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.     

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.     

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.     

Efficacy of Sarafloxacin Hydrochloride in Treating Naturally-Induced Edwardsiella ictaluri Infections in Channel Catfish,Ictalurus punctatus

Sarafin (sarafloxacin hydrochloride), a new antibacterial, was evaluated in the field on a naturally induced infection of Edwardsiella ictaluri in channel catfish, -Ictalurus punctatus. Healthy channel catfish (mean weight = 50 g) were stocked into nine cages at 200 fish per cage in a pond with an undergoing E. ictaluri infection. Seven days after stocking, dead fish were observed in the cages with clinical signs of enteric septicemia of channel catfish (ESC). After E. ictaluri was confirmed through isolative biochemical tests, medicated feed was applied for five consecutive days. During this period, fish in three control cages received a commercial 32% protein floating feed, three other cages of fish served as positive controls and were fed Romet, and three cages received the test feed with Sarafin. Both medicated feeds reduced the increase in cumulative percent mortality. In the control cages, cumulative percent mortalities continued to increase throughout the study period. Average daily mortality rates were significantly lower following both treatments of medicated feed, and treatments receiving Sarafin showed the greatest reduction in average daily mortality rates. Average daily mortality rates in the control did not change after the medicated feeding period. Toward the end of the study, temperatures reached 30°C, above the active range of ESC infections, and all mortalities ceased.     

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.     

Uptake and Clearance of Edwardsiella ictaluri in the Peripheral Blood of Channel Catfish Ictalurus punctatus Fingerlings during Immersion Challenge

Uptake and clearance of Edwardsiella ictaluri in the peripheral blood of channel catfish Ictalurus punctatus fingerlings were monitored for 216 h after exposure to E. ictaluri for 4 h and 8 h under static conditions. Most fish exposed to E. ictaluri developed bacteriemia 24 h post-exposure, and by 72 h post-exposure E. ictaluri was recovered from all the blood of all exposed fish. The number of E. ictaluri colony forming units (CFU) in the blood of moribund fish ranged between 1.7 × 10³ to 1.6 × 10⁵ CFU/50 μL whole blood. Clearance of bacteria from the blood was observed by 216 h post-exposure and all fish surviving bacterial exposure developed agglutinating antibody against E. ictaluri . The pathogenesis of the infection was accompanied by the shedding of viable E. ictaluri into the water which may serve as a mechanism by which fish to fish transmission occurs.     

Recovery and Viability of Edwardsiella ictaluri from Great Blue Herons Ardea herodias Fed E. ictaluri-Infected Channel Catfish Ictalurus punctatus Fingerlings

Feeding activities of great blue herons Ardea herodias in catfish ponds during outbreaks of enteric septicemia of catfish have been implicated as a mechanism for the transmission of the disease from infected to uninfected ponds. Although Edwardsiella ictaluri , the causative agent, has been identified in gastrointestinal tracts of great blue herons, the role of these birds as a vector of E. ictaluri is not well documented. The potential of these birds to contaminate catfish ponds with E. ictaluri was investigated by feeding captive herons over a 4-d period with catfish fingerlings injected intraperitoneally with live E. ictaluri . Daily fecal samples, throat and rectal swabs, and feather samples were collected, cultured and examined for E. ictaluri using both a selective media and a monoclonal indirect fluorescent antibody test specific for E. ictaluri . Gastrointestinal tracts sampled at the conclusion of the feeding trial were similarly examined. While E. ictaluri was detected using the indirect fluorescent antibody test, no viable E. ictaluri was cultured from either feces, gastrointestinal tracts or feathers. Growth of E. ictaluri was not observed at 40 C, the rectal temperature observed in captive great blue herons. Prior incubation at 40 C suppressed the growth of E. ictaluri at 24 C, an optimal temperature for growth of this bacterium. These results indicate that great blue herons appear to play little or no role in the transmission of E. ictaluri among catfish ponds.     

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.     

Vaccination of Mixed and Full-Sib Families of Channel Catfish Ictalurus punctatus Against Enteric Septicemia of Catfish With a Live Attenuated Edwardsiella ictaluri Isolate (RE-33)

These studies were conducted to evaluate the efficacy of a live attenuated Edwardsiella ictaluri vaccine against enteric septicemia of catfish. In one study channel catfish fingerlings (72 d of age post hatch) were immersed for 30 min in water containing E. ictaluri RE-33 at dosages of 1 × 10⁶, 1 × 10⁷ and 2 × 10⁷ CFU/ML of water. No mortalities were observed following vaccination. Following exposure to virulent Edwardsiella ictaluri the cumulative mortality of fish vaccinated with dosages of at least 1 × 10⁷ CFU/mL were significantly lower than that of non-vacccinated fish in both laboratory and field challenges. Vaccination with 1 × 10⁶ CFU RE-33mL provided some protection during the laboratory challenge but failed to protect fish under field conditions. In a second study, vaccination of 6 full-sib families of channel catfish at a vaccine dosage of 1 × 10⁷ CFU/mL resulted in a relative percent survival among families ranging from 67.1 to 100%. Significant differences in mortality were found among families and between vaccinated and unvaccinated groups, but there was no family by vaccine interaction. Families with the highest mortality after vaccination were also shown to have the highest mortality without vaccination (r = 0.82; P = 0.04).     

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.     

A Comparison of Immersion, Immersion/Oral Combination and Injection Methods for the Vaccination of Channel Catfish Ictalurus punctatus Against Edwardsiella ictaluri

Abstract.— Four different vaccination regimes, including non-vaccinated controls (C), immersion/oral combination (IO). late immersion (LI), and injection (IJ), were evaluated for antibody production and protection from challenge in channel catfish Ictalurus punctatus that were hatched and raised in recirculating culture systems over a 28-wk period. For IO, fry were vaccinated by immersion on day 12 post-hatch and given an oral booster during week 10 post-hatch. The LI and IJ delivered vaccine was also administered at week 10 post-hatch. No titers were detected in the IO group prior to the oral booster at week 10. Mean titers rose sharply after the week 10 vaccinations in all treatment groups, maintained a high level for about 8 wk, and slowly declined over the next 4 wk. Injected fish had significantly higher titers during the primary response period than either LI or IO fish. There was no statistical difference in titers between LI and IO fish during the primary response. After an oral booster delivered during week 22, titers in LI and IO fish increased significantly at week 25, while titers in IJ fish declined despite the booster delivery. An immersion booster delivered during week 25 resulted in significant increases in titers in all vaccinated groups. Of multiple challenges, the only one not marred by concurrent columnaris infections revealed a low, but significant level of protection in IJ fish at 12 wk post-vaccination.     

Influence of the Dietary Level of Iron from Iron Methionine and Iron Sulfate on Immune Response and Resistance of Channel Catfish to Edwardsiella ictaluri

Channel catfish Ictalurus punctatus fingerlings were fed purified diets supplemented with iron at levels of 0, 20, 60, and 180 mg/kg from iron sulfate (FeS) or 5, 10, 20, 60, and 180 mg/kg from iron methionine (FeM) in triplicate tanks for 8 wk. Fish were then divided into two groups and subjected to different assays to measure disease resistance and individual immune functions. Representative fish from each dietary treatment were challenged by bacterial immersion with virulent Edwardsiella ictaluri , and mortality due to enteric septicemia was recorded. Other fish were immunized with 0.2–mL formalin-killed E. ictaluri and boosted 21 d post-immunization. Antibody response was determined by FAST-ELISA. Chemiluminescent and chemotaxis assays were performed using peritoneal macrophages. Supplementation of the diet with various levels of iron from FeS or FeM did not significantly affect antibody production. Chemotactic migration by macrophages was depressed in iron-deficient fish and a level of 60 mgkg from either FeS or FeM provided the highest chemotactic indexes. A deficiency of dietary iron was found to increase mortality of channel caffish due to enteric septicemia of catfish (ESC). However, more studies should be conducted to better understand the effects of sources and levels of dietary iron on immune responses and disease resistance in channel caffish.     

Influence of Dietary Levels of Magnesium on Growth, Tissue Mineral Content, and Resistance of Channel Catfish Ictalurus punctatus Challenged with Edwardsiella ictaluri

Juvenile channel catfish were fed purified diets supplemented with magnesium (Mg) from Mg sulfate at levels of 0, 200, 400, 600, 800, and 1,000 mg/kg and 0, 200, 400, 600, and 800 mg/kg in two separate feeding studies. In study I, the effect of dietary levels of Mg on growth response, vertebral mineral content, and macrophage chemotaxis were evaluated. Study II had similar objectives except that whole body mineral content was measured, and resistance of channel catfish to Edwardsiella ictaluri challenge was also determined. Fish with an average weight of 10.89 g were stocked at a rate of 50 fish/110‐L aquarium (study I). In study II, fish with an average weight of 4.14 g were stocked at rates of 40 fish/110‐L aquarium. Prior to stocking, each batch of fish was acclimated to laboratory conditions and fed the basal diet for 2 wk. The concentration of Mg in rearing water was 1.8 mg/L. Each diet was fed to fish in quadruplicate and triplicate aquaria to apparent satiation for 10 wk for studies I and II, respectively. Fish fed the basal diet started to die as early as 3 d after the study began (17 d of feeding the diet without Mg supplementation). In both studies, weight gain, survival, and feed efficiency were lowest for fish fed the basal diet but increased with increasing dietary levels of Mg. However, the differences between the values of each of these parameters for fish fed diets containing supplemental Mg were not always significant. Magnesium‐deficiency signs observed were anorexia, sluggishness, convulsions, deformed snout, vertebral curvature, muscle flaccidity, and high mortality. Vertebral and whole body ash concentrations were high, but Mg content was low for fish fed the basal and the 200‐mg Mg diets. Bone Ca content did not differ among fish fed different diets (study I), but whole body Ca tended to increase for fish fed the basal diet, suggesting the possibility of calcification of soft tissues. Macrophage chemotaxis in the presence of exoantigen was highest for fish fed diets supplemented with Mg at 400 and 200 mgkg for studies I and II, respectively. When expressed in terms of chemotaxis index, however, maximum or near maximum value was observed at a dietary Mg level of 400 mg/kg. Thus, a dietary level of Mg of 400 mg/kg from Mg sulfate was required for optimum growth and survival, maintaining high tissue levels of Mg, prevention of muscle flaccidity and skeletal deformity, and stimulating macrophage chemotaxis. Dietary levels of Mg had no effect on the resistance of juvenile channel catfish to Edwarsiella. ictaluri challenge.     

Growth Response and Resistance to Edwardsiella ictaluri of Channel Catfish,Ictalurus punctatus,Fed Diets Containing Distiller’s Dried Grains with Solubles

A study was conducted to examine the effect of dietary levels of distiller’s dried grains with solubles (DDGS) on growth, body composition, hematology, immune response, and resistance of channel catfish, Ictalurus punctatus, to Edwardsiella ictaluri challenge. Five diets containing 0, 10, 20, 30, and 40% DDGS with supplemental lysine (Diets 1–5) as partial replacements of a combination of soybean meal and cornmeal on an equal protein basis were fed to juvenile catfish (13.33 ± 0.25 g) for 12 wk. Growth performance and feed utilization efficiency were similar for fish in all treatments. Body lipid and moisture increased and decreased, respectively, in fish feed DDGS‐containing diets relative to the control group. Dietary treatment had no effect on red and white blood cell counts. Hemoglobin and hematocrit were significantly higher in fish fed diets containing DDGS than in those fed the control diet. Fish fed 20–40% DDGS diets had increased serum total immunoglobulin, and those fed the 30% DDGS diet had significantly increased antibody titers 21 d following E. ictaluri challenge. Other immune variables evaluated were not affected by dietary treatments. Preliminary results on bacterial challenge showed an increased resistance against E. ictaluri in fish fed DDGS‐containing diets (Diets 2–5).     

Immune Response and Resistance to Stress and Edwardsiella ictaluri Challenge in Channel Catfish, Ictalurus punctatus, Fed Diets Containing Commercial Whole-Cell Yeast or Yeast Subcomponents

Dietary supplementation of yeast or yeast subcomponents (YYS) as commercial preparations of β‐glucan (MacroGard^®; Biotec‐Mackzymal, Tromsø, Norway; and Betagard A^®; Aqua‐In‐Tech, Inc., Seattle, WA, USA), mannan oligosaccharide (Bio‐Mos? Aqua Grade; Alltech, Nicholasville, KY, USA), or whole‐cell Saccharomyces cerevisiae (Levucell SB20^®; Lallemand Animal Nutrition, Milwaukee, WI, USA) at the manufacturer’s recommended levels was evaluated on the physiological performance of juvenile channel catfish, Ictalurus punctatus. Fish were fed YYS diets for 4 wk, followed by 2 wk of control diet. Fish were sampled at the end of each feeding period (4 and 6 wk) to measure hematological and immune parameters and growth and to determine the effects of dietary β‐glucan on resistance to Edwardsiella ictaluri infection and to low‐water stress (6 wk). Supplementation of YYS in diets did not affect growth performance, hematology, or immune function. Survival from E. ictaluri infection was from 5 to 17.5% higher in fish fed YYS diets than in the control group, but the increases were not significant. Some improvement in stress resistance was observed in YYS‐fed catfish after exposure to low‐water stress. Stress reduction in fish fed diets supplemented with yeast subcomponents has been reported previously, but thus far, no explanation has been proposed for this effect. The present study and the previously published research suggest that dietary YYS supplementation does not appear to improve resistance of channel catfish to E. ictaluri.     

Effect of Dietary Total Protein and Animal Protein on Growth and Feed Efficiency of Juvenile Channel Catfish Ictalurus punctatus and Their Response to Edwardsiella ictaluri Challenge

A laboratory study was conducted to evaluate effects of dietary total protein and animal protein source and concentration on growth and feed efficiency of juvenile channel catfish Ictalurus punctutus and their response to Edwardsiellu ictuluri challenge. Eight diets evaluated were: three diets containing either 28, 32, or 36% crude protein with 6% menhaden fish meal and 6% meat and bonehlood meal and five diets containing 32% crude protein with either no animal protein, 68 or 12% menhaden fish meal, or 6% or 12% meat and bonehlood meal, respectively. Twenty channel catfish with an average weight of 6.6 g/fish were stocked into each of forty 110-L flow-through aquaria (five aquaridtreatment). Fish were fed to approximate satiation twice daily for 9 wk. Fish in each tank were then exposed to E. ictaluri . There were no differences in feed consumption, weight gain, feed efficiency, and survival before and after challenge among fish fed diets containing 28, 32, or 36% protein with 6% menhaden fish meal and 6% meat and bone/ blood meal. Fish fed a 32% all-plant protein diet had weight gain and feed efficiency similar to fish fed diets containing 12% menhaden fish meal, but had a higher weight gain than fish fed a 32% protein diet containing 6% meat and bonehlood meal. No significant differences were observed in survival after E. ictuluri challenge among fish fed diets containing the various levels of animal proteins. Results indicate that dietary protein levels varying from 28% to 36% do not appear to affect growth, feed efficiency. and E. icraluri resistance or susceptibility in fingerling channel cattish fed to satiation and raised from approximately 7 to 56 g under laboratory conditions. Data also demonstrate that a 32% all-plant protein diet can be fed to small fingerling channel catfish without adversely affecting growth, feed efficiency, or resistance to E. ictuluri .     

Genetic Effects for Response to Live Edwardsiella ictaluri, Killed E. ictaluri, and Stress in Juveniles from All Crosses Among USDA 103, USDA 102, and Norris Channel Catfish Ictalurus punctatus Strains

Juveniles from all possible crosses among USDA 102. USDA 103, and Norris channel catfish Ictalurus punctatus strains were compared for: 1) survival and mix-Edwardsiella ictaluri antibody after exposure to live E. ictaluri bacterium (isolate 597-458); 2) antibody level after injection with formalin-killed E. ictaluri (597-458); and 3) pre-stress. post-stress, and stress-recovery serum Cortisol levels. Purebred and crossbred USDA 102 strain fish had higher survival (mean of five genetic groups = 87%) and lower anti- E.ictaluri antibody (mean optical density (OD) of five genetic groups = 0.167) 30 d after live E. ictaluri challenge than purebred Norris and USDA 103 strains and their crosses (means of four genetic groups = 60% survival and 0.210 OD antibody level). Significant general combining ability, line effects, and heterosis indicated that the USDA 102 strain contributed additive and dominance genetic effects for increased survival and lower antibody level after live E. ictaluri challenge. Antibody response to formalin-killed, intra-peritoneally injected E. ictaluri was not different among genetic groups (overall mean = 0.198 OD). Serum Cortisol was measured prior to (pre-stress), immediately after (post-stress), and 2 h after (stress-recovery) a standard stressor. Serum Cortisol level was highest in post-stress fish (35.8 ng/mL), intermediate in stress-recovery fish (10.9 ng/mL), and lowest in pre-stress fish (6.5 ng/mL), but was not different among genetic groups within a stress time period. Results indicate diat differences exist among genetic groups of channel catfish for survival and antibody production after live E. ictaluri challenge, but these differences were not related to antibody response to killed E. ictaluri or serum Cortisol levels.     

Isolation and Characterization of Edwardsiella ictaluri from Southern Catfish,Silurus soldatovi meridionalis, (Chen) Cultured in China

In April 2011, there was an outbreak of an infectious disease in southern catfish, Silurus soldatovi meridionalis, (Chen) (15–20 g) in Sichuan Province, China. Two isolates, LW101 and LW102, were isolated from kidney and liver of the sick fish on brain‐heart infusion (BHI) agar and were considered to be the cause of this disease based on experimental challenges. The morphological and physiological characteristics as well as the biochemical tests of the two isolates were the same and similar to Edwardsiella ictaluri. Furthermore, the sequencing of the 16S rRNA gene and the gryB gene revealed that the isolates were highly homogeneous with E. ictaluri. On the basis of the phenotypic characteristics and phylogenetic analysis of these genes, both isolates were identified as E. ictaluri. Susceptibility of the isolates to 22 antibiotics was tested using the disc diffusion method. Both isolates showed a similar antibiotic susceptibility, which was characterized by resistance to acetylspiramycin, ampicillin, clarithromycin, penicillin, oxytetracycline, and sinomin (SMZ/TMP); the strains were susceptible to amikacin, chloramphenicol, florfenicol, roxithromycin, ciprofloxacin, norfloxacin, doxycycline, and tenemycin. To our knowledge, this is the first report of E. ictaluri infection in southern catfish.