Investigação sobre os métodos de assepsia e equipamentos de proteção individual utilizados por manicures e pedicures / Research on asepsy methods and personal protective equipment used by manicurs and pedicurers / Investigación sobre métodos de asepsia y equipos de protección individual utilizados por manicuras y pedicuras

Introdução: As atividades laborais de manicures e pedicures geram riscos de exposições acidentais aos patógenos vinculados ao sangue como vírus, fungos e bactérias, devido à falta de adesão às medidas de biossegurança, e procedimentos de esterilização acarretando riscos à saúde dos profissionais e clientes. Objetivo:Verificar os conhecimentos das manicures e pedicures sobre os métodos de limpeza e esterilização de materiais, o uso de equipamentos de proteção individual e doenças relacionadas à profissão. Metodologia:Estudo transversal de abordagem quantitativa, realizado entre Outubro de 2017 a Janeiro de 2018, com 22 participantes finais. Incluídos os maiores de 18 anos, ambos os sexos, experiência mínima de um ano e residir em Nova Floresta. A técnica para seleção da amostra foi snowball. Resultados: A limitação de conhecimento sobre os riscos laborais na exposição de agentes infectantes, a baixa adesão no uso de equipamentos de proteção individuale a falta de padronização no processo de descontaminação, limpeza, desinfecção e esterilização dos instrumentais estão associados ao potencial risco de contaminação por agentes infecciosos, e ao acometimento de doenças infectocontagiosas por manicures, pedicures e clientela. Conclusões:Existem fragilidades no conhecimento destes profissionais,a maioria expõe métodos de antissepsia precários, devido a falta de sistematização e padronização dos procedimentos de limpeza, descontaminação dos instrumentais e o uso de equipamentos de proteção individual, além do conhecimento da profissão ter sido adquirido informalmente (AU).

Introduction: The work activities of manicurists and pedicurists generate risks of accidental exposure to pathogens linked to the blood such as viruses, fungi and bacteria, due to the lack of adherence to biosafety measures, and sterilization procedures causing risks to the health of professionals and clients. Objective:To verify the knowledge of manicurists and pedicurists on methods of cleaning and sterilizing materials, the use of personal protective equipment and diseases related to the profession. Methodology: Cross-sectional study with a quantitative approach, carried out between October 2017 and January 2018, with 22 final participants. Included are those over 18 years of age, both sexes, with at least one year's experience and residing in Nova Floresta. The technique for sample selection was snowball.Results:Limited knowledge about occupational risks in the exposure of infectious agents, low adherence to the use of personal protective equipmentand the lack of standardization in the process of decontamination, cleaning, disinfection and sterilizationof instruments are associated with the potential risk of contamination by infectious agents, and the spread of infectious diseases by manicurists, pedicurists and clientele. Conclusions:There are weaknesses in the knowledge of these professionals, most expose precarious antisepsis methods, due to the lack of systematization and standardization of cleaning procedures, decontamination of instruments and the use of personal protective equipment, in addition to the knowledge of the profession having been acquired informally (AU).

Introducción: Las actividades laborales de manicuras y pedicuras generan riesgos de exposiciónaccidental a patógenos ligados a la sangre como virus, hongos y bacterias, debido al incumplimiento de medidas de bioseguridad y procedimientos de esterilización generando riesgos a la salud de profesionales y clientes. Objetivo:Verificar los conocimientos de manicuras y pedicuras sobre métodos de limpieza y esterilización de materiales, el uso de equipos de protección personal y enfermedades relacionadas con la profesión. Metodología:Estudio transversal de abordaje cuantitativo, realizado entre octubre de 2017 y enero de 2018, con 22 participantes finales. Se incluyen los mayores de 18 años, de ambos sexos, con experiencia mínima de un año y vivir en Nova Floresta. La técnica para la selección de la muestra fue la de bola de nieve. Resultados:El conocimiento limitado sobre los riesgos laborales en la exposición de agentes infecciosos, la baja adherencia al uso de equipo de protección personal y la falta de estandarización en el proceso de descontaminación, limpieza, desinfección y esterilización de instrumentos se asocian con el riesgo potencial de contaminación por agentes infecciosos, y la propagación de enfermedades infecciosas entre manicuras, pedicuras y clientela. Conclusiones:Existen debilidades en el conocimiento de estos profesionales, la mayoría expone métodos de antisepsia precarios, debido a la falta de sistematización y estandarización de los procedimientos de limpieza, descontaminación de los instrumentos y el uso de equipo de protección Personal,además de haber adquirido los conocimientos de la profesión informalmente (AU).

Desinfección y esterilización en odontología frente al COVID-19 / Disinfection and sterilization in dentistry against COVID-19 / Desinfecção e esterilização em odontologia contra a COVID-19

A partir de la declaración de la Organización Mundial de la Salud del comienzo de la pandemia COVID-19 causada por el virus SARS-CoV-2 en marzo de 2020, los profesionales de la salud se vieron expuestos a esta enfermedad altamente contagiosa y potencialmente mortal que generó múltiples desafíos a toda la comunidad científica. Provocando cambios de paradigmas en la atención de los pacientes y en el uso de las barreras de protección personal. A nivel mundial se crearon múltiples protocolos para la atención odontológica a medida que se iba desarrollando e investigando el comportamiento del virus. Esta revisión bibliográfica resume las indicaciones y recomendaciones basadas en las evidencias disponibles para disminuir las posibilidades de contaminación ante la exposición a este virus, incluyendo medidas a utilizar desde el ingreso del paciente, los métodos de protección personal, la descontaminación y esterilización del material, así como también la desinfección del área de trabajo. Aunque se ha hecho un gran esfuerzo por mejorar los procesos de bioseguridad a nivel científico tecnológico, hay evidencias de que el factor humano sigue siendo el eslabón más débil de esta cadena.

Since the declaration by the World Health Organization of the beginning of the COVID-19 pandemic caused by the SARS-CoV-2 virus in March 2020, health professionals were exposed to this highly contagious and potentially fatal disease that generated multiple challenges to the entire scientific community. It caused paradigm shifts in patient care and in the use of personal protective barriers. Multiple protocols for dental care were created worldwide as the behavior of the virus was developed and investigated. This bibliographic review summarizes the indications and recommendations based on the available evidence to reduce the possibilities of contamination when exposed to this virus, including measures to be used from patient admission, personal protection methods, decontamination and sterilization of material, as well as disinfection of the work area. Although a great effort has been made to improve biosafety processes at the scientific and technological level, there is evidence that the human factor continues to be the weakest link in this chain.

Desde a declaração pela Organização Mundial da Saúde do início da pandemia de COVID-19 causada pelo vírus SARS-CoV-2 em março de 2020, os profissionais de saúde foram expostos a essa doença altamente contagiosa e potencialmente fatal, que criou vários desafios para toda a comunidade científica. Ela causou mudanças de paradigma no atendimento ao paciente e no uso de barreiras de proteção individual. Em todo o mundo, vários protocolos para atendimento odontológico foram criados à medida que o comportamento do vírus foi desenvolvido e pesquisado. Esta revisão da literatura resume as indicações e recomendações baseadas em evidências para reduzir a probabilidade de contaminação por exposição a esse vírus, incluindo medidas a serem usadas desde a admissão do paciente, métodos de proteção individual, descontaminação e esterilização de equipamentos, bem como desinfecção da área de trabalho. Embora muitos esforços tenham sido feitos para melhorar os processos de biossegurança em nível científico e tecnológico, há evidências de que o fator humano continua sendo o elo mais fraco dessa cadeia.

Analysis of alternate material Onyx™ for total knee arthroplasty instrumentation sets.

CONTEXT: A 25-pound weight limit is currently set on containerized instrumentation sets by the Association of periOperative Registered Nurses (AORN) and the Association for the Advancement of Medical Instrumentation (AAMI), in order to reduce strain on the staff and ensure that the sets are not too crowded in order to preserve sterilization and drying of the instruments. This is pushing companies to reduce the weight and number of instrumentation sets for the operating room. One solution has been to explore the viability of new, lighter materials such as Onyx. OBJECTIVES: The goal of this study is to evaluate the novel material Onyx as a viable material utilized in reusable total knee arthroplasty (TKA) instrumentation sets utilizing traditional steam sterilization as the sterilization method. METHODS: Mechanical and biocompatibility tests according to the American Society for Testing Materials (ASTM) and International Organization for Standardization (ISO) 10,993 were run to evaluate the Onyx Material to see if it would be a viable alternative to the stainless and martensitic steel that is currently being utilized. RESULTS: Gross warping and cracking after 10 rounds of sterilization was observed. This was qualitatively worse in the Onyx without a carbon fiber component. The Onyx material did not meet biocompatibility standards for its application. CONCLUSIONS: Onyx was determined to not be a viable material for TKA instruments regarding multiple high-pressure and -temperature sterilizations and cytotoxic cell testing.

Maintenance of sterility in SMS packaging in dental environments: concern with biosafety

Healthcare services must be guided by biosafety practices and microbial control. This control is highly influenced by humidity, which directly impacts the maintenance of sterility of the materials used in the appointments. High concentration of moisture, in the form of aerosol, splashes and spills, is caused during dental care. During the COVID-19 times the contamination by aerosol and droplets worries greatly. Considering that it could cause harm to the sterility of an autoclaved material, especially in dental environments, the objective was to evaluate the behavior of SMS sterilization packages (Spunbonded / Meltblown / Spunbonded) against microbial penetration in an aqueous vehicle. SMS of three brands were challenged, equally divided into two groups: virgin and processed (subjected to a single autoclaving cycle). Each specimen was aseptically deposited on Macconkey agar. Subsequently, 5 µL of Escherichia coliATCC 25922 saline solution [108CFU mL-1] was deposited in center of the SMS specimen and the dish incubated at36°C/ 48h. Reading was performed by the presence or absence of bacterial growth typical of the species under the SMS, observed on the back of Petri dish. The lowest penetration rate observed was 60% for one of the brands in the virgin condition, and 75% for two brands in the processed condition. Statistical analysis showed an association between bacterial penetration and the evaluated group, this association being valid only in the virgin condition. The different SMS behave similarly in terms of resistance to bacterial penetration after being processed. The data show that moisture can assist in bacterial transport through sterilized SMS. Therefore, SMS packages are not able to prevent bacterial penetration, and possibly other microorganisms, when in aqueous vehicles, offering a potential risk of breaking the aseptic chain. Thus, care must be taken in routines for handling and storage sterile packaging.

Investigation of SARS-CoV-2 inactivation using UV-C LEDs in public environments via ray-tracing simulation.

This paper proposes an investigating SARS-CoV-2 inactivation on surfaces with UV-C LED irradiation using our in-house-developed ray-tracing simulator. The results are benchmarked with experiments and Zemax OpticStudio commercial software simulation to demonstrate our simulator's easy accessibility and high reliability. The tool can input the radiant profile of the flexible LED source and accurately yield the irradiance distribution emitted from an LED-based system in 3D environments. The UV-C operating space can be divided into the safe, buffer, and germicidal zones for setting up a UV-C LED system. Based on the published measurement data, the level of SARS-CoV-2 inactivation has been defined as a function of UV-C irradiation. A realistic case of public space, i.e., a food court in Singapore, has been numerically investigated to demonstrate the relative impact of environmental UV-C attenuation on the SARS-CoV-2 inactivation. We optimise a specific UV-C LED germicidal system and its corresponding exposure time according to the simulation results. These ray-tracing-based simulations provide a useful guideline for safe deployment and efficient design for germicidal UV-C LED technology.

Assessment of laparoscopic instrument reprocessing in rural India: a mixed methods study.

BACKGROUND: Laparoscopy is a minimally-invasive surgical procedure that uses long slender instruments that require much smaller incisions than conventional surgery. This leads to faster recovery times, fewer post-surgical wound infections and shorter hospital stays. For these reasons, laparoscopy could be particularly advantageous to patients in low to middle income countries (LMICs). Unfortunately, sterile processing departments in LMIC hospitals are faced with limited access to equipment and trained staff which poses an obstacle to safe surgical care. The reprocessing of laparoscopic devices requires specialised equipment and training. Therefore, when LMIC hospitals invest in laparoscopy, an update of the standard operating procedure in sterile processing is required. Currently, it is unclear whether LMIC hospitals, that already perform laparoscopy, have managed to introduce updated reprocessing methods that minimally invasive equipment requires. The aim of this study was to identify the laparoscopic sterile reprocessing procedures in rural India and to test the effectiveness of the sterilisation equipment. METHODS: We assessed laparoscopic instrument sterilisation capacity in four rural hospitals in different states in India using a mixed-methods approach. As the main form of data collection, we developed a standardised observational checklist based on reprocessing guidelines from several sources. Steam autoclave performance was measured by monitoring the autoclave cycles in two hospitals. Finally, the findings from the checklist data was supported by an interview survey with surgeons and nurses. RESULTS: The checklist data revealed the reprocessing methods the hospitals used in the reprocessing of laparoscopic instruments. It showed that the standard operating procedures had not been updated since the introduction of laparoscopy and the same reprocessing methods for regular surgical instruments were still applied. The interviews confirmed that staff had not received additional training and that they were unaware of the hazardous effects of reprocessing detergents and disinfectants. CONCLUSION: As laparoscopy is becoming more prevalent in LMICs, updated policy is needed to incorporate minimally invasive instrument reprocessing in medical practitioner and staff training programmes. While reprocessing standards improve, it is essential to develop instruments and reprocessing equipment that is more suitable for resource-constrained rural surgical environments.

Escovas de limpeza em centrais de materiais e esterilização: características e cuidados / Cleaning brushes in Central Materials and Sterilization

A limpeza é a primeira e mais essencial etapa antes que qualquer processo de desinfecção ou esterilização possa ocorrer. Nesta etapa ocorre a fricção manual dos materiais com escovas apropriadas a fim de eliminar sujidades. A depender das características, estas escovas podem ter um custeio elevado para os serviços de saúde, especialmente em países com recursos limitados como o Brasil. Elucidar, em tempo oportuno, o que traz a literatura sobre as características necessárias às escovas de limpeza em Centrais de Materiais e Esterilização (CME), especialmente sobre a necessidade de serem autoclavadas.

Electron Beam Sterilization of Poly(Methyl Methacrylate)-Physicochemical and Biological Aspects.

Electron beam (E-beam) irradiation is an attractive and efficient method for sterilizing clinically implantable medical devices made of natural and/or synthetic materials such as poly(methyl methacrylate) (PMMA). As ionizing irradiation can affect the physicochemical properties of PMMA, understanding the consequences of E-beam sterilization on the intrinsic properties of PMMA is vital for clinical implementation. A detailed assessment of the chemical, optical, mechanical, morphological, and biological properties of medical-grade PMMA after E-beam sterilization at 25 and 50 kiloGray (kGy) is reported. Fourier transform infrared spectroscopy, thermogravimetric analysis, and differential scanning calorimetry studies indicate that E-beam irradiation has minimal effect on the chemical properties of the PMMA at these doses. While 25 kGy irradiation does not alter the mechanical and optical properties of the PMMA, 50 kGy reduces the flexural strength and transparency by 10% and 2%, respectively. Atomic force microscopy demonstrates that E-beam irradiation reduces the surface roughness of PMMA in a dose dependent manner. Live-Dead, AlamarBlue, immunocytochemistry, and complement activation studies show that E-beam irradiation up to 50 kGy has no adverse effect on the biocompatibility of the PMMA. These findings suggest that E-beam irradiation at 25 kGy may be a safe and efficient alternative for PMMA sterilization.

Performance evaluation of chemical, biological and physical indicators in the process of sterilization under the effect of non-condensable gases.

BACKGROUND: The risk concerning the presence of non-condensable gases (NCGs) has already been demonstrated, but routine monitoring still requires further research to be implemented in each sterilization cycle. AIM: Performance evaluation of the physical, chemical and biological indicators used in monitoring in comparison with a sterilizer integrated detector for NCG in the Sterilization Process. METHODS: Chemical indicators (type 2 Bowie-Dick test, type 5 and type 6 models), self-contained biological indicators and physical indicators (temperature, pressure, thermal qualification and a patented integrated air detector) were used to monitor the steam sterilization process in two situations of controlled failure: chamber leakage and door seal failure. This controlled failure was obtained by the presence of a known amount of air: 0-30 L/min for chamber leakage and 0-30% for the door seal failure. Evaluation tests were carried out with and without the use of process challenge devices (PCDs). FINDINGS: In both studies, the Bowie-Dick Test showed different results, according to the manufacturer. The biological, physical or chemical indicators without a PCD were unable to detect small volumes of NCGs in both simulations. CONCLUSION: The integrated air detector can be considered an option for the detection of NCGs in each cycle.

A comparative study of four Bowie-Dick test under the condition of pressure steam sterilizer simulating gas leakage.

We aimed to understand the evaluation of different Bowie-Dick test (B-D test) on the performance of pressure steam sterilization equipment in the case of simulated gas leakage, and we selected a pulsating vacuum steam sterilizer to set 4 different gas leakage levels: 1.1, 1.3, 1.5, and 1.7 mbar/min during the B-D test phase. In terms of methods, 4 different brands of B-D test kits (devices) were tested at 4 different leakage rates, and a total of 48 experiments were conducted. The results from univariate analysis revealed that there are statistically significant differences in the judgment of test results among different personnel and brands. The results from multivariate logistic regression analysis displayed that the difference between different personnel was statistically significant (χ = 45.34, P < .001); the difference between different products was statistically significant (χ = 129.37, P < .001); and there was no statistically significant difference between different degree of leakage (χ = 6.99, P > .05). Result judgments of brand 1 and brand 2 are susceptible to subjective factors. The judgment of brand 3 is intuitive and consistent with the evaluation result of brand 4. In conclusion, the order of capacity to evaluate air leakage from best to worst is brand 4→brand 3→brand 1→brand 2.

Evaluation of Low-Temperature Sterilization using Hydrogen Peroxide Gas Containing Peracetic Acid.

In low-temperature sterilization for the medical field, hydrogen peroxide sterilization is widely used for its safety. However, its low penetrability and residual amount of sterilant are major concerns. Recently, the combination of hydrogen peroxide and peracetic acid has been found to enforce sporicidal effect, with low concentration in hydrogen peroxide. The application of this finding in medical sterilization is still very limited. To elucidate the combination effect, we compare peracetic acid containing hydrogen peroxide gas sterilizer and conventional hydrogen peroxide gas (plasma) sterilizers. The sterilant penetrability was examined in hollow load process challenge devices with inner diameters of 1 and 2 mm and lengths of 1, 2, and 3 m. As a result, peracetic acid containing hydrogen peroxide gas sterilizer demonstrated total inactivation with all diameters and lengths and achieved the highest sterilant penetrability in this study. The amount of residual sterilant on the surface of the sterilized object was 4.2 µg/cm2, which corresponds to half amount of those of conventional hydrogen peroxide gas sterilizers. These results suggest that the addition of peracetic acid to hydrogen peroxide gas sterilizer can enhance sterilization efficiency and safety.

Inactivation of Infectious Bacteria Using Nonthermal Biocompatible Plasma Cabinet Sterilizer.

Nonthermal, biocompatible plasma (NBP) is a promising unique state of matter that is effective against a wide range of pathogenic microorganisms. This study focused on a sterilization method for bacteria that used the dielectric barrier discharge (DBD) biocompatible plasma cabinet sterilizer as an ozone generator. Reactive oxygen species play a key role in inactivation when air or other oxygen-containing gases are used. Compared with the untreated control, Escherichia coli(E. coli), Staphylococcus aureus (S. aureus), and Salmonella typhimurium (sepsis) were inhibited by approximately 99%, or were nondetectable following plasma treatment. Two kinds of plasma sterilizers containing six- or three-chamber cabinets were evaluated. There was no noticeable difference between the two configurations in the inactivation of microorganisms. Both cabinet configurations were shown to be able to reduce microbes dramatically, i.e., to the nondetectable range. Therefore, our data indicate that the biocompatible plasma cabinet sterilizer may prove to be an appropriate alternative sterilization procedure.

Low-cost biofuel-powered autoclaving machine for use in rural health care centres.

Surgical site infections (SSIs) in developing countries have been linked to inadequate availability of sterilising equipment. Existing autoclaves are mostly unaffordable by rural healthcare practitioners, and when they managed to procure them, the electricity supply to power the autoclaves is epileptic. The solar-powered autoclave alternatives are too bulky with a very high initial cost. Hence, low-cost biofuel-powered autoclave becomes an attractive option, and this study sought to present the design, development and clinical evaluation of the device performance. With the global drive for the adoption of green energy, biofuel will not only reduce greenhouse gas emission but also provide revenue for local producers and reduce biomass associated health complications. The theoretical energy requirement for the sterilisation process was calculated. The standard pressure and temperature needed for sterilisation were tested to be 121 °C and 15 psi. The device was also clinically tested with Staphylococcus aureus bacteria obtained from the Department of Medical Microbiology and Parasitology, University of Ilorin Teaching Hospital using Brain heart Infusion Broth, MacConkey and Blood agar as cultured media. No bacteria growth was observed when the samples containing the bacteria colony were autoclaved by the designed autoclave and incubated at 37 °C for 2 d. Hence, the device met the mechanical and biological validation standards for effective sterilisation.

Gravity steam reprocessing in healthcare facilities for the reuse of N95 respirators.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has significantly impacted the health of millions of people around the world. The shortage of personal protective equipment, including N95 respirators, in hospital facilities has put frontline healthcare professionals at high risk for contracting this virus. AIM: To develop a reproducible and safe N95 respirator reprocessing method that satisfies all presented regulatory standards and that can be directly implemented by hospitals using existing available equipment. METHODS: A non-toxic gravity steam reprocessing method has been developed for the reuse of N95 respirators consisting of 30 min of steam treatment at 121°C followed by 30 min of heat drying. Samples of model number 1860, 1860s, 1870+, and 9105 N95 respirators were either collected from hospitals (for microbiology testing) or purchased new (for functionality testing), with all functionality tests (i.e. filter efficiency, fit evaluation, and strap integrity) performed at the Centers for Disease Control and Prevention using standard procedures established by the National Institute for Occupational Safety and Health. FINDINGS: All tested models passed the minimum filter efficiency of 95% after three cycles of gravity steam reprocessing. The 1870+ N95 respirator model is the most promising model for reprocessing based on its efficient bacterial inactivation coupled with the maintenance of all other key functional respirator properties after multiple reprocessing steps. CONCLUSIONS: The gravity steam method can effectively reprocess N95 respirators over at least three reprocessing cycles without negatively impacting the functionality requirements set out by regulators. Enabling the reuse of N95 respirators is a crucial tool for managing both the current pandemic and future healthcare crises.

Portable ozone sterilization device with mechanical and ultrasonic cleaning units for dentistry.

A portable device for cleaning and ozone sterilization of small-sized delicate dental instruments that cannot withstand the high heat and humidity of standard autoclaving has been developed. The device contains a remote unit for magnetic mechanical washing, an ultrasonic bath for pre-cleaning treatment, and a container for ozone sterilization with a reactor based on dielectric barrier discharge. The maximum ozone concentration in water reached 8.5 mg/l for 10 min of operation at a water temperature of 19 °C. The results of inactivation of Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus are presented. Distilled water with such seeded micro-organisms was bubbled with an ozone-air mixture at an ozone concentration of 5.8 mg/l at 17 °C for 5 min, 10 min, 20 min, and 30 min. The maximum bactericidal effect was manifested for Pseudomonas aeruginosa cells with their total elimination within 10 min. Inactivation of Escherichia coli bacteria was monitored after 20 min of treatment, and Staphylococcus aureus-after 30 min of treatment.

Using heat to kill SARS-CoV-2.

The current coronavirus pandemic has reached global proportions and requires unparalleled collective and individual efforts to slow its spread. One critically important issue is the proper sterilization of physical objects that have been contaminated by the virus. Here, we review the currently existing literature on thermal inactivation of coronavirus (SARS-CoV-2) and present preliminary guideless on temperatures and exposure durations required to sterilize. We also compare these temperatures/exposure durations with potential household appliances that may be thought capable of performing sterilization.

Survival of Escherichia coli O157 in autoclaved and natural sandy soil mesocosms.

While the soil microbiome may influence pathogen survival, determining the major contributors that reduce pathogen survival is inconclusive. This research was performed to determine the survival of E. coli O157 in autoclaved and natural (unautoclaved) sandy soils. Soils were inoculated with three different E. coli O157 strains (stx1+/stx2+, stx1-/stx2-, and stx1-/stx2+), and enumerated until extinction at 30°C. There was a significant difference in the survival of E. coli O157 based on soil treatment (autoclaved versus natural) at 30°C on days 1 (P = 0.00022), 3, (P = 2.53e-14), 7 (P = 5.59e-16), 14 (P = 1.072e-12), 30 (P = 7.18e-9), and 56 (P = 0.00029), with greater survival in autoclaved soils. The time to extinction (two consecutive negative enrichments) for all three strains was 169 and 84 days for autoclaved and natural soils, respectively. A separate E. coli O157 trial supplemented with 16S rRNA gene sequencing of the soil microbiome was performed at 15°C and 30°C on days 0, 7, 14, and 28 for each soil treatment. Greater species richness (Chao1, P = 2.2e-16) and diversity (Shannon, P = 2.2e-16) was observed in natural soils in comparison with autoclaved soils. Weighted UniFrac (beta-diversity) showed a clear distinction between soil treatments (P = 0.001). The greatest reduction of E. coli O157 was observed in natural soils at 30°C, and several bacterial taxa positively correlated (relative abundance) with time (day 0 to 28) in these soils (P < 0.05), suggesting that the presence of those bacteria might cause the reduction of E. coli O157. Taken together, a clear distinction in E. coli O157 survival, was observed between autoclaved and natural soils along with corresponding differences in microbial diversity in soil treatments. This research provides further insights into the bacterial taxa that may influence E. coli O157 in soils.

Test Process and Results of Potential Masking of Sterilizing-Grade Filters.

Pre-use/post-sterilization integrity testing (PUPSIT) has been a widely debated topic for the last several years. To a large extent, the debate is because of the fact that scientific data were not available to provide additional clarity that could inform appropriate risk-based judgements and commensurate actions. To gain clarity, the Parenteral Drug Association (PDA) and BioPhorum Organizations Group (BioPhorum) formed the Sterile Filtration Quality Risk Management (SFQRM) consortium in late 2017. The consortium goals have been to fill existing gaps in scientific data as adequately as possible with studies and industry guidance that would provide professionals and license holders with the ability to make informed decisions about appropriate risk-management strategies. This paper is one in a series of publications that are the result of the collaboration, and these should be considered together and viewed holistically in order to determine the best course of action with regard to PUPSIT. In total, the four papers cover the following areas: (1) data mining to determine the influence of fluid properties on integrity test values, (2) filter masking studies and results (this publication), (3) risk assessment and management from filter production to end use, and (4) points to consider in the best practice of the use of PUPSIT. In total, 25 manufacturers and filter suppliers have contributed to the work of the Consortium, deploying their filtration experts and pooling their collective knowledge and applied science experience to address these questions. This effort has also been supported by many independent experts currently available who have contributed to and driven the Filtration Interest Group in the PDA for many years. Both PDA and BioPhorum have prioritized this program and combined their approaches to deliver this comprehensive body of work. We hope that collectively the publications aid decision making and create greater certainty and confidence and above all alignment between suppliers, manufacturers, and regulators alike on these important questions.

Data Mining to Determine the Influence of Fluid Properties on the Integrity Test Values.

Eudralex volume 4, Annex 1, the European Union Good Manufacturing Practice for sterile products, requires that "The integrity of the sterilised filter should be verified before use" (1). Implicit in this requirement for a PUPSIT is the rationale that the sterilizing filter could sustain damage during sterilization or use (i.e., subsequent to any pre-use test conducted prior to sterilization), causing a defect which would not be detected by the post-use integrity ("masked" during filtration). To assess whether a filter defect could be masked by partial filter plugging, we evaluated the impact of the bacterial challenge test (BCT) on the bubble point (BP) of the test filters. The BP tests that are conducted before and after the BCT have been collected and compared for 2086 filters (1571 × test filters and 515 × control filters), representing 531 BCTs on 518 different pharmaceutical products, buffers, and in-process fluids. These tests comprise a cross section of fluids from multiple firms spanning the pharmaceutical and biotechnology industry. A posttest to pretest BP ratio was calculated for each filter and the distribution of these ratios examined to determine whether there were cases of elevation of the BP because of bacterial loading to the point where masking of a filter defect could occur; that is, if a defective filter could pass integrity testing due to apparent reduction in filter pore size because of the bacteria retained during the BCT. Ratios were averaged across all tests for the same test fluid. The mean average ratio was 1.00 ± 0.15, indicating that on the average, elevation of the BP does not occur. To assess the risk of masking a filter defect, observed BP ratios were compared to the ratio of the minimum BP specification of a 0.2 µm filter to that of a 0.45 µm filter of the same membrane type. The lowest such ratio for any membrane type was 1.33. A BP ratio equal to or higher than this ratio was considered a risk for masking, because a 0.45 µm filter could appear to meet the specifications of a 0.2 µm filter. Out of 518 average BP ratios, only eight fluids (1.5%) produced BP ratios meeting this criterion for a masking risk. Potential risk factors associated with these cases are discussed. We conclude that filtration processes producing BP changes sufficient to present a risk of masking defects are not common, and are detectable during the routine BCT. The BP ratios observed during routine BCT are one means to assess the potential of a given filtration process to mask defects and can be considered when determining whether a PUPSIT should be implemented.

Long-term Effects of 222-nm ultraviolet radiation C Sterilizing Lamps on Mice Susceptible to Ultraviolet Radiation.

Germicidal lamps that emit primarily 254 nm ultraviolet radiation (UV) are routinely utilized for surface sterilization but cannot be used for human skin because they cause genotoxicity. As an alternative, 222-nm UVC has been reported to exert sterilizing ability comparable to that of 254-nm UVC without producing cyclobutane pyrimidine dimers (CPDs), the major DNA lesions caused by UV. However, there has been no clear evidence for safety in chronic exposure to skin, particularly with respect to carcinogenesis. We therefore investigated the long-term effects of 222-nm UVC on skin using a highly photocarcinogenic phenotype mice that lack xeroderma pigmentosum complementation group A (Xpa-) gene, which is involved in repairing of CPDs. CPDs formation was recognized only uppermost layer of epidermis even with high dose of 222-nm UVC exposure. No tumors were observed in Xpa-knockout mice and wild-type mice by repetitive irradiation with 222-nm UVC, using a protocol which had shown to produce tumor in Xpa-knockout mice irradiated with broad-band UVB. Furthermore, erythema and ear swelling were not observed in both genotype mice following 222-nm UVC exposure. Our data suggest that 222-nm UVC lamps can be safely used for sterilizing human skin as far as the perspective of skin cancer development.