Field Efficacy of Florfenicol for Control of Mortality in Channel Catfish, Ictalurus punctatus (Rafinesque), Caused by Infection With Edwardsiella ictaluri

A field study to assess the efficacy of florfenicol (FFC) against enteric septicemia of catfish (ESC) was conducted with pond‐reared channel catfish fingerlings held in 0.1‐acre earthen ponds. Fish were challenged with Edwardsiella ictaluri in a natural pond outbreak or by cohabitation with E. ictaluri‐infected fish held in netpens. Fourteen ponds were assigned in equal number to two treatment groups, that is, either treated (with 10 mg FFC/kg body weight in medicated feed) or not treated (control) for 10 consecutive d. The threshold for enrollment into the study was 0.3% cumulative mortality attributed to ESC. Treatment was initiated on different dates for each pond because each pond was enrolled when 33 fish/pond were diagnosed with ESC based on clinical signs, lesions, or positive cultures. Mortality was monitored during the 10‐d treatment period and during a 14‐d posttreatment observation period. At the end of the 14‐d posttreatment observation period, all fish were euthanized, and 20 fish from each pond were examined by gross necropsy and evaluated for the presence of E. ictaluri by bacterial culture. The odds of a mortality in the control group were 2.20 times the odds of a mortality in the FFC‐treated group. Significantly fewer (P≤ 0.05) FFC‐medicated catfish died in comparison to unmedicated catfish. The minimum inhibitory concentration of FFC for this strain of E. ictaluri was 0.25 μg/mL in all fish that were assayed. The mean zone of inhibition (Kirby Bauer) was 36.8 mm from E. ictaluri isolates of test fish. There were no FFC treatment‐related lesions seen on gross pathology. FFC was efficacious and safe for control of mortality from E. ictaluri infection in catfish.     

Efficacy of a Live‐attenuated Edwardsiella ictaluri Oral Vaccine in Channel and Hybrid Catfish

This study evaluated the efficacy of an oral live‐attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish (ESC) in channel and hybrid catfish. The vaccine was delivered one time orally by feeding fish a diet coated with an attenuated E. ictaluri isolate at four doses to deliver between 4 × 10⁶ to 3.2 × 10⁷ viable vaccine cells/g wet feed. Thirty‐five days postvaccination, control and vaccinated fish were challenged with virulent E. ictaluri and mortality was examined for 30 d postchallenge. Mortality of nonvaccinated hybrids (85%) and nonvaccinated channel catfish (73%) was similar but significantly greater than all groups of vaccinated fish. In channel catfish, a trend toward increasing mortality with decreasing dose was observed. Mortality of channel catfish vaccinated with the lowest dose (26.6%) was significantly higher than fish vaccinated with the highest dose (14.1%) but similar to fish vaccinated at the intermediate doses (17.5 and 19.4%). In contrast, mortality of four doses of vaccinated hybrid catfish was similar and ranged between 10.4 and 14.0%. The data demonstrate that the attenuated E. ictaluri vaccine at all four doses tested is effective at reducing ESC‐related mortalities in hybrid and channel catfish.     

A Non-Nutritive,Ambient Temperature-Processed Floating Pellet as a Carrier of Antibiotics for Channel Catfish

A non-nutritive, ambient temperature-processed floating pellet (ATFP) was used as a medium to deliver apramycin (Eli Lilly and Co., Greenfield, Indiana), 52% active, and oxytetracycline hydrochloride (Pfizer, Inc., Lee's Summit, Missouri) (91% active) to channel catfish, Ictalurus punctatus, experimentally infected with Edwardsiella ictaluri. The drugs were fed at 50 mg of active antibiotic per kilogram of fish per day. Two medicated feeding trials were run for 10 consecutive days in troughs with channel catfish, beginning 24 hours after exposure to E. ictaluri. Mortality in the non-infected control population in Trial 1 was 7.6%, significantly (P < 0.05) lower than the 88.3% of the infected, non-medicated fish. Fish fed diets containing apramycin experienced 62.5% mortality, while fish fed diets containing oxytetracycline hydrochloride had 63.3% mortality. Mortality of the two medicated groups were significantly (P < 0.05) different from either control group. In Trial II, the 23% mortality in the non-infected group was significantly lower than the 48% mortality in the infected, non-medicated group and the 41% mortality in the oxytetricycline hydrochloride-medicated group, but not different than fish fed diets containing apramycin (28%). Mortality of the two medicated groups in Trial II were significantly different from each other. The presence of thc apramycin and oxytetracycline hydrochloride in the ATFP was demonstrated by the antibiotic diffusion method on agar plates swabbed with E. ictaluri.     

Studies on Vaccination of Channel Catfish,Ictalurus punctatus,Against Edwardsiella ictaluri

Enteric septicemia of cattish (ESC), caused by the bacterium Edwardsiella ictaluri, has become the most significant disease problem affecting the commercial channel catfish, Ictalurus punctatus, industry in the United States. Although antibiotics are used extensively for the control of ESC, there are inherent problems associated with their use. Consequently, experiments were initiated to evaluate the effectiveness of vaccination program that used immersion and oral delivery methods to administer a killed E. ictaluri vaccine to fry and fingerling channel cafish. In a preliminary pond study with laboratory challenge, mortality in a group vaccinated with a combination of immersion and oral procedures was only 5.0% in both high- and low-dose challenges. This was significantly different (P c 0.01) from non-vaccinated controls, which had 46.7%mortality in the lowdose challenge and the 6 1.7% mortality in the highdose challenge. This corresponds to relative percent survival (RPS) values of 89.3 and 91.9 respectively. Subsequent field trials further indicated the efficacy of a vaccination program for the prevention of ESC in channel catfish. In 1987-1988, a field study was conducted using 12 commercial ponds, with three replicates of four treatments. The four treatments included vaccination by immersion only, oral only, a combination of both immersion and oral procedures, and non-vaccinated conwols. Relative percent survival was 57.4 for the immersion only treatment, 50.3 for the oral only treatment, and 53.5 for the combination immersion and oral treatment. In 1989-1990, no significant difference was found between vaccinated and non-vaccinated fish. However, in 1989-1990, a vaccine-oil emulsion was topcoated on a floating feed, rather than incorporating vaccine in a sinking pellet. In 1990-1991, overall mortality in vaccinated fish was significantly less (P < 0.05) than non-vaccinated fish, with 41.2% mortality in vaccinates compared to 63.5% in non-vaccinated fish, for an RPS of 35.1. In examining RPS values for individual farms, two farms had excellent results, with RPS values of 81.3 and 76.9; two farms had only moderate success, with RPS values of 26.6 and 15.4; and one location had greater mortality in the vaccinated fish than in the non-vaccinated fish. However, that farm had only two ponds in the study and experienced significant losses to proliferative gill disease in the pond with vaccinated fish.     

Vaccination of Full‐sib Channel Catfish Families Against Enteric Septicemia of Catfish with an Oral Live Attenuated Edwardsiella ictaluri Vaccine

This study evaluated the efficacy of an oral live attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish in 20 full‐sib fingerling channel catfish families. The vaccine was delivered orally by feeding fish a diet coated with an attenuated E. ictaluri isolate. Sixty‐nine days postvaccination, control and vaccinated fish were challenged with virulent E. ictaluri and mortality was examined for 21 d postchallenge. Vaccinated fish had significantly lower mortality than nonvaccinated fish following challenge (P < 0.001). Mortality of vaccinated fish was 1.7 ± 1.4% as opposed to 47.8 ± 28.7% in nonvaccinated fish. Relative percent survival ranged from 87.7 to 100% with an average of 95.2 ± 4.0% (±SE) among the 20 families of fish. There were significant differences in mortality among families in nonvaccinated fish (P < 0.01) while there were no differences among vaccinated families of fish. Results indicate that the live attenuated E. ictaluri vaccine is effective at reducing mortality in channel catfish exposed to virulent E. ictaluri. These data demonstrate that genetic differences among healthy families of channel catfish are not major considerations in developing an effective vaccination program utilizing the oral vaccination platform described in this study.     

Determination of Dose Rate of Florfenicol in Feed for Control of Mortality in Channel Catfish Ictalurus punctatus (Rafinesque) Infected with Edwardsiella ictaluri, Etiological Agent of Enteric Septicemia

–A dose titration study was conducted to determine the appropriate dosage of florfenicol in feed to control mortality in channel catfish Ictalurus punctatus associated with enteric septicemia of catfish caused by Edwardsiella ictaluri. Six tanks (20 fish/ tank) were assigned to each of the following treatment: 1) not challenged with E. ictaluri and fed unmedicated feed; 2) challenged with E. ictaluri and fed unmedicated feed; 3) challenged and fed 5-mg florfenicol/kg body weight (kg bw); 4) challenged and fed 10-mg florfenicol/kg bw; or 5) challenged and fed 15-mg florfenicol/kg bw. Treatment was initiated the day after inoculation, and feed was administered by hand at 2.5% body weight for 10 consecutive days. Feeding activity was scored for all groups and was noted to be significantly less than the challenged, unmedicated group. Cumulative mortality in the challenged untreated group was 60%. The mortality in the unchallenged untreated group was 0%, and in die 5-, 10-, 15-mg florfenicol/kg bw group was 2.5%, 0.8%, and 2.5%, respectively. The mortality in each challenged, treated group and the non-challenged control group was significantly less than the challenged, unmedicated controls (P < 0.0001 for each contrast). There were no pairwise statistically significant contrasts among the florfenicol treated groups and the non-challenged control group. All 600 fish in the study were necropsied, cultured for bacteria, and examined by gross pathology. No specific lesions that could be associated with the antibiotic were observed. The efficacy of the 10 mg/kg dosage was confirmed in a separate dose confirmation study. In this study, fish in 30 tanks (20 fish/ tank) were infected with E. icraluri by immersion. Two days post-inoculation, fish in 15 tanks were hand-fed unmedicated feed, and 15 tanks were hand-fed medicated feed at a dosage of 10-mg florfenicol/kg bw at 2.5% body weight for 10 d. Feeding activity was scored and was noted to be significantly less than the challenged, unmedicated group. Cumulative mortality in the florfenicol group (14%) was significantly less than cumulative mortality in the untreated group (87.3%) (P < 0.0001). All 600 fish were submitted for bacterial culture, necropsied. and examined for gross pathology, and once again, no specific lesions that could be associated with the antibiotic were observed. The minimum inhibitory concentration of florfenicol against E. ictaluri in both studies was 0.25 ug/mL. Florfenicol was palatable, safe, and efficacious for control of mortality due to infection by E. ictaluri in catfish.     

Effect of Dietary Vitamin C on Weight Gain,Tissue Ascorbate Concentration,Stress Response,and Disease Resistance of Channel Catfish Ictalurus punctatus1

Juvenile channel catfish Ictalurus punctatus (average initial weight, 6.5 g/fish) were fed twice daily to apparent satiation with practical-type diets containing 0, 50, 150, or 250 mg supplemental vitamin C/kg from L-ascorbyl-2-polyphosphate for 10 wk under laboratory conditions. At the end of the feeding period, one half of the fish were stressed for 2 h by confinement and both stressed and nonstressed fish were exposed to a virulent strain of Edwardsiella ictaluri. Weight gain and feed conversion efficiency were lower for fish fed the basal diet than those fed diets containing supplemental vitamin C. No differences were observed in weight gain and feed conversion among fish fed diets containing supplemental vitamin C. There were no differences in feed consumption and survival (prior to experimental infection) among treatments. No vitamin C deficiency signs except reduced weight gain were observed in fish fed the basal diet. Serum cortisol concentrations were higher in stressed fish than in non-stressed fish. Dietary vitamin C level had no effect on serum cortisol concentration. As dietary vitamin C increased, ascorbate concentration in serum and liver increased. Confinement stress had no effect on serum and liver ascorbate concentrations. Cumulative mortality of channel catfish 21 d subsequent to experimental infection with E. ictaluri was higher for stressed fish than for nonstressed fish. Regardless of stress or nonstress, overall mortality for fish fed the basal diet was lower than the fish fed diets containing supplemental vitamin C. There were no differences in post-infection antibody levels among treatments or between stressed and nonstressed fish. Results from this study indicate that channel catfish require no more than 50 mg/kg dietary vitamin C for normal growth, stress response, and disease resistance.     

Establishing bacterial infectivity models in striped Catfish Pangasianodon hypophthalmus (Sauvage) with Edwardsiella ictaluri

A bacterial infectivity challenge model of Edwardsiella ictaluri in striped catfish was developed. All experiments were conducted using a bacterial isolate of E. ictaluri that had been recovered during a natural outbreak of bacillary necrosis of Pangasianodon (BNP) in farmed striped catfish Pangasianodon hypophthalmus in Vietnam. Time of immersion in 10⁷ CFU.ml⁻¹ had a significant effect on mortality. The immersion bacterial dose of 10⁷ CFU/ml for 30 s resulted in a cumulative percentage mortality of 63%. Three to four days post-bacterial challenge, fish showed gross clinical signs of natural BNP and E. ictaluri was recovered and identified from these fish. Moreover, a cohabitation challenge was evaluated as an alternative challenge method, although the mortalities among the infected fish were lower at around 15%–40%. This study confirmed the horizontal transmission of E. ictaluri in striped catfish and elucidated that cohabitation challenge could be used in reproducing the disease under controlled conditions.     

Enhanced Disease Resistance to Enteric Speticemia in Channel Catfish,Ictalurus punctatus,Administered Lytic Peptide

Lytic peptides have been shown in vitro to be toxic to a wide range of fish bacterial pathogens, including Edwardsiella ictaluri, the causative agent for enteric septicemia. Fingerling channel catfish, Ictalurus punctatus, were challenged with an injection of Edwardsiella ictaluri, and lytic peptide was administered with a single intraperitoneal injection 24 hours later. In a second group, lyptic peptide was also administered by osmotic pump 7 days before the bacterial challenge. A range of peptide concentrations (20-400µg/g fish) was used. Mortalities were recorded for 7 days after injection of bacteria, and E. icraluri was isolated and quantified by standard plate counts from livers. Various tissues were examined for abnormalities due to bacteria and peptide treatment. Fish injected with lytic peptide at 400 µg/g had up to a 95% reduction in bacteria and a 65% reduction in mortality compared to complete mortality in bacteria-injected controls. Fish implanted with osmotic pumps had 99.3% reduction in bacterial counts and had 6% mortality. Tissue samples from infected fish showed changes typical of reaction to immunologic stimuli and infection, while control fish injected with saline or peptide had normal gills, spleens, and kidneys. This study shows the potential for enhancing channel catfish disease resistance to E. ictaluri and other diseases by transferring the gene coding for the lytic peptide into channel catfish.     

Oral administration of Escherichia coli lipopolysaccharide enhances the immune system of striped catfish,Pangasianodon hypophthalmus (Sauvage)

Lipopolysaccharide (LPS), a component of Gram negative bacteria, was reported as important immunostimulant for fish. In this study, striped catfish were fed diets containing different Escherichia coli LPS concentrations (0%, 0.01% and 0.05%) for 2 weeks and then fed control feed (0% LPS) for 4 weeks. Plasma cortisol and glucose were rather low and did not differ significantly among treatments (P > 0.05). The respiratory burst activity, lysozyme, complement, total of antibody as well as mortality in fish challenged with Edwardsiella ictaluri were recorded every 2 weeks (W2, W4 and W6). The lysozyme activity significantly increased in fish treated with LPS (P < 0.05) in W2, W4 and W6. The highest values of respiratory burst activity were observed at week 4 in fish fed 0.01% LPS. There were significant differences in total of antibody between fish fed LPS (0.01%) and control in W2, W4. The challenge test with Edwardsiella ictaluri showed that fish fed 0.01% LPS had lower cumulative mortality (40%, 33% and 42%) compared with the fish fed 0.05% LPS (50%, 40% and 47%) and control fish (40%, 57% and 53%) in the three difference sampling times respectively. These results suggest that feed supplemented with 0.01% LPS could enhance immunity of striped catfish after 2 weeks of oral administration and fish could be protected against bacterial infection during the following 4 weeks.     

Isolation and characterization of Edwardsiella ictaluri strains as pathogens from diseased yellow catfish Pelteobagrus fulvidraco (Richardson) cultured in China

Yellow catfish Pelteobagrus fulvidraco (Richardson) is a commercially important fish generally distributed in Southeast Asian countries. The well‐known aetiological agent of enteric septicaemia of catfish, Edwardsiella ictaluri, was isolated from diseased yellow catfish P. fulvidraco (Richardson) reared at two commercial fisheries in China. The economic losses due to the high mortalities (about 50%) caused by this bacterium have been increasing annually. The affected fish presented two different, typical symptoms: pale gills, slight exophthalmia and a ‘hole in the head’, and haemorrhage on the opercula, in the skin under the jaw, creating a ‘hole under the jaw’. These diseases were found frequently in cultured yellow catfish throughout China. The isolates from both outbreaks were all Gram negative, facultatively anaerobic and short rod. Morphological and biochemical tests and phylogenetic analysis based on the 16S rDNA sequences all strongly indicated that these yellow catfish isolates were highly identical to the known E. ictaluri. In addition, the isolates possessed the typical plasmid profile of E. ictaluri. Experimental infection assays were conducted and pathogenicity (by an intraperitoneal injection) was demonstrated in yellow catfish and channel catfish Ictalurus punctatus. The results showed that yellow catfish isolates were quite conservative phenotypically and genetically, and were able to cause two different, typical symptoms in this fish under unknown conditions and mechanism.     

Palatability of Sarafloxacin HCl-Medicated Feed to Channel Catfish

Two experiments were conducted to evaluate the palatability of sarafloxacin HCl to channel catfish, Ictalurus punctatus. Fingerling channel catfish were raised in 110-1 glass aquaria supplied with flowing well water at a rate of 1 l/min. Fish were fed to satiation once daily either a purified or a practical diet formulated to contain 0, 500, 1,000, 2,000, or 4,000 mg active sarafloxin HCl/kg of diet. Feed consumption was quantified daily for 5 days. All fish were weighed at the beginning and at the end of each experiment. There was a significant decrease in feed consumption of fish fed purified diets with increasing levels of sarafloxacin HCl. However, feed consumption was adequate to provide the proposed dosage regardless of drug concentration. Fingerling channel catfish readily consumed practical diets containing sarafloxacin HCl; feed consumption and weight gain of fish fed medicated and non-medicated practical diets did not differ. There were no apparent palatability problems that would negate use of sarafloxacin HCl in commercial channel catfish feeds.     

Effect of Sublethal Hypoxia on the Immune Response and Susceptibility of Channel Catfish, Ictalurus punctatus, to Enteric Septicemia

The effect of sublethal hypoxia exposure on stress and immune responses and susceptibility to Edwardsiella ictaluri infection in juvenile channel catfish, Ictalurus punctatus, was investigated. Fish were monitored for temporal changes in glucose and cortisol concentrations before, during, and after 2 h exposure to sublethal hypoxia (<2 mg/L dissolved oxygen [DO]) and when maintained under normoxic conditions (6.0 ± 0.3 mg/L DO). Both blood glucose and plasma cortisol increased significantly in response to hypoxic conditions. Fish exposed to hypoxic or normoxic conditions were challenged with a high dose (1.3 × 10⁷ colony‐forming units [CFU]/mL) or a low dose (1.3 × 10⁵ CFU/mL) of E. ictaluri or sterile culture broth by 30‐min immersion bath. Approximately 1% of fish in both the normoxic and the hypoxic groups died when challenged with the low dose of E. ictaluri. However, when challenged with the high dose of E. ictaluri, catfish exposed to hypoxic conditions had significantly higher cumulative mortality (36 ± 12.1%) than those maintained under normoxic conditions (12 ± 1.1%). Total hemolytic complement and bactericidal activities and antibody response were lower in hypoxia‐exposed channel catfish, indicating that increased susceptibility of channel catfish to E. ictaluri may be the result of the immunosuppressive effects of the stress response to hypoxia.     

Application of Compensatory Growth to Enhance Production in Channel Catfish Ictalurus punctatus

Four treatment groups that received repeating cycles of fixed feed deprivation for either 0, 1, 2, or 3 d (control, treatment 1, treatment 2, and treatment 3, respectively), followed by periods of refeeding with a 36% protein commercial catfish feed once daily as long as the active phase of compensatory growth (CG) persisted, were assessed in flow-through aquaria. No-feed periods elicited the CG state and were immediately followed by days of ad libatum refeeding. At the end of 10 wk, average growth rate (AGR) of fish was higher ( P < 0.05) than the control by 40%, 180%, and 191% for treatment 1, treatment 2, and treatment 3, respectively. The average weight of fish in treatment 3 was heavier ( P < 0.05) than the average control group at the end of the study period. Mean daily feed consumption was 3.91%, 5.03%, 5.36%, and 5.98% for control, treatment 1, treatment 2, and treatment 3, respectively. Mean feed consumption per fish per day was 24%, 71.3%, and 70.7% higher than the control in treatment 1, treatment 2, and treatment 3, respectively. Restricted feeding is one of the effective methods to contain ESC-related losses in commercial channel catfish fingerling operations. The mean cumulative survival of treatment groups registered higher ( P < 0.05) survival to Edwardsiella ictaluri infection compared to the daily fed control fish. Results from this study show that compensatory growth response triggered by periodic non-feeding days can improve growth rate, feed consumption, and improved survival to ESC infections in channel catfish fingerlings.     

Mortality and pathology of hybrid catfish,Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), associated with Edwardsiella ictaluri infection in southern Thailand

Enteric septicaemia of catfish (ESC) caused by Edwardsiella ictaluri is becoming an increasing problem in aquaculture and has been reported worldwide in a variety of fish species. This study reports ESC in hybrid catfish, Clarias macrocephalus (Günther) × Clarias gariepinus (Burchell), cultured in southern Thailand. The bacteria were identified as E. ictaluri by conventional and rapid identification systems, as well as by genetic and phylogenetic characterization. Analysis of 16S rRNA indicated 100% homology to the 16S rRNA sequence of several E. ictaluri strains in GenBank. Plasmid profiles demonstrated 4.0‐ and 5.6‐kb plasmids, compared with the 4.8‐ and 5.6‐kb plasmids in the US isolates, and representative genes of three of the four known pathogenicity islands of US isolates were present. Serologically, lipopolysaccharide (LPS) purified from the Thai isolates was not recognized by a monoclonal antibody against the LPS of US isolates. Fish experimentally infected with E. ictaluri showed 23–100% mortality within 14 days with a 168‐h LD₅₀ of 6.92 × 10⁷ CFU mL^?1 by immersion and a 96‐h LD₅₀ of 1.58 × 10⁶ CFU fish^?1 by intraperitoneal injection. Examination of tissue sections obtained from both naturally and experimentally infected fish indicated that infection of hybrid catfish with E. ictaluri produced lesions in several organs including liver, kidney, spleen, heart and brain. Histopathology findings included cellular necrosis, focal haemorrhage, infiltration of lymphocytes and multifocal granulomatous inflammation in the infected organs.     

Effects of Co‐stocking Smallmouth Buffalo,Ictiobus bubalus,with Channel Catfish,Ictalurus punctatus

Proliferative gill disease (PGD) in catfish is caused by the myxozoan Henneguya ictaluri. The complex life cycle requires Dero digitata as the oligochaete host. Efforts to control PGD by eradicating D. digitata have been unsuccessful. Smallmouth buffalo, Ictiobus bubalus, (SMB) are opportunistic bottom feeders and a putative option for controlling D. digitata. In 2011, 15 ponds (0.4 ha) were stocked with 5000 channel catfish, Ictalurus punctatus; 7 of these 15 ponds were also stocked with 300 SMB fingerlings. There were no differences in benthic invertebrate numbers or water quality variables between ponds with or without SMB. At harvest, there were no differences in percent survival, total weight, or catfish feed conversion ratio. In the second year, 18 ponds (0.4 ha) were stocked with 6000 channel catfish. Half the ponds were also stocked with 300 SMB. Sentinel fish were used to estimate disease severity, and pond water was collected for molecular estimation of H. ictaluri actinospore concentrations. Similar to the first year, there were no differences between treatments in any variable tested, including PGD severity in sentinel fish and parasite concentrations in pond water. Under these study conditions, presence of SMB did not have a measureable effect on PGD incidence, parasite density, or overall catfish production.     

A rapid bioassay for bactericides against the catfish pathogens Edwardsiella ictaluri and Flavobacterium columnare

The most common bacterial diseases in pond‐raised channel catfish Ictalurus punctatus (Rafinesque) are enteric septicemia of catfish and columnaris, caused by Edwardsiella ictaluri and Flavobacterium columnare respectively. Medicated feed containing antibiotics is one management approach that catfish producers use in the treatment of bacterial diseases. However, the future use of all types of medicated feed in catfish aquaculture is uncertain. To discover effective alternatives to antibiotics, a rapid 96‐well microplate bioassay utilizing E. ictaluri and F. columnare to evaluate natural compounds and extracts was developed. In this bioassay, bacterial growth is determined by absorbance measurements of microplate wells after 24 h incubation and then confirmed by detecting cell viability after the addition of 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide with additional incubation for 24 h. The minimum inhibitory concentration, minimum bactericidal concentration and 50% inhibition concentration (IC50) are determined by graphing the absorbance data. The 24 h IC50 results of test compounds are compared with the 24 h IC50 results of the drug controls oxytetracycline and florfenicol. Among the antibiotics evaluated, doxycycline and tetracycline appear more effective against E. ictaluri and F. columnare than either drug control. This bioassay is rapid, reproducible and economical for evaluating a large number of compounds and extracts.     

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.     

Effects of Gender and Sex Hormones on Disease Susceptibility of Channel Catfish to Edwardsiella ictaluri

The use of monosex populations for aquaculture is becoming widely used for several species. The current studies determined if there were any differences between male and female channel catfish, Ictalurus punctatus, in disease susceptibility to Edwardsiella ictaluri, one of the most important pathogenic bacteria in catfish culture. Disease challenge experiments were carried out on fingerling channel catfish fed 17β‐estradiol or testosterone before the challenge, and on all male and on sibling all female fingerlings. All male populations were produced by mating YY males with normal XX females. Sibling females were produced by hormonally sex reversing a subpopulation of six of the all male families. Weight gain or specific growth rate did not differ in fish fed testosterone or estrogen. Fish fed the highest dose of estrogen (50 mg/kg) had a significant higher mortality (P < 0.05), while mortality was similar in catfish fed 10 and 50 mg/kg of testosterone compared to controls. There were no differences in mortality between sibling males and females. These data indicate no increased disease susceptibility to E. ictaluri between males and females or due to exogenous sex hormones. Production of all male catfish for culture can proceed without concern for disease susceptibility to E. ictaluri.     

Effect of Feeding Frequency on Feed Consumption,Growth, and Feed Efficiency in Aquarium‐reared Norris and NWAC103 Channel Catfish (Ictalurus punctatus)*

Abstract.— A 6‐wk feeding study was conducted to determine the effect of feeding frequency on growth rate of juvenile Norris and NWAC103 channel catfish, Ictalurus punctatus, reared under laboratory conditions. Four replicate groups of Norris and NWAC103 catfish (average weight of 4.0 ± 0.2 g/fish, SEM) were fed to visual satiety at different feeding frequencies (one, two, or three times daily). The percent of total daily food consumed for Norris catfish fed three times daily (0800, 1200, and 1600 h) was 44.4, 27.7, and 27.9%, respectively, while NWAC103 catfish consumed 42.9, 26.7, and 30.4%, respectively. Specific growth rate and feed intake were higher in fish fed three times daily compared to fish fed once and twice daily for both strains. Feed efficiency was reduced in NWAC103 fed three times daily compared to fish fed once or twice, while feed efficiency was similar among the Norris treatments. Gastrointestinal (GI) tract index ([weight of GI tract/weight of fish] × 100) decreased in NWAC103 catfish as feeding frequency increased, while a similar nonsignificant trend was also observed in Norris catfish. The results of this study demonstrate that aquarium‐reared Norris and NWAC103 catfish fed three times a day consume more feed and gain more weight than catfish fed once or twice a day. The observed decrease in the GI index as a result of feeding more frequently demonstrates that the size of the GI tract increases, relative to body weight, when catfish are fed only once a day. Under laboratory conditions, the number of times a day the fish are fed should be considered when trying to maximize growth of NWAC103 and Norris strains of catfish.