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Columnaris disease generates substantial losses of many freshwater fish species; one is the hybrid striped bass. The ubiquitous aquatic bacterium Flavobacterium columnare can be highly effective in biofilm formation on fish skin and gills. Previous research showed a difference between columnaris disease susceptibility of hybrid striped bass (Morone saxatilis × M. chrysops) and white bass (M. chrysops). To understand these differential susceptibilities and possible mucosal relationship, we assessed total bacterial growth and biofilm formation with mucus derived from each moronid parental species: white bass and striped bass (M. saxatilis). Differential susceptibility was confirmed of the other parent species, the striped bass (M. saxatilis). In addition to intraspecies investigations, individual hybrid striped bass mucosal affects were also studied for deferential responses to bacterial growth and biofilm formation. Species- and concentration-dependent differences were detected in the total growth of the bacteria to host mucus. Our data suggest that bass mucus can significantly affect biofilm formation with the F. columnare isolate tested. There appears to be a correlation between the bacteria's response of growth and biofilms and bass species susceptibility. This study provides insight into our understanding of the host-pathogen interaction between F. columnare and moronids.

Columnaris disease is a major cause of mortality in tilapia hatcheries and commonly occurs during the summer season in Thailand. One way of reducing the problem is by selective breeding for increased disease resistance. The objective of this study was to estimate quantitative genetic parameters for resistance against columnaris in the Chitralada 4 strain of Nile tilapia. Data from 43 full-sib families (2,580 records) of fry (age = 32 ± 4 days post-hatch) were used in the analyses. Initially, fry were subjected to bath challenge with Flavobacterium columnare (LD50 concentration = 1.2 × 106 CFU/ml) for 14 days. Disease resistance was defined as the number of days from challenge until death (DD) or as a binary trait (dead/alive) on day 14. Linear animal and sire-dam models were used for DD, while threshold animal, threshold sire-dam, binary linear animal and binary linear sire-dam models were used for binary outcomes. Covariate effect of age, fixed effect of challenge day and random effects of the individual animals or sires and dams were included in the models. Mean survival was 32.4 ± 11.6%, and survival rates of the best and poorest families were 70% and 8%, respectively. The highest estimate of heritability (0.30 ± 0.025) was obtained under the threshold sire-dam model. Heritability estimates for DD (0.16 ± 0.034 and 0.17 ± 0.046) were comparable to those obtained from the threshold animal (0.15 ± 0.031) and the binary linear (0.14 ± 0.045 and 0.15 ± 0.044) models. The linear animal and sire-dam models for DD and the threshold sire-dam models performed equally with similar values of rEBV (0.629, 0.628 and 0.627) and accuracy of selection (0.793, 0.793 and 0.791). This study reveals the potential of selective breeding to increase disease resistance to F. columnare in the studied population of Nile tilapia.

Flavobacterium columnare causes columnaris disease of farmed and wild freshwater fish. Skin mucus is an important factor in early stages of columnaris pathogenesis, albeit little studied. Our objectives were to (a) characterize the terminal glycosylation pattern (TGP) of catfish mucus, (b) determine the growth of F. columnare in formulated water (FW)-containing channel catfish (Ictalurus punctatus) or hybrid catfish (Ictalurus punctatus X Ictalurus furcatus) mucus and (c) examine extracellular protease activity of two F. columnare isolates differing in virulence. The TGP of catfish mucus by lectin binding was as follows: alpha-D-mannose/alpha-D-glucose >N-acetyl-beta-D-glucosamine >N-acetyl-beta-D-glucosamine/N-acetylneuraminic acid >N-acetyl-D-galactosamine >alpha-D-galactose/N-acetyl-alpha-D-galactosamine >beta-D-galactose = alpha-L-fucose. Virulence studies demonstrated isolate AL-02-36 was highly virulent in channel catfish fry (0.1 g) with cumulative mortality of 90%-100% versus 60% for isolate ALG-00-530 at equivalent doses (~3 × 106 CFU/ml); a similar result was observed in larger (0.7 g) catfish. In multiple experiments, F. columnare replicated (2-3 logs) and survived (28 days) in formulated water-containing catfish mucus. Highly virulent isolate AL-02-36 possessed at least 2.5- to fivefold higher protease activity following growth in mucus than the less virulent ALG-00-530. Flavobacterium columnare utilized catfish mucus as a nutrient source and mucus presence modulated extracellular protease production.

Columnaris disease, caused by the bacterium Flavobacterium columnare, is currently the most frequently reported bacterial disease affecting farm-raised channel catfish in the USA. Common treatments against the disease include the use of medicated feed that has led to emergent antibiotic resistant strains of F. columnare. Nigella sativa (Black cumin) is a medicinal herb commonly used by many cultures as a natural remedy for numerous disorders. Recently, we have discovered the antibacterial activity of N. sativa and its oil extract against F. columnare. In this study, we showed N. sativa oil (NSO) strongly inhibited the growth of all of the strains of F. columnare tested and yielded significantly larger zones of inhibition than those produced by oxytetracyclin. We tested the protective effect against columnaris disease in vivo by incorporating NSO (5%) or N. sativa seeds (NSS) (5%) into fish feeds. Fishes (Ictalurus punctatus and Danio rerio) fed amended diets displayed significantly lower mortality than those fed control diets. Per cent mortalities in control groups ranged from 77% to 44% and from 70% to 18% in zebrafish and channel catfish, respectively. A dose study using different NSS concentrations showed that 5% NSS offered the most protection against columnaris disease in channel catfish.

Flavobacterium columnare is the causative agent of columnaris disease in diverse fish species worldwide. Although columnaris is an important disease, the antimicrobial susceptibility pattern of F. columnare is not well studied. Thus, the purpose of this study was to test the in vitro antimicrobial susceptibility of 97 F. columnare isolates collected worldwide between 1987 and 2011 from 17 fish species. The broth microdilution technique was utilized for reliable testing of these fastidious organisms. None of the isolates displayed acquired resistance to florfenicol, gentamicin, ormetoprim-sulfadimethoxine and trimethoprim-sulfamethoxazole. Acquired resistance to chloramphenicol was detected in 1%, to nitrofuran in 5%, to oxytetracycline in 11% and to enrofloxacin, flumequine and oxolinic acid in 10%, 16% and 16% of the isolates, respectively, as reflected by a bimodal or trimodal distribution of their minimum inhibitory concentrations (MICs). One isolate showed acquired resistance towards several antimicrobial agents including erythromycin. Another isolate revealed acquired resistance towards - amongst others - ampicillin. The isolates displaying acquired resistance originated from ornamental fish species or Vietnamese catfish, except for two isolates coming from wild channel catfish in which acquired resistance was encountered towards oxytetracycline only. Fifty per cent of the resistant isolates from ornamental fish were shown to have acquired resistance against three classes of antimicrobial agents, assigning these isolates as multiple resistant. These data might indicate less prudent use of antimicrobials especially in ornamental fish species.

Columnaris disease was induced in channel catfish, Ictalurus punctatus (Rafinesque), by bath exposure to four highly virulent isolates of Flavobacterium columnare. In untreated controls, mortality began 20 h after exposure and reached 100% by 48 h. Mortality in channel catfish given antibiotic treatments with oxytetracycline or a combination of sulphadimethoxine and ormetoprim in feed prior to bacterial challenge was zero with all four strains of F. columnare. Diquat® (Zeneca Agricultural Products, Wilmington, DE, USA) was the most effective bath treatment; mortality with all four strains was zero. With potassium permanganate, chloramine-T, hydrogen peroxide and copper sulphate, bath treatment efficacy varied significantly among strains (P = 0.0346) and among treatments (P = 0.0033). Bath treatments with chloramine-T and potassium permanganate significantly reduced (P

Channel catfish, Ictalurus punctatus (Rafinesque), goldfish, Carassius auratus (L.), striped bass, Morone saxatilis (Walbaum), and Gulf sturgeon, Acipenser oxyrinchus desotoi Vladykov, were acclimatized to fresh water or salinities of 9.0& or less and then exposed to Flavobacterium columnare (formerly known as Flexibacter columnaris), the bacterial pathogen that causes columnaris disease. None of the fish acclimatized to 3.0 or 9.0& salinity died, and all deaths in lower salinities occurred between 1 and 5 days after exposure to F. columnare. Mortality was 97.7% in fresh water and 67.1% in 1.0& salinity for channel catfish (model SE, 1.8) and 66.5% in fresh water and 40.8% in 1.0& salinity for goldfish (model SE, 1.2); and 96.9% in fresh water and 61.7% in 1.0& salinity for striped bass (model SE, 1.8). After exposure to F. columnare, none of the Gulf sturgeon died. Flavobacterium columnare was isolated from the skin and gills of all fish dying during the experiments, but was not isolated from survivors in fresh water and 1.0& salinity 21 days after bacterial exposure. In vitro growth of bacteria was significantly higher in 1.0 or 3.0& salinity than in control medium (0.3& salinity). However, in vitro adhesion of bacteria was reduced with increasing salinity, which could explain the lower mortality of fish at higher salinities.

The ability of Flavobacterium columnare (Flexibacter columnaris) to attach to the gills of common carp, Cyprinus carpio L., was evaluated using a gill perfusion model. A comparison between a high and a low virulence strain of F. columnare was made and evaluated in comparison to results obtained previously with an in vivo model. The ion composition of the water of the organ bath in which the gills were suspended was varied and the influence on adhesion processes assessed. Experiments were carried out to examine the influence of water quality (i.e. nitrite and organic matter) and temperature on the capacity of the bacteria to adhere. It was found that the high virulence strain adhered more readily than the low virulence strain, as was found during the in vivo experiments. Moreover, it was observed that adhesion of the high virulence strain was enhanced by a number of factors. These were immersion of the gill in bivalent, ion-rich water, the presence of nitrite or organic matter, and high temperatures.

A trial was performed to evaluate the efficacy of diquat (6,7-dihydrodipyrido[1,2-a:2',1'-c]pyrazinediium dibromide) against an acute experimental infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus. Diquat is an Environmental Protection Agency-approved herbicide and has the potential to be legally and practically used against columnaris. Channel catfish were challenged, by cutaneous abrasion, and waterborne exposure to F. columnare and treated once at 22-h post-challenge with 2.5, 5.0, 10.0 and 15 mg L(-1) of diquat active ingredient for 6 h. At the conclusion of the trial, 21-day post-challenge, diquat at 5.0, 10.0 and 15 mg L(-1) significantly (P0.05) reduced the mortality of infected fish from 95% in the challenged non-treated fish to 68%, 59% and 49%, respectively. In vitro, the minimum inhibitory concentration (MIC) of 23 isolates of F. columnare was assayed. The majority of the isolates had an MIC value of 5 microg mL(-1) (15 of the 23 isolates). Infected fish exhibited acute clinical signs similar to a natural infection. The skin had severe ulcerative necrotizing dermatitis and the muscles had severe necrotizing myositis. The gills had severe multifocal necrotizing branchitis. The results demonstrate that diquat would reduce mortalities caused by an acute columnaris infection.

During Autumn 1990, the possible presence of microbial pathogens was investigated in two discrete populations of anaemic (rainbow trout fry syndrome; RTFS) rainbow trout fry. Dense pure culture growth of three types of yellow pigmentcd bacteria were recovered from moribund and dead animals on low nutrient media. The cultures were identified as coryneforms. Cytophaga columnaris and Janthinobacterium sp. All taxa induced clinical disease (with similarities to RTFS) in rainbow trout fry.

Columnaris disease, Flexibacter columnaris infection, was first described in North America. Recent publications indicate that its distribution is worldwide. Both warmwater and coldwater fish are infected with F. columnaris. All freshwater fish are probably susceptible to the disease under environmental conditions favourable to the bacterium and stressful to the fish. Flexibacter columnaris cannot tolerate sea water. Generally, outbreaks occur when the average water temperatures reach 15°C. The bacterium apparently enters the fish through the skin and gills. The swarms of bacteria involved give the lesions a characteristic yellow pigmentation. The external lesions destroy the skin, muscle and gill tissues. It is relatively easy to isolate the bacteria from the internal organs of the diseased fish, but there is no evidence of internal tissue damage. In this short review, emphasis is placed on several factors relative to water environment which play a role in the development of outbreaks of columnaris disease: 1 The optimum temperature for an outbreak of F. columnaris infection is thought to be between 20 and 30°C. Mortalities seldom occur at temperatures below 15°C, although highly virulent strains can kill fish at lower temperatures. 2 Flexibacter columnaris can survive longer in water of normal to high hardness, as well as in water containing small amounts of sodium, potassium, calcium or magnesium salts, but it cannot live in sea water. 3 Flexibacter columnaris grows well on paniculate fish feeds so that such deposits in the water may act as reservoirs of infection. 4 An important source of infection is feral fish. In the Columbia River or the Fraser River, resident coarse fish such as suckers, Catostomus spp., are thought to transmit the infection to returning adult salmon. 5 Flexibacter columnaris fails to invade fish in the presence of competitive bacteria like Citrobacter freundii when its initial numbers are approximately one in a 100 less than those of the competitors. 6 Crowding seems to be not only a stressor for fish, but also a factor that increases the chance of adhesion of F. columnaris to the external tissues of fish.

Infectivity of Flexibator columnaris to loach, Misgurnus anguillicaudatus (Blecker, Gunther), was studied with variation conditions and the presence of other bacteria in the water medium. Invasion of fish by the poathogen varied with different water conditions. The highest prevalence of infection was observed in an experimentally formulated water containing 0·03% NaCl 0·01% KCl, 0·002% CaCl22H2O and 0·004% MgCI2 6H2O. The percentage of infection in tap water was similar to that of the formulated water. The infeetion rate was low in the individual salt solutions, and no infection occurred in distilled water. Even in the formulated water, the pathogen failed to infect the fish when Citrobacter freundii, Aeromonas hydrophila or Pseudomonas fluorescens were present at densities 100 times that of F. columnaris. However, in such cases, the numbers of F. columnaris in the mucus of the fish body surface did not increase much throughout the experimental period. In contrast, F. columnaris suceessfully invaded the fish in the presence of either Streptococcus sp., Flavobacterium flavescens or Flavobacterium fuscum, even when the numbers of these bacteria were 1000 times that of F. columnaris. In this ease, F. columnaris gradually increased its numbers on the fish body surface.

The influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied experimentally in the laboratory and at a fish farm. In experiment I, the effect of parasitic infection on columnaris disease was studied using F. columnare carrier fish. The fish were exposed to Diplostomum spathaceum cercariae and a set of other stressors in order to induce clinical columnaris infection. Parasitic infection and other stressors failed to induce the disease. Disease occurred when the fish were challenged with F. columnare, but D. spathaceum infection did not enhance the severity of the infection. In experiment II, the influence of rearing density and water temperature was studied. Overall mortality was highest in fish at normal rearing density with high temperature (+23 degrees C). At low temperature (+18 degrees C) mortality was not affected by rearing density, but the transmission of columnaris disease was faster at normal rearing density at both temperatures. This supports the view that reduction of fish density could be used in prevention of columnaris disease especially if water temperature is high. Because the lower rearing density can also decrease the transmission of ectoparasites and penetrating endoparasites, it could be an efficient tool in ecological disease management.

A recurring disease known as ‘black patch necrosis’ (BPN) has been the cause of severe mortalities in 0-group and 1-group Dover sole during the period 1974-1978 at the Sea Fish Industry Authority's Marine Cultivation Unit at Hunter-ston, Ayrshire, Scotland. Samples of diseased and healthy Dover sole tissue were examined microbiologically and a long thin filamentous Gram-negative organism, strongly resembling Flexibacter columnar's, was repeatedly isolated from diseased tissue but was not found in healthy tissue. This organism was pathogenic to Dover sole, producing 100% mortality in 96 h at 17.5°C (±2°C) and the organism was reisolated from all the infected fish. The pathogenic isolate and reisolate were examined and compared with reference strain NCMB 1038 of Flexibacter columnarts, formerly known as Chondrococcus columnaris or Cytophaga columnaris. The isolate, reisolate and type culture were identical with respect to cell morphology, oxidase test, catalase production, production of H2S, casein hydrolysis, gelatin liquefaction, hydrolysis of Tween 20, degradation of tyrosine and starch hydrolysis. All bacterial types showed similar patterns of sensitivity to a range of antibiotics. It is proposed that a Flexibacter columnari's-like organism is associated with the condition in farmed Dover sole known as ‘black patch necrosis’.

Heavy mortalities of underyearling Atlantic salmon parr have occurred at the Mactaquac Fish Culture Station in New Brunswick as the result of ‘saddleback’ disease. The organism causing this lesion was isolated and identified as Flexibacter columnaris. This bacterium was transmitted to other salmon parr, where it produced the typical saddle lesion from which it was re-isolated. The condition was more readily transmitted at 20°C than at 15°C. Transmission required entrance to the dermis through a break in the epidermal surface and was unsuccessful in intact fish. The morphology of the saddleback isolate and its histopathological effect on the fish tissues is described. F. columnaris appeared to be restricted to the dermis and underlying muscle of the saddle area when histological techniques were used, although it was also isolated from the kidneys of some affected fish. The gills, kidney and other organs of the fish did not seem to be affected by the presence of F. columnaris.