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. 2020 Jun 19:12:29.
doi: 10.1186/s13099-020-00367-4. eCollection 2020.

Development of a highly effective low-cost vaporized hydrogen peroxide-based method for disinfection of personal protective equipment for their selective reuse during pandemics

Affiliations

Development of a highly effective low-cost vaporized hydrogen peroxide-based method for disinfection of personal protective equipment for their selective reuse during pandemics

Vikram Saini et al. Gut Pathog. .

Abstract

Background: Personal Protective Equipment (PPE) is required to safely work with biological agents of bacterial (i.e. Mycobacterium tuberculosis) or viral origin (Ebola and SARS). COVID-19 pandemic especially has created unforeseen public health challenges including a global shortage of PPE needed for the safety of health care workers (HCWs). Although sufficient stocks of PPE are currently available, their critical shortage may develop soon due to increase in demand and depletion of existing supply lines. To empower our HCWs and ensure their continued protection, proactive measures are urgently required to develop procedures to safely decontaminate the PPEs to allow their "selective reuse" during contingency situations.

Methods: Herein, we have successfully developed a decontamination method based on vaporized hydrogen peroxide (VHP). We have used a range of concentration of hydrogen peroxide to disinfect PPE (coveralls, face-shields, and N-95 masks). To ensure a proper disinfection, we have evaluated three biological indicators namely Escherichia coli, Mycobacterium smegmatis and spores of Bacillus stearothermophilus, considered as the gold standard for disinfection processes. We next evaluated the impact of repeated VHP treatment on physical features, permeability, and fabric integrity of coveralls and N-95 masks. Next, we performed Scanning Electron Microscopy (SEM) to evaluate microscopic changes in fiber thickness of N-95 masks, melt blown layer or coverall body suits. Considering the fact that any disinfection procedure should be able to meet local requirements, our study included various regionally procured N-95 masks and coveralls available at our institute All India Institute of Medical Sciences (AIIMS), New Delhi, India. Lastly, the practical utility of VHP method developed herein was ascertained by operationalizing a dedicated research facility disinfecting used PPE during COVID-19.

Results: Our prototype studies show that a single VHP cycle (7-8% Hydrogen peroxide) could disinfect PPE and PPE housing room of about 1200 cubic feet (length10 ft × breadth 10 ft × height 12 ft) in less than 10 min, as noted by a complete loss of B. stearothermophilus spore revival. The results are consistent and reproducible as tested in over 10 cycles in our settings. Further, repeated VHP treatment did not result in any physical tear, deformity or other appreciable change in the coverall and N-95 masks. Our permeation tests evaluating droplet penetration did not reveal any change in permeability post-VHP treatments. Also, SEM analysis indeed revealed no significant change in fiber thickness or damage to fibers of coveralls or melt blown layer of N-95 masks essential for filtration. There was no change in user comfort and experience following VHP treatment of PPE. Based on results of these studies, and parameters developed and optimized, an institutional research facility to disinfect COVID-19 PPE is successfully established and operationalized with more than 80% recovery rate for used PPE post-disinfection.

Conclusions: Our study, therefore, successfully establishes the utility of VHP to effectively disinfect PPE for a possible reuse as per the requirements. VHP treatment did not damage coveralls, cause physical deformity and also did not alter fabric architecture of melt blown layer. We observed that disinfection process was successful consistently and therefore believe that the VHP-based decontamination model will have a universal applicability and utility. This process can be easily and economically scaled up and can be instrumental in easing global PPE shortages in any biosafety facility or in health care settings during pandemic situation such as COVID-19.

Keywords: Bacillus stearothermophilus spores; COVID-19; Personal protective equipment; Vaporized hydrogen peroxide.

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Conflict of interest statement

Competing interestsNone declared.

Figures

Fig. 1
Fig. 1
Comparison of heat- and alcohol- based disinfection. aE. coli and bM. smegmatis bacterial cultures were treated with heat (70 °C and 80 °C for 5 and 10 min), ethanol (75% and 85% for 0.5 and 1 min) and propan-2-ol (75% and 85% for 0.5 and 1 min), followed by CFU plating on respective agar media. Untreated culture controls were plated in parallel [****p < 0.0001 obtained from one-way ANOVA with Bonferroni post-test correction]. c Representative image of B. stearothermophilus spore strips exposed to heat (90 °C/30 min), 85% ethanol (1 min) and 85% propan-2-ol (1 min) inoculated in BHI media. Each experiment was performed at least two times with three biological replicates
Fig. 2
Fig. 2
Effect of vaporized hydrogen peroxide on survival of biological indicators. a Pictorial depiction of the areas on the coverall inoculated with 107E. coli or 107 M. smegmatis or B. stearothermophilus spores. Blue circles indicate area of high propensity of exposure during clinical examination. After a cycle of VHP-based disinfection, bacteria were retrieved from the coveralls. CFU/ml are shown for the untreated and treated sets of bE. coli and cM. smegmatis plated on respective media. Swabs were taken from random surfaces in the room and plated. Data is represented as an average of at least 8 to 24 biological replicates. d Representative image of E. coli, M. smegmatis and B. stearothermophilus spores untreated vs VHP exposed, plated and inoculated on respective growth media. (N = 10). [****p < 0.0001 obtained from one-way ANOVA with Bonferroni post-test correction]
Fig. 3
Fig. 3
Analysis of integrity of selected brands of coveralls and N-95 masks following multiple cycles of VHP exposure by liquid permeation test. Using water droplets of different volume, we investigated the change in permeability status of coveralls arising due to multiple cycles of VHP exposure. a, b Coveralls. c, d N-95 masks. 50 µL (a, c) and 100 µL (b, d) water was seeded on selected brands of coveralls and N-95 masks. Hard non-permeable surface (petri plate surface) was used as control for evaporation. The time taken for water droplet disappearance from PPE surfaces was normalized against droplet evaporation time from the petri dish (i.e. natural evaporation time). Time comparable to or slightly higher to evaporation control in tested coveralls and N-95 masks indicate no permeation associated loss of droplet indicating integrity of PPE. Lack of availability of some suits limited a full range of analysis on some datasets. [Statistical evaluation done by Two-way ANOVA, with Bonferroni post-test correction]. Clearly, VHP treatment did not result into significant changes in permeability of PPE
Fig. 4
Fig. 4
Analysis of integrity of selected brands of coveralls and N-95 masks following multiple cycles of VHP exposure by scanning electron microscopy. a Representative SEM photomicrograph of a brand of coverall at 200X and 1000X magnification pre- and post-multiple cycles of VHP exposure. b Comparison of fibre width of selected brand of coverall, before and after VHP exposure. c Representative SEM photomicrograph of a brand of N-95 mask outer layer at 200X and 1000X magnification pre- and post-5 cycles of VHP exposure. d Comparison of fiber width of the outer layer of selected brands of N-95 masks, before and after VHP exposure. e Representative SEM photomicrograph of a N-95 mask’s melt blown layer at 200X and 1000X magnification, pre- and post- 5 cycles of VHP exposure. f Comparison of fiber width of the melt blown layer of selected brands of N-95 masks, pre- and post- VHP exposure [Statistical evaluation done by two-way ANOVA, with Bonferroni post-test]. No significant differences observed due to VHP treatment either in external layer of mask or in the melt blown layer of the N-95 mask. Similarly, there were no discernable changes observed in integrity of coveralls

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