Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria
Main Author: | |
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Publication Date: | 2021 |
Other Authors: | , , , , |
Format: | Article |
Language: | eng |
Source: | Repositório Institucional da UNESP |
Download full: | http://dx.doi.org/10.1111/jam.15080 http://hdl.handle.net/11449/207523 |
Summary: | Aims: This study aimed to evaluate the in vitro cytotoxicity and efficacy of synthetic host defence peptides (HDPs), alone or in combination with florfenicol (FFC), oxytetracycline (OTC) or thiamphenicol (TAP), against different pathogenic bacteria isolated from diseased fish. Methods and Results: Solid-phase synthesis, purification and characterization of several HDPs were performed manually, using the fluorenylmethyloxycarbonyl protecting group in different resins and via high-performance liquid chromatography-mass spectrometry, respectively. The in vitro cytotoxicity and antimicrobial activity of HDPs, FFC, OTC and TAP against Nile tilapia red blood cells (RBCs) and relevant fish pathogenic bacteria (Aeromonas, Citrobacter, Edwardsiella, Streptococcus, Lactococcus and Vibrio) was determined using the haemolysis assay and broth microdilution method, respectively. The checkerboard assay was used to evaluate the synergy between the most active HDPs and other antimicrobials against the tested strains. MUC 7 12-mer, FFC, OTC and TAP were not cytotoxic to Nile tilapia RBCs, in all tested concentrations. LL-37, (p-BthTX-I)2 and Hylin-a1 were not cytotoxic at concentrations up to 78·13, 19·53 and 9·77 μg ml−1, respectively. HDPs demonstrated potent antimicrobial activity (minimum inhibitory concentration ≤31·25 µg ml−1) against Aeromonas jandaei (KR-12-a5), Citrobacter freundii (Kr-12-a5; (p-BthTX-I)2; LL-37; and Hylin a1), Streptococcus agalactiae (Hylin a1; (p-BthTX-I)2 and LL-37), Lactococcus garviae (Hylin a1), and Vibrio fluvialis (KR-12-a5). The combinations of (p-BthTX-I)2 with TAP and LL-37 with FFC showed synergistic activity against C. freundii (fractional inhibitory concentration index of 0·25 and 0·50, respectively). Conclusions: Synthetic HDPs have the potential as a good treatment option for bacterial diseases in aquaculture. Significance and Impact of the Study: The in vivo effectiveness of synthetic HDPs such as KR-12-a5; LL-37; (p-BthTX-I)2 and Hylin a1 can be tested alone or in combination with conventional antimicrobials as a treatment option to reduce the use of antimicrobials in aquaculture. |
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Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteriaantimicrobial activitycytotoxicityfish pathogenic bacteriasynergysynthetic peptidesAims: This study aimed to evaluate the in vitro cytotoxicity and efficacy of synthetic host defence peptides (HDPs), alone or in combination with florfenicol (FFC), oxytetracycline (OTC) or thiamphenicol (TAP), against different pathogenic bacteria isolated from diseased fish. Methods and Results: Solid-phase synthesis, purification and characterization of several HDPs were performed manually, using the fluorenylmethyloxycarbonyl protecting group in different resins and via high-performance liquid chromatography-mass spectrometry, respectively. The in vitro cytotoxicity and antimicrobial activity of HDPs, FFC, OTC and TAP against Nile tilapia red blood cells (RBCs) and relevant fish pathogenic bacteria (Aeromonas, Citrobacter, Edwardsiella, Streptococcus, Lactococcus and Vibrio) was determined using the haemolysis assay and broth microdilution method, respectively. The checkerboard assay was used to evaluate the synergy between the most active HDPs and other antimicrobials against the tested strains. MUC 7 12-mer, FFC, OTC and TAP were not cytotoxic to Nile tilapia RBCs, in all tested concentrations. LL-37, (p-BthTX-I)2 and Hylin-a1 were not cytotoxic at concentrations up to 78·13, 19·53 and 9·77 μg ml−1, respectively. HDPs demonstrated potent antimicrobial activity (minimum inhibitory concentration ≤31·25 µg ml−1) against Aeromonas jandaei (KR-12-a5), Citrobacter freundii (Kr-12-a5; (p-BthTX-I)2; LL-37; and Hylin a1), Streptococcus agalactiae (Hylin a1; (p-BthTX-I)2 and LL-37), Lactococcus garviae (Hylin a1), and Vibrio fluvialis (KR-12-a5). The combinations of (p-BthTX-I)2 with TAP and LL-37 with FFC showed synergistic activity against C. freundii (fractional inhibitory concentration index of 0·25 and 0·50, respectively). Conclusions: Synthetic HDPs have the potential as a good treatment option for bacterial diseases in aquaculture. Significance and Impact of the Study: The in vivo effectiveness of synthetic HDPs such as KR-12-a5; LL-37; (p-BthTX-I)2 and Hylin a1 can be tested alone or in combination with conventional antimicrobials as a treatment option to reduce the use of antimicrobials in aquaculture.Laboratory of Microbiology and Parasitology of Aquatic Organisms São Paulo State University (UNESP) Aquaculture Center of Unesp (Caunesp)Faculdade de Ciências Agrárias Universidade Zambeze (UniZambeze)Department of Biochemistry and Technological Chemistry Institute of Chemistry São Paulo State University (UNESP)Registro Experimental Campus São Paulo State University (UNESP)Graduate Program in Agricultural and Livestock Microbiology São Paulo State University (UNESP) School of Agricultural and Veterinarian SciencesLaboratory of Microbiology and Parasitology of Aquatic Organisms São Paulo State University (UNESP) Aquaculture Center of Unesp (Caunesp)Department of Biochemistry and Technological Chemistry Institute of Chemistry São Paulo State University (UNESP)Registro Experimental Campus São Paulo State University (UNESP)Graduate Program in Agricultural and Livestock Microbiology São Paulo State University (UNESP) School of Agricultural and Veterinarian SciencesUniversidade Estadual Paulista (Unesp)Universidade Zambeze (UniZambeze)Assane, I. M. [UNESP]Santos-Filho, N. A. [UNESP]de Sousa, E. L. [UNESP]de Arruda Brasil, M. C.O. [UNESP]Cilli, E. M. [UNESP]Pilarski, F. [UNESP]2021-06-25T10:56:36Z2021-06-25T10:56:36Z2021-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1111/jam.15080Journal of Applied Microbiology.1365-26721364-5072http://hdl.handle.net/11449/20752310.1111/jam.150802-s2.0-85103187577Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Applied Microbiologyinfo:eu-repo/semantics/openAccess2024-04-09T15:43:25Zoai:repositorio.unesp.br:11449/207523Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-04-09T15:43:25Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
title |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
spellingShingle |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria Assane, I. M. [UNESP] antimicrobial activity cytotoxicity fish pathogenic bacteria synergy synthetic peptides |
title_short |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
title_full |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
title_fullStr |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
title_full_unstemmed |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
title_sort |
Cytotoxicity and antimicrobial activity of synthetic peptides alone or in combination with conventional antimicrobials against fish pathogenic bacteria |
author |
Assane, I. M. [UNESP] |
author_facet |
Assane, I. M. [UNESP] Santos-Filho, N. A. [UNESP] de Sousa, E. L. [UNESP] de Arruda Brasil, M. C.O. [UNESP] Cilli, E. M. [UNESP] Pilarski, F. [UNESP] |
author_role |
author |
author2 |
Santos-Filho, N. A. [UNESP] de Sousa, E. L. [UNESP] de Arruda Brasil, M. C.O. [UNESP] Cilli, E. M. [UNESP] Pilarski, F. [UNESP] |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Universidade Zambeze (UniZambeze) |
dc.contributor.author.fl_str_mv |
Assane, I. M. [UNESP] Santos-Filho, N. A. [UNESP] de Sousa, E. L. [UNESP] de Arruda Brasil, M. C.O. [UNESP] Cilli, E. M. [UNESP] Pilarski, F. [UNESP] |
dc.subject.por.fl_str_mv |
antimicrobial activity cytotoxicity fish pathogenic bacteria synergy synthetic peptides |
topic |
antimicrobial activity cytotoxicity fish pathogenic bacteria synergy synthetic peptides |
description |
Aims: This study aimed to evaluate the in vitro cytotoxicity and efficacy of synthetic host defence peptides (HDPs), alone or in combination with florfenicol (FFC), oxytetracycline (OTC) or thiamphenicol (TAP), against different pathogenic bacteria isolated from diseased fish. Methods and Results: Solid-phase synthesis, purification and characterization of several HDPs were performed manually, using the fluorenylmethyloxycarbonyl protecting group in different resins and via high-performance liquid chromatography-mass spectrometry, respectively. The in vitro cytotoxicity and antimicrobial activity of HDPs, FFC, OTC and TAP against Nile tilapia red blood cells (RBCs) and relevant fish pathogenic bacteria (Aeromonas, Citrobacter, Edwardsiella, Streptococcus, Lactococcus and Vibrio) was determined using the haemolysis assay and broth microdilution method, respectively. The checkerboard assay was used to evaluate the synergy between the most active HDPs and other antimicrobials against the tested strains. MUC 7 12-mer, FFC, OTC and TAP were not cytotoxic to Nile tilapia RBCs, in all tested concentrations. LL-37, (p-BthTX-I)2 and Hylin-a1 were not cytotoxic at concentrations up to 78·13, 19·53 and 9·77 μg ml−1, respectively. HDPs demonstrated potent antimicrobial activity (minimum inhibitory concentration ≤31·25 µg ml−1) against Aeromonas jandaei (KR-12-a5), Citrobacter freundii (Kr-12-a5; (p-BthTX-I)2; LL-37; and Hylin a1), Streptococcus agalactiae (Hylin a1; (p-BthTX-I)2 and LL-37), Lactococcus garviae (Hylin a1), and Vibrio fluvialis (KR-12-a5). The combinations of (p-BthTX-I)2 with TAP and LL-37 with FFC showed synergistic activity against C. freundii (fractional inhibitory concentration index of 0·25 and 0·50, respectively). Conclusions: Synthetic HDPs have the potential as a good treatment option for bacterial diseases in aquaculture. Significance and Impact of the Study: The in vivo effectiveness of synthetic HDPs such as KR-12-a5; LL-37; (p-BthTX-I)2 and Hylin a1 can be tested alone or in combination with conventional antimicrobials as a treatment option to reduce the use of antimicrobials in aquaculture. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-06-25T10:56:36Z 2021-06-25T10:56:36Z 2021-01-01 |
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/10.1111/jam.15080 Journal of Applied Microbiology. 1365-2672 1364-5072 http://hdl.handle.net/11449/207523 10.1111/jam.15080 2-s2.0-85103187577 |
url |
http://dx.doi.org/10.1111/jam.15080 http://hdl.handle.net/11449/207523 |
identifier_str_mv |
Journal of Applied Microbiology. 1365-2672 1364-5072 10.1111/jam.15080 2-s2.0-85103187577 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Applied Microbiology |
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 |
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1797790276850286592 |