Cross-protection elicited in channel catfish (Ictalurus punctatus Rafinesque) immunized with a low dose of virulent Edwardsiella ictaluri strains

Cross-protection of channel catfish ( Ictalurus punctatus Rafinesque) immunized with a low dose of virulent Edwardsiella ictaluri and challenged with six E. ictaluri strains was examined in four trials. The relative per cent survival among low-dose immunized and then challenged fish ranged from 27.7% to 100%. Significant protection ( P <0.05), with the exception of strain ATCC-33202, was conferred by immunization with a given E. ictaluri strain challenged either with a homologous or a heterologous strain. Antibody titres of pooled serum collected on day 22 from surviving fish examined by enzyme-linked immunosorbent assay (ELISA) ranged from 1:40 to 1:320, but no differences were apparent among different vaccinated groups. The protein profiles of six E. ictaluri strains examined by Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a relatively homogeneous pattern. Immuno-blots probed with pooled serum from immunized and challenged fish showed a pattern similar to LPS-reaction patterns observed with E. ictaluri in other studies. Since the present studies further corroborate that E. ictaluri is a clonal bacterial species with no apparent antigenic differences, it is possible that immunization with a single E. ictaluri field strain should confer protection against any other strain.     

Susceptibility of channel catfish, Ictalurus punctatus (Rafinesque), to Edwardsiella ictaluri challenge following copper sulphate exposure

Channel catfish, Ictalurus punctatus (Rafinesque), with or without a preliminary 24 h exposure to 2 mg copper sulphate L(-1), were challenged with 7.5 x 10(6) colony forming units L(-1) of Edwardsiella ictaluri to determine the effect of copper sulphate on disease resistance. Catfish previously exposed to copper sulphate were significantly more resistant to the bacterial challenge than those not exposed. Catfish not exposed to copper sulphate suffered 35.5% mortality while catfish exposed to copper sulphate experienced 14.1% mortality. Copper concentrations were the same in tank waters of both exposed and control fish at the time of challenge, eliminating the possibility that copper in the water may have been toxic to bacteria. Copper concentrations in freeze dried and ground tissues of unexposed, exposed, and purged channel catfish were highest in fish before copper sulphate exposures suggesting that elevated tissue levels of copper were not responsible for the increased resistance to bacterial challenge. Competition for sites of bacterial attachment to gill or epithelial cells may account for the reduction in mortality; although this is not supported by the low copper content of fish tissue after copper exposure.     

The effect of feeding diets containing deoxynivalenol contaminated corn on channel catfish (Ictalurus punctatus) challenged with Edwardsiella ictaluri

Channel catfish were fed practical corn‐soybean meal diets for 10 weeks that contained various weighed amounts of ground, dried field corn contaminated with 20 mg deoxynivalenol (DON) kg⁻¹. Weighed amounts of DON corn were blended with weighed amounts of ground, clean corn that contained no DON (0 mg kg⁻¹) to yield five diets that had 0, 2.5, 5.0, 7.5 and 10.0 mg DON kg⁻¹ of diet. Results show that catfish fed diets that contained DON for 7 weeks did not experience lower weight gains or poorer feed conversion ratios that were significantly (P > 0.05) different from control‐fed fish. Mortality of catfish during the 21‐day post‐challenge period indicate that catfish fed diets containing DON‐contaminated corn that provided at least 5.0 mg DON kg⁻¹ of diet had significantly (P < 0.05) lower mortality than catfish fed the control diet or the diet that provided 2.5 mg DON kg⁻¹ of diet. The presence of DON‐contaminated corn in the experimental diets did not significantly (P > 0.05) alter fish body weight gains and appeared to provide a protective effect for channel catfish challenged with the pathogenic bacterium Edwardsiella ictaluri.     

嗜水气单胞菌3种疫苗对斑点叉尾免疫原性比较研究

将经福尔马林灭活的嗜水气单胞菌(Aeromonas hydrophila)、菌体脂多糖(LPS)和菌体外膜蛋白(OMP)作为免疫原,分别接种斑点叉尾(Ictalurus punctatus Rafinsque)后,通过测定受免鱼的凝集抗体效价,头肾和血液中吞噬细胞的吞噬活性和用A.hydrophila活菌攻毒的方法,探讨了斑点叉尾对A.hydrophila的3种疫苗的免疫应答状况和对活菌攻毒的免疫保护效果。试验结果显示:对斑点叉尾经腹腔注射接种3种疫苗均能刺激受免鱼产生较强的免疫应答,3种疫苗的受免鱼均产生了特异性凝集抗体,接种F-Ah灭活菌苗的试验鱼最高,接种OMP疫苗的试验鱼其次,而接种LPS疫苗的试验鱼血清中凝集抗体效价最低;与未接种疫苗的对照鱼相比,受免鱼头肾和血液中吞噬细胞的吞噬活性明显上升,头肾和血液中吞噬细胞活性由高到低依次是LPS、OMP和F-Ah免疫接种的斑点叉尾;活菌攻毒的结果证明接种3种疫苗的受免斑点叉尾A.hydrophila的感染产生了不同程度的相对免疫保护力(RPS),以OMP免疫接种后的斑点叉尾RPS最高,达到72.5%,接种LPS的斑点叉尾稍差,RPS为62.5%,而RPS最低的是接种F-Ah的免疫组,RPS只有24.2%。     

Assay performance during validation of freezing channel catfish Ictalurus punctatus (Rafinesque) infected with a Gram‐negative bacterium

Recovery of bacteria from infected fish during population sampling can be affected by factors including the type of assay, method of specimen preservation and concentration of bacteria present. Consequently, before use in field sampling, methods should be validated. The three objectives of this study were, first, to determine whether a channel catfish Ictalurus punctatus (Rafinesque) fingerling classified as positive for Gram‐negative Edwardsiella ictaluri infection according to bacterial culture before freezing was also classified as positive after freezing, second, to determine how direct culture from the kidney (DIRECT), culture of homogenate (HOMOG) and standard PCR (PCR) agree with bacterial culture in terms of classifying fish as positive or negative and third, to estimate diagnostic sensitivity (dSe) and diagnostic specificity (dSp) for DIRECT, HOMOG and PCR. In fresh and frozen fish, as bacterial concentration decreased, the ability of each assay to detect positive fish also decreased, especially when there were <10⁴ colony‐forming units per gram (CFU g^?1) tissue. HOMOG proved to be the most reliable at correctly classifying catfish, whether they were subclinically or clinically infected. PCR assay was the least reliable. Overall, values for this study population for dSe were 0.66, 0.92 and 0.43, and for dSp were 0.86, 0.91 and 0.95, for DIRECT, HOMOG and PCR respectively.     

Experimental challenge studies in Vietnamese catfish,Pangasianodon hypophthalmus (Sauvage), exposed to Edwardsiella ictaluri and Aeromonas hydrophila

The two main diseases in the pangasius catfish industry are bacillary necrosis of Pangasianodon (BNP) and motile aeromonas septicaemia (MAS), where the aetiological agents have been identified as Edwardsiella ictaluri and Aeromonas hydrophila, respectively. In this study, apparently healthy Pangasianodon hypophthalmus were exposed to E. ictaluri, A. hydrophila or both bacterial species by intraperitoneal injection or immersion. There were 20 fish per treatment group, and the bacterial isolates used for the study were recovered from natural infections of BNP or MAS in farmed Vietnamese P. hypophthalmus. The results of the experimental infections mimicked the natural disease outbreaks reported from these pathogens in P. hypophthalmus. Furthermore, it was clearly demonstrated that E. ictaluri was only recovered from the fish exposed to the bacterium and not recovered from the animals receiving A. hydrophila.     

Serological detection of Edwardsiella ictaluri Hawke lipopolysaccharide antibody in serum of channel catfish, Ictalurus punctatus Rafinesque

Abstract. Bacterial agglutination, passive haemagglutination, complement-dependent passive haemolysis, indirect immunofluorescence, agar gel immunodiffusion and agglutination with fractions of immunized fish serum were compared for detecting humoral antibody to the lipopolysacchande (LPS) of Edwardsiella icialuri Hawke in channel catfish. Bacterial agglutination titres averaged 1: 672; passive haemagglutination titres averaged 1: 1152; and complement-dependent haemolysis titres averaged 1: 2360. Serum from non-vaccinated fish ranged from 0 to 1:32. Indirect fluorescence and immunodiffusion demonstrated positive reactions to the LPS antibody. Fractionation of immune sera produced three fractions, one of which strongly haemagglutinated E. ictaluri but the other two did not. All six serological techniques were sensitive to E. ictaluri LPS antibody.     

Vaccination of channel catfish, Ictalurus punctatus (Rafinesque), by immersion and oral booster against Edwardsiella ictaluri

Abstract. A commercially prepared vaccine against Edwardsiella ictaluri was used to vaccinate 12-day-old channel catfish fry by immersion, or by immersion plus an oral booster 2 months later. One month after the fish were fed the booster vaccine, they were challenged by waterborne exposure to 2·1 × 10⁶ cells ml⁻¹ of E. ictaluri. Immersion only vaccinated fish suffered 6·7% mortality and immersion plus oral-boosted fish had a 3·3% mortality. Mortality among non-vaccinated controls was 96·7% and was significantly ( P < 0·01) above the vaccinated mortality. The relative per cent survival for the immersion-only fish was 93·1, while it was 96·6 for the immersion plus oral-boosted fish. Agglutinating antibody titres of the vaccinated fish were significantly ( P < 0·05) higher than the control fish. When the ponds were drained 6 months after stocking, 42·7% of non-vaccinated, 56·3% of immersion-only and 70·8% of immersion plus oral-boosted fish were harvested. Survival of immersion plus orally-boosted fish was significantly ( P < 0·05) higher than the controls of immersion-only fish. Duplicate populations of immersion plus oral-booster-vaccinated fish grew 34% and 56% faster, respectively, on an average daily gain than the control fish, while immersion-only fish in one pond grew 20% less per day and fish in the second pond grew 48% faster.     

Comparative Susceptibility of Channel Catfish,Ictalurus punctatus; Blue Catfish,Ictalurus furcatus; and Channel (♀) × Blue (♂) Hybrid Catfish to Edwardsiella piscicida,Edwardsiella tarda,and Edwardsiella anguillarum

Members of the genus Edwardsiella are important pathogens of cultured and wild fish globally. Recent investigations into the phenotypic and genotypic variation of Edwardsiella tarda have led to the segregation of E. tarda into three distinct taxa: E. tarda, Edwardsiella piscicida, and Edwardsiella anguillarum. In catfish aquaculture in the southeastern USA, E. piscicida has been more commonly associated with disease than E. tarda or E. anguillarum, and recent research has demonstrated E. piscicida to be more pathogenic in channel catfish than E. tarda or E. anguillarum. Anecdotal reports from industry suggest an increased prevalence of E. piscicida associated with the culture of channel (♀) × blue (♂) hybrid catfish. This work investigated the comparative susceptibility of channel catfish, blue catfish, and their hybrid cross to molecularly confirmed isolates of E. tarda, E. piscicida, and E. anguillarum. There was significantly higher mortality in hybrid catfish compared to channel catfish following intracoelomic injection of E. piscicida. To our knowledge, E. piscicida is the first bacterial pathogen to demonstrate increased pathogenicity in hybrid catfish compared to channel catfish.     

Pathology of experimental enteric septicaemia in channel catfish, Ictalurus punctatus (Rafinesque), following immersion-exposure to Edwardsiella ictaluri

Abstract. Fingerling channel catfish, Ictalurus punctatus (Rafinesque), were exposed experimentally to Edwardsiella ictaluri by immersion for 1 h in water containing 5 × 10⁸ colony-forming units (CFU) of the bacterium per ml. Ninety per cent of the fish developed lesions typical of enteric septicaemia of catfish (ESC), 93% of affected fish developed the acute form of ESC and 7% developed chronic ESC. Acute disease was characterized grossly by cutaneous haemorrhage and ulceration, and microscopically by enteritis, olfactory sacculitis, hepatitis and dermatitis. The earliest lesions of acute ESC, i.e. enteritis and olfactory sacculitis, were observed microscopically at 2 days post-exposure (PE); gross lesions, primarily mild subcutaneous haemorrhage at the base of fins, were first apparent at 4 days PE. Chronic ESC, seen most commonly 3–4 weeks PE, was characterized by dorsocranial swelling and ulceration, granulomatous olfactory neuritis/perineuritis, and meningoencephalitis involving the olfactory bulbs, olfactory tracts and olfactory lobes of the brain. Gross and microscopic lesions in the acute and chronic forms of experimental ESC were similar to the lesions reported in naturally occurring ESC. Definitive pathogenesis of acute and chronic ESC remains to be determined.     

Apparent digestibility of alternative plant‐protein feedstuffs for channel catfish,Ictalurus punctatus (Rafinesque)

A study was conducted with channel catfish, Ictalurus puntatus to determine apparent digestibility/availability coefficients of protein, amino acids, lipid and energy for alternative plant‐protein feedstuffs: corn gluten feed, corn germ meal, distillers dried grains with solubles, and canola meal, compared with those for soybean meal. A 32%‐protein, chemically defined diet containing 8% fish meal was used as the reference diet. Test diets consisted of 70:30 ratio of reference diet to test ingredient with chromic oxide (0.75% of the diet) as an inert indicator. Apparent digestibility/availability coefficients of protein, essential amino acids and energy in alternative protein feedstuffs tested were generally lower than those in soybean meal by channel catfish. Apparent digestibility coefficients of protein ranged from 75 to 87% and those of energy ranged from 52 to 59% for alternative feedstuffs. Lysine in alternative feedstuffs was 67–79%, methionine was 69–85% and cystine was 73–82% available to channel catfish. There were no significant differences in lipid ADCs among test feedstuffs (91.9–96.8%). Results from the present study can be useful in formulating cost‐effective catfish feeds using these alternative feedstuffs.