Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil

Detalhes bibliográficos
Autor(a) principal: de Abreu Reis Ferreira, Daniel [UNESP]
Data de Publicação: 2022
Outros Autores: Assane, Inácio Mateus [UNESP], Vaneci-Silva, Daiane [UNESP], do Vale Oliveira, André [UNESP], Tamashiro, Geovana Dotta [UNESP], Hashimoto, Diogo Teruo [UNESP], Pilarski, Fabiana [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1016/j.aquaculture.2022.738486
http://hdl.handle.net/11449/241221
Resumo: Flavobacterium columnare is an important pathogen of freshwater fish that has severe impacts on the commercial aquaculture sector worldwide. In this study, we investigated the morpho-molecular characteristics, pathogenicity against pacu, Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent F. columnare isolated from Nile tilapia cultured in Brazil. Diseased Nile tilapia having ulcerations on the skin were collected from a commercial fish farm located in Porto Ferreira, São Paulo, Brazil for the isolation of pathogenic bacteria. Isolates (n = 7) were identified by morphological, biochemical, and molecular analyses as F. columnare. Bacteria molecular characteristics were determined by restriction fragment length polymorphism (RFLP) using the restriction enzyme Hae III and phylogenetic analysis based on the partial sequence of the 16S rRNA gene. Susceptibility to enrofloxacin (ENRO), florfenicol (FFC), oxytetracycline (OTC), thiamphenicol (TAP), and their combinations were assessed by broth microdilution and chequerboard methods, respectively. The pathogenicity against pacu was analysed by experimental infection by immersion challenge with one of the clinical isolates (DV001, GeneBank Accession Number MW186708). The molecular analyses revealed that the clinical isolate belongs to F. columnare genetic group 4 and genomovar II and differs from strains previously isolated from cultured fish in Brazil. The clinical isolate was classified as wild-type for all tested antimicrobials, except for ENRO. Synergistic, additive, and antagonistic activities were reported when the combination of TAP and ENRO (ΣFIC = 0.5), FFC and TAP (ΣFIC = 0.75), and OTC and ENRO (ΣFIC = 3) were tested, respectively. The disease was reproduced in the experimental infection with F. columnare DV001 at 1 × 102, 1 × 104, and 1 × 105 colony-forming units (CFU) mL−1, and the cumulative mortality was 50%, 75%, and 87.5% within 15 days, respectively. The strain was highly virulent against pacu, and the LD50, LD90, and LD99 were established in 1.16 × 102, 4.6 × 104, and 1.8 × 107 CFU mL−1, respectively. This is the first report of F. columnare pathogenicity against pacu. Although the results of antimicrobial susceptibility testing suggest that available antimicrobials can be effective against the pathogen, the resistance against ENRO and pathogenicity against pacu has several implications. These findings will hopefully be useful in the development of effective vaccines and therapeutic strategies for the prevention and control of columnaris disease.
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spelling Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in BrazilAntimicrobial resistanceBacterial infectionFlavobacterium columnarePathogenicityPiaractus mesopotamicusRestriction fragment length polymorphismFlavobacterium columnare is an important pathogen of freshwater fish that has severe impacts on the commercial aquaculture sector worldwide. In this study, we investigated the morpho-molecular characteristics, pathogenicity against pacu, Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent F. columnare isolated from Nile tilapia cultured in Brazil. Diseased Nile tilapia having ulcerations on the skin were collected from a commercial fish farm located in Porto Ferreira, São Paulo, Brazil for the isolation of pathogenic bacteria. Isolates (n = 7) were identified by morphological, biochemical, and molecular analyses as F. columnare. Bacteria molecular characteristics were determined by restriction fragment length polymorphism (RFLP) using the restriction enzyme Hae III and phylogenetic analysis based on the partial sequence of the 16S rRNA gene. Susceptibility to enrofloxacin (ENRO), florfenicol (FFC), oxytetracycline (OTC), thiamphenicol (TAP), and their combinations were assessed by broth microdilution and chequerboard methods, respectively. The pathogenicity against pacu was analysed by experimental infection by immersion challenge with one of the clinical isolates (DV001, GeneBank Accession Number MW186708). The molecular analyses revealed that the clinical isolate belongs to F. columnare genetic group 4 and genomovar II and differs from strains previously isolated from cultured fish in Brazil. The clinical isolate was classified as wild-type for all tested antimicrobials, except for ENRO. Synergistic, additive, and antagonistic activities were reported when the combination of TAP and ENRO (ΣFIC = 0.5), FFC and TAP (ΣFIC = 0.75), and OTC and ENRO (ΣFIC = 3) were tested, respectively. The disease was reproduced in the experimental infection with F. columnare DV001 at 1 × 102, 1 × 104, and 1 × 105 colony-forming units (CFU) mL−1, and the cumulative mortality was 50%, 75%, and 87.5% within 15 days, respectively. The strain was highly virulent against pacu, and the LD50, LD90, and LD99 were established in 1.16 × 102, 4.6 × 104, and 1.8 × 107 CFU mL−1, respectively. This is the first report of F. columnare pathogenicity against pacu. Although the results of antimicrobial susceptibility testing suggest that available antimicrobials can be effective against the pathogen, the resistance against ENRO and pathogenicity against pacu has several implications. These findings will hopefully be useful in the development of effective vaccines and therapeutic strategies for the prevention and control of columnaris disease.National Council for Scientific ResearchGraduate Program in Agricultural and Livestock Microbiology São Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences, JaboticabalPostgraduate Program in Aquaculture Laboratory of Microbiology and Parasitology of Aquatic Organisms São Paulo State University (Unesp) Aquaculture Center of Unesp, JaboticabalFaculdade de Ciências Agrárias Universidade Zambeze (UniZambeze), TetePostgraduate Program in Aquaculture Laboratory of Aquaculture Genetics and Conservation São Paulo State University (Unesp) Aquaculture Center of Unesp, JaboticabalGraduate Program in Agricultural and Livestock Microbiology São Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences, JaboticabalPostgraduate Program in Aquaculture Laboratory of Microbiology and Parasitology of Aquatic Organisms São Paulo State University (Unesp) Aquaculture Center of Unesp, JaboticabalPostgraduate Program in Aquaculture Laboratory of Aquaculture Genetics and Conservation São Paulo State University (Unesp) Aquaculture Center of Unesp, JaboticabalNational Council for Scientific Research: 131070/2019-3Universidade Estadual Paulista (UNESP)Universidade Zambeze (UniZambeze)de Abreu Reis Ferreira, Daniel [UNESP]Assane, Inácio Mateus [UNESP]Vaneci-Silva, Daiane [UNESP]do Vale Oliveira, André [UNESP]Tamashiro, Geovana Dotta [UNESP]Hashimoto, Diogo Teruo [UNESP]Pilarski, Fabiana [UNESP]2023-03-01T20:52:20Z2023-03-01T20:52:20Z2022-11-15info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.aquaculture.2022.738486Aquaculture, v. 560.0044-8486http://hdl.handle.net/11449/24122110.1016/j.aquaculture.2022.7384862-s2.0-85132751215Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengAquacultureinfo:eu-repo/semantics/openAccess2024-04-09T15:10:46Zoai:repositorio.unesp.br:11449/241221Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-04-09T15:10:46Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
title Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
spellingShingle Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
de Abreu Reis Ferreira, Daniel [UNESP]
Antimicrobial resistance
Bacterial infection
Flavobacterium columnare
Pathogenicity
Piaractus mesopotamicus
Restriction fragment length polymorphism
title_short Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
title_full Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
title_fullStr Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
title_full_unstemmed Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
title_sort Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
author de Abreu Reis Ferreira, Daniel [UNESP]
author_facet de Abreu Reis Ferreira, Daniel [UNESP]
Assane, Inácio Mateus [UNESP]
Vaneci-Silva, Daiane [UNESP]
do Vale Oliveira, André [UNESP]
Tamashiro, Geovana Dotta [UNESP]
Hashimoto, Diogo Teruo [UNESP]
Pilarski, Fabiana [UNESP]
author_role author
author2 Assane, Inácio Mateus [UNESP]
Vaneci-Silva, Daiane [UNESP]
do Vale Oliveira, André [UNESP]
Tamashiro, Geovana Dotta [UNESP]
Hashimoto, Diogo Teruo [UNESP]
Pilarski, Fabiana [UNESP]
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (UNESP)
Universidade Zambeze (UniZambeze)
dc.contributor.author.fl_str_mv de Abreu Reis Ferreira, Daniel [UNESP]
Assane, Inácio Mateus [UNESP]
Vaneci-Silva, Daiane [UNESP]
do Vale Oliveira, André [UNESP]
Tamashiro, Geovana Dotta [UNESP]
Hashimoto, Diogo Teruo [UNESP]
Pilarski, Fabiana [UNESP]
dc.subject.por.fl_str_mv Antimicrobial resistance
Bacterial infection
Flavobacterium columnare
Pathogenicity
Piaractus mesopotamicus
Restriction fragment length polymorphism
topic Antimicrobial resistance
Bacterial infection
Flavobacterium columnare
Pathogenicity
Piaractus mesopotamicus
Restriction fragment length polymorphism
description Flavobacterium columnare is an important pathogen of freshwater fish that has severe impacts on the commercial aquaculture sector worldwide. In this study, we investigated the morpho-molecular characteristics, pathogenicity against pacu, Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent F. columnare isolated from Nile tilapia cultured in Brazil. Diseased Nile tilapia having ulcerations on the skin were collected from a commercial fish farm located in Porto Ferreira, São Paulo, Brazil for the isolation of pathogenic bacteria. Isolates (n = 7) were identified by morphological, biochemical, and molecular analyses as F. columnare. Bacteria molecular characteristics were determined by restriction fragment length polymorphism (RFLP) using the restriction enzyme Hae III and phylogenetic analysis based on the partial sequence of the 16S rRNA gene. Susceptibility to enrofloxacin (ENRO), florfenicol (FFC), oxytetracycline (OTC), thiamphenicol (TAP), and their combinations were assessed by broth microdilution and chequerboard methods, respectively. The pathogenicity against pacu was analysed by experimental infection by immersion challenge with one of the clinical isolates (DV001, GeneBank Accession Number MW186708). The molecular analyses revealed that the clinical isolate belongs to F. columnare genetic group 4 and genomovar II and differs from strains previously isolated from cultured fish in Brazil. The clinical isolate was classified as wild-type for all tested antimicrobials, except for ENRO. Synergistic, additive, and antagonistic activities were reported when the combination of TAP and ENRO (ΣFIC = 0.5), FFC and TAP (ΣFIC = 0.75), and OTC and ENRO (ΣFIC = 3) were tested, respectively. The disease was reproduced in the experimental infection with F. columnare DV001 at 1 × 102, 1 × 104, and 1 × 105 colony-forming units (CFU) mL−1, and the cumulative mortality was 50%, 75%, and 87.5% within 15 days, respectively. The strain was highly virulent against pacu, and the LD50, LD90, and LD99 were established in 1.16 × 102, 4.6 × 104, and 1.8 × 107 CFU mL−1, respectively. This is the first report of F. columnare pathogenicity against pacu. Although the results of antimicrobial susceptibility testing suggest that available antimicrobials can be effective against the pathogen, the resistance against ENRO and pathogenicity against pacu has several implications. These findings will hopefully be useful in the development of effective vaccines and therapeutic strategies for the prevention and control of columnaris disease.
publishDate 2022
dc.date.none.fl_str_mv 2022-11-15
2023-03-01T20:52:20Z
2023-03-01T20:52:20Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1016/j.aquaculture.2022.738486
Aquaculture, v. 560.
0044-8486
http://hdl.handle.net/11449/241221
10.1016/j.aquaculture.2022.738486
2-s2.0-85132751215
url http://dx.doi.org/10.1016/j.aquaculture.2022.738486
http://hdl.handle.net/11449/241221
identifier_str_mv Aquaculture, v. 560.
0044-8486
10.1016/j.aquaculture.2022.738486
2-s2.0-85132751215
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Aquaculture
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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