Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand

Suphot Wattanaphansak; Carlos Eduardo Real Pereira; Wenika Kaenson; Pornchalit Assavacheep; Rachod Tantilertcharoen; Talita Pilar Resende; Javier Alberto Barrera-Zarate; Juliana Saes Vilaça De Oliveira-Lee; Ulrich Klein; Connie Jane Gebhart; Roberto Maurício Carvalho Guedes

doi:10.1186/s12866-019-1397-7

Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand

Suphot Wattanaphansak, Carlos Eduardo Real Pereira, Wenika Kaenson, Pornchalit Assavacheep, Rachod Tantilertcharoen, Talita Pilar Resende, Javier Alberto Barrera-Zarate, Juliana Saes Vilaça De Oliveira-Lee, Ulrich Klein, Connie Jane Gebhart, Roberto Maurício Carvalho Guedes

Veterinary and Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

Background: Lawsonia intracellularis is an obligate intracellular bacterium which cannot be cultured by conventional bacteriological methods. Furthermore, L. intracellularis needs enriched medium and a unique atmosphere for isolation, cultivation and propagation. Because of this,there are only a few isolates of L. intracellularis available and few studies in vitro demonstrating the susceptibility of this bacterium to antimicrobial agents. The objectives of this study were to isolate South American and Southeast Asia strains of L.intracellularis and to determine the in vitro antimicrobial activity against these isolates. Tested antimicrobials included: chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin(against both Brazilian and Thailand strains) and additionally, amoxicillin, zinc-bacitracin, carbadox, enrofloxacin, gentamicin, sulfamethazine, trimethoprim, spectinomycin and a combination (1:1) of spectinomycin and lincomycin were also tested against the Thai isolates. The minimum inhibitory concentration (MIC) was determined by the antimicrobial activity that inhibited 99% of L. intracellularis growth in a cell culture as compared to the control (antimicrobial-free). Results: Two strains from Brazil and three strains from Thailand were successfully isolated and established in cell culture. Each antimicrobial was evaluated for intracellular and extracellular activity. Pleuromutilin group (valnemulin and tiamulin) and carbadox were the most active against L. intracellularis strains tested. Tylosin showed intermediate activity, chlortetracycline had variable results between low and intermediate activity, as well as spectinomycin, spectinomycin and lincomycin, amoxicillin, sulfamethazine and enrofloxacin. L. intracellularis was resistant to lincomycin, gentamicin, trimethoprim, colistin and bacitracin in in vitro conditions. Conclusions: This is the first report of isolation of L. intracellularis strains from South America and Southeast Asia and characterization of the antimicrobial susceptibility patterns of these new strains.

Original language	English (US)
Article number	27
Journal	BMC microbiology
Volume	19
Issue number	1
DOIs	https://doi.org/10.1186/s12866-019-1397-7
State	Published - Jan 31 2019

Bibliographical note

Funding Information:
RMCG has a research fellowship from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). SW was partially supported by a research fellowship from Ratchadapiseksompotch Fund, Chulalongkorn University. TPR has PhD funding from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil. The authors are grateful for partial funding from Novartis Animal Health (now Elanco Animal Health), Fapemig, Capes, and CNPq.

Funding Information:
Though the project had been partially funded by Novartis Animal Health, the company was not involved in conducting the study.

Publisher Copyright:
© 2019 The Author(s).

Keywords

Antimicrobial susceptibility
Ileitis
Lawsonia intracellularis
MIC
Pigs
Proliferative enteropathy

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Wattanaphansak, S., Pereira, C. E. R., Kaenson, W., Assavacheep, P., Tantilertcharoen, R., Resende, T. P., Barrera-Zarate, J. A., De Oliveira-Lee, J. S. V., Klein, U., Gebhart, C. J., & Guedes, R. M. C. (2019). Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand. BMC microbiology, 19(1), Article 27. https://doi.org/10.1186/s12866-019-1397-7

Wattanaphansak, S, Pereira, CER, Kaenson, W, Assavacheep, P, Tantilertcharoen, R, Resende, TP, Barrera-Zarate, JA, De Oliveira-Lee, JSV, Klein, U, Gebhart, CJ & Guedes, RMC 2019, 'Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand', BMC microbiology, vol. 19, no. 1, 27. https://doi.org/10.1186/s12866-019-1397-7

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abstract = "Background: Lawsonia intracellularis is an obligate intracellular bacterium which cannot be cultured by conventional bacteriological methods. Furthermore, L. intracellularis needs enriched medium and a unique atmosphere for isolation, cultivation and propagation. Because of this,there are only a few isolates of L. intracellularis available and few studies in vitro demonstrating the susceptibility of this bacterium to antimicrobial agents. The objectives of this study were to isolate South American and Southeast Asia strains of L.intracellularis and to determine the in vitro antimicrobial activity against these isolates. Tested antimicrobials included: chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin(against both Brazilian and Thailand strains) and additionally, amoxicillin, zinc-bacitracin, carbadox, enrofloxacin, gentamicin, sulfamethazine, trimethoprim, spectinomycin and a combination (1:1) of spectinomycin and lincomycin were also tested against the Thai isolates. The minimum inhibitory concentration (MIC) was determined by the antimicrobial activity that inhibited 99% of L. intracellularis growth in a cell culture as compared to the control (antimicrobial-free). Results: Two strains from Brazil and three strains from Thailand were successfully isolated and established in cell culture. Each antimicrobial was evaluated for intracellular and extracellular activity. Pleuromutilin group (valnemulin and tiamulin) and carbadox were the most active against L. intracellularis strains tested. Tylosin showed intermediate activity, chlortetracycline had variable results between low and intermediate activity, as well as spectinomycin, spectinomycin and lincomycin, amoxicillin, sulfamethazine and enrofloxacin. L. intracellularis was resistant to lincomycin, gentamicin, trimethoprim, colistin and bacitracin in in vitro conditions. Conclusions: This is the first report of isolation of L. intracellularis strains from South America and Southeast Asia and characterization of the antimicrobial susceptibility patterns of these new strains.",

keywords = "Antimicrobial susceptibility, Ileitis, Lawsonia intracellularis, MIC, Pigs, Proliferative enteropathy",

author = "Suphot Wattanaphansak and Pereira, {Carlos Eduardo Real} and Wenika Kaenson and Pornchalit Assavacheep and Rachod Tantilertcharoen and Resende, {Talita Pilar} and Barrera-Zarate, {Javier Alberto} and {De Oliveira-Lee}, {Juliana Saes Vila{\c c}a} and Ulrich Klein and Gebhart, {Connie Jane} and Guedes, {Roberto Maur{\'i}cio Carvalho}",

note = "Funding Information: RMCG has a research fellowship from Conselho Nacional de Desenvolvimento Cient{\'i}fico e Tecnol{\'o}gico (CNPq). SW was partially supported by a research fellowship from Ratchadapiseksompotch Fund, Chulalongkorn University. TPR has PhD funding from Coordena{\c c}{\~a}o de Aperfei{\c c}oamento de Pessoal de N{\'i}vel Superior (CAPES), Brazil. The authors are grateful for partial funding from Novartis Animal Health (now Elanco Animal Health), Fapemig, Capes, and CNPq. Funding Information: Though the project had been partially funded by Novartis Animal Health, the company was not involved in conducting the study. Publisher Copyright: {\textcopyright} 2019 The Author(s).",

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T1 - Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand

AU - Wattanaphansak, Suphot

AU - Pereira, Carlos Eduardo Real

AU - Kaenson, Wenika

AU - Assavacheep, Pornchalit

AU - Tantilertcharoen, Rachod

AU - Resende, Talita Pilar

AU - Barrera-Zarate, Javier Alberto

AU - De Oliveira-Lee, Juliana Saes Vilaça

AU - Klein, Ulrich

AU - Gebhart, Connie Jane

AU - Guedes, Roberto Maurício Carvalho

N1 - Funding Information: RMCG has a research fellowship from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). SW was partially supported by a research fellowship from Ratchadapiseksompotch Fund, Chulalongkorn University. TPR has PhD funding from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil. The authors are grateful for partial funding from Novartis Animal Health (now Elanco Animal Health), Fapemig, Capes, and CNPq. Funding Information: Though the project had been partially funded by Novartis Animal Health, the company was not involved in conducting the study. Publisher Copyright: © 2019 The Author(s).

PY - 2019/1/31

Y1 - 2019/1/31

N2 - Background: Lawsonia intracellularis is an obligate intracellular bacterium which cannot be cultured by conventional bacteriological methods. Furthermore, L. intracellularis needs enriched medium and a unique atmosphere for isolation, cultivation and propagation. Because of this,there are only a few isolates of L. intracellularis available and few studies in vitro demonstrating the susceptibility of this bacterium to antimicrobial agents. The objectives of this study were to isolate South American and Southeast Asia strains of L.intracellularis and to determine the in vitro antimicrobial activity against these isolates. Tested antimicrobials included: chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin(against both Brazilian and Thailand strains) and additionally, amoxicillin, zinc-bacitracin, carbadox, enrofloxacin, gentamicin, sulfamethazine, trimethoprim, spectinomycin and a combination (1:1) of spectinomycin and lincomycin were also tested against the Thai isolates. The minimum inhibitory concentration (MIC) was determined by the antimicrobial activity that inhibited 99% of L. intracellularis growth in a cell culture as compared to the control (antimicrobial-free). Results: Two strains from Brazil and three strains from Thailand were successfully isolated and established in cell culture. Each antimicrobial was evaluated for intracellular and extracellular activity. Pleuromutilin group (valnemulin and tiamulin) and carbadox were the most active against L. intracellularis strains tested. Tylosin showed intermediate activity, chlortetracycline had variable results between low and intermediate activity, as well as spectinomycin, spectinomycin and lincomycin, amoxicillin, sulfamethazine and enrofloxacin. L. intracellularis was resistant to lincomycin, gentamicin, trimethoprim, colistin and bacitracin in in vitro conditions. Conclusions: This is the first report of isolation of L. intracellularis strains from South America and Southeast Asia and characterization of the antimicrobial susceptibility patterns of these new strains.

AB - Background: Lawsonia intracellularis is an obligate intracellular bacterium which cannot be cultured by conventional bacteriological methods. Furthermore, L. intracellularis needs enriched medium and a unique atmosphere for isolation, cultivation and propagation. Because of this,there are only a few isolates of L. intracellularis available and few studies in vitro demonstrating the susceptibility of this bacterium to antimicrobial agents. The objectives of this study were to isolate South American and Southeast Asia strains of L.intracellularis and to determine the in vitro antimicrobial activity against these isolates. Tested antimicrobials included: chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin(against both Brazilian and Thailand strains) and additionally, amoxicillin, zinc-bacitracin, carbadox, enrofloxacin, gentamicin, sulfamethazine, trimethoprim, spectinomycin and a combination (1:1) of spectinomycin and lincomycin were also tested against the Thai isolates. The minimum inhibitory concentration (MIC) was determined by the antimicrobial activity that inhibited 99% of L. intracellularis growth in a cell culture as compared to the control (antimicrobial-free). Results: Two strains from Brazil and three strains from Thailand were successfully isolated and established in cell culture. Each antimicrobial was evaluated for intracellular and extracellular activity. Pleuromutilin group (valnemulin and tiamulin) and carbadox were the most active against L. intracellularis strains tested. Tylosin showed intermediate activity, chlortetracycline had variable results between low and intermediate activity, as well as spectinomycin, spectinomycin and lincomycin, amoxicillin, sulfamethazine and enrofloxacin. L. intracellularis was resistant to lincomycin, gentamicin, trimethoprim, colistin and bacitracin in in vitro conditions. Conclusions: This is the first report of isolation of L. intracellularis strains from South America and Southeast Asia and characterization of the antimicrobial susceptibility patterns of these new strains.

KW - Antimicrobial susceptibility

KW - Ileitis

KW - Lawsonia intracellularis

KW - MIC

KW - Pigs

KW - Proliferative enteropathy

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SN - 1471-2180

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JO - BMC microbiology

JF - BMC microbiology

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ER -

Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand

Abstract

Bibliographical note

Keywords

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