Representativeness of EN 1040/13727 Assay Conditions for Evaluating In Vitro the Bactericidal Activity of a Chlorhexidine Digluconate and Benzalkonium Chloride Antiseptic Preparation

The representativeness of the mandatory bacterial strains specified in European standards for in vitro assay of the bactericidal activity of antiseptics was evaluated by testing the activity of an antiseptic combining chlorhexidine digluconate 0.2% and benzalkonium chloride 0.5% against 21 additional bacterial strains, and the positive interaction between these two biocidal agents was assessed. Methods and Results: The bactericidal activity of the antiseptic solution used pure or diluted was assessed according to the European standards EN 1040 and EN 13727. The contact time was 1 min at 20°C. Interfering substances used in the EN 13727 assay were bovine serum albumin and sheep erythrocytes, simulating “dirty” conditions, and hard water. A reduction of colony-forming units by ≥5 log10 was deemed to meet the requirements to conclude bactericidal activity. Under “basic” conditions, the bactericidal activity of the antiseptic was observed against all four mandatory strains specified in the standards as well as against nearly all the additional strains tested, including most of those with acquired antibiotic-resistance. The positive interaction between the two biocidal agents was also confirmed. Under “dirty” conditions, the bactericidal activity of the antiseptic solution was maintained against all the mandatory strains and was reduced against only four of the additional strains tested. Conclusions: With regard to the antiseptic tested and under the experimental conditions described, bactericidal activity evidenced against the mandatory strains appeared to be representative of that manifested against a wide range of the main pathogenic bacteria. Reduced bacterial activity against some of the additional strains tested (e.g. Enterobacteriaceae) was observed under “dirty” conditions. Significance and Impact of the Study: EN 13727 with some experimental adjustments represents an additional appropriate standard that needs to be considered for mucocutaneous antiseptic assessment. However, it may be worth including other specific bacterial strains to those specified in the standard, when evaluating antiseptics intended for use in certain clinical situation

Bactericidal activity of 3 cutaneous/mucosal antiseptic solutions in the presence of interfering substances: improvement of the NF EN 13727 European Standard ?

Objective: there is no standard protocol for the evaluation of antiseptics used for skin and mucous membranes in the presence of interfering substances. Our objective was to suggest trial conditions adapted from the NF EN 13727 standard, for the evaluation of antiseptics used in gynecology and dermatology.Methods: three antiseptic solutions were tested in vitro: a chlorhexidine-benzalkonium (CB) combination, a hexamidine-chlorhexidine-chlorocresol (HCC) combination, and povidone iodine (P). The adaptation of trial conditions to the standard involved choosing dilutions, solvent, and interfering substances. The activity of solutions was assessed on the recommended strains at concentrations of 97% (pure solution), 50%, and 10% (diluted solution), and 1%. A logarithmic reduction ≥ 5 was expected after 60seconds of contact, to meet requirements of bactericidal activity. Results: HCC did not present any bactericidal activity except on P. aeruginosa at a concentration of 97%. P was not bactericidal on E. hirae at any concentration and on S. aureus at 97%. CB had the most homogeneous bactericidal activity with a reduction>5 log on the 4 bacterial strains at concentrations of 97%, 50% and 10%. Conclusion: adapting the NF EN 13727 standard allowed assessing the 3 tested solutions: only CB was bactericidal in dirty conditions. This study proved the possibility of validating antiseptic choice in vitro, in current practice conditions, for adjunctive treatment of skin and mucous membranes disorders, primarily of bacterial origin or with a potential of superinfection

Antimicrobial activity of metal oxide microspheres: an innovative process for homogeneous incorporation into materials

AIMS:To investigate the potent control of microbial surface contamination of an innovative process which consists in incorporating metal oxide microspheres homogeneously into materials. Spherical microspheres containing zinc and magnesium oxides synthesized via a one-step manufacturing process (Pyrolyse Pulvérisée® ) and incorporated into different plastic matrices were evaluated for their antimicrobial activity according to JIS Z 2801 standard. A significant activity was observed for microsphere-added polyethylene coupons with a reduction of all tested bacteria populations, including Gram negative and Gram positive even expressing acquired antibiotic resistance (Escherichia coli ESBL, Staphylococcus aureus metiR). An antiviral activity higher than 2 log of reduction was also observed on H1N1 and HSV-1 viruses. This antimicrobial effect was dose-dependent and time-dependent for both polyethylene and polypropylene matrices. Antimicrobial activity was maintained after exposition to disinfectants and totally preserved 50 months after the preparation of the coupons. CONCLUSIONS:Incorporated into plastic matrices, metal oxide microspheres showed significant antibacterial and antiviral activities. SIGNIFICANCE AND IMPACT OF STUDY: This is, to our knowledge, the first report on an original process incorporating metal oxide microspheres, which have specific physico-chemical and antimicrobial properties, into materials that could be used for surface contamination prevention

A Myrtus communis extract enriched in myrtucummulones and ursolic acid reduces resistance of Propionibacterium acnes biofilms to antibiotics used in acne vulgaris

Recent works present evidence of Propionibacterium acnes growing as a biofilm in cutaneous follicles. This formation of clusters is now considered as an explanation for the in vivo resistance of P. acnes to the main antimicrobials prescribed in acne vulgaris. Purpose : Our objective was to explore this hypothesis and propose a new therapeutic approach focusing on anti-biofilm activity of Myrtacine® New Generation (Mediterranean Myrtle extract–Botanical Expertise P. Fabre) alone or combined with antibiotics. Methods/Results : Using in vitro models able to promote the growth of adhered bacteria, the loss of sensitivity of P. acnes biofilms (48 h) towards erythromycin and clindamycin was checked considering either sensitive or resistant strains. In the same time, the activity of Myrtacine® New Generation against biofilm formation and mature biofilm (48 h) was evaluated. Using a dynamic model of biofilm formation, we noted an inhibition of biofilm formation (addition of Myrtacine® New Generation at T 0) and a significant effect on mature biofilm (48 h) for 5 min of contact. This effect was also checked using the static model of biofilm formation for Myrtacine® New Generation concentrations ranging from 0.03% to 0.0001%. A significant, dose-dependent anti-biofilm effect was observed and notable even at a concentration lower than the active concentration on planktonic cells, i.e. 0.001%. Finally, the interest of the combination of Myrtacine® New Generation with antibiotics was explored. An enhanced efficacy was noted when erythromycin (1000 mg/l) or clindamycin (500 mg/l) was added to 0.001% Myrtacine®, leading to significant differences in comparison to each compound used alone

Antimicrobial activity of metal oxide microspheres: an innovative process for homogeneous incorporation into materials

International audienceAIMS:To investigate the potent control of microbial surface contamination of an innovative process which consists in incorporating metal oxide microspheres homogeneously into materials. Spherical microspheres containing zinc and magnesium oxides synthesized via a one-step manufacturing process (Pyrolyse Pulvérisée® ) and incorporated into different plastic matrices were evaluated for their antimicrobial activity according to JIS Z 2801 standard. A significant activity was observed for microsphere-added polyethylene coupons with a reduction of all tested bacteria populations, including Gram negative and Gram positive even expressing acquired antibiotic resistance (Escherichia coli ESBL, Staphylococcus aureus metiR). An antiviral activity higher than 2 log of reduction was also observed on H1N1 and HSV-1 viruses. This antimicrobial effect was dose-dependent and time-dependent for both polyethylene and polypropylene matrices. Antimicrobial activity was maintained after exposition to disinfectants and totally preserved 50 months after the preparation of the coupons. CONCLUSIONS:Incorporated into plastic matrices, metal oxide microspheres showed significant antibacterial and antiviral activities. SIGNIFICANCE AND IMPACT OF STUDY: This is, to our knowledge, the first report on an original process incorporating metal oxide microspheres, which have specific physico-chemical and antimicrobial properties, into materials that could be used for surface contamination prevention

Demonstrating the In Vitro and In Situ Antimicrobial Activity of Oxide Mineral Microspheres: An Innovative Technology to Be Incorporated into Porous and Nonporous Materials

International audienceThe antimicrobial activity of surfaces treated with zinc and/or magnesium mineral oxide microspheres is a patented technology that has been demonstrated in vitro against bacteria and viruses. This study aims to evaluate the efficiency and sustainability of the technology in vitro, under simulation-of-use conditions, and in situ. The tests were undertaken in vitro according to the ISO 22196:2011, ISO 20473:2013, and NF S90-700:2019 standards with adapted parameters. Simulation-of-use tests evaluated the robustness of the activity under worst-case scenarios. The in situ tests were conducted on high-touch surfaces. The in vitro results show efficient antimicrobial activity against referenced strains with a log reduction of >2. The sustainability of this effect was time-dependent and detected at lower temperatures (20 ± 2.5 °C) and humidity (46%) conditions for variable inoculum concentrations and contact times. The simulation of use proved the microsphere’s efficiency under harsh mechanical and chemical tests. The in situ studies showed a higher than 90% reduction in CFU/25 cm2 per treated surface versus the untreated surfaces, reaching a targeted value of <50 CFU/cm2. Mineral oxide microspheres can be incorporated into unlimited surface types, including medical devices, to efficiently and sustainably prevent microbial contamination