Biosafety In Microbiological And Biomedical Laboratories Research Articles

Safeguards for Using Viral Vector Systems in Human Gene Therapy: A Resource for Biosafety Professionals Mitigating Risks in Health Care Settings.

Health care workers who work daily with human body fluids and hazardous drugs are among those at the highest risk of occupational exposure to these agents. The Occupational Safety and Health Administration's (OSHA) Bloodborne Pathogens Standard (29 CFR 1910.1030) prescribes safeguards to protect workers against health hazards related to bloodborne pathogens. Similarly, the United States Pharmacopeia General Chapter 800 (USP <800>), a standard first published in February 2016 and implementation required by December 2019, addresses the occupational exposure risks of health care workers at organizations working with hazardous drugs. With emerging technologies in the field of gene therapy, these occupational exposure risks to health care workers now extend beyond those associated with bloodborne pathogens and hazardous drugs and now include recombinant DNA. The fifth edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) and the National Institutes of Health Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines) mostly govern work with biohazardous agents and recombinant DNA in a laboratory research setting. When gene therapy products are utilized in a hospital environment, health care workers have very few resources to identify and reduce the risks associated with product use during and after the administration of treatments. At the Abigail Wexner Research Institute at Nationwide Children's Hospital, a comprehensive gap analysis was executed between the research and health care environment to develop a program for risk mitigation. The BMBL, NIH Guidelines, World Health Organization Biosafety Manual, OSHA Bloodborne Pathogens Standard, and USP <800> were used to develop a framework for the gap analysis process. The standards and guidelines for working with viral vector systems in a research laboratory environment were adapted to develop a program that will mitigate the risks to health care workers involved in the preparation, transportation, and administration of gene therapies as well as subsequent patient care activities. The gap analysis identified significant differences in technical language used in daily operations, work environment, training and education, disinfection practices, and policy development between research and health care settings. These differences informed decisions and helped the organization develop a collaborative framework for risk mitigation when a gene therapy product enters the health care setting. With continuing advances in the field of gene therapy, the oversight structure needs to evolve for the health care setting. To deliver the best outcomes to the patients of these therapies, researchers, Institutional Biosafety Committees, and health care workers need to collaborate on training programs to safeguard the public trust in the use of this technology both in clinical trials and as FDA-approved therapeutics.

Biosafety and Biosecurity Challenges Facing Veterinary Diagnostic Laboratories in Lower-Middle Income Countries in Southeast Asia: A Case Study of Thailand.

Introduction:Global concerns over emerging and transboundary infectious zoonotic diseases have increased disease diagnostic demands, especially in the veterinary sector. In developing or newly developed countries where the sector often works under limited capacity, biosafety and biosecurity are unlikely to be high-priority issues. A recent development program supported by the Biological Threat Reduction Program of the Defense Threat Reduction Agency funded by the US government aimed to increase biosafety and biosecurity measures of government veterinary diagnostic and research laboratories in Thailand.Objective:The purpose of this article is to identify biosafety and biosecurity challenges, opportunities, and recommendations.Methods:Eleven government laboratory centers were assessed against the Biosafety in Microbiological and Biomedical Laboratories (BMBL) biosafety level 2 (BSL-2) requirements checklist. The BMBL assessment outcomes were then combined with the outcomes of discussion sessions, and the results of pre- and post-test questionnaires conducted during biosafety assessment workshops and self-evaluation reports using the Food and Agriculture Organization Biosafety Laboratory Mapping Tool of each laboratory center were reviewed and summarized.Results:Despite established national policies on laboratory biosafety and biosecurity, major challenges included (1) harmonization and enforcement of these policies, especially at the regional level, and (2) engagement of personnel in implementations of biosafety and biosecurity measures.Conclusion:Consistent biosafety policy and allocated resources together with regular training are required to develop sustainable biosafety and biosecurity at the national level. Collaboration between regional countries, international organizations, and donors is essential for improving biosafety and biosecurity on a global scale through setting regional priorities, enacting regulatory standards, and providing technical and financial support.

The BMBL and Biosafety Levels

Scientists began developing and publishing a series of best practices to mitigate laboratory risks in the 1970’s. These biosafety guidelines are disseminated by the Department of Health and Human Services in the publication Biosafety in Microbiological and Biomedical Laboratories (BMBL).

A Practical Guide to Implementing a BSL-2+ Biosafety Program in a Research Laboratory

Biosafety Level Two Plus (BSL-2+) is a term frequently used to describe laboratories where work with microorganisms is conducted in a BSL-2 laboratory with biosafety practices and procedures typically found at BSL-3. The risk assessment process may determine that safety practices over and above those required at BSL-2 are needed for a research project, yet a more complex BSL-3 laboratory facility is not necessary. This hybrid approach has been used for many years; however, many research institutions still find it challenging to decide when to use this approach and which BSL-3 practices to use. This is due to the fact that it is not a recognized containment level in biosafety guidance documents such as the Centers for Disease Control and Prevention's (CDC) Biosafety in Microbiological and Biomedical Laboratories (BMBL) (U.S. Department of Health and Human Services, 2009) or the National Institutes of Health's (NIH) Guidelines for Recombinant DNA Research (the Guidelines) (NIH, 2011). This article aims to assist environmental health and safety and biosafety professionals by detailing a practical approach to implementing BSL-2+, including how to identify what projects may benefit from BSL-3 practices and how to modify BSL-3 practices and selected facility requirements for the BSL-2 environment based on institutional needs. In addition, strategies to ensure researchers are properly trained in the use of BSL-3 practices in a BSL-2 laboratory and for developing a project-specific Standard Operating Procedure (SOP) are detailed. Implementing BSL-2+ can be successful when there is a collaboration among the Principal Investigator, Biosafety Officer, Institutional Biosafety Committee, and laboratory personnel.

Biosafety Guidelines for Handling Microorganisms in the Teaching Laboratory: Development and Rationale

The safe handling of microorganisms in the teaching laboratory is a top priority. However, in the absence of a standard set of biosafety guidelines tailored to the teaching laboratory, individual educators and institutions have been left to develop their own plans. This has resulted in a lack of consistency, and differing levels of biosafety practices across institutions. Influenced by the lack of clear guidelines and a recent outbreak of Salmonella infections that was traced back to teaching laboratory exposures, the Education Board of the American Society for Microbiology charged a task force to develop a uniform set of biosafety guidelines for working with microorganisms in the teaching laboratory. These guidelines represent best practices for safely handling microbes, based on the safety requirements found in the Centers for Disease Control and Prevention’s (CDC’s) Biosafety in Microbiological and Biomedical Laboratories (BMBL). Guidelines for safely handling microbes at both biosafety level 1 (BSL1) and biosafety level 2 (BSL2) were developed. The guidelines are brief by design for ease of use and are accompanied by an extensive appendix containing explanatory notes, sample documents, and additional resources. These guidelines provide educators with a clear and consistent way to safely work with microorganisms in the teaching laboratory.

Aseptic Laboratory Techniques: Plating Methods

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the streak-plating method. ● Use pour-plating and spread-plating methods to determine the concentration of bacteria. ● Perform soft agar overlays when working with phage. ● Transfer bacterial cells from one plate to another using the replica-plating procedure. ● Given an experimental task, select the appropriate plating method.

Aseptic Laboratory Techniques: Plating Methods

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the streak-plating method. ● Use pour-plating and spread-plating methods to determine the concentration of bacteria. ● Perform soft agar overlays when working with phage. ● Transfer bacterial cells from one plate to another using the replica-plating procedure. ● Given an experimental task, select the appropriate plating method.

Standard ANSI Z9.14: Testing and performance verification methodologies for ventilation systems for Biological Safety Level 3 (BSL-3) and animal Biological Safety Level 3 (ABSL-3) facilities

The American Industrial Hygiene Association (AIHA) and the American National Standards Institute (ANSI) are developing a national standard titled “Testing and performance verification methodologies for ventilation systems for Biological Safety Level 3(BSL-3) and animal Biological Safety Level 3 (ABSL-3) laboratories” known as ANSI/AIHA Z9.14. The ANSI Z9.14 standard will focus on performance verification of engineering controls related specifically to ventilation system features of BSL-3/ABSL-3 facilities. Currently the design of these facilities is largely guided by the criteria defined in successive versions of the Biosafety in Microbiological and Biomedical Laboratories (BMBL),1 while facilities funded by the National Institutes of Health (NIH) follow BMBL as well as the NIH Design Requirements Manual (DRM).2 Among professionals such as architects, engineers, contractors, commissioning agents and owners who are asked to specify or perform tests for performance of ventilation systems in high containment facilities, there is a consensus that there is no comprehensive methodology based on a risk assessment of each individual facility. An extensive literature review was conducted to determine if there are any regulations, standards or guidance available that provide a “methodology” to verify that the ventilation systems in such facilities are performing appropriately for the current or potential future use. This ‘Gap and Needs Analysis’ provides evidence that there is no single resource for a comprehensive testing methodology that can be used uniformly from one facility to another to verify that the ventilation systems in such facilities are performing appropriately. ANSI Z9.14 can provide one component of a more extensive graduated, risk-based approach to reaching containment goals appropriate to the risk of the agent and the laboratory activity.

Comparative Analysis of the Fourth and Fifth Editions of Biosafety in Microbiological and Biomedical Laboratories, Vertebrate Animal Biosafety Level Criteria (ABSL1-4)

We have developed a matrix of changes between the Vertebrate Animal Biosafety Level Criteria sections of the current and former editions of Biosafety in Microbiological and Biomedical Laboratories (BMBL). Citations containing multiple statements were subdivided into individually-addressable statements and statements with similar/identical scope were aligned, thereby allowing a precise comparative analysis. In addition, statements were categorized for further analysis based on the subject of the change. Results are presented in a change matrix tool that uses a basic Microsoft Excel filter function to allow the user to sort the data based on the animal biosafety level and subject. The tool also contains a side-by-side comparison of animal biosafety levels one through three (ABSL1-3) in the fifth edition of BMBL. In this report, we present a brief summary of major changes.

Comparative Analysis of the Fourth and Fifth Editions of Biosafety in Microbiological and Biomedical Laboratories Section IV (BSL2-4)

We developed a matrix of changes between Section IV of the current and former editions of Biosafety in Microbiological and Biomedical Laboratories (BMBL), focusing on biosafety levels two, three, a...

Comparative Analysis of Biosafety Guidelines of the USA, WHO, and Russia (Organizational and Controlling, Medical and Sanitary—Antiepidemiological Aspects)

Carrying out research for the purpose of prophylactic medicine with dangerous, highly infectious causative agents of disease requires appropriate safety conditions. In addition, the need for appropriate biological safety and security considerations is acquiring an ever-growing national and international importance given the existing threat of the use of these agents by terrorists. Because of these facts, the harmonization of national safety guidelines regulating the organization and work with Hazard Group I-IV pathogens in different countries is gaining more importance and relevance. This paper presents a comparative analysis of safety guidelines and regulations from the United States contained in the Centers for Disease Control and Prevention-National Institutes of Health Biosafety in Microbiological and Biomedical Laboratories (BMBL), the World Health Organization Laboratory Biosafety Manual (WHO LBM) and Russia's Sanitary Rules (SRs).