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The operator protection factor (OPF) of four biological safety cabinets (BSCs) has been measured under standard and suboptimal conditions.The OPF for one BSC1, two BSC2, and an acid-fast bacilli staining station (AFBSS) was measured using the potassium iodide method for in situ testing of BSCs (CEN12469) over a range of inflow velocities under standard conditions and with common interfering factors (fans, opening doors, and walk pasts).The BSC1 and the AFBSS gave a high level of protection under standard test conditions at all airflows (down to 0.3 and 0.38 m/s, respectively). During interfering processes, the BSC1 and AFBSS gave a high level of protection (OPF10Although BSC2s are capable of giving a high level of performance, this is design dependent and the BSC1 and AFBSS give a more predictable level of performance due to their simpler design. In environments where BSC certification is not possible, they may provide more robust and sustainable primary containment.

Introduction: Fluorescent-activated cell sorting (FACS) is often the most appropriate technique to obtain pure populations of a cell type of interest for downstream analysis. However, aerosol dropl...

Most existing biocontainment units (BCUs) in U.S. hospitals are designed to care for a limited number of patients infected with epidemiologically significant pathogens. The COVID-19 pandemic presented substantial challenges to hospital preparedness and operations because of its high incidence rate and the high risk of transmission to staff members. This article describes a novel practice innovation: a hospital-wide deployment of nurses on a trained BCU team to support hospital staff in safely caring for patients with COVID-19. Their responsibilities included assisting in the development of guidelines and providing training on safety protocols and the appropriate use of personal protective equipment. The authors show how this deployment contributed significantly to staff education and support during the pandemic.

Genetically modified organisms (GMOs) have emerged as an integral component of a sustainable bioeconomy, with an array of applications in agriculture, bioenergy, and biomedicine. However, the rapid development of GMOs and associated synthetic biology approaches raises a number of biosecurity concerns related to environmental escape of GMOs, detection thereof, and impact upon native ecosystems. A myriad of genetic safeguards have been deployed in diverse microbial hosts, ranging from classical auxotrophies to global genome recoding. However, to realize the full potential of microbes as biocatalytic platforms in the bioeconomy, a deeper understanding of the fundamental principles governing microbial responsiveness to biocontainment constraints, and interactivity of GMOs with the environment, is required. Herein, we review recent analytical biotechnological advances and strategies to assess biocontainment and microbial bioproductivity, as well as opportunities for predictive systems biodesigns towards securing a viable bioeconomy.

In engineered living materials (ELMs) non-living matrices encapsulate microorganisms to acquire capabilities like sensing or biosynthesis. The confinement of the organisms to the matrix and the prevention of overgrowth and escape during the lifetime of the material is necessary for the application of ELMs into real devices. In this study, a bilayer thin film hydrogel of Pluronic F127 and Pluronic F127 acrylate polymers supported on a solid substrate is introduced. The inner hydrogel layer contains genetically engineered bacteria and supports their growth, while the outer layer acts as an envelope and does not allow leakage of the living organisms outside of the film for at least 15 days. Due to the flat and transparent nature of the construct, the thin layer is suited for microscopy and spectroscopy-based analyses. The composition and properties of the inner and outer layer are adjusted independently to fulfil viability and confinement requirements. We demonstrate that bacterial growth and light-induced protein production are possible in the inner layer and their extent is influenced by the crosslinking degree of the used hydrogel. Bacteria inside the hydrogel are viable long term, they can act as lactate-sensors and remain active after storage in phosphate buffer at room temperature for at least 3 weeks. The versatility of bilayer bacteria thin-films is attractive for fundamental studies and for the development of application-oriented ELMs.

Published by Elsevier Ltd., Background: Differences in the distribution of individual-level clinical risk factors across regions do not fully explain the observed global disparities in COVID-19 outcomes. We aimed to investigate the associations between environmental and societal factors and country-level variations in mortality attributed to COVID-19 among people with rheumatic disease globally. Methods: In this observational study, we derived individual-level data on adults (aged 18-99 years) with rheumatic disease and a confirmed status of their highest COVID-19 severity level from the COVID-19 Global Rheumatology Alliance (GRA) registry, collected between March 12, 2020, and Aug 27, 2021. Environmental and societal factors were obtained from publicly available sources. The primary endpoint was mortality attributed to COVID-19. We used a multivariable logistic regression to evaluate independent associations between environmental and societal factors and death, after controlling for individual-level risk factors. We used a series of nested mixed-effects models to establish whether environmental and societal factors sufficiently explained country-level variations in death. Findings: 14 044 patients from 23 countries were included in the analyses. 10 178 (72·5%) individuals were female and 3866 (27·5%) were male, with a mean age of 54·4 years (SD 15·6). Air pollution (odds ratio 1·10 per 10 μg/m3 [95% CI 1·01-1·17]; p=0·0105), proportion of the population aged 65 years or older (1·19 per 1% increase [1·10-1·30]; p, MAG is supported by the National Institute of Arthritis and Musculoskeletal and Skin Diseases (grant numbers K01 AR070585 and K24 AR074534 [JY]). KDW is supported by the Department of Veterans Affairs and the Rheumatology Research Foundation Scientist Development award. JAS is supported by the National Institute of Arthritis and Musculoskeletal and Skin Diseases (grant numbers K23 AR069688, R03 AR075886, L30 AR066953, P30 AR070253, and P30 AR072577), the Rheumatology Research Foundation (K Supplement Award and R Bridge Award), the Brigham Research Institute, and the R. Bruce and Joan M. Mickey Research Scholar Fund. NJP is supported by the National Institute of Arthritis and Musculoskeletal and Skin Diseases (T32-AR-007258). AD-G is supported by grants from the Centers for Disease Control and Prevention and the Rheumatology Research Foundation. RH was supported by the Justus-Liebig University Giessen Clinician Scientist Program in Biomedical Research to work on this registry. JY is supported by grants from the National Institutes of Health (K24 AR074534 and P30 AR070155).

The health care systems in resource limited countries are facing major challenges in dealing with Coronavirus disease (COVID-19). In Bangladesh, a steady increase in the number of COVID-19 cases since its first report on March 8, 2020, has led to an increased demand for COVID-19 detection facilities throughout the country. The detection of severe acute respiratory syndrome (SARS-CoV-2), the causative organism of COVID-19 and a highly infectious group 3(three) organism, requires a high biocontainment laboratory with a certain standard prerequisite infrastructure. This study describes the necessary steps for establishing and running a COVID-19 laboratory under resource constraint settings. Our experience indicates that, with collaborative efforts, funding, and technical support from locally available expertise, it is feasible to set up an optimally functional biocontainment facility with an acceptable quality performance despite several short comings. BSMMU J 2021; 14 (COVID -19 Supplement): 45-50

INTRODUCTION: Existing biosafety guidelines and biosecurity policies do not adequately address the Increased deployment of mobile high-containment biological laboratories (MBSLs) to manage disease outbreak response demonstrates potentials for rapid diagnosis of risk group 3 and 4 pathogens and allows for research under the biosafety level (BSL)-4/3 environment. OBJECTIVES: The primary objective of this study was to evaluate limitations in current biosafety and biosecurity guidelines to address the unique operational environment and new and varied international stakeholders involved in these deployments. RESULTS: Current biosafety best practices guidelines and biosecurity policy and regulatory directives are crafted mostly toward conventional high-containment biological facilities and may not be readily applicable to the MBSLs. Although the pilot MBSLs reported in published literature have adopted multiple safety precautions to reduce exposure risks and have modified process flows in the constrained space to basically meet the proximate requirements for epidemic response, there are no systematic risk assessments for these sophisticated BSL-4/3 mobile laboratory infrastructure—deployed in locations with poor public health infrastructure backup and security problems and handling highly pathogenic agents in constrained environments—with potentials for failure, the nature and extent of which are yet to be determined. The probability of potential occupational exposure and risks to the local general environment may require a reassessment of the biosafety best practice management procedures for MBSLs to deal with these contingencies.

Introduction:Animal carcasses differ in composition from other types of solid waste, and through prior testing it was determined that cycle parameters applied to general, solid biohazardous waste did not ensure proper sterilization of ferret carcasses.Objectives:The goals of this study were to develop and validate an autoclave cycle that would ensure the decontamination of infectious animal carcasses before removal from an animal biosafety level 2/3 containment suite for downstream disposal and to test different ways to prepare and package animal carcasses for autoclaving.Methods:Intact ferret carcasses were implanted with biological indicators, and the carcasses were placed in biohazard bags, then into metal pans. To test the efficacy of the autoclave cycle on larger biomasses, 1, 2, or 4 ferret carcasses were placed in a biohazard bag. A total of 4 carcasses were placed in each pan. An autoclave cycle was created to begin the study. After initial tests, minor modifications to the initial test cycle parameters were made, and a new cycle was validated for ferret carcasses up to 2 kg each. Parameters for the validated cycle were as follows: sterilization time 240 minutes, temperature 125°C, 5 prevacuum pulses, and chamber pressure 15 psi.Results:The results of this study indicate that an extended sterilization time is required to successfully decontaminate animal carcasses compared with regular, solid, and biohazardous waste.Conclusions:This study demonstrates that it is possible to sterilize multiple intact ferret carcasses per load under validated autoclave cycle conditions.

Genetically modified microorganisms (GMMs) can enable a wide range of important applications including environmental sensing and responsive engineered living materials. However, containment of GMMs to prevent environmental escape and satisfy regulatory requirements is a bottleneck for real-world use. While current biochemical strategies restrict unwanted growth of GMMs in the environment, there is a need for deployable physical containment technologies to achieve redundant, multi-layered, and robust containment. We developed a hydrogel-based encapsulation system that incorporates a biocompatible multilayer tough shell and an alginate-based core. This DEployable Physical COntainment Strategy (DEPCOS) allows no detectable GMM escape, bacteria to be protected against environmental insults including antibiotics and low pH, controllable lifespan, and easy retrieval of genomically recoded bacteria. To highlight the versatility of DEPCOS, we demonstrated that robustly encapsulated cells can execute useful functions, including performing cell-cell communication with other encapsulated bacteria and sensing heavy metals in water samples from the Charles River.

Biocontainment systems have been developed to mitigate the concerns regarding biosafety and environmental risk because of the possible escape of genetically modified organisms into the environment following large-scale outdoor cultivation. Here, we present a biocontainment system entailing genetically modified Synechococcus elongatus PCC 7942, also engineered for α-farnesene production using a de-evolutionary strategy. In this approach, the gene cluster encoding the β-carboxysome and the associated carbon concentrating mechanism (CCM) were deleted in the α-farnesene-producing cyanobacteria, resulting in no cell growth and no α-farnesene production at ambient CO2 concentrations (100% air bubbling). However, cell growth and α-farnesene production were detected in the CCM-deficient strains at high CO2 concentrations (5% CO2 [v/v], 10% CO2 [v/v]), albeit at levels lower than those of the parental control. To overcome this limitation, the overexpression of carbonic anhydrase and bicarbonate transporter genes in the CCM-deficient strains restored cell growth and the production level of α-farnesene (5.0 ± 0.6 mg/L) to that of the parental control. The production of α-farnesene in the later strains strictly depended on CO2 concentration in the photobioreactor and did not rely on a chemical induction process. Thus, next generation bio-solar cell factories could be promoted with the suggested biocontainment system.

Research animals models infected with Biosafety Level-3 (BSL-3) agents need to be housed in specialized biocontainment caging. Most of these specialized cages have input and exhaust that is high efficiency particulate air filtered and sealed to prevent escape of the BSL-3 agent. An alternative to the use of the above BSL-3 biocontainment caging is the use of a flexible film or modified semi-rigid plastic film isolator that has its own high efficiency particulate air–filtered input and exhaust and is sealed with respect to the animal room environment, thus preventing BSL-3 agent escape. Standard caging can be housed within such an isolator. Computational fluid dynamics was used to evaluate the integrity of modified semi-rigid isolators for containment of aerosolized BSL-3 agents. Three isolators were located inside an animal BSL-3 room to provide an extra tier of protection and to permit different infectious studies within the same room while reducing or eliminating the risk of cross-contamination. The isolators were sized to house caging for rabbits and smaller non-human primates such as marmosets, African greens, and macaques. Multiple case studies of failure scenarios were investigated, including isolator breaches through the plastic membrane seam separation and rips, and exhaust fan failure. Breaching the level of containment provided by the isolators required the improbable simultaneous event of a plastic membrane rip in addition to the rare malfunction of the back-up exhaust fans. Each isolator was equipped with 2 blower motors connected in parallel to a common exhaust plenum and a battery backup. Even with this rare double (simultaneous) event, the animal BSL-3 room air exhaust system was able to contain the few droplets released in the simulated computational fluid dynamics breach. The modified semi-rigid isolators with negative airflow proved safe and effective for aerosol studies using BSL-3 agents, even in the unlikely event of a breach in containment.

Objectives, Purposes, or Aim: To identify design strategies utilized in airborne infection isolation and biocontainment patient rooms that improve infection control potential in an alternative care environment. Background: As SARS-CoV-2 spreads and health care facilities near or exceed capacity, facilities may implement alternative care sites (ACSs). With COVID-19 surges predicted, developing additional capacity in alternative facilities, including hotels and convention centers, into patient care environments requires early careful consideration of the existing space constraints, infrastructure, and modifications needed for patient care and infection control. Design-based strategies utilizing engineering solutions have the greatest impact, followed by medical and operational strategies. Methods: This article evaluates infection control and environmental strategies in inpatient units and proposes system modifications to ACS surge facilities to reduce infection risk and improve care environments. Results: Although adequate for an acute infectious disease outbreak, existing capacity in U.S. biocontainment units and airborne infection isolation rooms is not sufficient for widespread infection control and isolation during a pandemic. To improve patients’ outcomes and decrease infection transmission risk in the alternative care facility, hospital planners, administrators, and clinicians can take cues from evidence-based strategies implemented in biocontainment units and standard inpatient rooms. Conclusions: Innovative technologies, including optimized air-handling systems with ultraviolet and particle filters, can be an essential part of an infection control strategy. For flexible surge capacity in future ACS and hospital projects, interdisciplinary design and management teams should apply strategies optimizing the treatment of both infectious patients and minimizing the risk to health care workers.

Recent advances in genetic engineering and synthetic biology have enabled a variety of genetically modified microorganisms (GMOs) designed for open environmental applications. The use of GMOs in these applications, however, raised concerns about the spread of artificial biological matters into the natural environment. To address the issue, genetic biocontainment system has emerged as a way to prevent unauthorized propagation of GMOs and genetic materials into the ecosystem. In this review, we introduce biocontainment systems that control the proliferation of microorganisms and the release of genetic materials, focusing primarily on genetic means to eliminate cells and DNA. We outline each biocontainment system with representative examples, highlighting its advantages and disadvantages. We also discuss future challenges that genetic biocontainment systems should overcome for practical uses.

New technologies have allowed researchers to better design, build, and analyze complex consortia. These developments are fueling a wider implementation of consortium-based bioprocessing by leveraging synthetic biology, delivering on the field's multitudinous promises of higher efficiencies, superior resiliency, augmented capabilities, and modular bioprocessing. Here we chronicle current progress by presenting a range of screening, computational, and biomolecular tools enabling robust population control, efficient division of labor, and programmatic spatial organization; furthermore, we detail corresponding advancements in areas including machine learning, biocontainment, and standardization. Additionally, we show applications in myriad sectors, including medicine, energy and waste sustainability, chemical production, agriculture, and biosensors. Concluding remarks outline areas of growth that will promote the utilization of complex community structures across the biotechnology spectrum.

Safety-by-design (SbD) is paramount for risk management in synthetic biology, with genetic safeguards being a key technology for its implementation. While attractive in theory, the integration of genetic safeguards into SbD strategies is rarely exercised in practice, despite technological advances. Here we question why and what might be done about it.

Biocontainment systems are needed to neutralize genetically modified organisms (GMOs) that pose ecological threats outside of controlled environments. In contrast, benign selection markers complement GMOs with reduced fitness. Benign selection agents serve as alternatives to antibiotics, which are costly and risk spread of antibiotic resistance. Here, we present a yeast biocontainment strategy leveraging engineered fluoride sensitivity and DNA vectors enabling use of fluoride as a selection agent. The biocontainment system addresses the scarcity of platforms available for yeast despite their prevalent use in industry and academia. In the absence of fluoride, the biocontainment strain exhibits phenotypes nearly identical to those of the wildtype strain. Low fluoride concentrations severely inhibit biocontainment strain growth, which is restored upon introduction of fluoride-based vectors. The biocontainment strategy is stringent, easily implemented, and applicable to several eukaryotes. Further, the DNA vectors enable genetic engineering at reduced costs and eliminate risks of propagating antibiotic resistance., Non-antibiotic selection systems could also serve as biocontainment strategies. Here the authors present a fluoride sensitivity selection system for use in yeast.

In response to the Ebola outbreak of 2014–2016, the US Office of the Assistant Secretary for Preparedness and Response (ASPR) established 10 regional treatment centers, called biocontainment units (BCUs), to prepare and provide care for patients infected with high-consequence pathogens. Many of these BCUs were among the first units to activate for coronavirus disease 2019 (COVID-19) patient care. The activities of the Johns Hopkins BCU helped prepare the Johns Hopkins Health System for COVID-19 in the 3 domains of containment care: (1) preparedness planning, education and training, (2) patient care and unit operations, and (3) research and innovation. Here, we describe the role of the JH BCU in the Hopkins COVID-19 response to illustrate the value of BCUs in the current pandemic and their potential role in preparing healthcare facilities and health systems for future infectious disease threats.

Two theories on the origins of COVID-19 have been widely circulating in China and the West respectively, one blaming the United States and the other a highest-level biocontainment laboratory in Wuhan, the initial epicentre of the pandemic. Both theories make claims of biological warfare attempts. According to the available scientific evidence, these claims are groundless. However, like the episodes of biological warfare during the mid-twentieth century, the spread of these present-day conspiracy theories reflects a series of longstanding and damaging trends in the international scene which include deep mistrust, animosities, the power of ideologies such as nationalism, and the sacrifice of truth in propaganda campaigns. Also, the threats associated with biological warfare, bioterrorism, and the accidental leakage of deadly viruses from labs are real and growing. Thus, developing a better global governance of biosafety and biosecurity than exists at present is an urgent imperative for the international community in the broader context of a looming Cold War II. For such a governance, an ethical framework is proposed based upon the triple ethical values of transparency, trust, and the common good of humanity.

DNA barcodes in small packages Under adverse environmental conditions, some microorganisms form spores that provide robust protection for genetic material. Qian et al. developed a system in which DNA barcodes are encapsulated inside nongerminating microbial spores and can be dispersed on objects or in the environment (see the Perspective by Nivala). These barcoded spores provide a durable, specific marker that can be read out quickly with simple equipment. When applied to soil, the spores can be transferred to and from objects around them, enabling tracking at meter-scale resolution. On plant leaves, the spores are not readily transferred, and the authors demonstrate a potential use for tracking agricultural products. Science , this issue p. 1135 ; see also p. 1058

The biocontainment unit at Johns Hopkins Hospital is a specially designed, inactive high-level isolation unit designated to care for patients infected with high-consequence pathogens. The unit team designed a facility-specific readiness scale and checklist that focus on infrastructure, consumable supplies, and staffing to assess activation readiness of the biocontainment unit. Over a period of 50 days and 14 days, these tools were used as part of a routine risk assessment to first identify barriers and then tier the impact of these barriers into activation categories of "Ready," "Ready with Considerations," and "Not Ready." The assessment identified the greatest risks to activation readiness were staffing and waste management capabilities. Assessing threats to activation readiness and the risk of not being ready should be a priority for maintaining facility, regional, and national capacity to safely isolate and care for patients infected with high-consequence pathogens while maintaining healthcare worker safety.

Biocontaining was one way that Western, affluent, allopathic cultures tended to respond and make meaning during the 2013-2015 Ebola virus disease (EVD) pandemic. It became a pathway to restore trust in biomedicine itself, which had been shaken by unease across the globe when the EVD threat was at its height. Yet biocontaining barely qualifies as a public health measure. Successful public health efforts rely on trust, which is difficult to maintain during a pandemic. Such efforts require balancing the need to be close to patients to care well for them against the need to remain distant from a virulent pathogen. Biocontainment tries to navigate this tension and, in so doing, simultaneously frustrates and supports public trust. This article suggests 5 things clinicians and health professions students should consider about the project of biocontainment that could affect their orientation to their public health duties.

The study of many arthropod-borne pathogens requires high biosafety considerations, including the use of specialized facilities and equipment for arthropod containment. Mosquito- and tick-borne viruses such as yellow fever, West Nile, and Crimean Congo hemorrhagic fever viruses require facilities that are suitable for housing vertebrates. Multidisciplinary studies that incorporate the vector, vertebrate, and pathogens are essential for a complete understanding of the interactions between these transmission cycle components, especially if they aim to evaluate and model relative susceptibilities of different arthropods and vertebrates to infection and transmission between these. Under laboratory conditions, these studies can be relatively simple, for example, involving colonized arthropods, small animals, and attenuated viruses. Other studies are complex with large animals, high-biocontainment pathogens, and field-collected arthropods. These require a higher level of containment and special design considerations. Both of these types of experiments have their relative merits. A thorough understanding of the issues related to these types of studies and the benefits and drawbacks to using various challenge models will enable the researcher to develop realistic goals for various experiments. This review examines the varied issues that should be considered prior to starting these experiments and covers the basics from the procurement of various arthropods, rearing, high-containment facilities and operational issues specific to work with arthropods, types of infection experiments, and specific issues with arthropod and animal experiments in biosafety levels 3 and 4.

The National Bioforensic Analysis Center has been central to the nation's biodefense strategy since its inception in 2004 and provides ISO 17025 accredited bioforensic capabilities at BSL-0 through BSL-4 for the FBI and other federal partners. NBFAC is continually expanding to maintain cutting edge capabilities in the face of expanding threat landscapes and rapidly evolving technology, and it works with partners in academia, industry, and government to ensure that federal bioterror/biocrime investigations have the best possible scientific support.

Purpose: There is a lack of information regarding the physiological effects of healthcare workers performing patient care while wearing enhanced personal protective equipment (PPE). This study aimed to provide foundational data necessary to maintain the health and safety of healthcare workers while performing tasks wearing enhanced PPE related to patient care in a biocontainment unit.Design and method: This study is a descriptive study evaluating the physiological effects of conducting patient care while wearing enhanced PPE. Participant vital signs (blood pressure, body temperature, and oxygen saturation) were monitored pre-donning, throughout the transport, and post-doffing.Results: There is no statistically significant difference in the physiological measurements of any of the participants. The results consisted of: blood pressure (systolic p = 0.0659; diastolic p = 0.94345); oxygen saturation (p = 0.6233); and orally derived body temperature (p = 0.1093). Physical balance was subjectively measured by both the participant and the trained observer during the donning and doffing process. Results show no visible difference in balance between the donning period and the doffing period. Conclusion: This study demonstrated there were no physiological changes to the healthcare workers in this study performing patient care activities during patient transport and in a biocontainment unit while wearing the provided enhanced PPE. However, limitations of this study include: (1) it was conducted at one location within the Midwest; (2) with a small number of participants; and (3) only the enhanced ensemble of PPE was worn during testing. Therefore, the results may not be generalizable to other locations or levels of PPE.Clinical relevance: This study will help to guide assessment standards for healthcare workers providing care while wearing enhanced level PPE.

Biosafety laboratory accidents are a normal part of laboratory science, but the frequency of such accidents is unclear due to current reporting standards and processes. To better understand accident reporting, a survey was created, with input from ABSA International, which included a series of questions about standards, requirements, and likely motivations for reporting or nonreporting. A total of 60 biosafety officers completed the survey. Respondents reported working with more than 5,000 people in laboratories, including more than 40 biosafety level 3 or animal biosafety level 3 laboratories, which work with higher-risk pathogens. Most of the respondents were located in the United States, Canada, or New Zealand, or did not identify their location. Notable results included that 97% of surveyed biosafety officers oversee laboratories that require reporting exposure to at least some pathogens. However, 63% relayed that the reports are not usually sent outside of the institution where they occurred. A slight majority (55%) stated that paper reports were used, with the rest reporting they used a variety of computer systems. Even in laboratories that used paper-based reporting systems, 67% relayed that these reports were used alongside, or entered into, a digital system. While 82% of these biosafety officers agreed that workers understood the importance of reporting for their own safety, 82% also agreed that a variety of disincentives prevent laboratory workers from reporting incidents, including concerns about job loss and loss of funding.