Influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum).

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 de…

Immunostimulants in prevention of columnaris disease of rainbow trout, Oncorhynchus mykiss (Walbaum).

Freezing Induces Biased Results in the Molecular Detection of Flavobacterium columnare

ABSTRACT Specific PCR detection and electron microscopy of Flavobacterium columnare revealed the risk of false-negative results in molecular detection of this fish pathogen. Freezing and thawing destroyed the cells so that DNA was for the most part undetectable by PCR. The detection of bacteria was also weakened after prolonged enrichment cultivation of samples from infected fish.

Chondroitin AC lyase activity is related to virulence of fish pathogenic Flavobacterium columnare.

The virulence of eight Flavobacterium columnare strains was studied to find correlations between several virulence-related factors and virulence. Virulence was tested in vivo using rainbow trout, Oncorhynchus mykiss (Walbaum). Suggested virulence-related factors such as production of the degradative enzyme chondroitin lyase, plasmid occurrence and adhesion capability were studied in vitro. Infection with the four most virulent strains resulted in 95-100% mortality within 114 h. Chondroitin lyase activity was found to be significantly related to the virulence of the strains at 25 degrees C and it was also shown to be temperature-dependent, being higher at 25 degrees C than at 20 degrees C. V…

Reliability of mitochondrial DNA in an acanthocephalan: The problem of pseudogenes

The utility of mitochondrial DNA as a molecular marker for evolutionary studies is well recognized. However, several problems can arise when using mitochondrial DNA, one of which is the presence of nuclear mitochondrial pseudogenes, or Numts. Pseudogenes of cytochrome oxidase I were preferentially amplified from Acanthocephalus lucii (Acanthocephala) using a universal PCR approach. To verify the presence and abundance of pseudogenes, length heterogeneity analysis of the PCR fragments was performed. PCR products obtained with universal primers often contained fragments of different sizes. Cloned sequences from universal PCR products nearly always contained sequence abnormalities such as inde…

Flavobacterium columnare colony types: connection to adhesion and virulence?

Four different colony morphologies were produced by Flavobacterium columnare strains on Shieh agar plate cultures: rhizoid and flat (type 1), non-rhizoid and hard (type 2), round and soft (type 3), and irregularly shaped and soft (type 4). Colonies produced on AO agar differed from these to some extent. The colony types formed on Shieh agar were studied according to molecular characteristics [Amplified Fragment Length Polymorphism (AFLP), Automated Ribosomal Intergenic Spacer Analysis (ARISA), and whole cell protein SDS-PAGE profiles], virulence on rainbow trout fingerlings, and adhesion on polystyrene and fish gills. There were no molecular differences between colony types within one strai…

Temporal variability in detritus resource maintains diversity of bacterial communities

Competition theory generally predicts that diversity is maintained by temporal environmental fluctuations. One of the many suggested mechanisms for maintaining diversity in fluctuating environments is the gleaner-opportunist trade-off, whereby gleaner species have low threshold resource levels and low maximum growth rates in high resource concentration while opportunist species show opposite characteristics. We measured the growth rates of eight heterotrophic aquatic bacteria under different concentrations of chemically complex plant detritus resource. The growth rates revealed gleaner-opportunist trade-offs. The role of environmental variability in maintaining diversity was tested in a 28-…

Molecular diversity and growth features of Flavobacterium columnare strains isolated in Finland

ABSTRACT: Columnaris disease caused by Flavobacterium columnare is a problem in fish farmingworldwide. During the last 15 yr, outbreaks have started to emerge in Finland. Flavobacteriumcolumnare Type Strain NCIMB 2248 T and 30 Finnish F. columnare isolates were studied usinganalysis of 16S rDNA by restriction-fragment length polymorphism (16S RFLP), length heterogeneityanalysis of polymerase chain reaction (LH-PCR) products, automated ribosomal intergenic spaceranalysis (ARISA), and 16S rDNA sequence analysis. All isolates fell into RFLP Genomovar I and hadthe same length in the LH-PCR analysis. Based on ARISA, 8 genetically different strains wereselected for further analyses. The growth of…

Intensive fish farming and the evolution of pathogen virulence: the case of columnaris disease in Finland.

Ecological changes affect pathogen epidemiology and evolution and may trigger the emergence of novel diseases. Aquaculture radically alters the ecology of fish and their pathogens. Here we show an increase in the occurrence of the bacterial fish disease Flavobacterium columnare in salmon fingerlings at a fish farm in northern Finland over 23 years. We hypothesize that this emergence was owing to evolutionary changes in bacterial virulence. We base this argument on several observations. First, the emergence was associated with increased severity of symptoms. Second, F. columnare strains vary in virulence, with more lethal strains inducing more severe symptoms prior to death. Third, more vir…

Effect of Pseudomonas sp. MT5 baths on Flavobacterium columnare infection of rainbow trout and on microbial diversity on fish skin and gills

Use of Pseudomonas sp. strain MT5 to prevent and treat Flavobacterium columnare infection was studied in 2 experiments with fingerling rainbow trout Oncorhynchus mykiss. In the first experiment, length heterogeneity analysis of PCR-amplified DNA fragments (LH-PCR) was used to assess the effect of antagonistic baths on the microbial diversity of healthy and experimentally infected fish. In the 148 samples studied, no difference was found between bathed and unbathed fish, and 3 fragment lengths were detected most frequently: 500 (in 75.7% of the samples), 523 (62.2%) and 517 bp (40.5%). The species contributing to these fragment sizes were Pseudomonas sp., Rhodococcus sp. and F. columnare, re…

Saprophytism of a fish pathogen as a transmission strategy

Fish farming creates conditions where disease transmission is enhanced and antibiotic treatments are commonly used to cure bacterial diseases to prevent severe losses due to infections. Ability to persist in such an environment has been suggested to lead to the evolution of high virulence. Columnaris disease caused by Flavobacterium columnare is a growing problem in freshwater fish farming. Transmission of the disease is poorly known, and survival of F. columnare in the rearing environment has not been studied. This paper addresses both transmission of columnaris disease and survival strategy of F. columnare. Saprophytic activity of F. columnare was studied by infecting rainbow trout finger…

A link between ectoparasite infection and susceptibility to bacterial disease in rainbow trout

Rainbow trout, Oncorhynchus mykiss, were infected concomitantly with Argulus coregoni and Flavobacterium columnare and their survival was compared with that of fish infected with either the parasite or the bacterium alone. The mortality of fish challenged with A. coregoni was negligible while infection with F. columnare alone led to significantly lower survival. However, compared with single infections, the mortality was significantly higher and the onset of disease condition was earlier among fish, which were concomitantly infected by A. coregoni and F. columnare. This data presents, for the first time, experimental support for the hypothesis that an ectoparasite infection increases suscep…

Treatment of columnaris disease of rainbow trout: low pH and salt as possible tools?

The impact of salt and low pH on columnaris disease of fish was studied. Survival of Flavobacterium columnare after exposure to either 4% NaCl (pH 7.2) or pH 5.0, pH 4.86 or pH 4.6 for 15 min or 1 h was studied in vitro. All conditions significantly reduced the numbers of viable bacterial cells. The effects of salt (4 and 2%) and acidic baths (pH 4.6) were studied in 2 experiments in vivo with rainbow trout Oncorhynchus mykiss infected with F. columnare. Both salt and acidic baths failed to prevent fish mortality; the overall mortality reached 100% in all groups. However, according to survival analysis, the mortality rate was lower in fish treated with 4% salt baths compared to a control gr…

The efficacy of two immunostimulants against Flavobacterium columnare infection in juvenile rainbow trout (Oncorhynchus mykiss).

Abstract Bacterium Flavobacterium columnare is the causative agent of columnaris disease in many wild and farmed fish species. Immunostimulants are used with success in aquaculture against many pathogens, but the ability to improve innate resistance to columnaris disease has not been studied. Fingerling rainbow trout were treated with two immunostimulants, yeast β-glucan and β-hydroxy-β-methylbutyrate (HMB). Selected innate immune function parameters, the production of reactive oxygen species (ROS) by whole blood and by isolated head kidney leukocytes, plasma lysozyme activity and complement bacteriolytic activity, were determined to assess the immune status of fish. The fish were then bath…