The Use of PCR Targeted 16S rDNA for Identification of Genomovars of Flavobacterium columnare.

Triyanto,; Kumamaru, Atsuro; Wakabayashi, Hisatsugu

doi:10.3147/jsfp.34.217

Cited by 13 publications

(9 citation statements)

References 3 publications

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Section: Discussionsupporting

confidence: 94%

“…Among these were 28 Michigan genomovar I F. columnare isolates originally recovered from salmonids. This is in agreement with the findings of others, who have shown that F. columnare isolates recovered from salmonids most frequently belong to genomovar I (Triyanto & Wakabayashi 1999a, 1999bMichel, Messiaen & Bernardet 2002;Arias et al 2004;Darwish & Ismaiel 2005;Schneck & Caslake 2006;Suomalainen et al 2006;Olivares-Fuster et al 2007a,b;Avendaño-Herrera et al 2011) and have led some to postulate that this genomovar has a tropism for salmonids (Lafrentz et al 2012). In addition, the F. columnare isolates recovered from smallmouth bass and muskellunge also fell into genomovar I. Interestingly, F. columnare isolates previously recovered from largemouth bass Micropterus salmoides (Lacapede) and bluegill Lepomis macrochirus (Rafinesque), which belong in the same family as smallmouth bass (i.e.…”

Section: Discussionsupporting

confidence: 91%

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Isolation and characterization of Flavobacterium columnare strains infecting fishes inhabiting the Laurentian Great Lakes basin

Faisal

Diamanka

Loch

et al. 2016

Journal of Fish Diseases

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Flavobacterium columnare, the aetiological agent of columnaris disease, causes significant losses in fish worldwide. In this study, the prevalence of F. columnare infection was assessed in representative Great Lakes fish species. Over 2000 wild, feral and hatchery-propagated salmonids, percids, centrarchids, esocids and cyprinids were examined for systemic F. columnare infections. Logistic regression analyses showed that the prevalence of F. columnare infection varied temporally and by the sex of the fish, whereby females had significantly higher prevalence of infection. A total of 305 isolates of F. columnare were recovered. Amplification of the near complete 16S rRNA gene from 34 representative isolates and subsequent restriction fragment length polymorphism analyses demonstrated that all belonged to F. columnare genomovar I. Phylogenetic analysis of near complete 16S rRNA gene sequences also placed the isolates in genomovar I, but revealed some intragenomovar heterogeneity. Together, these results suggest that F. columnare genomovar I is widespread in the Great Lakes Basin, where its presence may lead to mortality.

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Section: Discussionsupporting

confidence: 94%

Section: Discussionsupporting

confidence: 91%

Isolation and characterization of Flavobacterium columnare strains infecting fishes inhabiting the Laurentian Great Lakes basin

Faisal

Diamanka

Loch

et al. 2016

Journal of Fish Diseases

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“…Specific primers Col 72F and Col 1260R were selected from published sequences for PCR amplification of the bacterial gene encoding ribosomal 16S rRNA (Triyanto, Kumamaru & Wakabayashi 1999). Amplification was performed with final volumes of 50 lL, using 0.4 lL of Taqpolymerase Gibco (5 u lL À1 ), 2 lL of dNTPs 10 mm (Invitrogen, Cergy, Pontoise, France), 2 lL of primers 20 mm, and 5 lL of template DNA samples in relevant buffers.…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Muscle infections in imported neon tetra, Paracheirodon innesi Myers: limited occurrence of microsporidia and predominance of severe forms of columnaris disease caused by an Asian genomovar of Flavobacterium columnare

Michel

Messiaen

Bernardet

2002

Journal of Fish Diseases

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Serious and repeated mortality recently experienced in imported neon tetra, Paracheirodon innesi Myers, by French ornamental fish traders and empirically ascribed to the microsporidian Pleistophora hyphessobryconis on the basis of clinical signs, was investigated. Although Pleistophora sp. spores were observed in a few cases, laboratory results demonstrated that similar clinical signs were generally caused by the pathogenic bacterium Flavobacterium columnare. In all cases, muscle was the main target tissue, and the most noticeable external signs were limited to fading of skin colouration and the development of white areas of necrosis. Three isolates were studied and typed by bacteriological tests and molecular techniques. Although their phenotypic characteristics were in accordance with F. columnare descriptions, except for higher optimal growth temperatures (18–30 °C), they all appeared to differ genetically from common European and American isolates and to be similar to Asian isolates recently assigned to a new genomovar by Japanese workers. Experimental infections suggested the isolates were highly virulent for ornamental species. The use of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) for identification and detection of the agent in tissue samples, and the implications of this finding for health control management of imported fish and domestic species are discussed.

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“…Using a species-specific primer for F. columnare Col-72F (5′-GAAGGAGCTTGTTCCTTT-3′) and Col-1260R (5′-GCCTACTTGCGTAGTG-3′) as describe by Triyanto et al (1999). A PCR reaction was performed in final volume of 50 μL, using 1 μL of Q5 ® High-Fidelity DNA Polymerase (M0491), 10 μL Q5 Reaction Buffer, 10 μL Q5 High GC Enhancer, 1 μL of dNTPs 10 mM, 2.5 μL of primers 10 mM, and 3 μL of template DNA samples.…”

Section: Methodsmentioning

confidence: 99%

Walleye Autochthonous Bacteria as Promising Probiotic Candidates against Flavobacterium columnare

et al. 2017

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Walleye (Sander vitreus) is the second most fished freshwater species in Canada. While much sought by anglers, walleye also supports substantial commercial fisheries. To cope with the recent decline of wild walleye populations, fish farmers produce juveniles for lake stocking. However, walleye breeding is particularly tedious, mostly due to high disease susceptibility at larval and juvenile developmental stages. The main threat is the columnaris disease, which is caused by Flavobacterium columnare, an opportunistic bacteria. As F. columnare strains exhibit increasing antibiotic resistance, there is a strong need to develop efficient and sustainable alternative strategies to control columnaris disease. Bacterial probiotics have been shown to mitigate infections either by enhancing host immune response or by inhibiting pathogen growth. Being successfully assessed in many fish/pathogen combinations, we developed a tailored probiotic strategy for walleye to prevent and treat columnaris disease. Thirty-seven endogenous bacterial strains were isolated from healthy walleye’s skin and gut, were tested in vitro against F. columnare. Significant antagonistic effect against F. columnare was measured for 2 out of 37 endogenous strains. These two probiotic strains were identified as Pseudomonas fluorescens. The antagonistic effect of these two successful probiotics was further validated in vivo during a 2-month stress trial: groups receiving probiotic treatments showed on average 53.74% survival improvement.

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The Use of PCR Targeted 16S rDNA for Identification of Genomovars of Flavobacterium columnare.

Cited by 13 publications

References 3 publications

Isolation and characterization of Flavobacterium columnare strains infecting fishes inhabiting the Laurentian Great Lakes basin

Isolation and characterization of Flavobacterium columnare strains infecting fishes inhabiting the Laurentian Great Lakes basin

Muscle infections in imported neon tetra, Paracheirodon innesi Myers: limited occurrence of microsporidia and predominance of severe forms of columnaris disease caused by an Asian genomovar of Flavobacterium columnare

Walleye Autochthonous Bacteria as Promising Probiotic Candidates against Flavobacterium columnare

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