Cytochrome oxidase gene sequencing reveals channel catfish ovary cell line is contaminated with brown bullhead cells

The channel catfish (Ictalurus punctatus, Rafinesque) ovary (CCO) cell line is the standard cell line used for channel catfish diagnostics. Next‐gen sequencing studies of a virus cultured in the CCO cells revealed mitochondrial sequences matching those of brown bullhead (Ameiurus nebulosus, Lesueur). Therefore, we systematically performed partial cytochrome oxidase 1 gene sequencing of several sources of the CCO cell line and all matched the brown bullhead and not the channel catfish.     

Detection of channel catfish virus DNA in acutely infected channel catfish, Ictalurus punctatus (Rafinesque), using the polymerase chain reaction

Channel catfish virus (CCV) causes an acute haemorrhagic disease in channel catfish, Ictalurus punctatus (Rafinesque), fry and fingerlings. The present study describes a polymerase chain reaction (PCR)-based assay for detection of CCV DNA in the tissues of acutely infected juvenile catfish. The assay is rapid, sensitive and specifically detects CCV DNA derived from epidemiologically distinct viral isolates. The use of two independent PCR primers sets, each specific for particular CCV genes (open reading frames 8 and 59), provides a means to confirm the results and minimize false-positive results. The method identifies CCV DNA in several tissues of acutely infected fish, including the brain, blood, intestine, kidney and liver. The CCV PCR assay is useful for the diagnosis of acute CCV disease and for studies to investigate the molecular basis of CCV pathogenesis.     

斑点叉尾鮰病毒囊膜蛋白ORF6在昆虫细胞中的表达

为构建基于杆状病毒表达系统的CCV新型亚单位疫苗,将CCV的囊膜蛋白基因ORF6克隆至杆状病毒转座载体pFastBacTM 1质粒中,并将阳性重组转座质粒转化进含穿梭载体Bacmid的感受态细胞DH10Bac中,获得重组子rBacmid-ORF6。在脂质体介导下将该重组子转染sf9昆虫细胞,获得重组杆状病毒AC-ORF6。AC-ORF6感染的sf9昆虫细胞,经超薄切片电镜观察,可见重组杆状病毒呈多粒包埋,经间接免疫荧光、Western-blotting检测,CCV的ORF6蛋白可以在感染了AC-ORF6的sf9细胞中表达。研究表明,获得了插入ORF6基因的重组杆状病毒,并且该基因可以在重组杆状病毒介导下在昆虫细胞中表达,从而为基于CCV ORF6的杆状病毒亚单位疫苗研究奠定了基础。     

Comparison of susceptibility of various fish species to experimental infection with channel catfish virus

Channel catfish virus (CCV) disease is an acute haemorrhagic disease in juvenile channel catfish ( Ictalurus punctatus ). To date channel catfish is the only species affected by natural outbreaks of the CCV but juvenile large mouth bass ( Micropterus salmoides ) and silurus ( Silurus meriaionalis ) have suffered high mortalities in recent years in China. Histopathological phenomenon of sick fish is similar to CCV disease, and the identified virus was CCV. In this report, the pathogenicity of infectious CCV was examined by infection trials on the first known host species, the channel catfish and other teleosts. Our results indicated that there were higher detection rates of CCV from large mouth bass and silurus fish. Channel catfish virus did not induce mortality in other cypriniformes, but histopathological studies revealed that carp might be infected by both bathing and intraperitoneal infection. No deaths, clinical or histopathological signs, were found in the six other species exposed by immersion or injection. Experimental infection studies confirm that CCV infect not only channel catfish but also other species (large mouth bass, silutus and carp). The outbreaks of CCV disease only occurred when the cultured temperature was above 25 °C.     

Histopathology, electron microscopy and isolation of channel catfish virus in experimentally infected European catfish, Silurus glanis L.

Abstract Two groups of European catfish, Silurus glanis L., fingerlings were infected with channel catfish virus (CCV) by either intraperitoneal injection with 10⁵ TCID₅₀ of CCV, or bathing in water containing 10⁵ TCID₅₀ of CCV per 1·0 ml. The virus was isolated from spleen, intestine and brain of CCV-injected fish at day 1 and the titres ranged from 10^2·1 to 10^3·3 TCID₅₀/g. However, the tissue distribution of CCV was irregular and no virus was isolated after day 3 post-exposure. In CCV-bathed fish, the virus was isolated only from the liver of one specimen at day 3 post-exposure. No clinical signs of CCV disease developed in any of the fish. Specimens in each regime from all sampling periods showed some minor histopathological changes, but there were no differences between treatments. Lesions included oedema and focal haemorrhage in the liver and the spleen was congested. Electron micrographs of tissue samples showed the presence of a few virus particles around the nuclei of kidney, spleen and intestinal cells, and in or around a myelinated nerve within the optic lobes of infected fish during the first 4 days of infection.     

A Length–Weight Relationship for Pond‐raised Hybrid Catfish Fingerlings

A length–weight relationship was developed for pond‐raised hybrid catfish fingerlings, Ictalurus punctatus × Ictalurus furcatus. A total of 4663 fish were sampled ranging from 20 to 240 mm total length. Predicted weight (g) from length (mm) is based on the equation: Y = 0.000017311X^2.868474309 (r² = 0.996) where Y is the estimated individual fish weight and X is measured total length. The predicted values generated by this model differ from, but follow the same general trends seen with channel catfish, I. punctatus, fingerlings and can be used as a baseline for the development of a length–weight table for hybrid catfish fingerling production.     

Adherence of channel catfish virus to sperm and leukocytes

Although channel catfish virus (CCV) is thought to be vertically transmitted, no mechanism has been demonstrated for such a process. When contained in small volumes of saline, CCV associated rapidly with channel catfish leukocytes and sperm, associated slowly with a continuous cell line of channel catfish cells and did not adhere to channel catfish eggs. This virus-sperm adherence appeared to be a mechanism by which eggs might become infected with virus. However, virus-sperm adherence was not detected when sperm were added to 250 ml of CCV-containing water, indicating that sperm are probably not involved with transmitting CCV into eggs from water or ovarian fluid. Egg infections in channel catfish, if they occur, must be accomplished by mechanisms different from those thought to be involved in the infection of salmonid eggs by infectious hematopoietic necrosis virus.     

Detection of channel catfish virus in channel catfish,Ictalurus punctatus (Rafinesque): use of a nucleic acid probe*

A nucleic acid probe for channel catfish virus (CCV) was constructed using recombinant DNA techniques. This probe consisted of a specific viral DNA fragment generated by digestion of CCV DNA with the restriction enzyme EcoRI. The probe was used to examine DNA isolated from tissues of fish that had been injected with CCV. Viral DNA was detected in some tissues of various injected fish. The sensitivity limit of detection was determined to be one viral DNA per cell.     

Chironomid abundance and consumption by juvenile channel catfish in plastic‐lined and earthen culture ponds

In 2004, research was conducted to compare chironomid larvae populations and their use by channel catfish (Ictalurus punctatus) fingerlings in two different culture systems. Over a 4‐month culture period, chironomid larvae densities in plastic‐lined ponds were significantly less than those in earthen ponds. The consumption of chironomid larvae by channel catfish fingerlings was related to chironomid abundance in earthen ponds. The significance of these findings is the possible relationship among pond type, initial consumption of commercial diets and subsequent survival rates of fingerling channel catfish.     

Detection of channel catfish virus in asymptomatic adult channel catfish,Ictalurus punctatus (Rafinesque)*

Abstract. Two populations of channel catfish were examined for the presence of channel catfish virus (CCV) by use of a nucleic acid probe. In one population of 22 fish with no history of CCV, viral DNA was found in every liver. These fish had previously been examined by a technique involving co-cultivation of their leucocytes with catfish tissue culture cells. The co-cultivation method had identified virus in 10 of these fish. The second fish population consisted of 14 adults that had survived a CCV outbreak in 1980. Of the 14 fish, 11 showed positive indication of CCV DNA. The tissue distribution of the CCV differed from fish to fish. All fish from the first group and one fish from the second group showed some alterations in the DNA banding patterns expected from pure CCV DNA. This might be indicative of modifications in the genomic structure of the CCV DNA when the virus is latent in a fish.     

Effect of dietary calcium on growth and survival of silver catfish fingerlings, Rhamdia quelen (Heptapteridae), exposed to different water pH

The purpose of this study was to verify the effect of dietary Ca²⁺ on the growth and survival of silver catfish fingerlings (Rhamdia quelen) exposed to different water pH (5.5, 7.5 and 9.0). Silver catfish fingerlings were randomly placed in a thermoregulated water re‐use system with twelve 250 L‐tanks, two 1000 L‐biofilters and a 2000 L‐reservoir with a medium flow of 3.84 L min^?1 tank. Stocking density was 0.16 fingerlings L^?1. To prepare the treatment diets, the control diet (0.8 g kg^?1 Ca²⁺) was supplemented with CaCO₃ to yield experimental diets with 6.4, 9.5 and 23.9 g kg^?1 Ca²⁺. There were three replicates/treatments. Survival was more than 93.9% in all treatments. Exposure of silver catfish fingerlings to alkaline or acid water reduced growth, and this effect was not ameliorated by dietary Ca²⁺ supplementation. Moreover, when fingerlings were maintained in water with pH 7.5, the best dietary Ca²⁺ range for silver catfish fingerling growth was 0.8–6.4 g kg^?1.     

Cathelicidins enhance protection of channel catfish,Ictalurus punctatus,and channel catfish ♀ × blue catfish,Ictalurus furcatus ♂ hybrid catfish against Edwarsiella ictaluri infection

Cathelicidins are a class of antimicrobial peptides (AMPs) known to possess rapid and direct antimicrobial activities against a variety of microorganisms. Recently identified cathelicidins derived from alligator and sea snake were found to be more effective in inhibiting microbial growth than other AMPs previously characterized. The ability of these two cathelicidins along with the peptides, cecropin and pleurocidin, to protect channel catfish (Ictalurus punctatus, Rafinesque) and hybrid catfish (I. punctatus ♀ × blue catfish, Ictalurus furcatus, Valenciennes ♂) against Edwardsiella ictaluri, one of the most prevalent pathogens affecting commercial catfish industry, was investigated. Cathelicidin-injected fish (50 µg ml⁻¹ fish⁻¹) that were simultaneously challenged with E. ictaluri through bath immersion at a concentration of ~1 × 10⁶ CFU/ml had increased survival rates compared with other peptide treatments and the infected control. Bacterial numbers were also reduced in the liver and kidney of channel catfish and hybrid catfish in the cathelicidin treatments 24 hr post-infection. After 8 days of challenge, serum was collected to determine immune-related parameters such as bactericidal activity, lysozyme, serum protein, albumin and globulin. These immune-related parameters were significantly elevated in fish injected with the two cathelicidins as compared to other peptide treatments. These results indicate that cathelicidins derived from alligator and sea snake can stimulate immunity and enhance the resistance to E. ictaluri infection in channel catfish and hybrid catfish.     

Pond farming of Nile tilapia, Oreochromis niloticus (L.)j in northern Cameroon. Controlling a sexing error of 1% in hand-sexed male tilapia monosex culture by African catfish, Clarias gariepinus (Burchell)

Catfish (mean W₀ 189 g) were added to ponds (525 m^J each) stocked with 230 hand-sexed, male tilapia (W_u163 g), at 0.04, 0.10 and 0.15 catfish m^?2. In each pond, two female tilapias were introduced, thereby creating a sexing error of less than 1%. Feeding was fixed throughout the experiment at 2.5 kg of cottonseed cake per day per pond 6 days per week (mean feeding rate R = 41 kg ha^?1 day-¹). Rearing time was 125 days. Average net pond production per treatment (ranging between 7.5 and 7.9 t ha^?1 year^?1) and marketable production were not different between treatments but net tilapia production was significantly lower at the highest catfish density. Both catfish and tilapia growth were negatively correlated with catfish density due to feed competition near the end of the experiment. It was concluded that catfish efficiency in controlling tilapia recruitment was strongly reduced by the availability of supplementary high-protein feed. Large catfish competed with the parent tilapia for the cottonseed cake but apparently did not exploit the tilapia recruits. Yield of tilapia recruits was lowest at the highest catfish fingerling density, although this was not significant. The number of catfish fingerlings was significantly higher at the lowest catfish density, which indicated that large catfish preyed on catfish fingerlings.     

Production Characteristics of Channel Catfish,Ictalurus punctatusStocked at Two Densities in the Partitioned Aquaculture System

Abstract

This study was conducted to determine if channel catfish could be cultured at an increased density in the Partitioned Aquaculture System (PAS) without significantly affecting performance. Channel catfish fingerlings (36.53±6.76 g; mean ±SD) were initially stocked into six 9.15 m³ sections at 3,461±317 fish per section, twice the designed carrying capacity, and fed twice daily to satiation. After 75 days, the density of three units was reduced by approximately 50% and all six units were fed for another 97 days. There was no significant difference (P > 0.05) in mean growth rate, feed conversion ratio, or production between fish grown at the low- and high-density treatments. Also, length variation (CV) and condition factors (K) were similar (P > 0.05) between fish grown at the low and high-density. Increasing density by twice the designed carrying capacity did not affect performance of channel catfish in the PAS.     

Evaluation of the Economics of Multi-Batch Channel Catfish Ictalurus punctatus Production Fed an Industry Standard and Three Alternative Diets

The purpose of this study was to determine the feasibility of growing marketable channel catfish, Ictalurus punctatus, from pond-run fingerlings (15.9 g/fish) using low stocking densities (7,413 or 14,826 fish/ha) and electrified bug lights to enhance natural forage available to fish. Even at low stocking densities, fish only averaged 0.2 kg at the end of the growing season. Because marketable sizes of fish were not reached over the growing season, stocking small fingerlings at these rates would not be practical under most commercial production scenarios. Nutritionally, captured insects from electrified bug lights were near a complete diet for catfish, but bug lights did not capture sufficient quantities of insects to affect fish production in either stocking density. Stocking small fingerlings at low stocking rates does not produce market‐sized catfish during one growing season; commercially available bug lights did not provide adequate amounts of natural forage to affect production variables.     

Effect of Dietary Protein Concentration on Growth and Processing Yield of Channel Catfish,Ictalurus punctatus Raised from Advanced Fingerlings to Large Marketable Size

Abstract

This study evaluated the effects of dietary protein concentration (26, 28, and 32%) and an all-plant protein diet (28% protein) on growth, feed efficiency, processing yield, and body composition of channel catfish, Ictalurus punctatus raised from advanced fingerlings to large marketable size (about 800 to 900 g/fish) for two growing seasons. Fingerling channel catfish (average weight = 56 g/fish) were stocked into twenty 0.04-ha ponds at a density of 18,525 fish/ha. Fish were fed once daily to satiation during the two growing seasons and fed according to recommended winter feeding schedules during the winter. There were no differences in diet consumption, weight gain, feed conversion ratio, survival, processing yields (carcass, shank fillet, and nugget), or fillet composition (moisture, protein, fat, and ash) among fish fed the various diets. These results indicate that a 26% protein diet containing plant and animal proteins or a 28% all-plant protein diet is adequate for channel catfish raised in ponds from advanced fingerlings to large marketable size without adversely affecting weight gain, feed efficiency, processing yield, or body composition. Large marketable-size channel catfish appear to use diets less efficiently but give higher processing yields compared to small marketable-size fish.     

Use of monoclonal antibodies in the indirect fluorescent antibody technique (IFA) for the diagnosis of Edwardsiella ictaluri*

Abstract. Forty-five channel catfish, Ictalurus pitnctaius (Rafinesque), fingerlings were inoculated with Edwardsiella ictaluri to determine the usefulness of monoclonal antibodies for indirect fluorescent antibody techniques for confirming clinical diagnosis of enteric septicaemia of catfish. Bacteriological and indirect fluorescent antibody results were compared statistically and found to correlate in 90·3% of the cases. A method for incorporating monoclonals for indirect fluorescent antibody into the speciation of Edwardsiella ictaluri and the use of monoclonal antibodies in diagnosis is described.     

The Effect of Varying Biomass of Carryover Fish in Multiple‐batch Catfish,Ictalurus punctatus,Pond Production

The presence of carryover (fish >350 g stocked the previous year but not yet market size) channel catfish, Ictalurus punctatus, in multiple‐batch production ponds has been shown to affect overall production performance and costs. However, little attention has been paid to effects of varying biomasses of carryover fish in ponds. Twelve 0.1‐ha earthen ponds were stocked March 20, 2007, with 15,000 catfish fingerlings per ha (mean weight 31 g), and carryover fish at either 726, 1460, or 2187 kg/ha (mean weight 408 g, range 204–703 g) to compare the effect of three different biomasses of carryover catfish on the production performance of understocked fingerlings. Gross and net yields increased with increasing biomass of carryover fish. Growth and mean weight at harvest of fingerlings were significantly greater at the lowest biomass of carryover fish (<1460 kg/ha), but there was no difference between the medium and high carryover density treatments. Net returns were highest with the highest biomass of carryover fish, but fell by $688/ha in Year 2 because of slower growth of fingerlings in Year 1.     

Salt in the Food and Water as a Supportive Therapy for Ichthyophthirius multifiliis Infestation on Silver Catfish, Rhamdia quelen, Fingerlings

This study verified the efficacy of common salt (NaCl) in the food and water as a supportive therapy for Ichthyophthirius multifiliis and growth of silver catfish, Rhamdia quelen, fingerlings. After appearance of the first white spots following experimental exposure, indicating infestation by this parasite, fish were transferred to eighteen 40‐L polypropylene tanks and kept for 30 d. Experimental diets were prepared with five different levels of salt supplementation: 0.0, 1.2, 2.5, 5.0, and 6.0% for 30 d. In addition, another group of fish were fed a diet without salt supplementation but exposed to 4 g of salt/L of water. Fingerlings were collected at different times for analyses of whole‐body Na⁺, K⁺, and Cl^?. Dietary salt supplementation provoked a transitory increase in whole‐body Na⁺, K⁺, and Cl^? in infested catfish fingerlings, but the best stabilization of these ion levels was observed in specimens exposed to salt in the water. Silver catfish fed a diet containing 1.2% salt showed higher weight and biomass after 30 d compared to those fed with 6% salt. The use of 4 g of salt/L of water improved survival of infested silver catfish, but dietary salt supplementation was ineffective as a therapy for ichthyophthiriasis.