparison of the virulence of epizootic haematopoietic necrosis virus (EHNV) to European stocks of redfin perch, Perca fluviatilis L., and rainbow trout, Oncorhynchus mykiss (Walbaum), was tested. The model investigated intraperitoneal (IP), bath and cohabitation routes at 10, 15 and 20 °C for 5–6 g fish and 15 °C for 20 g perch. In the IP challenges of perch, significant mortality occurred at 15 °C and 20 °C. In challenge trials for rainbow trout, significant mortalities were observed in IP and bath challenges at 20 °C. The mortality observed in IP challenged 20 g perch was not significantly different from that recorded for 6 g fish challenged IP. No significant mortality was observed in any other treatment groups. Re-isolation of ranavirus was confirmed by IFAT and was consistently associated with dead or moribund fish in the trial groups challenged with EHNV. The findings indicate that EHNV does not pose a high risk for wild perch and trout populations in Europe by natural exposure. Mortality appears to be primarily a function of environmental factors, with temperature playing an important role, and not just the presence of the virus in the fish.     

Reduction of stress response in carp,Cyprinus carpioL., held under deteriorating environmental conditions, by oral administration of bovine lactoferrin

Bovine lactoferrin (LF) was evaluated for its ability to suppress stress reactions in carp, Cyprinus carpio L., held under deteriorating environmental conditions. Three test diets containing different levels of LF (0.01, 0.1 and 1%) were fed at 2% of fish body weight per day, i.e. LF at 2, 20 and 200 mg kg(-1) body weight day(-1) was administered orally to carp for 14 days at 20 °C. In the fish reared at a density of 2 individuals 10 l(-1) , plasma cortisol, adrenaline and noradrenaline decreased in the groups fed with 0.1 and 1% LF content diets. In the fish held at a density of 10 individuals 10 l(-1) for 3 days, plasma cortisol, adrenaline, noradrenaline, dopamine and glucose were higher than those at 2 individuals 10 l(-1) in both the control and LF-treated groups. The extent to which these parameters were elevated in the fish held at the same density was reduced with increasing LF content of the diet. Twenty-four hours after exposure to hypoxic conditions (dissolved oxygen level, 2.5 mg l(-1) ), haemoglobin, haematocrit, plasma cortisol, adrenaline, noradrenaline and dopamine increased significantly in all groups. The responses of hypoxic fish fed with 0.1 and 1% LF diets for 14 days were smaller than those of the controls. These results suggest that LF has an ability to reduce the stress response of carp held under deteriorating environmental conditions.     

广西野生鲤与建鲤的遗传多样性分析

利用10个微卫星标记,对2代群体选育的建鲤(Cyprinus carpio var.Jian)和封闭水体的广西野生鲤鱼进行遗传多样性分析,共检测到53个等位基因,等位基因位点数在4~7个之间,平均等位基因数5.3个,片段长度在118~284 bp之间,有效等位基因在1.167 5~1.520 3,平均为1.360 2,广西野生鲤群体的有效等位基因数1.562,大于建鲤的1.388;位点观测杂合度在0.397~0.762之间,平均为0.562 6,期望杂合度0.301~0.629之间,平均0.463 3,广西野生鲤群体的期望杂合度0.581,大于建鲤的0.473;微卫星位点多态性信息含量(PIC)在0.289 7~0.685 0之间,平均多态信息含量0.438 8,为中度多态性,广西野生鲤群体多态信息含量0.685,大于建鲤的0.424,两个群体平均基因分化系数为0.106 9,广西本地野生鲤与建鲤的遗传距离为0.1463,相似性系数为0.828 4,表明广西野生鲤与建鲤之间存在明显的遗传异质性。     

三峡水库高、低水位下开县汉丰湖鱼类资源变化特征

2012年10-11月三峡水库高水位期以及2013年5-6月三峡水库低水位期调查了汉丰湖的鱼类资源,研究三峡水库高、低水位下汉丰湖鱼类种类组成、优势种类、生物多样性以及群落相似性,并对不同水位下汉丰湖鱼类群落结构的扰动特征和增殖放流情况进行了辨识和探讨。共采集到鱼类17 553尾,42种,隶属于4目7科31属。其中三峡水库低水位时采集到鱼类33种,高水位时采集到鱼类34种。汉丰湖优势种类在三峡水库低水位时为银鮈Squalidus argentatus、似鳊Pseudobrama simoni、歺又鱼Hemiculter leucisculus、蒙古鲌Culter mongolicus mongolicus和鲤Cyprinus(Cyprinus)carpio;在三峡水库高水位时为似鳊、鲤、鲫Carassius auratus、张氏歺又鱼Hemiculter tchangi和鲇Silurus asotus。汉丰湖鱼类Shannon-wiener多样性指数(H’)、Simpson优势度指数(D)、Pielous均匀度指数(E)和Margalef种类丰富度指数(R)在三峡水库低水位时分别为1.63、0.67、0.46和3.52;在三峡水库高水位时分别为2.15、0.80、0.61和4.34。三峡水库低、高水位时汉丰湖鱼类群落结构的Bray-Curtis相似性仅为26.52%,喜缓、静水生境的银鮈、张氏歺又鱼、鲤、歺又鱼、鲫、鲇、似鳊和蒙古鲌是群落结构差异的主要种类,累积差异贡献率为90.67%。汉丰湖鱼类群落在高水位时受到中等程度的干扰,而在低水位时受到严重干扰。建议每年均放流鲢、鳙、草鱼并进一步加大放流规模。     

镜鲤与建鲤生长性状共享 QTL 标记及优势基因型

本研究以1个镜鲤(Cyprinus carpio L.)全同胞家系(190个个体)构建的微卫星遗传图谱(992个标记)为基础,从体重、体长、体高和体厚的QTL区间内发掘了54个标记,其与性状具有显著相关性,进而通过对不同基因型性状间的比较,筛选出83个优势基因型。在此基础上,用54个镜鲤QTL标记分析了建鲤(Cyprinus carpio var.jian)作图群体,其中40个标记在建鲤中表现出多态性,比例为74.07%;相关性分析结果显示,其中22个标记与建鲤家系的体重、体长、体高或体厚性状具有显著相关性(P0.05),占多态标记的55.00%;镜鲤与建鲤共享的22个QTL标记中,18个标记与至少1个相同的性状具有显著相关性(P0.05),从中筛选出建鲤性状具有优势的基因型30个,可用于指导建鲤的选育。品种间共享QTL的发掘能够扩展QTL标记的使用空间,减少新品种重新构建图谱进行QTL标记定位的工作量和成本。     

A note on the effects of L-carnitine on the energy metabolism of individually reared carp, Cyprinus carpio L.

Twelve carp, Cyprinus carpio L., were grown from 30 to 120 g live mass on four diets, control (405 g kg⁻¹ crude protein, 20.6 MJ kg⁻¹ gross energy) and control + 200, 400 or 600 mg kg⁻¹ L -carnitine. The diets were designated as CONT, C200, C400 and C600, respectively. Each diet was randomly assigned three fish kept in individual open-circuit respirometers. Fish that ate diets C400 and C600 grew faster than the controls, expended less energy and retained more energy per unit of food energy intake. Average energetic efficiency, expressed as energy retained per unit of energy expended, was 30% higher in the C400 group than in the controls. However, owing to the small number of fish in each group and the large variations in all these parameters, especially within the control group, none of these differences reached statistical significance at the P = 0.1 level.