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Biosafety devices to control the spread of potentially contaminated dispersion particles. New associated strategies for health environments.

ABSTRACT: Dental procedures produce a large amount of spatter and aerosols that create concern for the transmission of airborne diseases, such as Covid-19. This study established a methodology with the objective of evaluating new associated strategies to reduce the risk of cross-transmission in a health environment by simulating spread of potentially contaminated dispersion particles (PCDP) in the environment. This crossover study, was conducted in a school clinic environment (4 clinics containing 12 dental chairs each). As a positive control group (without barriers), 12 professionals activated at the same time the turbine of dental drill, for one minute, with a bacterial solution (Lactobacillus casei Shirota, 1.5x108 CFU/mL), which had been added in the cooling reservoir of the dental equipment. In the experimental groups, the professionals made use of; a) an individual biosafety barrier in dentistry (IBBD) which consists of a metal support covered by a disposable PVC film barrier; b) a Mobile Unit of Disinfection by Ultraviolet-C, consisting of 8 UV lamps-C of 95W, of 304μW/cm2 of irradiance each, connected for 15 minutes (UV-C) and; c) the association between the two methods (IBBD + UV-C). In each clinic, 56 Petri dishes containing MRS agar were positioned on the lamps, benches and on the floor. In addition, plates were placed prior to each test (negative control group) and plates were also placed in the corridor that connects the four clinics. In the groups without barrier and IBBD + UV-C the passive air microorganisms in Petri dishes was also evaluated at times of 30, 60, 90 and 120 minutes after the end of the dental's drill activation. The mean (standard deviation) of CFU of L. casei Shirota for the positive control group was 3905 (1521), while in the experimental groups the mean using the IBBD was 940 (466) CFU, establishing a reduction on average, of 75% (p<0.0001). For the UV-C group, the mean was 260 (309) CFU and the association of the use of IBBD + UV-C promoted an overall average count of 152 (257) CFU, establishing a reduction on average of 93% and 96%, respectively (p<0.0001). Considering these results and the study model used, the individual biosafety barrier associated with UV-C technology showed to be efficient strategies to reduce the dispersion of bioaerosols generated in an environment with high rate of PCDP generation and may be an alternative for the improvement of biosafety in different healthy environment.

SUBMITTER: Montalli VAM

PROVIDER: S-EPMC8389494 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Biosafety devices to control the spread of potentially contaminated dispersion particles. New associated strategies for health environments.

Montalli Victor Angelo Martins VAM Freitas Patrícia Rejane de PR Torres Milenna de Figueiredo MF Torres Junior Oscar de Figueiredo OF Vilhena Dienne Hellen Moutinho De DHM Junqueira José Luiz Cintra JLC Napimoga Marcelo Henrique MH

PloS one 20210826 8

Dental procedures produce a large amount of spatter and aerosols that create concern for the transmission of airborne diseases, such as Covid-19. This study established a methodology with the objective of evaluating new associated strategies to reduce the risk of cross-transmission in a health environment by simulating spread of potentially contaminated dispersion particles (PCDP) in the environment. This crossover study, was conducted in a school clinic environment (4 clinics containing 12 dent ...[more]

PMID: 34437589

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Project description:BackgroundContamination of vertebrate cell lines with animal retroviruses has been documented repeatedly before. Although such viral contaminants can be easily identified with high sensitivity by PCR, it is impossible to screen for all potential contaminants. Therefore, we explored two novel methods to identify viral contaminations in cell lines without prior knowledge of the kind of contaminant.ResultsThe first hint for the presence of contaminating retroviruses in one of our cell lines was obtained by electron microscopy of exosome-like vesicles released from the supernatants of transfected 293T cells. Random amplification of particle associated RNAs (PAN-PCR) from supernatant of contaminated 293T cells and sequencing of the amplicons revealed several nucleotide sequences showing highest similarity to either murine leukemia virus (MuLV) or squirrel monkey retrovirus (SMRV). Subsequent mass spectrometry analysis confirmed our findings, since we could identify several peptide sequences originating from monkey and murine retroviral proteins. Quantitative PCRs were established for both viruses to test currently cultured cell lines as well as liquid nitrogen frozen cell stocks. Gene fragments for both viruses could be detected in a broad range of permissive cell lines from multiple species. Furthermore, experimental infections of cells negative for these viruses showed that both viruses replicate rapidly to high loads. We decided to further analyze the genomic sequence of the MuLV-like contaminant virus. Surprisingly it was neither identical to MuLV nor to the novel xenotropic MuLV related retrovirus (XMRV) but showed 99% identity to a synthetic retrovirus which was engineered in the 1980s.ConclusionThe high degree of nucleotide identity suggests unintended spread of a biosafety level 2 recombinant virus, which could also affect the risk assessment of gene-modified organisms released from contaminated cell cultures. The study further indicates that both mass spectrometry and PAN-PCR are powerful methods to identify viral contaminations in cell lines without prior knowledge of the kind of contaminant. Both methods might be useful tools for testing cell lines before using them for critical purposes.

Project description:IntroductionTo ensure the sterility of cell products that cannot undergo conventional sterilization processes, it is imperative to establish and maintain a clean room environment, regulated through environmental monitoring, including particle counts. Nevertheless, the impact of particles generated by operators as potential contaminants remains uncertain. Thus, in this study, we conducted an accelerated test to assess the correlation between particles generated by operators and airborne bacteria, utilizing biosafety cabinets within a typical laboratory setting. These biosafety cabinets create a controlled environment with air conditioning and high-efficiency particulate air (HEPA) filters, offering fundamental data relevant to cell production.Materials and methodsWe conducted a simulation followed by real-time experiments involving human operations to explore the quantity of particles, particle sizes, and the percentage of bacteria within these particles. This investigation focused on conditions with heightened particle generation from operators within a biosafety cabinet. The experiment was conducted on operators wearing textile and non-woven dustless clothing within biosafety cabinets. It entailed tapping the upper arms for a duration of 2 min.ResultsObservations under biosafety cabinet-off conditions revealed the presence of various particles and falling bacteria in textile clothing. In contrast, no particles or falling bacteria were detected in operators wearing dustless clothing within biosafety cabinets. Notably, a correlation between 5 μm particles and colony-forming units in textile clothing was identified through this analysis. The ratio of falling bacteria to the total number of particles within the biosafety cabinet was 0.8 ± 0.5 % for textile clothing, while it was significantly lower at 0.04 ± 0.2 % for dustless clothing.ConclusionThis study demonstrated that the number of particles and falling bacteria varied depending on the type of clothing and that quantitative data could be used to identify risks and provide basic data for operator education and evidence-based control methods in aseptic manufacturing areas. Although, this study aims to serve as an accelerated test operating under worst-case conditions, the results need to make sure the study range in general research.

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Biosafety devices to control the spread of potentially contaminated dispersion particles. New associated strategies for health environments.

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Biosafety devices to control the spread of potentially contaminated dispersion particles. New associated strategies for health environments.

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