High-level Biosafety Laboratories Research Articles

动物高等级生物安全实验室培训体系的构建

动物高等级生物安全实验室培训体系的构建

The Evolving Landscape of Biosafety and Biosecurity: A Review of International Guidelines and Best Practices

Biosafety and biosecurity encompass the intersection of bioengineering and biotechnology, along with the evaluation and control of risks to human, animal health, and the environment. This link encompasses the danger created by research and its use, as well as research and application to reduce risk via bioengineering and biotechnology. High-level biosafety laboratories provide a secure and reliable setting, integrating robust containment measures, well-educated staff, and precise biosafety protocols to safeguard scientists from infections while working with microbial pathogens. Simultaneously, they prevent the accidental release of these pathogens into the surrounding environment. In recent decades, there has been the construction and operation of labs with different tiers of protection, the establishment of legal regulations and a laboratory biosafety management system, and these functional labs have played a crucial role in addressing newly emerging and recurring infectious diseases. This review article offers useful insights into the complicated environment of biosafety and biosecurity at both the national and international levels. It advocates for more international cooperation among states, organizations, and stakeholders to ensure the effective implementation of biosafety and biosecurity measures, fostering a safer and more secure global biotechnology landscape.

Investigations of indoor environment of high level biosafety laboratory in China based on field measurements

The high-level biosafety laboratory is not only the basic support for infectious disease prevention and control, but also interrelated with key areas such as environmental security and social security, which has attracted increasing attention. A good indoor environment is the premise to ensure the smooth progress of the experiment and biological risk prevention and control. In order to better understand the indoor environment of high-level biosafety laboratories, 19 high-level biosafety laboratories in China (with a total of 65 main rooms) were carefully selected as the test objects from December 2020 to December 2022. According to the test methods specified in the Chinese standard GB 50346, the air change, cleanliness, static pressure difference, temperature, relative humidity, and illumination were tested and analyzed. The results showed that all the measured parameters met the requirements of the Chinese standard GB 50346, and the bio-safety performance was completely satisfactory. However, individual parameters showed some overlarge values: the proportion of main rooms with cleanliness levels of 7 and 8 exceeding 50% of the lower limit for air changes was 54.5% and 69.8%, respectively; the proportion of main rooms in BSL-3-b1 laboratories with atmospheric pressure differentials exceeding 50% of the standard lower limit was 94.7%; and the atmospheric pressure differential in the main rooms of BSL-3-b2 laboratories reached a maximum of nearly -160 Pa. On the premise of ensuring the cleanliness and pressure gradient of the main room, it may be possible to reduce the air change. This study, for the first time, reveals the environmental parameters of various types and levels of biosafety laboratories, which can provide reference for the design and operation of such facilities.

Discussion on whether circulating air can be used in high-level biosafety laboratories

Discussion on whether circulating air can be used in high-level biosafety laboratories

Application of Pseudotyped Viruses.

Highly pathogenic emerging and reemerging viruses have serious public health and socioeconomic implications. Although conventional live virus research methods can more reliably investigate disease pathogenicity and evaluate antiviral products, they usually depend on high-level biosafety laboratories and skilled researchers; these requirements hinder in vitro assessments of efficacy, as well as efforts to test vaccines and antibody drugs. In contrast, pseudotyped viruses (i.e., single-round infectious viruses that mimic the membrane structures of various live viruses) are widely used in studies of highly pathogenic viruses because they can be handled in biosafety level 2 facilities. This chapter provides a concise overview of various aspects of pseudotyped virus technologies, including (1) exploration of the mechanisms of viral infection; (2) evaluation of the efficacies of vaccines and monoclonal antibodies based on pseudovirion-based neutralization assay; (3) assessment of antiviral agents (i.e., antibody-based drugs and inhibitors); (4) establishment of animal models of pseudotyped virus infection in vivo; (5) investigation of the evolution, infectivity, and antigenicity of viral variants and viral glycosylation; and (6) prediction of antibody-dependent cell-mediated cytotoxic activity.

Cost-effective high-speed, three-dimensional live-cell imaging of HIV-1 transfer at the T cell virological synapse

The availability of cost-effective, highly portable, and easy to use high-resolution live-cell imaging systems could present a significant technological break-through in challenging environments, such as high-level biosafety laboratories or sites where new viral outbreaks are suspected. We describe and demonstrate a cost-effective high-speed fluorescence microscope enabling the live tracking of virus particles across virological synapses that form between infected and uninfected Tcells. The dynamics of HIV-1 proteins studied at the cellular level and the formation of virological synapses in living Tcells reveals mechanisms by which cell-cell interactions facilitate infection between immune cells. Dual-color 3D fluorescence deconvolution microscopy of HIV-1 particles at frames rates of 100 frames per second allows us to follow the transfer of HIV-1 particles across the Tcell virological synapse between living Tcells. We also confirm the successful transfer of virus by imaging Tcell samples fixed at specific time points during cell-cell virus transfer by super-resolution structured illumination microscopy.

Mitigation of the biosafety risks of SARS-CoV-2 at BSL-3 laboratories

Aim: The aim of this study is to reduce the biosafety risks of laboratory exhaust air due to SARS-CoV-2 at a biosafety level-3 laboratory used for vaccine development against COVID-19 pandemic. Materials and Methods: In this study, the infrastructure and the organisation of the containment laboratory, which aimed to be used to struggle with pandemic, was used for risk assessment. Assessment of the laboratory exhaust air procedure as a component of the heating-ventilation-air conditioning system, which is significant for high-level biosafety laboratories, was conducted through a risk assessment matrix.

The Use of Distinctive Monoclonal Antibodies in FMD VLP- and P1-Based Blocking ELISA for the Seromonitoring of Vaccinated Swine.

The serum neutralization (SN) test has been regarded as the “gold standard” for seroconversion following foot-and-mouth disease virus (FMDV) vaccination, although a high-level biosafety laboratory is necessary. ELISA is one alternative, and its format is constantly being improved. For instance, standard polyclonal antisera have been replaced by monoclonal antibodies (MAbs) for catching and detecting antibodies, and inactive viruses have been replaced by virus-like particles (VLPs). To the best of current knowledge, however, no researchers have evaluated the performances of different MAbs as tracers. In previous studies, we successfully identified site 1 and site 2 MAbs Q10E and P11A. In this study, following the established screening platform, the VLPs of putative escape mutants from sites 1 to 5 were expressed and used to demonstrate that S11B is a site 3 MAb. Additionally, the vulnerability of VLPs prompted us to assess another diagnostic antigen: unprocessed polyprotein P1. Therefore, we established and evaluated the performance of blocking ELISA (bELISA) systems based on VLPs and P1, pairing them with Q10E, P11A, S11B, and the non-neutralizing TSG MAb as tracers. The results indicated that the VLP paired with S11B demonstrated the highest correlation with the SN titers (R2 = 0.8071, n = 63). Excluding weakly positive serum samples (SN = 16–32, n = 14), the sensitivity and specificity were 95.65% and 96.15% (kappa = 0.92), respectively. Additionally, the P1 pairing with Q10E also demonstrated a high correlation (R2 = 0.768). We also discovered that these four antibodies had steric effects on one another to varying degrees, despite recognizing distinct antigenic sites. This finding indicated that MAbs as tracers could not accurately detect specific antibodies, possibly because MAbs are bulky compared to a protomeric unit. However, our results still provide convincing support for the application of two pairs of bELISA systems: VLP:S11B-HRP and P1:Q10E-HRP.

Evaluation and comparison of three virucidal agents on inactivation of Nipah virus

Modern human activity is profoundly changing our relationship with microorganisms with the startling rise in the rate of emerging infectious diseases. Nipah virus together with Ebola virus and SARS-CoV-2 are prominent examples. Since COVID-19 and the West African Ebola virus disease outbreak, different chemical disinfectants have been developed for preventing the direct spread of viruses and their efficacy has also been evaluated. However, there are currently no published efficacy studies for the chemical disinfection of Nipah virus. In this study, the virucidal efficacy of three disinfectants (Micro-Chem Plus detergent disinfectant cleaner, FWD and Medical EtOH) against Nipah virus was evaluated in quantitative suspension tests including. Our results showed that the > 4 log reduction achieved for all products in inactivating Nipah virus in 15 s. Even, 19% ethanol was able to inactivate Nipah virus when applied for at least 8 min contact time. Comparative analysis displayed virucidal efficacy of each of the evaluated disinfectants against SARS-CoV-2, Ebola virus and Nipah virus, with only minor differences in working concentrations and contact times required for complete inactivation. We expect that our study can assist in decontamination in healthcare settings and high level biosafety laboratories and can be beneficial to control for emerging enveloped viruses.

Study on contaminant distribution in a mobile BSL-4 laboratory based on multi-region directional airflow.

The outbreak of COVID-19 has caused increasing public attention to laboratory-acquired infections (LAIs), especially for a mobile Bio-Safety Level 4 Lab (BSL-4) with high potential of exposure. In this paper, the distribution and removal mechanism of bioaerosols in the biosafety laboratory were studied. A simulation model of airflow distribution in the opening and closing state of air-tight door was established and verified. The results showed that the airflow entrainment velocity during the opening of the door was approximately 0.12 m/s. It increased the probability of vortex generation in the laboratory. The deposition rate of particles was doubled when the air-tight door opening is compared with air-tight door closing. Besides, nearly 80% of the particles deposited on the surface of the wall and ceiling, increasing the possibility of LAIs. The findings of this paper could provide new scientific methods for high-level biosafety laboratories to avoid cross-infection. Moreover, future work regarding air-tight door rotation speed regulation and control should be emphasized.

Cost-Effective High-Speed, Three-Dimensional Live-Cell Imaging of HIV-1 Transfer at the T Cell Virological Synapse

The availability of cost-effective, highly portable and easy to use high resolution live-cell imaging systems could present a significant technological break-through in challenging environments, such as high-level biosafety laboratories or sites where new viral outbreaks are suspected. We describe and demonstrate a cost-effective high-speed fluorescence microscope enabling the tracking of virus particles across virological synapses that form between infected and uninfected T cells. The dynamics of HIV-1 proteins studied at the cellular level and the formation of live virological synapses reveals mechanisms by which cell-cell interactions facilitate infection between cells. Dual-color 3D fluorescence deconvolution microscopy of HIV-1 particles at frames rates of 100 frames per second allows us to follow the transfer of HIV-1 particles across the T cell virological synapse between living T cells. We confirm the successful transfer of virus by imaging T cell samples fixed at specific time points during cell-cell virus transfer by super-resolution structured illumination microscopy.

Basic requirements of dunk tank settings and verification of the disinfection effect in high-level biosafety laboratories

This study presents the structure and application of dunk tanks in high-level biosafety laboratories. The test methods and sterilization effects of dunk tanks were evaluated by using Bacillus subtilis as indicator. Results showed that sterilization effects of dunk tanks could be achieved by immersing the goods in 0.5% sodium hypochlorite disinfectant solution for 30 min at room temperature. Neutralizer sodium thiosulfate had no effect on the growth of Bacillus subtilis, and thus would not affect the disinfection results of sodium hypochlorite. Furthermore, suggestions for safe operation of dunk tanks are also provided in this study.

Laboratory biosafety in China: past, present, and future

The launch of the new journal, Biosafety and Health, presents me with a unique opportunity to recount the progress of laboratory biosafety (LB) in China and my contribution to this area over the past 30 years. Since the severe acute respiratory syndrome epidemic in 2003, China has constructed a primary network of high-level biosafety laboratories at different levels and established an expert team on LB. Furthermore, a series of LB management documents, including laws, regulations, standards, and guidelines, have been developed and published. This gradually maturing LB system has played a pivotal role in emerging infectious disease control and prevention, as well as in research, which in turn contributes to public health. In recent years, international collaboration between China and other countries has also been accelerated. Despite these achievements, we are still facing many challenges and opportunities in the field of LB. Sustainable LB development requires the joint efforts of the entire society and continuous international cooperation to safeguard global public health.

Studies on developing a safe-management standard system for Chinese biosafety laboratories

Standardization management is one of the key prerequisites to guarantee the effective and safe functioning of high-level biosafety laboratories. Since the outbreak of SARS, increasing numbers of biosafety laboratory levels have been established in China. Therefore, creating a specialized and standardized management system for biosafety laboratories has become necessary and is a core concern. This paper describes the current status of Chinese standards for biosafety management and analyzes its relation to other laboratory biosafety standard systems in developed countries. Based on the analysis on developing a biosafety laboratory, a laboratory biosafety standard system framework was developed, and a policy environment for building the system was suggested with the aim of promoting the promulgation of specific biosafety standards and securing the effective functioning of high-level biosafety laboratories.

Research and development of airtight biosafety containment facility for stainless steel structures

ObjectiveAirtightness of containment structures of a high-level biosafety laboratory is a critical parameter for preventing leakage of harmful bioaerosols. The aim of this study is to investigate the sealing technology of stainless steel structural airtight biosafety containment facilities. MethodsAn experimental study was conducted on a domestic high-level pathogenic microorganism model laboratory, considering the sealing process of the containment structure, including airtight doors, pass boxes, dunk tanks, through-wall pipeline sealing devices, and sealed floor drains. ResultsThe results indicate that the airtightness of the model laboratory containment structure meets the tightness requirements of a biosafety level-4 (BSL-4) laboratory. They also indicate that the construction technology of stainless steel enclosure structures used by the laboratory and the tightness performance and installation process of the developed airtight protective equipment meet the technical requirements of a BSL-4 laboratory. ConclusionsThis successful model laboratory indicates that China has the research and development capacity for stainless steel airtight containment structure manufacturing processes, airtight protective equipment, and technical capacity for independent construction of the highest-level pathogenic microorganism laboratories.

Monitoring and evaluation of the immune status of female Kunming mice maintained in different biosafety level laboratories.

ABSTRACTHigh-level biosafety laboratories (BSL), such as BSL-3 and BSL-4, which deal with high infectivity and virulence pathogens, have become indispensable. Mice are frequently used in animal BSL (ABSL) to establish animal models for infection and to evaluate in vivo immune responses. A project of monitoring and evaluation on the physiology and immune status of mice housed in different ABSL labs was performed in the ABSL-2/3/4 labs of Kunming National High-level Biosafety Research Center, China. Female Kunming mice were housed in the ABSL-2/3/4 labs for 1 month, and mouse behavior, body physiology/immune status, pulmonary immune status and respiratory bacteria composition were evaluated and compared among mice from the different labs. Mice settled in their new housing environment of the different labs after transfer and gained weight steadily. Blood hematology testing, serum cytokine/chemokine profiles and blood/spleen lymphocyte constitutions were comparable between the ABSL-2/3/4 labs. The numbers of different pulmonary leukocytes in the bronchoalveolar lavage fluid were at baseline levels in mice from the ABSL-2/3/4 labs. Diversity and dominance of mice respiratory bacteria were semblable among the ABSL-2/3/4 labs. Our results confirm the stability of physiology and immune status of Kunming mice maintained in different ABSL-2/3/4 labs for at least 1 month.

A blunt sampling probe developed for an in situ leak test of HEPA filters in high-level biosafety laboratory

High-efficiency air filtration is an important means of preventing harmful aerosol overflow in biosafety laboratories. For convenience in the in situ scanning leak test of a high-efficiency particulate air (HEPA) filter to ensure the filter's intactness, a cylindrical blunt sampling probe is developed. The sampling inlet is of the slot type and the inlet's length equal the width of the HEPA filter. Thus, the entire downstream face of the HEPA filter may be scanned. To evaluate the blunt sampling probe's ability to detect leaks, the scanning leak test performed on a HEPA filter unit was compared with a test using the thin-walled rectangular sampling probe that is recommended by relevant standards. Based on the comparison, the leak penetration determined using the blunt sampling probe was slightly lower than that determined using the reference sampling probe. Additionally, differences were observed according to the location that was tested. However, the blunt sampling probe could detect leaks through the comprehensive analysis of the local penetration of most-penetrating particle size (MPPS) and non-MPPS particles.

Advance in Environmental Risk Assessment of High Level Biosafety Laboratory

Abstract Environmental risk assessment of high level biosafety laboratory is one of important parts of biosafety research, which focused on the environmental effects of pathogenic microorganism leaked from laboratory, including pollution of waste air, sewage and solids. Environmental risk assessment was firstly studied in 1970s in the developed countries, especially in the United States. Ecological assessment and human health assessment have been widely researched, while the laboratory environmental risk assessment of pathogenic microorganism leakage was still not well studied. Pathogenic microorganism can be reproduced on the host body and different types of them have different life cycles in the environment. Therefore, the risks and their changes of pathogenic microorganism are complex in the environment, leading to no guidelines for microorganism risk assessment.

Environmental Risk and Assessment Management System of High-level Biosafety Laboratory.

How to avoid the potential risk from high-level biosafety laboratories (BSL-3, BSL-4) has been highly concerned by governments all over the world. Based on the study of construction status and environmental risk of high-level biosafety laboratories and on the study of the layout structure and bio risk measures of biosafety laboratory contaminants, this paper analyzed the key point of environmental risk, and proposed the risk management system of high-level biosafety laboratories. System framework and realization process is given in the paper in order to provide reference for domestic construction of high-level biosafety laboratories and the improvement of risk emergency system.

Development of a Multifunction Isolator System for the Care of Medium-Sized Laboratory Animals Harboring Infectious Diseases

The close phylogenetic relationship between humans and nonhuman primates makes nonhuman primates an irreplaceable model for the study of human infectious diseases. In this study, we describe the development of an automatic multifunctional isolation chamber for use with medium-sized laboratory animals carrying an infectious disease. The isolator system, including the transfer chain, disinfection chain, negative air pressure isolation system, animal welfare system, and the automated system, was designed to meet all biological safety standards. To create an internal chamber environment that was completely isolated from the exterior, variable frequency drive blowers were used in the air-intake and air-exhaust system, precisely controlling the filtered airflow and providing an air-barrier protection. To transfer the material between the interior of the isolation chamber and the outside, a double door transfer port was used. A peracetic acid sterilizer and its affiliated pipeline allowed for a complete disinfection of the isolation chamber. All of the isolator system parameters can be automatically controlled by a programmable computerized menu allowing for work with different animals in different sized cages depending on the research project. The multifunctional isolator system should be a useful safety device for working with medium-sized laboratory infectious animals in high-level biosafety laboratories.