Related Experiment Video
Updated: May 11, 2026

A Platform of Anti-biofilm Assays Suited to the Exploration of Natural Compound Libraries
Published on: December 27, 2016
Human methicillin-sensitive Staphylococcus aureus biofilms: potential associations with antibiotic resistance
Charlene Babra1, Jully Tiwari, Paul Costantino
1School of Biomedical Sciences, Faculty of Health Sciences, Curtin Health Innovation Research Institute, West Australian Biomedical Research Institute, Curtin University, Bentley Campus, Perth, WA, Australia.
Abstract:
The development of persistent antibiotic resistance by human methicillin-sensitive Staphylococcus aureus (MSSA) strains and substantial association with poly-N-acetyl glucosamine (PNAG) in biofilms is reported in this investigation. Sixteen of 31 MSSA strains under study were found to have developed resistance to one or more antibiotics, with four strains, two of which did not produce biofilms, showing resistance to cefoxitin, undetectable by mecA amplification. Antibiotic resistance displayed by 13/14 biofilm-forming S. aureus isolates remained persistent for 4 weeks prior to reverting back to the original antibiotic susceptibility, prompting a suggestion of determining antibiograms for clinical S. aureus isolates subcultured from biofilms developed in vitro as well as planktonic subcultures prepared from the site of infection. While there was correlation of antibiotic resistance with biofilm formation confirming previous reports, this is the first time that persistence of the biofilm-associated antibiotic resistance by S. aureus as planktonic cells is reported. Among the two methods used for assessment of biofilm formation, the tissue culture plate (TCP) method revealed that almost all strains were strong or moderate biofilm producers whereas only 19/31 strains were biofilm producers using the Congo Red agar (CRA) method indicating the superiority of the TCP method in detecting biofilm producers. We also observed no association between biofilm formation and major capsule types. However, substantial, although not absolute, association of biofilm formation with PNAG was observed, warranting continued identification of additional surface-associated polysaccharide and/or protein antigens associated with biofilm formation for development of an effective vaccine against S. aureus infections regardless of capsular phenotype.
Insights
Persistent antibiotic resistance in Staphylococcus aureus biofilms is linked to poly-N-acetyl glucosamine (PNAG). This study observed resistance persistence in biofilms and planktonic cells, highlighting the need for regular antibiogram testing.
Area of Science:
- Microbiology
- Infectious Diseases
- Molecular Biology
Background:
- Antibiotic resistance in Staphylococcus aureus (S. aureus) is a significant clinical concern.
- Biofilm formation by S. aureus is associated with persistent infections and treatment challenges.
- Poly-N-acetyl glucosamine (PNAG) has been implicated in S. aureus biofilm structure.
Purpose of the Study:
- To investigate the development and persistence of antibiotic resistance in methicillin-sensitive S. aureus (MSSA) strains within biofilms.
- To explore the association between biofilm formation, antibiotic resistance, and PNAG in S. aureus.
- To compare methods for detecting S. aureus biofilm formation.
Main Methods:
- Phenotypic antibiotic susceptibility testing of MSSA strains.
- Assessment of biofilm formation using tissue culture plate (TCP) and Congo Red agar (CRA) methods.
- Detection of mecA amplification for cefoxitin resistance.
- Observation of PNAG association with biofilm formation.
Main Results:
- Sixteen of 31 MSSA strains exhibited antibiotic resistance; four were cefoxitin-resistant without mecA.
- Antibiotic resistance in 13/14 biofilm-forming strains persisted for 4 weeks.
- The TCP method was superior to CRA for detecting biofilm production.
- A substantial association between biofilm formation and PNAG was observed.
Conclusions:
- Persistent antibiotic resistance in S. aureus biofilms, even in planktonic cells, is a critical finding.
- Regular antibiogram determination for isolates from biofilms and clinical sites is recommended.
- PNAG is significantly associated with S. aureus biofilm formation, suggesting its potential as a vaccine target.
Related Concept Videos
Clinical Significance of Antibiotic Resistance
Mechanism of Antibiotic Resistance in MRSA
Biofilms
Staphylococcal Skin Infections
Development of Antibiotic Resistance
Surface Appendages of Archaea
