recA mediated spontaneous deletions of the icaADBC operon of clinical Staphylococcus epidermidis isolates: a new mechanism of phenotypic variations

Nuryastuti, Titik; Mei, Henny C. van der; Busscher, Henk J.; Kuijer, Roel; Aman, Abu Tholib; Krom, Bastiaan P.

doi:10.1007/s10482-008-9249-8

Cited by 19 publications

(30 citation statements)

References 25 publications

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“…Furthermore, Nuryastuti et al (35) showed that deletion of the ica operon from S. epidermidis can be caused by the deregulation of RecA, which is involved in genetic deletions and rearrangements.…”

Section: Discussionmentioning

confidence: 99%

Biofilm Formation by Staphylococcus haemolyticus

et al. 2009

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Infections due to coagulase-negative staphylococci (CoNS) most frequently occur after the implantation of medical devices and are attributed to the biofilm-forming potential of CoNS. Staphylococcus haemolyticus is the second most frequently isolated CoNS from patients with hospital-acquired infections. There is only limited knowledge of the nature of S. haemolyticus biofilms. The aim of this study was to characterize S. haemolyticus biofilm formation. We analyzed the biofilm-forming capacities of 72 clinical S. haemolyticus isolates. A detachment assay with NaIO 4 , proteinase K, or DNase was used to determine the main biofilm components. Biofilm-associated genes, including the ica operon, were analyzed by PCR, and the gene products were sequenced. Confocal laser scanning microscopy (CLSM) was used to elucidate the biofilm structure. Fifty-three isolates (74%) produced biofilms after growth in Trypticase soy broth (TSB) with glucose, but only 22 (31%) produced biofilms after growth in TSB with NaCl. It was necessary to dissolve the biofilm in ethanol-acetone to measure the optical density of the full biofilm mass. DNase, proteinase K, and NaIO 4 caused biofilm detachment for 100%, 98%, and 38% of the isolates, respectively. icaRADBC and polysaccharide intercellular adhesin (PIA) production were found in only two isolates. CLSM indicated that the biofilm structure of S. haemolyticus clearly differs from that of S. epidermidis. We conclude that biofilm formation is a common phenotype in clinical S. haemolyticus isolates. In contrast to S. epidermidis, proteins and extracellular DNA are of functional relevance for biofilm accumulation, whereas PIA plays only a minor role. The induction of biofilm formation and determination of the biofilm mass also needed to be optimized for S. haemolyticus.

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Section: Discussionmentioning

confidence: 99%

Biofilm Formation by Staphylococcus haemolyticus

et al. 2009

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“…Clinical isolates of S. epidermidis were screened for the presence of icaA by PCR using the primers listed in Table 2 (19,32,40). Briefly, S. epidermidis strains were grown overnight at 37°C on a TSB agar plate.…”

Section: Bacterial Strains Sixteen Clinical Isolates Of S Epidermidismentioning

confidence: 99%

“…Biofilm formation in the presence of cinnamon oil and the effect on icaA expression. Biofilms were grown as described previously (7,32). Briefly, wells of a 96-well tissue culture polystyrene microtiter plate (Falcon; Becton Dickinson Labware, Franklin Lakes, NJ) were filled with 100 l of TSB containing cinnamon oil (twofold final concentrations) and subsequently inoculated with a 1:100 dilution of an overnight culture.…”

Section: Vol 75 2009 Effect Of Cinnamon Oil Against S Epidermidis mentioning

confidence: 99%

“…Total RNA was isolated from 24-h biofilm cultures grown with and without 0.01% cinnamon oil in 12-well tissue culture polystyrene plates (water contact angle, 59 degrees; Costar, Corning, NY) as described previously (32). Briefly, after resuspending the biofilms by pipetting, bacteria were pelleted by centrifugation and frozen at Ϫ80°C.…”

Section: Vol 75 2009 Effect Of Cinnamon Oil Against S Epidermidis mentioning

confidence: 99%

“…Chlorhexidine, triclosan, and gentamicin were used as positive controls in addition to examination of possible synergistic effects by combining cinnamon oil with any of these clinically used antimicrobials. (Table 1) were collected from Sardjito Hospital, Yogyakarta, Indonesia, and identified as reported previously in the Microbiology Department, Gadjah Mada University, Yogyakarta, Indonesia (32). Isolates were obtained from blood, cerebrospinal fluid, pus, and urine.…”

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Effect of Cinnamon Oil on icaA Expression and Biofilm Formation by Staphylococcus epidermidis

Nuryastuti

Mei

Busscher

et al. 2009

Appl Environ Microbiol

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131

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Staphylococcus epidermidis is notorious for its biofilm formation on medical devices, and novel approaches to prevent and kill S. epidermidis biofilms are desired. In this study, the effect of cinnamon oil on planktonic and biofilm cultures of clinical S. epidermidis isolates was evaluated. Initially, susceptibility to cinnamon oil in planktonic cultures was compared to the commonly used antimicrobial agents chlorhexidine, triclosan, and gentamicin. The MIC of cinnamon oil, defined as the lowest concentration able to inhibit visible microbial growth, and the minimal bactericidal concentration, the lowest concentration required to kill 99.9% of the bacteria, were determined using the broth microdilution method and plating on agar. A checkerboard assay was used to evaluate the possible synergy between cinnamon oil and the other antimicrobial agents. The effect of cinnamon oil on biofilm growth was studied in 96-well plates and with confocal laser-scanning microscopy (CLSM). Biofilm susceptibility was determined using a metabolic 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Real-time PCR analysis was performed to determine the effect of sub-MIC concentrations of cinnamon oil on expression of the biofilm-related gene, icaA. Cinnamon oil showed antimicrobial activity against both planktonic and biofilm cultures of clinical S. epidermidis strains. There was only a small difference between planktonic and biofilm MICs, ranging from 0.5 to 1% and 1 to 2%, respectively. CLSM images indicated that cinnamon oil is able to detach and kill existing biofilms. Thus, cinnamon oil is an effective antimicrobial agent to combat S. epidermidis biofilms.

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Ica‐expression and gentamicin susceptibility of Staphylococcus epidermidis biofilm on orthopedic implant biomaterials

Nuryastuti

Krom

Aman

et al. 2010

J Biomedical Materials Res

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Ica-expression by Staphylococcus epidermidis and slime production depends on environmental conditions such as implant material and presence of antibiotics. Here, we evaluate biofilm formation and ica-expression of S. epidermidis strains on biomaterials involved in total hip- and knee arthroplasty [polyethylene (PE), polymethylmethacrylate (PMMA), stainless steel (SS)]. Ica-expression, assayed using real-time RT-PCR, was highest on PE as confirmed using confocal laser scanning microscopy. Yet biofilm formation by S. epidermidis was most extensive on SS, with less slime production. Ica-expression and slime production were minimal on PMMA. After 3 h of continued growth of 24 h old biofilms in the presence of gentamicin, biofilms on PE showed lower susceptibility to gentamicin, relative to the other materials, presumably as a result of the stronger ica-expression. A higher gentamicin concentration further decreased metabolic activity on all biomaterials. It is concluded that the level of biomaterial-induced ica-expression does not correlate with the amount of biofilm formed, but initially aids bacteria in surviving antibiotic attacks. Once antibiotic treatment has started however, also the antibiotic itself induces slime production and only if its concentration is high enough, killing results. Results suggest that biomaterial-associated infections in orthopedics by S. epidermidis on PE may be more difficult to eradicate than on PMMA or SS.

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recA mediated spontaneous deletions of the icaADBC operon of clinical Staphylococcus epidermidis isolates: a new mechanism of phenotypic variations

Cited by 19 publications

References 25 publications

Biofilm Formation by Staphylococcus haemolyticus

Biofilm Formation by Staphylococcus haemolyticus

Effect of Cinnamon Oil on icaA Expression and Biofilm Formation by Staphylococcus epidermidis

Ica‐expression and gentamicin susceptibility of Staphylococcus epidermidis biofilm on orthopedic implant biomaterials

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