Park K, Kim MN, Sung H. Evaluation of the efficacy of three medical device detergents on bacteria and yeast derived biofilm: a comparative study. Ann Clin Microbiol 2023;26:117-24.
Annals of Clinical Microbiology (Ann Clin Microbiol) 2023 December, Volume 26, Issue 4, pages 117-124. https://doi.org/10.5145/ACM.2023.26.4.117
Received on 26 October 2023, Revised on 01 December 2023, Accepted on 01 December 2023, Published on 20 December 2023.
Background: This study aimed to evaluate the efficacy of three medical detergents against bacteria and yeast-derived biofilms. Methods: The biofilm removal efficacy of EmpowerTM (Metrex, USA), CidezymeTM (Johnson and Johnson Medical Inc, USA), and Matrix mintTM (Whiteley Medical, Australia) were compared to that of chlorine bleach. Biofilms were produced using Staphylococcus aureus RN9120, Escherichia coli ATCC35218, Pseudomonas aeruginosa ATCC27853, Candida albicans ATCC14053, and clinical isolates of Enterococcus faecalis, E. coli, Klebsiella pneumoniae, Candida auris, and Trichosporon asahii. The organisms were suspended in tryptic soy broth (TSB) in 96-well microplates and cultured for 72 hours. They were treated with the detergents, and the residual biofilm mass was quantified using crystal violet staining followed by optical density measurements at 620 nm (OD620 ). Results: EmpowerTM and CidezymeTM significantly reduced the biofilm mass derived from all species by > 50% of OD620 at 37ºC except those from E. faecalis, T. asahii, and C. auris. Matrix mintTM had no effect on the biofilms under any condition. Conclusion: The culture conditions and the species of the biofilm-producing organism influenced the effectiveness of the detergent. Biofilms produced by E. faecalis, C. auris, and T. asahii were resistant to all detergent treatments under all conditions.
Biofilm, Detergent, Bacteria, Yeast, Efficacy
It is not a human population study; therefore, approval by the institutional review board or the obtainment of informed consent is not required.
The study detergents included two enzymatic cleaners, EmpowerTM (Metrex, Orange, CA, USA), Cidezyme TM (Johnson and Johnson Medical Inc, Arlington, TX, USA), one non-enzymatic cleaner Matrix mint TM (Whiteley Medical, Sydney, Australia). A chlorine bleach YuhanroxTM Regular (Yuhanclorox, Seoul, Korea) was also tested as a control (Table 1). Each biofilm-producing strain was cultured for 72 hr at the same condition of biofilm production to prepare a 96-well microplate coated with biofilm. The biofilmproducing plates prepared by each strain was tested with each detergent under five different conditions: treatment for 8 min once, two times of 8 min, three times of 8 min, and 30 min once at room temperature (RT) and 30 min once at 37℃. After the detergent treatment, OD620 was measured after elution with 100 μ L 95% ethanol in crystal violet-stained wells. The assay value of OD620 was determined by subtracting the value of blank control cultured with TSB medium only from the original value. All OD620 values smaller than the value of blank control were regarded as 0. A single 8-min treatment with DW at RT was taken as the reference for 0% biofilm removal efficacy.
For each condition, the mean and standard deviation of three replicates were assigned to OD620. Using an independent samples t-test (two tailed), biofilm production ability was compared between strains and removal efficacy was compared between detergents. Microsoft Excel (Microsoft Corp., Redmond, WA, USA) was used for statistical analysis.
P. aeruginosa ATCC27853, K. pneumoniae AMC244, E. coli ATCC35218, E. coli AMC6615, E. faecalis AMC98901, S. aureus RN9120, C. albicans ATCC14053, C. auris AMC1704, and T. asahii AMC11511 produced significant biofilm production (Fig. 1). S. aureus RN9120 showed an OD620 < 0.5 when cultured with TSB. Because biofilm production of this strain was significantly increased in 4% NaCl-TSB (P = 0.017), but not in 4% ethanol-TSB (P = 0.170) (Fig. 2), it was cultured in 4% NaCl-TSB in biofilm production step when used to test removal efficacy.
Yuhanrox showed consistent biofilm removal efficacy of > 70% at 37°C incubation for all study isolates. Two enzymatic detergents had biofilm removal rates of greater than 50% for biofilm derived from P. aeruginosa ATCC27853, E. coli ATCC35218, E. coli AMC6615, C. albicans ATCC14053, and S. aureus RN9120 at 37°C for 30 min; 66%, 82%, 52%, 82%, and 71% by Cidezyme, and 13%, 80%, 69%, 92%, and 71% by Empower, respectively. For K. pneumoniae AMC244-derived biofilm, Empower showed the removal rates of 42%, 75%, and 55%, respectively, with treatment for 1, 2, and 30 min at RT, while 5% only when 30 min-treatment at 37°C. None of the detergents were effective enough to remove > 30% of biofilms derived from E. faecalis AMC98901, C. auris AMC1704, and T. asahii AMC11511. Matrix did not show significant removal efficacy against biofilm under any strain or condition (Fig. 3).
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It is not a human population study; therefore, approval by the institutional review board or the obtainment of informed consent is not required.
No potential conflicts of interest relevant to this article were reported.
None.