Applied Biosafety最新文献

Introduction: Biosafety Level 2 (BSL-2) training programs were benchmarked at institutions of higher learning in the United States by surveying Biosafety Officers (BSOs). The goal of this survey was to not only compare Institutional Biosafety Committee (IBC) requirements for BSL-2 training but also compare delivery format, length of training, inclusion of a hands-on or interactive component, and requirement for refresher training.

Methods: Survey subjects were drawn from the National Institutes of Health (NIH) registry of BSOs and were limited to BSOs at institutions of higher learning in the United States. Twelve survey questions were developed in this study. The survey was sent to 324 BSOs, of which 108 responded with sufficient data to be included in the analysis.

Results: The survey found that BSL-2 training is required by the institutional and/or IBC policy at 88% of institutions, is optional at 5.6%, and not offered at 6.5%. More than half of the respondents (60.4%) offered BSL-2 training in an on-demand manner. Some institutions (34.5%) include a component of training that is in person. However, this training does not always include hands-on exercises or interactive activities. The survey indicated that most (96.6%) institutions offered BSL-2 training less than three hours in length, and many (58%) institutions required refresher training.

Conclusions: Most institutions of higher learning in the United States, which are registered with the NIH, require BSL-2 training for laboratory personnel involved in research and teaching.

Introduction: The Research Alliance for Veterinary Science and Biodefense BSL-3 Network (RAV3N) convened an "Applied Biorisk and Biosafety Gap Assessment Workshop" held February 9-10, 2023, in Baltimore, Maryland. As the global prevalence, complexity, and severity of infectious and transboundary veterinary diseases and emerging zoonotic diseases are increasing, there is growing recognition and concern that biorisk management data required to understand and counter these threats are lacking. With sponsorship from the U.S. Department of Agriculture and the U.S. Department of State, RAV3N partnered with Gryphon Scientific and ABSA International to organize, plan, and deliver this biosafety gap analysis workshop.

Methods: The workshop brought together U.S. and international subject matter experts on veterinary and agricultural biorisk management from seven different countries to methodically identify, categorize, and assess the most pressing biosafety or biocontainment evidence gaps related to research and diagnostic activities of agricultural and veterinary importance.

Results: The workshop findings aligned into the following categories: applied biorisk gap identified and required experiment proposed; research performed, but data not published or shared, mechanisms to share data required; risk assessment tools and process enhancements required; literature review and published summaries required; and additional survey, workshops, and "lessons learned" activities required. RAV3N members further analyzed the original dataset and report 25 prioritized applied biosafety research recommendations of importance to biocontainment facility and veterinary biorisk managers. These recommendations warrant consideration by agencies and policy makers considering funding research to ensure that biorisk management practices and regulations for biocontainment laboratories are evidence-based and built upon the best available science.

Background: The validation of high-efficiency particulate air (HEPA) housing decontamination efficacy is crucial for maintaining critical environments and is normally assessed by positioning biological indicators at the downstream port (filtered side) before the decontamination cycle. This method is favored for its significantly lower biohazard risk. HEPA housings and filters typically designed for high airflow face challenges during decontamination due to the diameters and positions of upstream and downstream ports, potentially impacting the distribution of decontamination agents.

Objective: This study investigates the effectiveness of this prevalent validation method, particularly focusing on the uniformity of decontamination agent dispersion across the HEPA filter.

Methods: Preparation: HEPA filters and housings were set up in a controlled environment. Placement of Biological Indicators: Indicators were positioned at multiple points, including the downstream port and various peripheral regions of the filter. Decontamination Cycle: The decontamination process was initiated using a standard protocol for hydrogen peroxide vapor or another selected agent. Measurement: The concentration of decontamination agents was measured at each indicator position using appropriate detection techniques. Analysis: Data were analyzed to assess the uniformity of decontamination agent distribution and identify any significant variations across different areas of the filter.

Results: Our findings reveal a notable variability in the concentration of decontamination agents across different areas of the HEPA filter, especially in peripheral regions. Such variability poses questions about the current method's ability to ensure comprehensive decontamination, given that the concentration at the exit port does not accurately reflect exposure across all filter surfaces. This discrepancy between expected and actual decontamination efficacy underscores the need for optimization in decontamination practices.

Conclusion: The discrepancy between expected and actual decontamination efficacy underscores the need for optimization in decontamination practices. This study highlights the importance of validating decontamination methods and sets the stage for future research aimed at enhancing the uniformity of agent exposure across the HEPA filter. This study advocates for further investigation into methods that could ensure more reliable and effective decontamination, which is vital for biorisk management and the prevention of pathogen contamination.

Introduction: Quantitative data informing biosafety practices have long been lacking. In this study, we describe the conduct of the first large-scale investigation into human reliability in the life sciences laboratory to estimate an error rate during routine biological experiments.

Methods: To generate these critical data, we conducted two sets of experiments: blinded experiments in clinical laboratories in Brazil, Jordan, and Tunisia, and volunteer experiments in training laboratories in two U.S. universities. In these experiments, GloGerm was used to indicate where spills occurred during laboratory manipulations. In the blinded clinical experiments, dummy samples were introduced into the normal workflow and workers processed them as they would a normal clinical sample. Surfaces were examined at the end of the shift for contamination. In the experiments in the United States, volunteers would repetitively pipette a solution of GloGerm into a 96-well plate and the work area was inspected after each plate.

Conclusion: The median volunteer is estimated to have an error rate of about 4 or 8 errors per 1,000 manipulations (for volunteers with significant laboratory experience vs. those with little laboratory experience, respectively). Estimated error rates from both experiments are comparable, suggesting that studies using volunteers who know they are working with nonhazardous materials can be used to replicate real laboratory conditions to provide critical data in biosafety. The volunteers were also asked to declare when they thought they made an error. By comparing true errors to those declared, we found that volunteers identified a maximum of 52% of their total mistakes, indicating that many mistakes go unnoticed.

Introduction: The transport of infectious substances is a highly regulated process. The European Agreement for International Carriage of Dangerous Goods by Road instructs that infectious specimens must be transported in a triple packaging system containing leakproof receptacles. Despite regulatory efforts, leakages occur. Current evidence for Parafilm usage is lacking. This study evaluates the use of Parafilm for preventing leakage from commonly used primary receptacles during ground transport.

Methods: Two test methods, manual inversion and a rocker, were used to simulate standard ground transport. Testing was performed with two common receptacles and three differing volumes of test solution. Each test was performed in quintuplicate. Fluorescein solution was used to aid in visual identification of leakage in addition to the weight of the receptacles before and after testing.

Results: Despite lids being correctly applied, 10% of full 30 mL universal tubes leaked as a result of the rocker testing (n = 40). Overall, 92.5% of full 30 mL universals tested with a cross-threaded lid experienced leakage. Two-milliliter Sarstedt tubes did not experience a single leak even while cross-threaded. Clockwise application of Parafilm decreased leakage from receptacles. Wrapping Parafilm over and under the receptacle provided no benefit compared with not using Parafilm.

Conclusion: Clockwise application of Parafilm may decrease leakage in the event of cross-threading. Despite this, receptacles must be opened in the correct containment, as spillage during unwrapping and lid removal is common. Further investigation is required to assess the effects of pressure on modes of transport such as air travel.

Introduction: High-containment biological laboratories (HCBLs) are specialized facilities designed for handling high-risk biological agents. The lack of data regarding the quantity, condition, and distribution of these laboratories in Brazil poses challenges for governmental strategic decisions.

Objective: This research sought to offer a comprehensive view of the present condition of Brazilian HCBLs, highlighting their quantity, geographical distribution, function, operational range, and commissioning and certification processes.

Method: A questionnaire was designed to collect data on the construction and operation of Brazilian HCBLs and was disseminated to facilities nationwide, identifying themselves as HCBLs.

Results: In this survey, 66 HCBLs were identified across Brazil, with 32 participating in the study. A majority of the laboratories were associated with public universities, predominantly in the Southeast region. The pathogens most frequently handled were SARS-CoV-2 and Mycobacterium tuberculosis. Among the responding laboratories, ∼85% were engaged in research activities, <50% reported being part of a national network, a mere 15.6% were connected to international laboratory networks, and ∼50% reported obtaining certification before operational launch.

Conclusion: Establishing a national regulatory framework in Brazil for the design, construction, commissioning, and certification of HCBLs is crucial to standardize and harmonize procedures across the country. Furthermore, implementing a national policy for laboratory biosafety and biosecurity, alongside establishing a "National HCBLs Network", is essential to foster multidisciplinary and collaborative efforts, thereby optimizing resource allocation. Such enhancements in biosafety and biosecurity protocols will significantly benefit both animal and public health domestically and contribute positively to global health outcomes.

Introduction: Positive pressure breathing-air-fed protective suits are used in biosafety level 4 (BSL-4) containment laboratories as personal protective equipment to protect workers from high-consequence pathogens. However, even with the use of primary containment devices, the exterior surfaces of these suits could potentially become contaminated with those pathogens and result in their inadvertent removal from containment. To address the risk of such pathogens escaping from containment via contaminated protective suits, these suits are decontaminated in a disinfectant chemical shower situated in an anteroom prior to exiting the BSL-4 laboratory. Properly diluted chemical disinfectants such as Micro-Chem Plus™ (MCP) or peracetic acid are used for this purpose. However, whether these suits are properly decontaminated during the chemical shower process needs to be validated.

Methods: The purpose of this study was to develop a suit decontamination validation method for the BSL-4 chemical showers using a risk group 2 (RG2) surrogate virus for the high consequence pathogens that are handled in the BSL-4 laboratories. Here, we evaluated the efficacy of a 5% MCP shower using coupons made from different parts of protective suits (suit fabric, visor, boot, vinyl tape) laden with a dried-on mixture of vesicular stomatitis virus in tripartite organic soil load.

Discussion: This validation study demonstrated that a chemical deluge shower procedure using 5% MCP for 2 min followed by a 3-min water rinse was successful in decontaminating the positive pressure suits that were experimentally contaminated with the live RG2 virus. This offers valuable insights into the rigor of the decontamination process being undertaken in the BSL-4 laboratory chemical showers.

Introduction: Artificial intelligence (AI) tools continue to be developed and used within the life sciences. The impact of these tools on the biosecurity landscape surrounding mail-order DNA synthesis and how to address the impacts have not been critically examined in the literature.

Methods: The impacts of AI-driven chatbots and biological design tools on the biosecurity landscape surrounding mail-order DNA synthesis were analyzed and described. The findings are informed by the authors' experience in the field.

Results: Generally, chatbots lower barriers to access of information that could be misused while biological design tools may provide new abilities to users with the intent of misuse. Six recommendations to the United States Government that attempt to maximize the benefits of these new technologies while mitigating risks are provided.

Conclusion: Mandating mail-order DNA synthesis providers to screen DNA synthesis orders is a critical safeguarding step that should be taken as soon as possible. Over time, biological design tools will reduce the effectiveness of such a regulation and actions should be taken now to limit the negative impacts in the future.