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1.
In response to a mortality event, seven Pangasius catfish (Pangasianodon hypophthalmus) were submitted to the University of the West Indies, School of Veterinary Medicine, Trinidad and Tobago, for diagnostic evaluation. These fish were part of a consignment that arrived from Kolkata two weeks earlier. Fish presented with perianal haemorrhage and blister‐like swellings on the skin which ruptured to leave ulcers. Edwardsiella ictaluri was consistently recovered from the brain and skin. Repetitive sequence‐mediated PCR analysis revealed genetic fingerprints consistent with E. ictaluri isolates from farm‐raised channel catfish in Mississippi, USA. Plasmid analysis of the case isolates identified two unique plasmids that differ slightly in conformation and content from the pEI1 and pEI2 plasmids described for E. ictaluri from other fish hosts. The case isolates were also PCR negative for several E. ictaluri virulence factors. The biological implications of these genetic differences are unclear and warrant further study. This is the first report and documentation of E. ictaluri infection in Trinidad and Tobago, suggesting the pathogen may have been introduced concurrently with the importation of fish. This report emphasizes the importance of adequate health screenings of imported lots to minimize the threat of introducing E. ictaluri to non‐endemic areas.  相似文献   

2.
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.  相似文献   

3.
The intraspecific variability of E. ictaluri isolates from different origins was investigated. Isolates were recovered from farm‐raised catfish (Ictalurus punctatus) in Mississippi, USA, tilapia (Oreochromis niloticus) cultured in the Western Hemisphere and zebrafish (Danio rerio) propagated in Florida, USA. These isolates were phenotypically homologous and antimicrobial profiles were largely similar. Genetically, isolates possessed differences that could be exploited by repetitive‐sequence‐mediated PCR and gyrB sequence, which identified three distinct E. ictaluri genotypes: one associated with catfish, one from tilapia and a third from zebrafish. Plasmid profiles were also group specific and correlated with rep‐PCR and gyrB sequences. The catfish isolates possessed profiles typical of those described for E. ictaluri isolates; however, plasmids from the zebrafish and tilapia isolates differed in both composition and arrangement. Furthermore, some zebrafish and tilapia isolates were PCR negative for several E. ictaluri virulence factors. Isolates were serologically heterogenous, as serum from a channel catfish exposed to a catfish isolate had reduced antibody activity to tilapia and zebrafish isolates. This work identifies three genetically distinct strains of E. ictaluri from different origins using rep‐PCR, 16S, gyrB and plasmid sequencing, in addition to antimicrobial and serological profiling.  相似文献   

4.
A molecular epidemiology study was conducted on 90 Edwardsiella ictaluri isolates recovered from diseased farmed freshwater catfish, Pangasianodon hypophthalmus, cultured in the Mekong Delta, Vietnam. Thirteen isolates of E. ictaluri derived from diseased channel catfish, Ictalurus punctatus, cultured in the USA were included for comparison. All the E. ictaluri isolates tested were found to be biochemically indistinguishable. A repetitive (rep)‐PCR using the single (GTG)5 primer was shown to possess limited discriminatory power, yielding two similar DNA profiles categorized as (GTG)5‐PCR group 1 or 2 among the Vietnam isolates and (GTG)5‐PCR group 1 within the USA isolates. Macrorestriction analysis identified 14 and 22 unique pulsotypes by XbaI and SpeI, respectively, among a subset of 59 E. ictaluri isolates. Numerical analysis of the combined macrorestriction profiles revealed three main groups: a distinct cluster formed exclusively of the USA isolates, and a major and minor cluster with outliers contained the Vietnam isolates. Antibiotic susceptibility and plasmid profiling supported the existence of the three groups. The results indicate that macrorestriction analysis may be regarded as a suitable typing method among the E. ictaluri species of limited intraspecific diversity. Furthermore, the findings suggest that E. ictaluri originating from Vietnam may constitute a distinct genetic group.  相似文献   

5.
Eight tetracycline resistant Edwardsiella ictaluri isolates obtained from diseased freshwater catfish (Pangasianodon hypophthalmus) in Vietnam, and showing different resistance phenotypes to other antimicrobial agents, were studied. The tet genes were determined using PCR. Conjugation experiments were performed to assess transferability of the tetracycline resistance determinant and the size and incompatibility group (Inc) of each tet-carrying plasmid were determined. PCR and sequencing were used for characterization of the co-transferred resistance genes. A tetA gene was demonstrated in the E. ictaluri isolates and for all of them, Escherichia coli transconjugants were obtained. All transconjugants contained high-molecular weight tetA-carrying plasmids (~ 140 kb) belonging to the incK group, as was shown with the PCR-based replicon typing method. The strA–strB, dhfr1 and sul 2 genes were detected on the tetA-carrying plasmids of the transconjugants showing resistance to streptomycin, trimethoprim and sulfonamides, respectively. The dhfr1 gene was found to be located in a class 1 integron as determined by PCR and sequencing. Interestingly, the 3′ CS region of class 1 integrons was not detected by PCR. This study shows the presence of incK plasmid-mediated tetracycline resistance among E. ictaluri isolates from diseased freshwater catfish in Vietnam.  相似文献   

6.
Abstract. Forty-five channel catfish, Ictalurus pitnctaius (Rafinesque), fingerlings were inoculated with Edwardsiella ictaluri to determine the usefulness of monoclonal antibodies for indirect fluorescent antibody techniques for confirming clinical diagnosis of enteric septicaemia of catfish. Bacteriological and indirect fluorescent antibody results were compared statistically and found to correlate in 90·3% of the cases. A method for incorporating monoclonals for indirect fluorescent antibody into the speciation of Edwardsiella ictaluri and the use of monoclonal antibodies in diagnosis is described.  相似文献   

7.
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.  相似文献   

8.
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.  相似文献   

9.
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.  相似文献   

10.
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.  相似文献   

11.
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.  相似文献   

12.
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.  相似文献   

13.
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.  相似文献   

14.
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.  相似文献   

15.
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.  相似文献   

16.
A disease outbreak in the danio (Danio devario) resulted in the isolation of Edwardsiella ictaluri as the etiologic agent. This isolate was compared with typical E. ictaluri and found to give identical reactions. It was also pathogenic for channel catfish. This is the second isolation of E. ictaluri from non-ictalurid fish.  相似文献   

17.
Mucins are large glycoproteins that cover epithelial surfaces of the body and play important roles in prevention of inflammatory and various infectious diseases. In this study, five membrane‐bound and seven secreted mucin genes in the channel catfish were identified. All these identified mucin genes possess at least one PTS, von Willebrand D (VWD) or SEA domains. The expression of the 12 mucin genes in channel catfish was first studied with infection of Edwardsiella ictaluri and Flavobacterium columnare. Expression difference in MUC13a, MUC13, MUC2 and MUC5b was found in the intestine after E. ictaluri infection. Eight mucin gene expressions (except MUC3a, MUC2, MUC4 and MUC5f) were up‐regulated at 4 hr and down‐regulated after 24 hr in the gill with F. columnare infection. Expression level of MUC2 gene was up‐regulated in the intestine with E. ictaluri infection without no significant change in the gill under the F. columnare infection, which indicate that MUC2 is tissue‐specific gene expression and has different immune respond to two bacterial challenge. Taken together, the study showed mucin from the gill by F. columnare challenge induced an obvious response than mucin from the intestine with E. ictaluri infection.  相似文献   

18.
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.  相似文献   

19.
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.  相似文献   

20.
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.  相似文献   

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