Enhanced susceptibility of channel catfish to the bacterium Edwardsiella ictaluri after parasitism by Ichthyophthirius multifiliis

Bacterium Edwardsiella ictaluri and parasite Ichthyophthirius multifiliis (Ich) are two common pathogens of cultured fish. The objective of this study was to evaluate the susceptibility of channel catfish Ictalurus punctatus to E. ictaluri and determine bacterial loads in different fish organs after parasitism by Ich. Fish received the following treatments: (1) infected by I. multifiliis at 5000 theronts/fish and exposed to E. ictaluri; (2) infected by I. multifiliis alone; (3) exposed to E. ictaluri alone; and (4) non-infected control. E. ictaluri in fish organs were quantified by quantitative real-time polymerase chain reaction and reported as genome equivalents per mg of tissue (GEs/mg). The results demonstrated that the Ich-parasitized catfish showed significantly (P<0.05) higher mortality (91.7%) when exposed to E. ictaluri than non-parasitized fish (10%). The bacterial loads in fish infected by 5000 theronts/fish ranged from 6497 to 163,898 GEs/mg which was between 40 and 2000 fold higher than non-parasitized fish (49-141 GEs/mg). Ich infection enhanced the susceptibility of channel catfish to bacterial invasion and increased fish mortality.     

The effect of high total ammonia concentration on the survival of channel catfish experimentally infected with Flavobacterium columnare

Ammonia concentrations in water can affect the severity of Flavobacterium columnare infections in fish. Two trials lasting 7 d each were conducted to determine the effect of a single immersion flush treatment of total ammonia nitrogen (TAN; 15 mg/L) on the survival of channel catfish Ictalurus punctatus infected with E columnare; the chemical was added while the water flowed continuously through the tanks. Both trials consisted of four treatments: (1) no ammonia exposure and no bacterial challenge (control), (2) ammonia exposure only, (3) bacterial challenge only, and (4) both ammonia exposure and bacterial challenge. Two hours after exposure to ammonia, the highest un-ionized ammonia level was 0.43 mg/L. The percent un-ionized ammonia is based on TAN, temperature, and pH. Caudal fins from three fish in each treatment were sampled at 24 h posttreatment to be analyzed by quantitative real-time polymerase chain reaction (qPCR). No significant difference in survival (mean +/- SE) was noted between the channel catfish in treatment 1 (95.2 +/- 1.2%) and those in treatment 2 (95.6 +/- 1.0%); however, survival in both treatments 1 and 2 differed significantly from that in treatments 3 (8.5 + 4.5%) and 4 (41.8 +/- 12.7%). Treatment 4 catfish had significantly higher survival than treatment 3 catfish. Quantitative PCR data showed that treatment 4 fish had significantly less F. columnare (7.6 x 10(5)) than did treatment 3 fish (1.2 x 10(7)), and treatment 2 fish (8.5 x 10(3)) had significantly less bacteria than did treatment 1 fish (6.9 x 10(4)), indicating that ammonia limited the F. columnare infection. The highest mean concentration of the bacteria (3.9 x 10(7)) was found on moribund fish. The ammonia concentrations tested did not negatively influence fish survival but interfered with the infection process. An in vitro assay was also conducted to evaluate the direct effects of ammonia on F columnare.     

Concomitant exposure of rainbow trout fry to Gyrodactylus derjavini and Flavobacterium psychrophilum: effects on infection and mortality of host

Although rainbow trout fry mortality syndrome caused by the bacterium Flavobacterium psychrophilum is widespread in fish farms it is difficult to reproduce infection of rainbow trout in the laboratory using immersion exposure with bacterial suspensions. It has therefore been speculated that ectoparasites could act as enhancers of bacterial infections under natural conditions. In the present study rainbow trout fry were exposed to infections with F. psychrophilum (immersion for 30 min or 10 h) alone, exposed to the ectoparasitic monogenean Gyrodactylus derjavini alone or exposed to both pathogens in combination. Infection levels and host mortality were subsequently monitored to elucidate if the ectoparasitic monogeneans could enhance infection of fish with the bacterium. Immersion of fish in bacterial suspensions alone did not result in infection. Only one fish became infected with the bacterium and this fish belonged to the combination exposure group. The parasite populations increased differently in the various groups and it was found that host mortality was correlated to gyrodactylid infection levels (r=0.94) but not to bacterial exposure. The results emphasise the pathogenicity of the parasite G. derjavini, the relative resistance of intact fish to direct exposure to F. psychrophilum but provide only a weak indication of a possible enhancement of bacterial invasion due to ectoparasitic infections. It cannot be excluded that higher parasite burdens and/or prolonged immersion (more than 10 h) in bacterial suspensions may result in bacterial invasion.     

Early Changes in Pigmented Macrophages in Head Kidney of Channel Catfish Infected with Aeromonas hydrophila

Abstract

Channel catfish Ictalurus punctatus with scarified skin were exposed to the bacterial pathogen Aeromonas hydrophila. Systemic infections developed in 80% of the exposed fish, and the remaining exposed fish had infections limited to the cutaneous lesion. Skin of control fish was injured in the same manner as for the exposed fish, but control fish were not exposed to A. hydrophila nor was A. hydrophila recovered from them. None of the fish died during the 3-d experiment, and gross lesions in head kidneys, including changes in the size or relative weight, were not found in infected fish. Macrophage aggregates in head kidney of systemically infected fish increased 68% in mean area, 28% in number, and 111% in total relative volume. Volume of lipofuscin, the predominate pigment found in channel catfish head kidney, in systemically infected fish was about twice that of control fish. Volume of hemosiderin was about four times as great and macrophages containing hemosiderin were about three times as numerous in systemically infected fish as in control fish. The amount of melanin in macrophages did not change in infected fish. Significant differences in macrophage aggregates and pigments were not found between controls and fish with only superficial infections. Microscopic lesions, other than changes in macrophage aggregates and pigments, were not found in the head kidney. Increases in macrophage aggregates, lipofuscin, and hemosiderin in head kidney of channel catfish were useful for quantification of injury caused by a systemic bacterial infection.     

The Effect of High Total Ammonia Concentration on the Survival of Channel Catfish Experimentally Infected with Flavobacterium columnare

Abstract

Ammonia concentrations in water can affect the severity of Flavobacterium columnare infections in fish. Two trials lasting 7 d each were conducted to determine the effect of a single immersion flush treatment of total ammonia nitrogen (TAN; 15 mg/L) on the survival of channel catfish Ictalurus punctatus infected with F. columnare; the chemical was added while the water flowed continuously through the tanks. Both trials consisted of four treatments: (1) no ammonia exposure and no bacterial challenge (control), (2) ammonia exposure only, (3) bacterial challenge only, and (4) both ammonia exposure and bacterial challenge. Two hours after exposure to ammonia, the highest un-ionized ammonia level was 0.43 mg/L. The percent un-ionized ammonia is based on TAN, temperature, and pH. Caudal fins from three fish in each treatment were sampled at 24 h posttreatment to be analyzed by quantitative real-time polymerase chain reaction (qPCR). No significant difference in survival (mean ± SE) was noted between the channel catfish in treatment 1 (95.2 ± 1.2%) and those in treatment 2 (95.6 ± 1.0%); however, survival in both treatments 1 and 2 differed significantly from that in treatments 3 (8.5 ± 4.5%) and 4 (41.8 ± 12.7%). Treatment 4 catfish had significantly higher survival than treatment 3 catfish. Quantitative PCR data showed that treatment 4 fish had significantly less F. columnare (7.6 × 10⁵) than did treatment 3 fish (1.2 × 10⁷), and treatment 2 fish (8.5 × 10³) had significantly less bacteria than did treatment 1 fish (6.9 × 10⁴), indicating that ammonia limited the F. columnare infection. The highest mean concentration of the bacteria (3.9 × 10⁷) was found on moribund fish. The ammonia concentrations tested did not negatively influence fish survival but interfered with the infection process. An in vitro assay was also conducted to evaluate the direct effects of ammonia on F. columnare.

Received September 15, 2010; accepted May 7, 2011     

Effects of variable periods of food deprivation on the development of enteric septicemia in channel catfish

Enteric septicemia of catfish (ESC), caused by the bacterium Edwardsiella ictaluri, is the most significant bacterial disease affecting channel catfish Ictalurus punctatus. Withholding feed during outbreaks of ESC is a widely accepted industry practice used to control losses from the disease. Scientific evidence concerning the validity of the practice is contradictory. Two studies were conducted to further evaluate the survival of channel catfish fingerlings following variable periods of feed deprivation before and after exposure to E. ictaluri in controlled aquarium experiments. In the first study, feed was withheld for varying time periods before bacterial challenge. After bacterial challenge, feed was either withheld or fish were fed daily. The second study utilized fish fed daily or fish deprived of feed 7 d before bacterial challenge. Daily feeding was resumed 4, 48, and 96 h after fish were exposed to E. ictaluri. In both experiments, the prechallenge feed treatments did not affect mortality. In contrast, withholding feed after bacterial challenge reduced mortalities by 52% in experiment 1 and by 45% in experiment 2. The highest mortality was observed when fish were fed immediately after immersion exposure and the lowest when fish were completely denied feed or fed daily starting 96 h after challenge. This reduction in mortality occurred when the concentration of E. ictaluri in aquarium water was negligible. These data suggest that when E. ictaluri is present in the water, feeding fish increases mortality by enhancing oral exposure to the pathogen.     

Responses of Blue Catfish and Channel Catfish to Environmental Nitrite

Abstract

In nitrite-exposure experiments, percent methemoglobin, plasma nitrite concentration, and plasma chloride ion concentration were compared between channel catfish Ictalurus punctatus and blue catfish I. furcatus exposed to sublethal levels of nitrite for 48 h at 25°C. In nitrite-recovery experiments, fish exposed to elevated environmental nitrite for 12 h were transferred to freshwater, and blood characteristics were monitored during the 24-h recovery period. Blue catfish appeared to be more resistant to environmental nitrite than channel catfish. Methemoglobin levels (percent of hemoglobin in methemoglobin form) were significantly lower in blue catfish than in channel catfish. Maximum plasma nitrite concentrations were 137 mg NO₂-/L plasma in blue catfish and 164 mg NO₂-/L plasma in channel catfish. Percent methemoglobin and plasma nitrite concentration were closely correlated. Plasma chloride decreased initially with exposure to nitrite but quickly returned to control levels. Blue catfish exposed to nitrite at 10°C required 1 week to recover when placed in nitrite-free water. The methemoglobin reductase enzyme apparently functioned at a slow rate in fish acclimated to cold temperatures.     

Dietary bovine lactoferrin induces changes in immunity level and disease resistance in Asian catfish Clarias batrachus

The effects of including bovine lactoferrin (Lf) in the diet of the Asian catfish (Clarias batrachus) on specific and non-specific immunity as well as disease resistance were investigated. The catfish were fed four different diets for 2 weeks: a commercial diet as control and the same diet supplemented with 50, 100 and 200mg bovine Lf/kg feed. After 1 and 2 weeks, serum bacterial agglutination titre against Aeromonas hydrophila as a measure of specific immunity; natural serum haemolysin titre, lysozyme activity and oxidative radical production by neutrophils as a measure of non-specific immunity as well as disease resistance against A. hydrophila challenge to vaccinated and non-vaccinated animals were evaluated. The results showed that Lf supplements, particularly at 100mg level, significantly (P<0.05) enhanced serum lysozyme level, oxidative radical production and level of protection against A. hydrophila challenge in non-vaccinated animals irrespective of length of exposure. The specific immunity was not influenced by Lf feeding as evidenced from the bacterial agglutination titre and level of protection in vaccinated animals. As Lf feeding at 100mg/kg for 1 week is able to enhance the non-specific immunity and disease resistance of catfish efficiently, these results support the possible use of Lf as an immunostimulant for farmed catfish.     

Effect of Indomethacin on the Development of Proliferative Gill Disease

Abstract

After parenteral treatment with the cyclooxygenase inhibitor indomethacin, channel catfish Ictalurus punctatus were exposed to mature spores of an Aurantiactinomyxon sp. demonstrated to be the etiological agent of proliferative gill disease (PGD). Fish that received indomethacin at a dose of 2.0 or 5.0 mg/kg body weight within 0.5 h before exposure to the myxozoan and again at 24 h postexposure had significantly (P < 0.05) less severe gill lesions 7 d after exposure than fish that received the drug vehicle alone. Fish that received 0.5 mg indomethacin/kg had moderately severe lesions. All fish were confirmed to be infected with the organism associated with PGD by microscopic examination of gills 4 or 7 d postexposure. These results suggest that products of the cyclooxygenase pathway (e.g., prostaglandins) participate in the pathophysiologic host response to PGD.     

Identification and expression profiles of multiple genes in Nile tilapia in response to bacterial infections

To understand the molecular mechanisms involved in response of Nile tilapia (Oreochromis niloticus) to bacterial infection, suppression subtractive cDNA hybridization technique was used to identify upregulated genes in the posterior kidney of Nile tilapia at 6h post infection with Aeromonas hydrophila. A total of 31 unique expressed sequence tags (ESTs) were identified from 192 clones of the subtractive cDNA library. Quantitative PCR revealed that nine of the 31 ESTs were significantly (p<0.05) upregulated in Nile tilapia at 6h post infection with A. hydrophila at an injection dose of 10(5)CFU per fish (≈ 20% mortality). Of the nine upregulated genes, four were also significantly (p<0.05) induced in Nile tilapia at 6h post infection with A. hydrophila at an injection dose of 10(6)CFU per fish (≈ 60% mortality). Of the four genes induced by A. hydrophila at both injection doses, three were also significantly (p<0.05) upregulated in Nile tilapia at 6h post infection with Streptococcus iniae at doses of 10(6) and at 10(5)CFU per fish (≈ 70% and ≈ 30% mortality, respectively). The three genes induced by both bacteria included EST 2A05 (similar to adenylate kinase domain containing protein 1), EST 2G11 (unknown protein, shared similarity with Salmo salar IgH locus B genomic sequence with e value of 0.02), and EST 2H04 (unknown protein). Significant upregulation of these genes in Nile tilapia following bacterial infections suggested that they might play important roles in host response to infections of A. hydrophila and S. iniae.     

Comparison of the hemagglutination of formalinized American channel catfish (Ictalurus punctatus) erythrocytes between formalinized and live Aeromonas hydrophila isolated from the catfish rearing in Kasumigaura lake in Japan

Formalinized Aeromonas (A.) hydrophila agglutinated loosely with the formalinized American channel catfish erythrocytes (FACCE), while live A. hydrophila agglutinated tightly with the FACCE. There was a significant difference on the number of attaching bacterial cells to the FACCE (p<0.01) (n=40 erythrocytes) between formalinized and live A. hydrophila. The other bacteria such as Salmonella (S.) Typhimurium ST-5, Escherichia (E.) coli V-517 and Staphylococcus (S.) hyicus ATCC1249 used in this experiment did not attach the FACCE.     

Enteric Septicemia Resistance in Blue Catfish and Three Channel Catfish Strains

Abstract

Seven full-sib families in each of three strains of channel catfish Ictalurus punctatus (mean weight, 10.7 g; SD, 2.6 g) and one family of blue catfish I. furcatus (mean, 10.9 g; SD, 0.4 g) were challenged by immersion with the bacterium Edwardsiella ictaluri, the causative agent of enteric septicemia, to evaluate variation in disease resistance. Blue catfish were resistant to infection and had only 0.7% mortality. Significant (P < 0.001) variation in channel catfish resistance was found among strains, families, and body weights. Red River strain channel catfish were the most resistant (14.9% mortality), followed by Mississippi-select fish (67.1%) and Mississippi-normal fish (72.3%). Mean family mortality of channel catfish ranged from 1.3% in a Red River family to 95.3% in a Mississippi-select family. Although weight had a significant effect on survival, the regression slope of survival on weight was ?0.10 (SE, 0.02). Sources of variation in channel catfish mortality adjusted for weight were 67.3% from strains, 29.7% from families, and 3.0% from replicate tanks. The mechanism for differential resistance presently is unknown. Results indicate considerable potential for reducing disease losses by using E. ictaluri-resistant catfish strains in aquaculture and for genetic improvement in resistance via selective breeding programs.     

Cool Weather Feeding Influences Responses of Channel Catfish to Edwardsiella ictaluri Challenge

Abstract

A pond study was conducted from November 1 through April 30 with young (age-0; average size, 43 g) and market-size (age-2; average size, 660 g) channel catfish Ictalurus punctatus to compare three management regimens: No feeding, partial feeding (no feeding in December, January, and February), and continuous feeding according to fish size and water temperature. Weight change, feed conversion ratio, and responses to experimental challenge with Edwardsiella ictaluri were evaluated. No significant difference in weight gain occurred between partially fed and continuously fed fish in either age-group; average weight increase for fed age-0 fish was 99% and for fed age-2 fish was 38%. The nonfed age-0 fish lost 12.3% and the nonfed age-2 fish lost 7% of their initial weight. Feed conversion ratios were significantly lower in partially fed fish than in continuously fed fish in both age-groups. Mortality from E. ictaluri challenge among age-0 fish was significantly higher in the nonfed fish, but among the age-2 fish, mortality was significantly lower in the nonfed fish. No difference in mortality rate occurred between partially fed and continuously fed fish in either age-group. Starvation induced lower antibody production against E. ictaluri antigen in the age-0 fish but higher antibody production in the age-2 fish. Phagocytic index was lower in nonfed fish than in fed fish from both age-groups. This study indicates no benefit from feeding age-0 and age-2 channel catfish during December, January, or February if feeding is reintroduced in March and continued through April. Although starvation was immunosuppressive in small channel catfish, it enhanced resistance to bacterial infection in larger fish. However, more research information is needed before reduced feeding can be recommended to enhance resistance of channel catfish to E. ictaluri.     

Dose Titration of Sarafloxacin (A-56620) against Edwardsiella ictaluri Infection in Channel Catfish

Abstract

Two dose-titration studies were performed with sarafloxacin (A-56620) on channel catfish Ictalurus punctatus infected with Edwardsiella ictaluri. Fish were infected with E. ictaluri by water-bath exposure and subsequently fed rations for 5 d to equal 0, 2, 6, 10, or 14 mg sarafloxacin/kg body weight per day. In trial I, mortality was significantly reduced from 69% for fish receiving no medication to 17% for those receiving 10 or 14 mg sarafloxacin/kg per day (P ≤ 0.01). In trial II, mortality was reduced from 33% of the nonmedicated infected group to 5 and 10% offish receiving 10 and 14 mg sarafloxacin/kg per day, respectively. Fish receiving 2 or 6 mg sarafloxacin/kg per day had intermediate mortality rates.     

Channel Catfish Response to Ultraviolet-B Radiation

Abstract

Fingerling channel catfish Ictalurus punctatus exposed to simulated ultraviolet-B radiation at an average daily dose of 2.9 J/cm² were quite sensitive to the radiation. After a 24-h exposure, thinning of the most dorsal epidermis frequently was accompanied by edema. Compared with epidermis of unexposed fish, mucous cells in exposed fish were less superficial and club cells were less numerous both dorsally and high on the lateral surface of the body. Sunburn cells with pyknotic nuclei were evident in the epidermis of exposed fish. Among fish exposed for 48 h, focal necrosis and sloughing of the outer epidermal layer were widespread. A methanol-extractable skin substance that is associated with resistance to sunburn in other fish species was not detected in channel catfish.     

Preliminary Assessment of the Tolerance and Efficacy of Florfenicol against Edwardsiella ictaluri Administered in Feed to Channel Catfish

Abstract

A tolerance study was conducted to determine the palatability of florfenicol to channel catfish Ictalurus punctatus. Four tanks of fish (20 fish/tank) were assigned to each of five treatments distinguished by the amount of florfenicol given in feed per kilogram of body weight, namely, 0, 10, 20, 40, or 100 mg. Fish were fed at a rate of 2.5% of body weight per day for 10 consecutive days. On day 11, all surviving fish were euthanatized, counted, and weighed as a group. Florfenicol- medicated feed was palatable to fish at doses of 10, 20, 40, and 100 mg for 10 consecutive days. All 400 fish were necropsied and examined by histopathology, and no treatment-related changes were observed. In a separate exploratory efficacy study, four tanks (20 fish/tank) were assigned to each of the following treatments: (1) not challenged with Edwardsiella ictaluri and fed unmedicated feed, (2) challenged with E. ictaluri and fed unmedicated feed, (3) challenged with E. ictaluri and fed florfenicol at 10 mg per kilogram of body weight, (4) challenged and fed florfenicol at 20 mg/kg, and (5) challenged and fed florfenicol at 40 mg/kg. Treatment was initiated the day after inoculation, and feed was administered by hand for five consecutive days at 2.5% of body weight. The cumulative mortality observed over a 17-d period in treatment groups 1–5 was 2.5, 57.5, 0, 1.25, and 1.25%, respectively. All 400 fish were necropsied and examined by histopathology. The results indicate that florfenicol was effective in controlling mortality from enteric septicemia of catfish with no adverse treatment-related tissue changes. Florfenicol was palatable, safe, and efficacious in controlling mortality due to infection by E. ictaluri in channel catfish.     

Evaluation of Two Aryl-Fluoroquinolones against Bacterial Pathogens of Channel Catfish

Abstract

The in vitro and in vivo efficacies of two aryl-fluoroquinolones, A-56619 and A-56620, against two bacterial pathogens of channel catfish Ictalurus punctatus were determined The minimum inhibitory concentrations (MIC) of A-56619, A-56620, oxytetracycline, nalidixic acid, spectinomycin, ampicillin, doxycycline, and ormetoprim-sulfadimethoxine against 10 isolates of both Edwardsiella ictaluri and Aeromonas sobria were determined by the agar-dilution method. The in vivo efficacies of A-56619 and A-56620 were determined by bacterial challenge of channel catfish with E. ictaluri. Fish were fed a prepared diet that supplied 12.5, 25.0, or 50.0 mg of active drug per kilogram body weight per day. In vitro tests showed that all strains of E. ictaluri and A. sobria were sensitive to A-56619 and A-56620. The two aryl-fluoroquinolones demonstrated a lower mean MIC against these organisms than any of the other antibiotics tested. Significant reductions in mortality occurred within all channel catfish groups treated with these two chemicals as compared with controls. Mortality from E. ictaluri infection of fish given feed medicated with A-56619 and A-56620 ranged from 4 to 12%. Mortality among unmedicated controls ranged from 20 to 68%. No significant difference in mortality among dosage levels was demonstrated.     

Effects of Dietary Zinc and Calcium on Select Immune Functions of Channel Catfish

Abstract

The effects of dietary zinc and calcium on immunocompetence of channel catfish Ictalurus punctatus (mean initial weight = 3.1 g/fish) were determined after a 10-week feeding trial. Fish were fed daily amounts equal to 3% of their body weight of one of six purified diets that were deficient in zinc (2 mg/kg of diet), calcium (0.02% of diet), or both; replete with zinc (20 mg/kg of diet) and calcium (0.5% of diet); or excessive in zinc (200 mg/kg of diet) or calcium (2.5% of diet). Immunocompetence of immunized and nonimmunized fish fed the various diets was assessed by measuring serum immunoglobulin M levels and mortality after an intraperitoneal injection of live Aeromonas hydrophila. Channel catfish fed diets deficient in zinc, calcium, or both had significantly (P ≤ 0.05) less weight gain and survival as compared with those fed diets replete with or excessive in zinc or calcium. Tissue mineral concentrations of fish indicated that various levels of zinc and calcium nutriture existed when immune functions were tested. Diet affected mortality of only nonimmunized fish in the bacterial-challenge test; zinc deficiency and calcium excess decreased mortality. Whereas growth and survival of fish were reduced by dietary zinc or calcium deficiency, the measured immunological characteristics were not equally impaired by these mineral deficiencies.     

Isolation and Identification of Aeromonas hydrophila from the Case of Channel Catfish Bowel Disease

To ensure the cause of suspected bacteria disease for channel catfish, the bacteria from clinical cases were isolated and cultured, then the biochemical and molecular biology assay, and animal regression test were used to identify the species of the isolated bacteria, bacteria medicine sensitive test was used to detect the bacterial drug resistance. The results showed that the bacteria were gram negative bacilli. The colony morphology and biochemical characteristics of the isolated bacteria were same as Aeromonas hydrophila. The sequencing test of the 16S rDNA also confirmed the isolated bacteria were Aeromonas hydrophila. The isolated bacteria could infect the healthy channel catfish and cause the same symptoms as the clinical case. The isolated bacteria were sensitive to floroquinolones and resistant to penicillin and cephalosporin drugs. The results showed that the pathogen of case catfish bowel disease was Aeromonas hydrophila, and the floroquinolones could use to prevent and treat the disease.     

斑点叉尾鮰套肠病病原嗜水气单胞菌的分离与鉴定

为确定疑似细菌感染的斑点叉尾鮰病原,本试验对临床病例进行细菌分离培养、生物化学鉴定和分子生物学鉴定、动物回归试验和病原菌药敏试验。结果显示,分离菌为革兰氏阴性杆菌,菌落形态和生化特性与嗜水气单胞菌相符;16S rDNA检测与测序分析结果进一步证实分离菌确为嗜水气单胞菌;动物回归试验显示,分离菌能引起与临床自然病例相似的症状;药敏试验显示,分离菌对喹诺酮类药物敏感,对青霉素类、头孢类药物产生耐药性。结果表明,疑似细菌感染斑点叉尾鮰病例为嗜水气单胞菌感染,可选择喹诺酮类药物进行病例的预防治疗。