Reactivation of koi herpesvirus infections in common carp Cyprinus carpio

Two co-habitation studies with common carp were conducted to determine whether latent infections of koi herpesvirus (KHV) exist. Fish were exposed to KHV using 2 different temperature profiles, which induced low and high initial mortality. Subsequently, certain groups of fish were co-habited with naive fish while others were not. Koi herpesvirus was reactivated in fish from 3 of the 5 experimental tanks. Reactivation of the virus occurred regardless of the initial mortality associated with the virus or whether fish were co-habited with naive fish. The reactivation of the virus in our experiments occurred several months after the initial exposure to KHV and appeared to be temperature dependent.     

Mass mortality associated with koi herpesvirus in wild common carp in Canada

Koi herpesvirus (KHV) was identified as being associated with more than one mortality event affecting common carp in Canada. The first was an extensive mortality event that occurred in 2007 in the Kawartha Lakes region, Ontario, affecting Lakes Scugog and Pigeon. Fish had branchial necrosis and hepatic vasculitis with an equivocal interstitial nephritis. Several fish also had branchial columnaris. Subsequent mortality events occurred in 2008 in additional bodies of water in south-central Ontario, such as Lake Katchewanooka and outside of Ontario in Lake Manitoba, Manitoba. Koi herpesvirus was detected in fish submitted for examination from all of these lakes by polymerase chain reaction (PCR), and sequence of the PCR product revealed 100% homology to KHV strains U and I. Real-time PCR analysis of KHV-infected wild carp revealed viral loads ranging from 6.02×10(1) to 2.4×10(6) copies μg(-1) host DNA. This is the first report of KHV in Canada.     

Seminoma in a koi carp Cyprinus carpio: histopathological and immunohistochemical findings

A spontaneous seminoma in a 3 yr old male koi carp Cyprinus carpio L. is described. The animal, presenting a symmetric abdominal enlargement, showed a celomatic multinodular, white-yellowish and firm mass that infiltrated the liver and the intestine wall. Histologically, the neoplasm was non-encapsulated and poorly demarcated, showed invasive growth and was characterized by a lobular architecture, subdivided by abundant fibro-connective septa. Large necrotic and calcified areas together with small aggregates of residual spermatids were present. We diagnosed a classical seminoma with a diffuse pattern. Neoplastic cells cross-reacted with vimentin, placental alkaline phosphatase, and c-KIT. An immunohistochemical phenotypization of the tumor was performed to exclude other celomatic neoplasms and to compare this seminoma with those reported in mammals and humans.     

Effect of ploidy on scale-cover pattern in linear ornamental (koi) common carp Cyprinus carpio

The effect of ploidy on scale-cover pattern in linear ornamental (koi) common carp Cyprinus carpio was investigated. To obtain diploid and triploid linear fish, eggs taken from a leather C. carpio female (genotype ssNn) and sperm taken from a scaled C. carpio male (genotype SSnn) were used for the production of control (no shock) and heat-shocked progeny. In heat-shocked progeny, the 2 min heat shock (40° C) was applied 6 min after insemination. Diploid linear fish (genotype SsNn) demonstrated a scale-cover pattern typical for this category with one even row of scales along lateral line and few scales located near operculum and at bases of fins. The majority (97%) of triploid linear fish (genotype SssNnn) exhibited non-typical scale patterns which were characterized by the appearance of additional scales on the body. The extent of additional scales in triploid linear fish was variable; some fish had large scales, which covered almost the entire body. Apparently, the observed difference in scale-cover pattern between triploid and diploid linear fish was caused by different phenotypic expression of gene N/n. Due to incomplete dominance of allele N, triploids Nnn demonstrate less profound reduction of scale cover compared with diploids Nn.     

Initial characteristics of koi herpesvirus and development of a polymerase chain reaction assay to detect the virus in koi,Cyprinus carpio koi

Since 1998, episodes of mass mortality have occurred in populations of common carp Cyprinus carpio carpio in Israel and in populations of koi Cyprinus carpio koi in Israel and the USA. A herpesvirus isolated from infected fish has been shown in experimental studies to induce disease and mortality similar to those observed in outbreaks at infected farms. Initial characteristics of the virus show that it is clearly different from Herpesvirus cyprini (CHV), the most commonly known herpesvirus from cyprinid fish. The koi herpesvirus (KHV) has 31 virion polypeptides. Twelve of the virion polypeptides of KHV have similar molecular weights to those of CHV and 10 are similar to those of channel catfish virus (CCV). Both virion polypeptide and restriction fragment length polymorphism analyses of genomic DNA showed that the first KHV isolates from Israel and the USA were identical. In contrast, the genomic DNA restriction fragments clearly distinguish KHV from CHV and CCV. A polymerase chain reaction (PCR) assay to detect the virus in koi tissues was developed with sequences obtained from 1 restriction fragment of KHV DNA. The PCR assay effectively detected a 484 base pair sequence from KHV but did not amplify genomic DNA from either CHV or CCV. The PCR assay detected as little as 1 pg of KHV DNA mixed with 100 ng of host DNA. Viral sequences were amplified from koi obtained from field collections and from koi that were experimentally exposed to 10(2) TCID50 ml(-1) of KHV via the waterborne route. All KHV exposed fish dying of infection between 8 and 10 d post exposure or surviving to 14 d post exposure were found to be positive by PCR, while unexposed control koi were all negative. The assay also showed the presence of KHV DNA in tissues of koi obtained from farms in Israel. The PCR assay should assist virus isolation procedures and histologic and electron microscopic analyses now commonly used to detect KHV infection. Current studies are examining the possibility of using the PCR to detect KHV DNA in live fish and the relative sensitivity and specificity of the KHV PCR assay compared with other diagnostic tests.     

Phenotypic variation and associated predation risk of juvenile common carp Cyprinus carpio

Juvenile common carp Cyprinus carpio were collected from 10 lakes with variable predator abundance over 4 months to evaluate if morphological defences increased with increasing predation risk. Cyprinus carpio dorsal and pectoral spines were longer and body depth was deeper when predators were more abundant, with differences becoming more pronounced from July to October. To determine if morphological plasticity successfully reduced predation risk, prey selection of largemouth bass Micropterus salmoides foraging on deep- and shallow-bodied C. carpio was evaluated in open and vegetated environments. Predators typically selected deep- over shallow-bodied phenotypes in open habitats and neutrally selected both phenotypes in vegetated habitats. When exposed to predators, shallow-bodied C. carpio phenotypes shoaled in open habitat, whereas deep-bodied phenotypes occupied vegetation. Although deep-bodied phenotypes required additional handling time, shallow-bodied phenotypes were more difficult to capture. These results suggest that juvenile C. carpio gradually develop deeper bodies and larger spines as predation risk increases. Morphological defences made it more difficult for predators to consume these prey but resulted in higher vulnerability to predation in some instances.     

Characterization of a novel cell line from the caudal fin of koi carp Cyprinus carpio

A continuous cell line (KF‐101) derived from the caudal fin of the koi carp Cyprinus carpio was established and characterized. The KF‐101 cell line multiplied abundantly in Leibovitz's L‐15 medium containing 10% foetal bovine serum at 25° C, and was subcultured for >90 passages over a period of 3 years. Immunocytochemistry revealed that the KF‐101 cells contain keratin, junction proteins connexin‐43 and occludin, and ectodermal stem‐cell marker Pax‐6, but not vimentin. Furthermore, the KF‐101 cells reacted with anti‐human DARPP‐32 and anti‐human GATA‐4 antibodies, and the labelling was regulated according to the cell cycle. The labels of the DARPP‐32 and GATA‐4 antibodies in the KF‐101 cells were the suggested phosphatase‐1 inhibitor‐1 and GATA‐3, respectively. In addition, the KF‐101 cells were susceptible to koi herpesvirus but were resistant to eel herpesvirus, iridovirus, grouper nodavirus and chum salmon (Oncorhynchus keta) virus. The results indicate that the KF‐101 cells are suitable materials for investigating biological and virological development.     

Reduced swimming abilities in fast-growing transgenic common carp Cyprinus carpio associated with their morphological variations

Critical swimming speeds ( U _crit) and morphological characters were compared between the F₄ generation of GH-transgenic common carp Cyprinus carpio and the non-transgenic controls. Transgenic fish displayed a mean absolute U _crit value 22·3% lower than the controls. Principal component analysis identified variations in body shape, with transgenic fish having significantly deeper head, longer caudal length of the dorsal region, longer standard length ( L _S) and shallower body and caudal region, and shorter caudal length of the ventral region. Swimming speeds were related to the combination of deeper body and caudal region, longer caudal length of the ventral region, shallower head depth, shorter caudal length of dorsal region and L _S. These findings suggest that morphological variations which are poorly suited to produce maximum thrust and minimum drag in GH-transgenic C. carpio may be responsible for their lower swimming abilities in comparison with non-transgenic controls.     

Coping strategies in a strongly schooling fish,the common carp Cyprinus carpio

Individual common carp Cyprinus carpio were screened repeatedly for risk taking (rate of exploration of a novel, potentially dangerous environment) and for competitive ability (success in gaining access to a spatially restricted food source). Marked differences in behaviour were evident, and significant consistency in individual responses across trials was found for both risk taking and competitive ability. In addition, there was a significant positive relationship between individual performance in these two contexts, with fish that explored more quickly in the novel environment tending to be among the first to gain access to restricted food. In two follow‐up studies, resting metabolic rate, blood lactate and glucose and the expression of the cortisol receptor gene in the head kidney and brain were compared in fish from the two extremes of the risk‐taking spectrum. Mass‐specific metabolic rate was significantly higher in risk‐taking than in risk‐avoiding fish, while plasma lactate and glucose concentrations and expression of the cortisol receptor gene were lower. It was concluded that a behavioural syndrome based on boldness and aggression exists in C. carpio, as it does in many other animals, and that this is associated with differences in metabolic and stress physiology (down to the genomic level) similar to those described in animals with different coping strategies.     

Efficacy of quinine against ichthyophthiriasis in common carp Cyprinus carpio

Ichthyophthiriasis, caused by Ichthyophthirius multifiliis, is an economically important worldwide parasitic disease that infects all freshwater fish. Since the banning of malachite green for use in food fish, there has been a great need for alternative therapeutants. The goal of this study was to investigate the efficacy of quinine against I. multifiliis. Parasite developmental stages from our laboratory-established life cycle in rainbow trout Oncorhynchus mykiss were exposed to quinine in vitro, and a dual fluorescent staining technique was used to allow a clear distinction between viable and damaged parasites. Furthermore, the effect of quinine was assessed in vivo by oral administration and intraperitoneal injections in common carp Cyprinus carpio. The results of the in vitro experiments proved quinine to be effective against the parasite. Quinine injected at a dosage of 60 mg kg(-1) body weight resulted in a significantly lower number of trophonts. In contrast, in-feed trials did not show a significant reduction of trophonts after treatment commencing 1 d after infection with concentrations of up to 20 g quinine kg(-1) feed for 3 d. After a 14-d treatment at concentrations of up to 10 g quinine kg(-1) feed prior to theront exposure, there was also no significant difference in parasite numbers between treated and control groups. The results of oral versus parenteral application of quinine indicate that the substance is not completely absorbed from the intestinal tract of common carp. However, medicated feed containing higher concentrations of quinine was less readily accepted by the fish, presumably due to the bitter taste.     

Spontaneous diploidization of maternal chromosome set in ornamental (koi) carp, Cyprinus carpio L.

An unusually high frequency of spontaneous diploidization in a maternal chromosome set (SDM) was discovered in one ornamental carp (koi) female in an experiment on these fish for induced gynogenesis. Spontaneous appearance of diploid embryos in the gynogenetic offspring (intact eggs × irradiated sperm) and appearance of triploids among control fish (intact eggs × intact sperm) obtained from koi female and males of edible carp indicated spontaneous diploidization of maternal chromosomes (SDM). Possible cytological processes causing this phenomenon are discussed.     

Effects of a parasite-induced nephritis on osmoregulation in the common carp Cyprinus carpio

Carp Cyprinus carpio infected with the haemoflagellate Trypanoplasma borreli undergo progressive nephritis associated with a destruction of approx. 40% of the nephric tubules. In an attempt to analyse the effect of the nephritis on the osmoregulation of affected carp, the clinical chemical properties of plasma and urine samples were analysed. Parasitised carp excreted greater amounts of electrolytes in their urine than uninfected carp which excreted highly diluted urine with an osmolality of about 10% of plasma osmolality. During the course of the infection, urine osmolality increased up to 26% of plasma osmolality by Day 21 post-infection (p.i.). The plasma:urine ratio of Na+ also increased, while concomitant losses of Mg2+, Ca2+, K+ and inorganic phosphate were less pronounced. Infected carp were able to maintain a normal solute balance in their plasma. Plasma hydration (indicated by decreased protein contents) occurred on Day 21 p.i. Our data indicate that in T. borreli-infected carp, reabsorption processes of the distal renal tubule were disturbed, while secretory and absorption processes in the proximal tubule appeared to be less affected. In addition, infected carp were able to compensate their increased ion losses, probably by (energy-consuming) active absorption processes. The energy budget of infected carp was additionally affected by a substantial direct consumption of plasma glucose by the parasite.     

Determination of the surface area of common carp,Cyprinus carpio L.

A wrap method adaptation combined with AutoCAD2005 and Scion Image for Windows were used to determine the surface area of a fish. Compared with the corresponding r² and F of many models, the most accurate formula: S = 752.15W^0.675 (r² = 0.999, F = 18362.94, P < 0.0001) for estimating the surface area of common carp was obtained. Similarly, the fin formula: S = 1834.12W^0.708 (r² = 0.992, F = 2690.47, P < 0.0001) was also obtained for the same purpose. It was proven that these two formulae gave good estimates of surface and fin areas of four strains of common carp: Yellow‐river carp, fancy carp, mirror carp and Xingguo red carp.     

Mark retention and fish survival associated with a low-cost marking technique for common carp Cyprinus carpio (Linnaeus, 1758)

Common carp (Cyprinus carpio, [Linnaeus, 1758]) have long been established in the United States and in some cases their populations can be very dense, leading biologists to expend significant effort towards reducing numbers of common carp in some populations. Estimating abundance of common carp prior to removal efforts would be useful in evaluating success of these removal efforts, but marking large numbers of fish can be expensive. Therefore, a low-cost tagging option is needed. In this study, we used tank and field experiments to determine the retention and longevity of hole-punch marks in the opercula of common carp. For the tank experiment, fish were double marked with a size-3 self-piercing tag and an operculum hole-punch mark (using a paper hole-punch tool with a hole diameter of 6.4 mm) on opposite sides of the fish. Over the entirety of the 180–day tank experiment, retention of the self-piercing tags and hole-punch marks was 100% and no marking mortality was observed. For the field experiment, 883 common carp were tagged at random in two groups, a double-marked group (n = 416, both self-piercing tags and hole-punch) and a single-mark group (n = 467, self-piercing tag). Fish were sampled monthly for 398 days. Because the length distribution sampled was bimodal, we evaluated tag retention of fish <330 mm TL (small fish, n = 273) and > 331 mm TL (large fish, n = 143), separately. Hole-punch mark retention was high for both size classes throughout most of the field experiment. For large fish, retention of hole-punch marks was 100% for the entire 398-day experiment. For small fish, retention of hole-punch marks was 100% through 184-day and remained above 93% through 328-days, but declined to 0% by day 398. Our results suggest that the operculum hole-punch mark is a valuable low-cost, long-term technique for tagging common carp.     

Ultrastructural identification of a herpes-like virus infection in common carp Cyprinus carpio in Korea

We report on a herpes-like virus, which was found to be associated with mass mortality of common carp Cyprinus carpio for the first time in Korea in 1998. The external signs of infection in moribund fish were darkened coloration and severe branchial necrosis in the gill. Transmission electron microscopy revealed the presence of herpes-like viruses in spleen tissue. Infected spleen cells showed hypertrophied nuclei and degeneration. Numerous nucleocapsids of about 82 nm in diameter were found within the nucleus and cytoplasm of infected cells, and extracellular enveloped particles were also observed. We conclude that this virus was a likely significant cause of the high mortality of common carp in Korea in 1998.     

Cadmium-binding to transferrin in the plasma of the common carp Cyprinus carpio

In this study, it was investigated by autoradiography with radioactive cadmium after Western blotting of two-dimensional electrophoresis gels, to which proteins cadmium is mainly bound in plasma of common carp Cyprinus carpio. The obtained results demonstrate that in carp plasma, cadmium is primarily bound to two high molecular weight proteins. Relative small amounts are bound to a protein with M(r) approximately 60000. The other metal-binding protein, with M(r) approximately 70000 and pI approximately 6.7 was identified as transferrin. The conditional equilibrium constants for the binding of cadmium ions to the two metal-binding sites of this protein were calculated as logK(1)=5.40+/-0.12 and logK(2)=4.66+/-0.21, which are comparable to those of human transferrin under the same experimental conditions. Transport of cadmium in plasma of carp was found to be different from that of brown trout Salmo trutta and man, where cadmium is mainly bound to albumin and transferrin. The prominent binding of cadmium to transferrin can be explained by the absence or at least the very low concentrations in which albumin is present in carp plasma.     

Global patterns and clines in the growth of common carp Cyprinus carpio

This review provides a meta‐analytical assessment of the global patterns and clines in the growth of Cyprinus carpio as measured by length‐at‐age (L_t) or von Bertalanffy growth function (VBGF) parameters, mass–length relationship (W–L_t) and condition factor, based on literature data. In total, 284 studies were retrieved spanning 91 years of research and carried out on 381 waterbodies–locations in 50 countries in all five continents. Although native C. carpio achieved larger (asymptotic) size relative to its non‐native counterpart, the latter grew faster during the first 7 years of life. Lentic populations (especially in natural lakes) also achieved larger sizes relative to lotic ones and the same was true for populations in cold and temperate v. arid climates. Unlike previous studies (on much more restricted datasets), only weak latitudinal clines in instantaneous growth rate, L_t at age 3 and mortality were observed globally and this was probably due to the presence of counter‐gradient growth variation at all representative age classes (i.e. 1–10 years). Slightly negative allometry was revealed by the W–L_t and the related form factor tended to distinguish the more elongated and torpedo‐shaped body typical of the wild form from the deeper body of feral‐domesticated C. carpio. Existing population dynamics models for C. carpio will benefit from the comprehensive range of waterbody type × climate class‐specific VBGF parameters provided in the present study; whereas, more studies are needed on the species' growth in tropical regions and to unravel the possibility of confounding effects on age estimation due to both historical and methodological reasons.     

Specific dynamic action and feeding metabolism in common carp,Cyprinus carpio L.

• 1.1. Oxygen uptake attributable to Specific Dynamic Action (SDA) was measured in common carp, Cyprinus carpio L. (63.6–84.0 g) fed on 20, 35 and 50% dietary protein at 0.40 to 1.00% ration levels at 28°C.
• 2.2. After feeding both SDA magnitude and mean peak oxygen consumption increased directly with dietary protein and ration levels. SDA duration was not significantly related to dietary protein but significantly increased with ration levels.
• 3.3. SDA coefficients were 8.99, 13.51 and 15.94% with 20, 35 and 50% dietary protein showing a direction relationship to the protein content. The SDA coefficient did not change with ration size.
• 4.4. SDA models resulting from this work are of great interest for the aquaculturist, as post-feeding oxygen requirements in an intensive fish culture can be predicted where dietary protein and ration levels are known.