Morpho-molecular identification, pathogenicity for Piaractus mesopotamicus, and antimicrobial susceptibility of a virulent Flavobacterium columnare isolated from Nile tilapia cultured in Brazil
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , |
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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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) |
repository.mail.fl_str_mv |
|
_version_ |
1799964614079807488 |