Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

Yuwana Podin, Derek Sarovich, Erin Price, Mirjam Kaestli, Mark Mayo, KingChing Hii, HieUng Ngian, See-Chang Wong, IngTien Wong, JinShyan Wong, Anand Mohan, Mong Ooi, TemLom Fam, Jack Wong, Apichai Tuanyok, Paul S Keim, Philip Giffard, Bart Currie

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Melioidosis is a potentially fatal disease caused by the saprophytic bacterium Burkholderia pseudomallei. Resistance to gentamicin is generally a hallmark of B. pseudomallei, and gentamicin is a selective agent in media used for diagnosis of melioidosis. In this study, we determined the prevalence and mechanism of gentamicin susceptibility found in B. pseudomallei isolates from Sarawak, Malaysian Borneo. We performed multilocus sequence typing and antibiotic susceptibility testing on 44 B. pseudomallei clinical isolates from melioidosis patients in Sarawak district hospitals. Whole-genome sequencing was used to identify the mechanism of gentamicin susceptibility. A novel allelic-specific PCR was designed to differentiate gentamicin-sensitive isolates from wild-type B. pseudomallei. A reversion assay was performed to confirm the involvement of this mechanism in gentamicin susceptibility. A substantial proportion (86%) of B. pseudomallei clinical isolates in Sarawak, Malaysian Borneo, were found to be susceptible to the aminoglycoside gentamicin, a rare occurrence in other regions where B. pseudomallei is endemic. Gentamicin sensitivity was restricted to genetically related strains belonging to sequence type 881 or its single-locus variant, sequence type 997. Whole-genome sequencing identified a novel nonsynonymous mutation within amrB, encoding an essential component of the AmrAB-OprA multidrug efflux pump. We confirmed the role of this mutation in conferring aminoglycoside and macrolide sensitivity by reversion of this mutation to the wild-type sequence. Our study demonstrates that alternative B. pseudomallei selective media without gentamicin are needed for accurate melioidosis laboratory diagnosis in Sarawak. This finding may also have implications for environmental sampling of other locations to test for B. pseudomallei endemicity. � 2014, American Society for Microbiology. All Rights Reserved.
    Original languageEnglish
    Pages (from-to)162-166
    Number of pages5
    JournalAntimicrobial Agents and Chemotherapy
    Volume58
    Issue number1
    DOIs
    Publication statusPublished - 2014

    Fingerprint

    Borneo
    Burkholderia pseudomallei
    Malaysia
    Macrolides
    Aminoglycosides
    Gentamicins
    Melioidosis
    Mutation
    Genome
    Multilocus Sequence Typing
    District Hospitals
    Clinical Laboratory Techniques
    Microbiology

    Cite this

    Podin, Yuwana ; Sarovich, Derek ; Price, Erin ; Kaestli, Mirjam ; Mayo, Mark ; Hii, KingChing ; Ngian, HieUng ; Wong, See-Chang ; Wong, IngTien ; Wong, JinShyan ; Mohan, Anand ; Ooi, Mong ; Fam, TemLom ; Wong, Jack ; Tuanyok, Apichai ; Keim, Paul S ; Giffard, Philip ; Currie, Bart. / Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides. In: Antimicrobial Agents and Chemotherapy. 2014 ; Vol. 58, No. 1. pp. 162-166.
    @article{255bc4cb980649fc913dbc68eac1dab9,
    title = "Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides",
    abstract = "Melioidosis is a potentially fatal disease caused by the saprophytic bacterium Burkholderia pseudomallei. Resistance to gentamicin is generally a hallmark of B. pseudomallei, and gentamicin is a selective agent in media used for diagnosis of melioidosis. In this study, we determined the prevalence and mechanism of gentamicin susceptibility found in B. pseudomallei isolates from Sarawak, Malaysian Borneo. We performed multilocus sequence typing and antibiotic susceptibility testing on 44 B. pseudomallei clinical isolates from melioidosis patients in Sarawak district hospitals. Whole-genome sequencing was used to identify the mechanism of gentamicin susceptibility. A novel allelic-specific PCR was designed to differentiate gentamicin-sensitive isolates from wild-type B. pseudomallei. A reversion assay was performed to confirm the involvement of this mechanism in gentamicin susceptibility. A substantial proportion (86{\%}) of B. pseudomallei clinical isolates in Sarawak, Malaysian Borneo, were found to be susceptible to the aminoglycoside gentamicin, a rare occurrence in other regions where B. pseudomallei is endemic. Gentamicin sensitivity was restricted to genetically related strains belonging to sequence type 881 or its single-locus variant, sequence type 997. Whole-genome sequencing identified a novel nonsynonymous mutation within amrB, encoding an essential component of the AmrAB-OprA multidrug efflux pump. We confirmed the role of this mutation in conferring aminoglycoside and macrolide sensitivity by reversion of this mutation to the wild-type sequence. Our study demonstrates that alternative B. pseudomallei selective media without gentamicin are needed for accurate melioidosis laboratory diagnosis in Sarawak. This finding may also have implications for environmental sampling of other locations to test for B. pseudomallei endemicity. � 2014, American Society for Microbiology. All Rights Reserved.",
    keywords = "amoxicillin, azithromycin, ceftazidime, clavulanic acid, cotrimoxazole, doxycycline, gentamicin, kanamycin, macrolide, meropenem, adolescent, adult, aged, antibiotic resistance, antibiotic sensitivity, article, bacterial strain, bacterium isolate, Borneo, Burkholderia pseudomallei, child, clinical article, controlled study, endemic disease, female, gene locus, gene mutation, gene sequence, geographic distribution, human, male, melioidosis, middle aged, multilocus sequence typing, polymerase chain reaction, preschool child, priority journal, public hospital, school child, Aminoglycosides, Anti-Bacterial Agents, Gentamicins, Macrolides, Malaysia, Microbial Sensitivity Tests",
    author = "Yuwana Podin and Derek Sarovich and Erin Price and Mirjam Kaestli and Mark Mayo and KingChing Hii and HieUng Ngian and See-Chang Wong and IngTien Wong and JinShyan Wong and Anand Mohan and Mong Ooi and TemLom Fam and Jack Wong and Apichai Tuanyok and Keim, {Paul S} and Philip Giffard and Bart Currie",
    year = "2014",
    doi = "10.1128/AAC.01842-13",
    language = "English",
    volume = "58",
    pages = "162--166",
    journal = "Antimicrobial Agents and Chemotherapy",
    issn = "0066-4804",
    publisher = "American Society for Microbiology",
    number = "1",

    }

    Podin, Y, Sarovich, D, Price, E, Kaestli, M, Mayo, M, Hii, K, Ngian, H, Wong, S-C, Wong, I, Wong, J, Mohan, A, Ooi, M, Fam, T, Wong, J, Tuanyok, A, Keim, PS, Giffard, P & Currie, B 2014, 'Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides', Antimicrobial Agents and Chemotherapy, vol. 58, no. 1, pp. 162-166. https://doi.org/10.1128/AAC.01842-13

    Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides. / Podin, Yuwana; Sarovich, Derek; Price, Erin; Kaestli, Mirjam; Mayo, Mark; Hii, KingChing; Ngian, HieUng; Wong, See-Chang; Wong, IngTien; Wong, JinShyan; Mohan, Anand; Ooi, Mong; Fam, TemLom; Wong, Jack; Tuanyok, Apichai; Keim, Paul S; Giffard, Philip; Currie, Bart.

    In: Antimicrobial Agents and Chemotherapy, Vol. 58, No. 1, 2014, p. 162-166.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

    AU - Podin, Yuwana

    AU - Sarovich, Derek

    AU - Price, Erin

    AU - Kaestli, Mirjam

    AU - Mayo, Mark

    AU - Hii, KingChing

    AU - Ngian, HieUng

    AU - Wong, See-Chang

    AU - Wong, IngTien

    AU - Wong, JinShyan

    AU - Mohan, Anand

    AU - Ooi, Mong

    AU - Fam, TemLom

    AU - Wong, Jack

    AU - Tuanyok, Apichai

    AU - Keim, Paul S

    AU - Giffard, Philip

    AU - Currie, Bart

    PY - 2014

    Y1 - 2014

    N2 - Melioidosis is a potentially fatal disease caused by the saprophytic bacterium Burkholderia pseudomallei. Resistance to gentamicin is generally a hallmark of B. pseudomallei, and gentamicin is a selective agent in media used for diagnosis of melioidosis. In this study, we determined the prevalence and mechanism of gentamicin susceptibility found in B. pseudomallei isolates from Sarawak, Malaysian Borneo. We performed multilocus sequence typing and antibiotic susceptibility testing on 44 B. pseudomallei clinical isolates from melioidosis patients in Sarawak district hospitals. Whole-genome sequencing was used to identify the mechanism of gentamicin susceptibility. A novel allelic-specific PCR was designed to differentiate gentamicin-sensitive isolates from wild-type B. pseudomallei. A reversion assay was performed to confirm the involvement of this mechanism in gentamicin susceptibility. A substantial proportion (86%) of B. pseudomallei clinical isolates in Sarawak, Malaysian Borneo, were found to be susceptible to the aminoglycoside gentamicin, a rare occurrence in other regions where B. pseudomallei is endemic. Gentamicin sensitivity was restricted to genetically related strains belonging to sequence type 881 or its single-locus variant, sequence type 997. Whole-genome sequencing identified a novel nonsynonymous mutation within amrB, encoding an essential component of the AmrAB-OprA multidrug efflux pump. We confirmed the role of this mutation in conferring aminoglycoside and macrolide sensitivity by reversion of this mutation to the wild-type sequence. Our study demonstrates that alternative B. pseudomallei selective media without gentamicin are needed for accurate melioidosis laboratory diagnosis in Sarawak. This finding may also have implications for environmental sampling of other locations to test for B. pseudomallei endemicity. � 2014, American Society for Microbiology. All Rights Reserved.

    AB - Melioidosis is a potentially fatal disease caused by the saprophytic bacterium Burkholderia pseudomallei. Resistance to gentamicin is generally a hallmark of B. pseudomallei, and gentamicin is a selective agent in media used for diagnosis of melioidosis. In this study, we determined the prevalence and mechanism of gentamicin susceptibility found in B. pseudomallei isolates from Sarawak, Malaysian Borneo. We performed multilocus sequence typing and antibiotic susceptibility testing on 44 B. pseudomallei clinical isolates from melioidosis patients in Sarawak district hospitals. Whole-genome sequencing was used to identify the mechanism of gentamicin susceptibility. A novel allelic-specific PCR was designed to differentiate gentamicin-sensitive isolates from wild-type B. pseudomallei. A reversion assay was performed to confirm the involvement of this mechanism in gentamicin susceptibility. A substantial proportion (86%) of B. pseudomallei clinical isolates in Sarawak, Malaysian Borneo, were found to be susceptible to the aminoglycoside gentamicin, a rare occurrence in other regions where B. pseudomallei is endemic. Gentamicin sensitivity was restricted to genetically related strains belonging to sequence type 881 or its single-locus variant, sequence type 997. Whole-genome sequencing identified a novel nonsynonymous mutation within amrB, encoding an essential component of the AmrAB-OprA multidrug efflux pump. We confirmed the role of this mutation in conferring aminoglycoside and macrolide sensitivity by reversion of this mutation to the wild-type sequence. Our study demonstrates that alternative B. pseudomallei selective media without gentamicin are needed for accurate melioidosis laboratory diagnosis in Sarawak. This finding may also have implications for environmental sampling of other locations to test for B. pseudomallei endemicity. � 2014, American Society for Microbiology. All Rights Reserved.

    KW - amoxicillin

    KW - azithromycin

    KW - ceftazidime

    KW - clavulanic acid

    KW - cotrimoxazole

    KW - doxycycline

    KW - gentamicin

    KW - kanamycin

    KW - macrolide

    KW - meropenem

    KW - adolescent

    KW - adult

    KW - aged

    KW - antibiotic resistance

    KW - antibiotic sensitivity

    KW - article

    KW - bacterial strain

    KW - bacterium isolate

    KW - Borneo

    KW - Burkholderia pseudomallei

    KW - child

    KW - clinical article

    KW - controlled study

    KW - endemic disease

    KW - female

    KW - gene locus

    KW - gene mutation

    KW - gene sequence

    KW - geographic distribution

    KW - human

    KW - male

    KW - melioidosis

    KW - middle aged

    KW - multilocus sequence typing

    KW - polymerase chain reaction

    KW - preschool child

    KW - priority journal

    KW - public hospital

    KW - school child

    KW - Aminoglycosides

    KW - Anti-Bacterial Agents

    KW - Gentamicins

    KW - Macrolides

    KW - Malaysia

    KW - Microbial Sensitivity Tests

    UR - http://www.scopus.com/inward/record.url?scp=84891516945&partnerID=8YFLogxK

    U2 - 10.1128/AAC.01842-13

    DO - 10.1128/AAC.01842-13

    M3 - Article

    VL - 58

    SP - 162

    EP - 166

    JO - Antimicrobial Agents and Chemotherapy

    JF - Antimicrobial Agents and Chemotherapy

    SN - 0066-4804

    IS - 1

    ER -