Evaluation of the Pathogenic-Mixed Biofilm Formation of Pseudomonas aeruginosa / Staphylococcus aureus and Treatment with Limonene on Three Different Materials by a Dynamic Model

Edvige Gambino, Angela Maione, Marco Guida, Luisa Albarano, Federica Carraturo, Emilia Galdiero and Valeria Di Onofrio
Additional contact information
Edvige Gambino: Department of Biology, University of Naples “Federico II”, 80126 Naples, Italy
Angela Maione: Department of Biology, University of Naples “Federico II”, 80126 Naples, Italy
Marco Guida: Department of Biology, University of Naples “Federico II”, 80126 Naples, Italy
Luisa Albarano: Department of Biology, University of Naples “Federico II”, 80126 Naples, Italy
Federica Carraturo: Department of Biology, University of Naples “Federico II”, 80126 Naples, Italy
Emilia Galdiero: Department of Biology, University of Naples “Federico II”, 80126 Naples, Italy
Valeria Di Onofrio: Department of Sciences and Technologies, University of Naples “Parthenope”, 80143 Naples, Italy

IJERPH, 2022, vol. 19, issue 6, 1-12

Abstract: Background: Biofilms have been found growing on implantable medical devices. This can lead to persistent clinical infections. The highly antibiotic-resistant property of biofilms necessitates the search for both potent antimicrobial agents and novel antibiofilm strategies. Natural product-based anti-biofilm agents were found to be as efficient as chemically synthesized counterparts with fewer side effects. In the present study, the effects of limonene as an antibiofilm agent were evaluated on Pseudomonas aeruginosa and Staphylococcus aureus biofilm formed on different surfaces using the CDC model system in continuous flow. The flgK gene and the pilA gene expression in P. aeruginosa, and the icaA gene and eno gene in S. aureus, which could be considered as efficient resistance markers, were studied. Methods: Mono- and dual-species biofilms were grown on polycarbonate, polypropylene, and stainless-steel coupons in a CDC biofilm reactor (Biosurface Technologies, Bozeman, MT, USA). To evaluate the ability of limonene to inhibit and eradicate biofilm, a sub-MIC concentration (10 mL/L) was tested. The gene expression of P. aeruginosa and S. aureus was detected by SYBR Green quantitative Real-Time PCR assay (Meridiana Bioline, Brisbane, Australia). Results: The limonene added during the formation of biofilms at sub-MIC concentrations works very well in inhibiting biofilms on all three materials, reducing their growth by about 2 logs. Of the same order of magnitude is the ability of limonene to eradicate both mono- and polymicrobial mature biofilms on all three materials. Greater efficacy was observed in the polymicrobial biofilm on steel coupons. The expression of some genes related to the virulence of the two microorganisms was differently detected in mono- and polymicrobial biofilm. Conclusions: These data showed that the limonene treatment expressed different levels of biofilm-forming genes, especially when both types of strains alone and together grew on different surfaces. Our findings showed that limonene treatment is also very efficient when biofilm has been grown under shear stress causing significant and irreversible damage to the biofilm structure. The effectiveness of the sanitation procedures can be optimized by applying antimicrobial combinations with natural compounds (e.g., limonene).

Keywords: biofilms; Pseudomonas aeruginosa; Staphylococcus aureus; mixed infections; CDC biofilm reactor; continuous flow; limonene; gene expression (search for similar items in EconPapers)
JEL-codes: I I1 I3 Q Q5 (search for similar items in EconPapers)
Date: 2022
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.mdpi.com/1660-4601/19/6/3741/pdf (application/pdf)
https://www.mdpi.com/1660-4601/19/6/3741/ (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:gam:jijerp:v:19:y:2022:i:6:p:3741-:d:776162

Access Statistics for this article

IJERPH is currently edited by Ms. Jenna Liu

More articles in IJERPH from MDPI
Bibliographic data for series maintained by MDPI Indexing Manager ().