Cutaneous antibodies in excised skin from channel catfish, Ictalurus punctatus Rafinesque, immune to Ichthyophthirius multifiliis

This study determined whether cutaneous antibodies were present in the excised skin of channel catfish, Ictalurus punctatus Rafinesque, immune to Ichthyophthirius multifiliis Fouquet (Ich). Theronts were immobilized on or near the excised skin from immune fish. The survival of immobilized theronts was significantly reduced after exposure for 8 h to the culture of excised skin from immune fish. Culture fluids from excised skin of immune fish immobilized theronts with a peak in the immobilization titre at 24 h post-exposure. Immobility of theronts in the culture fluid from immune skin was removed after immunoabsorption with theronts. Indirect immunofluorescent staining of theronts treated with culture fluid from excised skin of immune fish revealed strong and uniform fluorescence on the cilia and cell surface of theronts. Western blot analysis of the culture fluid from immune fish revealed a 70-kDa band which corresponded to the molecular weight of catfish immunoglobulin heavy chain. The results of this study show that cutaneous antibodies to Ich theronts were present in and released from the excised skin from fish immune to Ich. Immobilization and killing of the theronts are two characteristics of the antibody response that appear to prevent the successful invasion of theronts into excised skin.     

Protective effect of cutaneous antibody produced by channel catfish,Ictalurus punctatus (Rafinesque), immune to Ichthyophthirius multifiliis Fouquet on cohabited non-immune catfish

Fish which survive a sublethal ichthyophthiriasis acquire protective immunity against Ichthyophthirius multifiliis Fouquet (Ich). This study evaluated the protective effect of cutaneous antibody secreted by channel catfish, Ictalurus punctatus (Rafinesque), immune to Ich on cohabited non-immune catfish. Non-immune and immune fish controls were separately maintained and infected with theronts. The Ich infection was assessed by scoring 0, < 50, 50-100, and > 100 trophonts fish(-1) at 5 days post-infection. The results of infection showed that cohabited fish at the ratio of 15 non-immune to two immune fish had < 50 trophonts fish(-1). Eighty per cent of the cohabited fish at the ratio of 10 non-immune to two immune fish showed 0 or < 50 trophonts fish(-1). The 76% of control non-immune fish had more than 100 trophonts fish(-1). The control immune fish had 0 trophonts fish(-1). Anti-Ich antibody was detected using enzyme-linked immunosorbent assay in water samples taken from tanks containing immune fish after the water samples were concentrated 40-fold. The study suggests that immune fish cohabited with non-immune fish may protect non-immune fish against Ich infection.     

Induced cross-protection in channel catfish, Ictalurus punctatus (Rafinesque), against different immobilization serotypes of Ichthyophthirius multifiliis

Abstract Channel catfish, Ictalurus punctatus (Rafinesque), were immunized with Ichthyophthirius multifiliis (Ich) theronts and trophonts, and the immune response and host protection against both homologous and heterologous serotypes of Ich were evaluated. Immunizations were done with two immobilization serotypes (ARS4 and ARS6) of live theronts by bath immersion (trial I) and with sonicated trophonts by intraperitoneal (i.p.) injection (trial II). Cutaneous and serum antibody titres against Ich following immunization were measured and survival of catfish was determined after theront challenge. Theronts were immobilized by the antiserum from fish immunized with homologous theronts or trophonts, but not by the serum of fish immunized with the heterologous serotype. Serum from fish immunized by immersion with live theronts showed higher enzyme-linked immunosorbent assay titres against both homologous and heterologous serotypes than fish immunized by i.p. injection of trophonts. Channel catfish immunized by immersion with live theronts or by i.p. injection with sonicated trophonts developed an immune response against Ich and provided cross-protection against challenge from both serotypes (ARS4 and ARS6) of the parasite. Sonicated trophont antigens in aqueous solution by i.p. injection could stimulate an immune response in fish, but the immunity was of short duration.     

Immune responses and host protection of channel catfish, Ictalurus punctatus (Rafinesque), against Ichthyophthirius multifiliis after immunization with live theronts and sonicated trophonts

The humoral immune responses and host protection of channel catfish, Ictalurus punctatus (Rafinesque), against Ichthyophthirius multifiliis (Ich) were determined after immunization with live theronts and sonicated trophonts. Immunizations with live theronts or sonicated trophonts were carried out by both bath immersion and intraperitoneal (i.p.) injection. Cutaneous and serum immunoglobulin (Ig) levels and anti-Ich antibodies were measured 12 and 21 days post-immunization. The level of Ich infection and survival of catfish were determined after theront challenge. Cutaneous and serum anti-Ich antibodies were significantly higher (P < 0.05) in fish immunized with live theronts by immersion or i.p. injection, or with sonicated trophonts administered by i.p. injection, than in fish immunized with sonicated trophonts by immersion, with bovine serum albumin by i.p. injection, or non-immunized controls. Host protection was noted only in fish immunized with live theronts by immersion or i.p. injection or with sonicated trophonts by i.p. injection. There was a positive correlation between higher levels of anti-Ich antibodies and host survival in the immunized fish.     

Antibody mediated immune response against Ichthyophthirius multifiliis using excised skin from channel catfish, Ictalurus punctatus (Rafinesque), immune to Ichthyophthirius

This study investigated antibody mediated immune response against Ichthyophthirius multifiliis (Ich) by determining whether theronts would retain the potential for reinfection, both in vitro and in vivo, after treatment with the culture fluid of excised skin from channel catfish, Ictalurus punctatus , immune to Ich. The invasion was reduced significantly ( P  < 0.05) for theronts treated with the immune culture fluid compared with those treated with the culture fluid from naive fish. The treatment of theronts with the immune culture fluid greatly reduced the size and survival of trophonts compared with those treated with the culture fluid from naive fish. Fewer fish were infected and the infection density was less for fish exposed to theronts treated with immune culture fluid. The infection was severe for fish invaded by theronts treated with the culture fluid from naive fish, with a high number of infected fish and heavy density of trophonts per fish. All fish were infected by Ich when exposed to the theronts treated with the immunoadsorbed culture fluid. In summary, results of this study show that cutaneous antibodies in the culture fluid of excised skin from immune fish significantly reduces theront infectivity by immobilizing or weakening theronts.     

Apoptosis in Ichthyophthirius multifiliis is associated with expression of the Fas receptor of theronts

The expression of type I membrane Fas receptors on the surface of Ichthyophthirius multifiliis (Ich) theronts and the possible association between Fas expression and theront apoptosis induced by the immune antibody was examined. Fas receptors were detected on the theront surface using fluorescein isothiocyanate-conjugated mouse monoclonal antibody against Fas. Fas-positive theronts significantly increased with time during in vitro incubation and with increasing theront concentration. Furthermore, the immune cutaneous antibody induced theront apoptosis; however, Fas ligand did not. A highly significant correlation was noted between theront Fas expression and immune cutaneous antibody-induced theront apoptosis. Numbers of apoptotic theronts increased with increasing number of Fas-positive theronts. The data indicated that theront apoptosis induced by immune cutaneous antibody appears to be positively correlated with the expression of Fas on the surface of Ich theronts.     

Susceptibility of three strains of blue catfish,Ictalurus furcatus (Valenciennes), to Ichthyophthirius multifiliis

This study compared the susceptibility of three blue catfish strains (D&B, USDA 101 and USDA 102) to the parasite Ichthyophthirius multifiliis (Ich). In Trial I, a cohabitation study (all strains stocked communally) was conducted and fish were exposed to theronts at 0, 200, 1000, 5000 or 25 000 theronts fish^?1, respectively. All fish died when exposed to theronts at 5000 or 25 000 theronts fish^?1. When exposed to 1000 theronts fish^?1, USDA 102 strain of blue catfish showed significantly lower mortality (78.5%) compared to USDA 101 and D&B strains (92.7% and 100%). In Trial II, the same three strains of blue fish were evaluated for their susceptibility to Ich with strains challenged in separate tanks by adding Ich theronts at 0, 200 and 1000 theronts fish^?1, respectively. All D&B and USDA 101 blue catfish died; however, 42.3% of USDA 102 strain survived the infection when exposed to 1000 theronts per fish. The results indicate that there are differences among strains of blue catfish for susceptibility to Ich, and these differences will be useful in the development of improved catfish germplasm for commercial aquaculture.     

Evidence that channel catfish, Ictalurus punctatus (Rafinesque), mortality is not linked to ingestion of the hepatotoxin microcystin-LR

Catfish farms located in the south-eastern USA using brackish (3–5 g NaCl L⁻¹) well water experience sporadic fish kills sometimes with high mortality. An investigation of three catastrophic losses occurring in this region identified no involvement of infectious diseases or traditional water quality problems, including oxygen, ammonia or nitrite. The high mortality and time course of the problem was indicative of exposure to a toxin. Attempts by other workers to explain the cause of this unique syndrome (high chloride associated toxicosis of catfish, HCTC), suggested that the losses might be because of microcystin-producing blooms of Microcystis aeruginosa , but our investigations failed to support this conclusion . We found that (1) the liver histology of catfish experimentally exposed to pure microcystin-LR is very different from that of catfish sampled during outbreaks of HCTC; (2) measurements of microcystin-LR concentrations in the three cases were far lower than the concentration required to kill catfish by experimental immersion; (3) the HCTC toxin appears to have a short half-life, whereas microcystin-LR does not; (4) experimental gavage of catfish with massive amounts of microcystin-LR does not cause the acute mortality typical of HCTC; (5) outbreaks of HCTC appear to be associated with heavy blooms of Anacystis marina , a halophytic cyanobacteria, not with blooms of M. aeruginosa .     

Immunization of channel catfish, Ictalurus punctatus Rafinesque, against Ichthyophthirius multifiliis (Fouquet): killed versus live vaccines

Abstract. Fish surviving infection with the pathogenic ciliated protozoan, Ichthyophthirius multifiliis (Fouquet, 1876), become resistant to subsequent infection by the parasite. The acquired immunity suggests that development of a vaccine against the parasite may be possible. Because of the advantages of immunoprophylaxis for treatment of the disease, an effort has been made to determine whether fish exposed to killed parasite preparations can resist subsequent lethal challenge. Both the route of administration and the effects of stage specific antigens have been examined. Channel catfish vaccinated by intraperitoneal (i.p.) injection or bath immersion with killed I multifiliis tomites show 100% mortality following a standard challenge protocol. Similarly, 100% mortality was observed in test groups injected with tomite cilia. In both cases, a consistent difference in days to death between control and test group animals was observed. Although complete mortality was seen with fish injected with tomite preparations, fish vaccinated with killed trophonts (the feeding stage of the parasite) had a much greater degree of protection with approximately 50% of fish surviving an otherwise lethal challenge. Finally, animals injected intraperitoneally with live tomites showed nearly complete immunity and were identical in their response to fish which survive natural infection. The response of fish vaccinated with live parasites indicates that animals injected intraperitoneally can develop surface immunity and that i.p. injection is a suitable route of administration for potential I. multifiliis vaccines.     

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.     

Logit models for evaluating spawning performance of channel catfish, Ictalurus punctatus (Rafinesque)

Broodstock evaluations are often measured by variables such as spawning success, fecundity, fertilization and hatching rates, usually expressed as percentage values. Outcomes are generally analysed as continuous random variables, assuming that they follow a normal distribution. Ordinary linear regression models (e.g. analysis of variance) as well as χ² analysis are typically applied. However, these models may not be the most appropriate as a number of test criteria may not be met. For example, spawning success outcomes are inherently discrete and non‐negative data and hence their distribution is not likely to be normal. As these models may not be the most appropriate, a case study using logit analysis as an alternative method for the evaluation of this type of data is presented by considering the response as binary data (spawned versus did not spawn). An exact version of logit analysis was performed due to the sparseness of the data. The results demonstrate that appropriate statistical models provide better insight into the cause–effect relationships that exist between control variables and the dependent variable (likelihood of spawning in this case). As would be expected, each strain of fish responded somewhat differently to the test variables. Changing the protein level of the diet from 32% to 42% or increasing the feeding frequency from three to six times per week either did not influence spawning or negatively affected spawning respectively. Additionally, older fish performed better than younger fish and the early spawning period was better than the later spawning period, regardless of strain. These responses, however, were only detected using logit analysis, which is a more sensitive test and would thus be recommended for this type of data.     

Dietary protein concentration and daily feed allowance influence response of channel catfish, Ictalurus punctatus (Rafinesque), to ractopamine

Year-1 channel catfish, Ictalurus punctatus (Rafinesque) (average initial weight, 48 g) were fed diets containing 240 g kg^?1, 300 g kg^?1, or 360 g kg^?1 dietary protein with 0 or 20 mg kg^?1 ractopamine to satiation and diets containing 240 g kg^?1 and 360 g kg^?1 protein with 0 or 20 mg kg^?1 ractopamine at a restricted rate (60% of satiation) in aquaria at 29 ± 2°C for 8 weeks. Fish fed ractopamine at the 360 g kg^?1 protein level gained significantly more weight than controls when fed to satiation but not when the ration was restricted. Fish fed ractopamine at the 240 g kg^?1 protein level did not gain more weight than the controls at satiation or restricted feeding. Fish fed ractopamine in the diet containing 300 g kg^?1 protein gained significantly more weight than controls when fed to satiation. Ractopamine caused no significant differences in feed conversion at the 240 g kg^?1 or 360 g kg^?1 protein levels with restricted or satiation feeding. Muscle fat decreased when ractopamine was fed at all dietary protein levels and at both feeding rates. Muscle protein increased with ractopamine feeding in all treatments except the high-protein, satiation treatment. Weight gain response of year-1 channel catfish to ractopamine is dependent upon nutrient concentration of the diet and ration size; however, the effect of ractopamine on muscle composition of channel catfish is less dependent on nutrient concentration or amount of diet fed.