Impact of sealing method and layer number of autoclave bag on autoclave sterilization effect in the BSL-3 laboratory / 中华实验和临床病毒学杂志

Objective@#To evaluate, the method of different packing layer and sealing mode for the packaging of the infectious wastes in order to achieve better autoclave sterilization effect.@*Methods@#In the autoclave cycles P4 and P9, different packing layer (single, double and triple) and sealing mode (slightly folded, autoclave tape, nylon tape closure) were applied to the internal infectious waste sterilization, biological indicators were used to detect the effect of autoclave sterilization of the upper, middle and lower layers, respectively.@*Results@#1) In the autoclave cycle P4, the triple autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, the middle layer and the lower layer were respectively in the upper layer 100%, 0%, 0%, in the middle layer 67%, 20%, 0%, and in the lower layer was 0%. The double autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, the middle layer and the lower layer were respectively the in the upper layer 100%, 85%, 0%, the middle layer 45%, 0%, 0%, and the lower layer 10%, 0%, 0%. the single autoclave bag packaging sealing method wa based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were respectively the upper layer 100%, 100%, 0%, the middle layer 70%, 45%, 0%, and the lower layer 12.5%, 0%, 0%. (2) In the autoclave cycle P9, the triple autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, middle and lower layers were respectively the upper layer all 100%, the middle layer 100%, 100%, 70%, and the lower layer 100%, 100%, 30%. The double autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were respectively the upper layer all 100%, the middle layer all 100% and the lower layer is 100%, 100%, and 60%. The single autoclave bag packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were all 100%.@*Conclusions@#When autoclave is recommended for sterilization of infectious waste, the best effect was achieved with a single unsealed autoclave bag. If it is necessary to seal, the autoclave bag should not be completely sealed, avoiding incomplete penetration of steam into the interior space, otherwise the infectious waste can not be completely sterilized.

Establishment of pseudovirus infection mouse models for pharmacodynamics evaluation of filovirus entry inhibitors

Filoviruses cause severe and fatal viral hemorrhagic fever in humans. Filovirus research has been extensive since the 2014 Ebola outbreak. Due to their high pathogenicity and mortality, live filoviruses require Biosafety Level-4 (BSL-4) facilities, which have restricted the development of anti-filovirus vaccines and drugs. An HIV-based pseudovirus cell infection assay is widely used for viral entry studies in BSL-2 conditions. Here, we successfully constructed nine pseudo-filovirus models covering all filovirus genera and three pseudo-filovirus-infection mouse models using Ebola virus, Marburg virus, and Lloviu virus as representative viruses. The pseudo-filovirus-infected mice showed visualizing bioluminescence in a dose-dependent manner. A bioluminescence peak in mice was reached on day 5 post-infection for Ebola virus and Marburg virus and on day 4 post-infection for Lloviu virus. Two known filovirus entry inhibitors, clomiphene and toremiphene, were used to validate the model. Collectively, our study shows that all genera of filoviruses can be well-pseudotyped and are infectious . The pseudo-filovirus-infection mouse models can be used for activity evaluation of anti-filovirus drugs. This sequential and evaluation system of filovirus entry inhibitors provides a secure and efficient platform for screening and assessing anti-filovirus agents in BSL-2 facilities.

Factors affecting the fit of respirators in the biosafety level 3 laboratory / 中华实验和临床病毒学杂志

Objective@#To test the performance of two kinds of protective respirators (3M9332, 3M1860) used in Biosafety Level 3 Laboratory (BSL-3) to ensure whether it can fit well in different populations in order to reduce the risk of infection in the process of operating high-risk pathogenic microorganisms.@*Methods@#Using a 8038PortaCount®Pro+ respirator fit tester to examine the fitness of personnel wear 3M 1860 N95 and 9332 N99 respirators. The influence factors such as gender, age, shape of respirator were evaluated. Statistical analysis was performed by the fit factor(FF) of assigned actions.@*Results@#A total of 62 people conducted the respirator fit test, of which 45 people passed the test of 3M9332 respirators, the pass rate was 72.58%; only 6 people passed the test of 3M1860 respirators, the pass rate was 9.67%. The pass rate of two different types of respirators was analyzed statistically, P=0.000. The P value of assigned actions in deep breathing, head side to side, head up and down, talking, bending over were respectively 0.094, 0.076, 0.542, 0.000, 0.000.@*Conclusions@#The experimental results showed that the type of respirator had a significant effect on respirator fit in this study, but gender and age had no significant effect on factors. As to reduce the risk of leakage of the respirator, redundant actions should try to be avoided, especially loud talking and bending in the course of the experiment.

Avaliação dos conhecimentos e procedimentos em biossegurança de trabalhadores de laboratórios nível de biossegurança 3 / Knowledge of the procedures and evaluation in biosafety laboratory workers biosafety level 3

A Biossegurança constitui um campo do conhecimento com limites amplos e em constante construção. Entretanto, no Brasil, sua evolução teve fraco acompanhamento, especialmente em relação aos conhecimentos e procedimentos dos profissionais e infraestrutura dos laboratórios nível de Biossegurança 3, com poucos dados que possam indicar o grau de desenvolvimento e aplicabilidade de suas normas nas diversas instituições de saúde. Contudo, não bastam a adequação e funcionabilidade da infraestrutura predial, mas sim o conhecimento das necessidades dos usuários no sentido de propiciar aceitação das normas de Biossegurança por meio da conscientização dos riscos levando à mudança na conduta e no comportamento. Para isto, torna-se imprescindível a elaboração de treinamentos teóricos e práticos com atualizações constantes dos profissionais da área de saúde que deverão ser avaliados periodicamente para que haja melhorias nos programas de treinamentos oferecidos, uma vez que esses programas visam a garantir a qualidade e a segurança dos profissionais, dos ambientes internos e externos, à população em geral e a qualidade dos resultados pretendidos. Neste contexto, este trabalho teve, como objetivo principal, avaliar, a partir da pesquisa descritiva com abordagem qualitativa e exploratória, os dados coletados por questionários e entrevistas estruturadas, aplicadas aos 37 profissionais e nove gestores dos seis laboratórios NB3 participantes. A análise dos dados foi feita pela abordagem da multirreferencialidade, considerando os conhecimentos e procedimentos em Biossegurança e os possíveis impactos das estratégias dos últimos treinamentos oferecidos a esses profissionais, assim como as condições das instalações laboratoriais no país...

Biosafety is a field of knowledge with wide limits and constant construction. However, in Brazil, its evolution had weak monitoring, especially in relation to knowledge and professional procedures and infrastructure of Biosafety Level 3 laboratories, with little data that may indicate the degree of development and applicability of its norms in the various health institutions. However, it is not enough the adequacy and functionality of building infrastructure, but the knowledge of users' needs in order to provide accepted standards of biosafety through awareness of risks leading to change in the conduct and behavior. For this, it is essential the development of theoretical and practical trainings with constant updates of professionals in the health area who should be evaluated periodically with objectives of improvements in the training programs offered, since these programs aim to ensure the quality and safety of professionals, internal and external environments, the general population and the quality of intended results. In this context, this work has as main goal, to evaluate, from the descriptive and exploratory qualitative research approach, the data collected by questionnaires and structured interviews, applied to 37 professionals and nine managers of the six participating BSL3 laboratories. Data analysis was performed by the multi-referential approach, considering the knowledge and procedures on Biosafety and the possible impacts of the strategies of the last training offered to these professionals, as well as the conditions of laboratory facilities in the country...

Analysis on job stress assessment in BSL-3 laboratories / 中国比较医学杂志

Objective The staffs of biosafety level 3 laboratories (BSL-3) face with the stress of handling highly pathogenic microbs and special laboratory environment.The job stress may result in accidents in the laboratory as negative factor for the risk control.The research may provide support for the control of risk in biosafety laboratories.Methods In order to assess the job stress in the staff in BSL-3 laboratory, we modified “the Chinese simple job stress questionnaire”based on the theory of the JDC mode and ERI mode, and an investigation was carried out.The present study included the staffs (87 employees) from six BSL-3 laboratories located in five provinces ( Shanghai, Zhejiang, Jiangsu, Fujian and Wuhan) .Results Analysis of the data indicates that variables of age, working years, job duties, manipulating of animals, type of microorganisms and transmission route have a significant influence on the level of job stress in BSL-3 laboratory.Conclusion The BSL-3 laboratory staff in higher stress level have the characteristicses:20-39 years old, short work years, regular staff, operating on air-borne microbiology, manipulating of animals and operating on one more microbiology.