Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood

Soraya Mendoza-Olazarán, Rayo Morfín-Otero, Licet Villarreal-Treviño, Eduardo Rodríguez-Noriega, Jorge Llaca-Díaz, Adrián Camacho-Ortiz, Gloria M González, Néstor Casillas-Vega, Elvira Garza-González

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

OBJECTIVES: We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood.

METHODS: The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method.

RESULTS: Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7% (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91% (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66% (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol.

CONCLUSIONS: S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular matrix with similar composition of proteins, DNA and N-acetylglucosamine; and presents high frequency and low expression of icaD gene. Biofilm production is associated with increased antibiotic resistance.

Original languageEnglish
Pages (from-to)e0144684
JournalPLoS One
Volume10
Issue number12
DOIs
Publication statusPublished - 2015
Externally publishedYes

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Staphylococcus hominis
Biofilms
biofilm
Blood
antibiotics
Anti-Bacterial Agents
blood
Genes
cells
Chromosomes
Methicillin Resistance
Methicillin
methicillin
chromosomes
Linezolid
Multiplex Polymerase Chain Reaction
Microbial Sensitivity Tests
minimum inhibitory concentration
Chemical analysis
genetic relationships

Cite this

Mendoza-Olazarán, S., Morfín-Otero, R., Villarreal-Treviño, L., Rodríguez-Noriega, E., Llaca-Díaz, J., Camacho-Ortiz, A., ... Garza-González, E. (2015). Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood. PLoS One, 10(12), e0144684. https://doi.org/10.1371/journal.pone.0144684
Mendoza-Olazarán, Soraya ; Morfín-Otero, Rayo ; Villarreal-Treviño, Licet ; Rodríguez-Noriega, Eduardo ; Llaca-Díaz, Jorge ; Camacho-Ortiz, Adrián ; González, Gloria M ; Casillas-Vega, Néstor ; Garza-González, Elvira. / Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood. In: PLoS One. 2015 ; Vol. 10, No. 12. pp. e0144684.
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title = "Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood",
abstract = "OBJECTIVES: We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood.METHODS: The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method.RESULTS: Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7{\%} (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91{\%} (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66{\%} (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol.CONCLUSIONS: S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular matrix with similar composition of proteins, DNA and N-acetylglucosamine; and presents high frequency and low expression of icaD gene. Biofilm production is associated with increased antibiotic resistance.",
author = "Soraya Mendoza-Olazar{\'a}n and Rayo Morf{\'i}n-Otero and Licet Villarreal-Trevi{\~n}o and Eduardo Rodr{\'i}guez-Noriega and Jorge Llaca-D{\'i}az and Adri{\'a}n Camacho-Ortiz and Gonz{\'a}lez, {Gloria M} and N{\'e}stor Casillas-Vega and Elvira Garza-Gonz{\'a}lez",
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Mendoza-Olazarán, S, Morfín-Otero, R, Villarreal-Treviño, L, Rodríguez-Noriega, E, Llaca-Díaz, J, Camacho-Ortiz, A, González, GM, Casillas-Vega, N & Garza-González, E 2015, 'Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood', PLoS One, vol. 10, no. 12, pp. e0144684. https://doi.org/10.1371/journal.pone.0144684

Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood. / Mendoza-Olazarán, Soraya; Morfín-Otero, Rayo; Villarreal-Treviño, Licet; Rodríguez-Noriega, Eduardo; Llaca-Díaz, Jorge; Camacho-Ortiz, Adrián; González, Gloria M; Casillas-Vega, Néstor; Garza-González, Elvira.

In: PLoS One, Vol. 10, No. 12, 2015, p. e0144684.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood

AU - Mendoza-Olazarán, Soraya

AU - Morfín-Otero, Rayo

AU - Villarreal-Treviño, Licet

AU - Rodríguez-Noriega, Eduardo

AU - Llaca-Díaz, Jorge

AU - Camacho-Ortiz, Adrián

AU - González, Gloria M

AU - Casillas-Vega, Néstor

AU - Garza-González, Elvira

PY - 2015

Y1 - 2015

N2 - OBJECTIVES: We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood.METHODS: The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method.RESULTS: Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7% (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91% (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66% (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol.CONCLUSIONS: S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular matrix with similar composition of proteins, DNA and N-acetylglucosamine; and presents high frequency and low expression of icaD gene. Biofilm production is associated with increased antibiotic resistance.

AB - OBJECTIVES: We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood.METHODS: The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method.RESULTS: Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7% (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91% (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66% (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol.CONCLUSIONS: S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular matrix with similar composition of proteins, DNA and N-acetylglucosamine; and presents high frequency and low expression of icaD gene. Biofilm production is associated with increased antibiotic resistance.

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DO - 10.1371/journal.pone.0144684

M3 - Article

VL - 10

SP - e0144684

JO - PLoS One

JF - PLoS One

SN - 1932-6203

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Mendoza-Olazarán S, Morfín-Otero R, Villarreal-Treviño L, Rodríguez-Noriega E, Llaca-Díaz J, Camacho-Ortiz A et al. Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood. PLoS One. 2015;10(12):e0144684. https://doi.org/10.1371/journal.pone.0144684