Oxidative Stress Contributes to Bacterial Airborne Loss of Viability.

While the airborne decay of bacterial viability has been observed for decades, an understanding of the mechanisms driving the decay has remained elusive. The airborne transport of bacteria is often a key step in their life cycle and as such, characterizing the mechanisms driving the airborne decay of bacteria is an essential step toward a more complete understanding of microbial ecology. Using the Controlled Electrodynamic Levitation and Extraction of Bioaerosols onto a Substrate (CELEBS), it was possible to systematically evaluate the impact of different physicochemical and environmental parameters on the survival of Escherichia coli in airborne droplets of Luria Bertani broth. Rather than osmotic stress driving the viability loss, as was initially considered, oxidative stress was found to play a key role. As the droplets evaporate and equilibrate with the surrounding environment, the surface-to-volume ratio increases, which in turn increased the formation of reactive oxygen species in the droplet. These reactive oxygen species appear to play a key role in driving the airborne loss of viability of E. coli. IMPORTANCE The airborne transport of bacteria has a wide range of impacts, from disease transmission to cloud formation. By understanding the factors that influence the airborne stability of bacteria, we can better understand these processes. However, while we have known for several decades that airborne bacteria undergo a gradual loss of viability, we have not previously identified the mechanisms driving this process. In this work, we discovered that oxygen surrounding an airborne droplet facilitates the formation of reactive oxygen species within the droplet, which then gradually damage and kill bacteria within the droplet. This discovery indicates that adaptations to help bacteria deal with oxidative stress may also aid their airborne survival and be essential adaptations for bacterial airborne pathogens. Understanding the adaptations bacteria need to survive in airborne droplets could eventually lead to the development of novel antimicrobials designed to inhibit their airborne survival, helping to prevent the transmission of disease.

Preparedness of Military Public Health for Epidemic and Pandemic Recognition and Response.

Disease epidemics have threatened American military preparedness and operational capabilities since 1775. The ongoing Severe Acute Respiratory Syndrome Coronavirus 2 (COVID-19) pandemic, which began in 2019, again demonstrates the significant potential for infectious diseases to impact military units and threaten military readiness. We reviewed the historical and continuing threats to the U.S. Military from infectious disease outbreaks, as well as changes in U.S. Military capabilities for conducting meaningful surveillance and response. We concluded that a structured review of military public health and preventive medicine capabilities should be conducted to assess the response to the COVID-19 pandemic and determine the capabilities necessary for infectious disease surveillance and response to future threats.

Military Epidemics, Then and Now: Smallpox and COVID-19.

We compared the COVID-19 experience in the first year of the current pandemic in the US with the smallpox experience of the 18th century, focusing on the US military but recognizing civilian and military populations are not separate and distinct. Despite the epidemics being separated by 21/2 centuries and with great advancements in technology having occurred over that time, we observed similarities which led us to several conclusions: • Infectious disease outbreaks will continue to occur and novel agents, naturally occurring or manipulated by humans, will threaten military and civilian populations nationally and globally. • Infectious disease outbreaks can affect both military and civilian populations, persist for long periods, and be catastrophic to military peacetime and wartime operations. • Effective surveillance is a prerequisite for early identification and subsequent meaningful responses to novel and reemerging threat agents and diseases. • Socio-cultural, religious, or political factors may limit the implementation of effective interventions in military or civilian populations. Public health officials must assess impediments to implementation of interventions and develop plans to overcome them.

Effect of Relative Humidity on Transfer of Aerosol-Deposited Artificial and Human Saliva from Surfaces to Artificial Finger-Pads.

Surface to hand transfer of viruses represents a potential mechanism for human exposure. An experimental process for evaluating the touch transfer of aerosol-deposited material is described based on controlling surface, tribological, and soft matter components of the transfer process. A range of high-touch surfaces were evaluated. Under standardized touch parameters (15 N, 1 s), relative humidity (RH) of the atmosphere around the contact transfer event significantly influenced transfer of material to the finger-pad. At RH < 40%, transfer from all surfaces was <10%. Transfer efficiency increased markedly as RH increased, reaching a maximum of approximately 50%. The quantity of material transferred at specific RHs above 40% was also dependent on roughness of the surface material and the properties of the aerosol-deposited material. Smooth surfaces, such as melamine and stainless steel, generated higher transfer efficiencies compared to those with textured roughness, such as ABS pinseal and KYDEX® plastics. Pooled human saliva was transferred at a lower rate compared to artificial saliva, indicating the role of rheological properties. The artificial saliva data were modeled by non-linear regression and the impact of environmental humidity and temperature were evaluated within a Quantitative Microbial Risk Assessment model using SARS-CoV-2 as an example. This illustrated that the trade-off between transfer efficiency and virus survival may lead to the highest risks of fomite transmissions in indoor environments with higher humidity.

The Future of Military Occupational and Environmental Medicine in the Department of Defense.

OBJECTIVE: Transfer of military medical facilities to the Defense Health Agency is transforming the Military Health System. Our objective is to inform this transformation with respect to optimum application of occupational and environmental medicine (OEM) expertise. METHODS: We defined and analyzed the external influences on military OEM practice using a structured framework to identify key drivers. RESULTS: Key drivers are political and economic factors. These may change the size or military/civilian ratio of the specialty. Limited career development pathways should prompt consideration of making OEM a second or combined residency, and military-funded training of civilian physicians may be required. OEM specialist utilization should be reassessed. CONCLUSIONS: OEM is a highly adaptable specialty defined by the needs of its stakeholders. Comprehensive analysis of external influences can ensure that OEM practice remains in step with changing needs.

Training Occupational and Environmental Medicine (OEM) Residents at the Uniformed Services University.

OBJECTIVE: To develop an actionable plan to sustain and improve the quality of the Uniformed Services University of the Health Sciences (USU) Occupational and Environmental Medicine (OEM) Residency Program. METHODS: Program metrics were collected and analyzed to assess strengths, weaknesses, opportunities, and threats (SWOT analysis). RESULTS: Program strengths are stable funding, full-time faculty and large class size. Weaknesses are limited toxicology curriculum, and the lack of complex clinical cases. Opportunities include establishing an OEM referral clinic, collaborating with U.S. Department of Defense (DoD) toxicology programs, aligning OEM research priorities in DoD, and including DoD Civilian physicians in OEM residency training. Threats are Military Health System reorganization, budget, and personnel cuts. CONCLUSIONS: The USU OEM Residency is strong but must be flexible to adjust to personnel, fiscal, and organizational changes. Aggregating the SWOT analyses for all the OEM residency programs may help identify strategies to sustain OEM training in the United States.

Uniformed Services University Occupational and Environmental Medicine Residency Program.

In 2019, the Uniformed Services University of the Health Sciences (USU) F. Edward Hébert School of Medicine celebrated the 30th anniversary of its Occupational and Environmental Medicine (OEM) Residency program. This unique program is among the largest preventive medicine residency programs in the United States. Residents from the US Army, Navy, Air Force, other federal institutions, and the Canadian Forces come to Bethesda, Maryland, to become OEM specialists in a unique training program encompassing both military and civilian OEM settings. This publication describes the historical development and practice of OEM in the military leading to the development of the USU OEM Residency Program, along with the program's past accomplishments and current operation. Finally, the publication explores potential future directions for this relatively small but important preventive medicine specialty in the practice of military medicine, considering the impacts of reorganization of the Military Health System along with the opportunities this reorganization presents for the USU OEM Residency program.

Transformative Approach To Investigate the Microphysical Factors Influencing Airborne Transmission of Pathogens.

Emerging outbreaks of airborne pathogenic infections worldwide, such as the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, have raised the need to understand parameters affecting the airborne survival of microbes in order to develop measures for effective infection control. We report a novel experimental strategy, TAMBAS (tandem approach for microphysical and biological assessment of airborne microorganism survival), to explore the synergistic interactions between the physicochemical and biological processes that impact airborne microbe survival in aerosol droplets. This innovative approach provides a unique and detailed understanding of the processes taking place from aerosol droplet generation through to equilibration and viability decay in the local environment, elucidating decay mechanisms not previously described. The impact of evaporation kinetics, solute hygroscopicity and concentration, particle morphology, and equilibrium particle size on airborne survival are reported, using Escherichia coli MRE162 as a benchmark system. For this system, we report that (i) particle crystallization does not directly impact microbe longevity, (ii) bacteria act as crystallization nuclei during droplet drying and equilibration, and (iii) the kinetics of size and compositional change appear to have a larger effect on microbe longevity than the equilibrium solute concentration.IMPORTANCE A transformative approach to identify the physicochemical processes that impact the biological decay rates of bacteria in aerosol droplets is described. It is shown that the evaporation process and changes in the phase and morphology of the aerosol particle during evaporation impact microorganism viability. The equilibrium droplet size was found to affect airborne bacterial viability. Furthermore, the presence of Escherichia coli MRE162 in a droplet does not affect aerosol growth/evaporation but influences the dynamic behavior of the aerosol by processing the culture medium prior to aerosolization, affecting the hygroscopicity of the culture medium; this highlights the importance of the inorganic and organic chemical composition within the aerosolized droplets that impact hygroscopicity. Bacteria also act as crystallization nuclei. The novel approach and data have implications for increased mechanistic understanding of aerosol survival and infectivity in bioaerosol studies spanning the medical, veterinary, farming, and agricultural fields, including the role of microorganisms in atmospheric processing and cloud formation.

Developing a Personal Protective Equipment Selection Matrix for Preventing Occupational Exposure to Ebola Virus.

In response to the 2014 Ebola outbreak in West Africa and resulting cases in the United States, the Occupational Safety and Health Administration developed a personal protective equipment selection matrix to help employers protect workers from exposure to Ebola virus in the event of additional US cases. Now, the world's second largest Ebola outbreak on record continues to expand in the Democratic Republic of Congo, where more than 70 Ebola-infected healthcare workers serve as reminders of the importance of robust infection prevention measures in keeping infectious disease responders from becoming victims themselves. Toward facilitating preparedness for cases associated with the ongoing or future outbreaks, this article discusses the matrix of personal protective equipment recommendations. The matrix applies to a variety of job tasks in health care, laboratories, waste handling, janitorial services, travel and transportation, and other sectors where workers may be exposed to the Ebola virus during outbreak events. A discussion of the information sources and decision-making process for developing the matrix forms the basis of the recommendations. The article then emphasizes challenges and considerations for formulating the matrix, including identifying information sources to help characterize occupational exposures, aligning recommendations among stakeholders with varying viewpoints, and balancing worker protections with feasibility concerns. These considerations highlight issues that remain relevant for preparedness efforts ahead of future US cases of Ebola or other emerging infectious diseases.

Assessing the airborne survival of bacteria in populations of aerosol droplets with a novel technology.

The airborne transmission of infection relies on the ability of pathogens to survive aerosol transport as they transit between hosts. Understanding the parameters that determine the survival of airborne microorganisms is critical to mitigating the impact of disease outbreaks. Conventional techniques for investigating bioaerosol longevity in vitro have systemic limitations that prevent the accurate representation of conditions that these particles would experience in the natural environment. Here, we report a new approach that enables the robust study of bioaerosol survival as a function of relevant environmental conditions. The methodology uses droplet-on-demand technology for the generation of bioaerosol droplets (1 to greater than 100 per trial) with tailored chemical and biological composition. These arrays of droplets are captured in an electrodynamic trap and levitated within a controlled environmental chamber. Droplets are then deposited on a substrate after a desired levitation period (less than 5 s to greater than 24 h). The response of bacteria to aerosolization can subsequently be determined by counting colony forming units, 24 h after deposition. In a first study, droplets formed from a suspension of Escherichia coli MRE162 cells (108 ml-1) with initial radii of 27.8 ± 0.08 µm were created and levitated for extended periods of time at 30% relative humidity. The time-dependence of the survival rate was measured over a time period extending to 1 h. We demonstrate that this approach can enable direct studies at the interface between aerobiology, atmospheric chemistry and aerosol physics to identify the factors that may affect the survival of airborne pathogens with the aim of developing infection control strategies for public health and biodefence applications.

Evaluation of Occupational Exposure Limits for Heat Stress in Outdoor Workers - United States, 2011-2016.

Heat stress, an environmental and occupational hazard, is associated with a spectrum of heat-related illnesses, including heat stroke, which can lead to death. CDC's National Institute for Occupational Safety and Health (NIOSH) publishes recommended occupational exposure limits for heat stress (1). These limits, which are consistent with those of the American Conference of Governmental Industrial Hygienists (ACGIH) (2), specify the maximum combination of environmental heat (measured as wet bulb globe temperature [WBGT]) and metabolic heat (i.e., workload) to which workers should be exposed. Exposure limits are lower for workers who are unacclimatized to heat, who wear work clothing that inhibits heat dissipation, and who have predisposing personal risk factors (1,2). These limits have been validated in experimental settings but not at outdoor worksites. To determine whether the NIOSH and ACGIH exposure limits are protective of workers, CDC retrospectively reviewed 25 outdoor occupational heat-related illnesses (14 fatal and 11 nonfatal) investigated by the Occupational Safety and Health Administration (OSHA) from 2011 to 2016. For each incident, OSHA assessed personal risk factors and estimated WBGT, workload, and acclimatization status. Heat stress exceeded exposure limits in all 14 fatalities and in eight of 11 nonfatal illnesses. An analysis of Heat Index data for the same 25 cases suggests that when WBGT is unavailable, a Heat Index screening threshold of 85°F (29.4°C) could identify potentially hazardous levels of workplace environmental heat. Protective measures should be implemented whenever the exposure limits are exceeded. The comprehensive heat-related illness prevention program should include an acclimatization schedule for newly hired workers and unacclimatized long-term workers (e.g., during early-season heat waves), training for workers and supervisors about symptom recognition and first aid (e.g., aggressive cooling of presumed heat stroke victims before medical professionals arrive), engineering and administrative controls to reduce heat stress, medical surveillance, and provision of fluids and shady areas for rest breaks.

Risk Factors for Heat-Related Illness in U.S. Workers: An OSHA Case Series.

OBJECTIVE: The aim of this study was to describe risk factors for heat-related illness (HRI) in U.S. workers. METHODS: We reviewed a subset of HRI enforcement investigations conducted by the Occupational Safety and Health Administration (OSHA) from 2011 through 2016. We assessed characteristics of the workers, employers, and events. We stratified cases by severity to assess whether risk factors were more prevalent in fatal HRIs. RESULTS: We analyzed 38 investigations involving 66 HRIs. Many workers had predisposing medical conditions or used predisposing medications. Comorbidities were more prevalent in workers who died. Most (73%) fatal HRIs occurred during the first week on the job. Common clinical findings in heat stroke cases included multiorgan failure, muscle breakdown, and systemic inflammation. CONCLUSION: Severe HRI is more likely when personal susceptibilities coexist with work-related and environmental risk factors. Almost all HRIs occur when employers do not adhere to preventive guidelines.

Exacerbated grassland degradation and desertification in Central Asia during 2000-2014.

Grassland degradation and desertification is a complex process, including both state conversion (e.g., grasslands to deserts) and gradual within-state change (e.g., greenness dynamics). Existing studies hardly separated the two components and analyzed it as a whole based on time series vegetation index data, which cannot provide a clear and comprehensive picture for grassland degradation and desertification. Here we propose an integrated assessment strategy, by considering both state conversion and within-state change of grasslands, to investigate grassland degradation and desertification process in Central Asia. First, annual maps of grasslands and sparsely vegetated land were generated to track the state conversions between them. The results showed increasing grasslands were converted to sparsely vegetated lands from 2000 to 2014, with the desertification region concentrating in the latitude range of 43-48° N. A frequency analysis of grassland vs. sparsely vegetated land classification in the last 15 yr allowed a recognition of persistent desert zone (PDZ), persistent grassland zone (PGZ), and transitional zone (TZ). The TZ was identified in southern Kazakhstan as one hotspot that was unstable and vulnerable to desertification. Furthermore, the trend analysis of Enhanced Vegetation Index during thermal growing season (EVITGS ) was investigated in individual zones using linear regression and Mann-Kendall approaches. An overall degradation across the area was found; moreover, the second desertification hotspot was identified in northern Kazakhstan with significant decreasing in EVITGS , which was located in PGZ. Finally, attribution analyses of grassland degradation and desertification were conducted by considering precipitation, temperature, and three different drought indices. We found persistent droughts were the main factor for grassland degradation and desertification in Central Asia. Considering both state conversion and gradual within-state change processes, this study provided reference information for identification of desertification hotspots to support further grassland degradation and desertification treatment, and the method could be useful to be extended to other regions.

Aerobiology: Experimental Considerations, Observations, and Future Tools.

Understanding airborne survival and decay of microorganisms is important for a range of public health and biodefense applications, including epidemiological and risk analysis modeling. Techniques for experimental aerosol generation, retention in the aerosol phase, and sampling require careful consideration and understanding so that they are representative of the conditions the bioaerosol would experience in the environment. This review explores the current understanding of atmospheric transport in relation to advances and limitations of aerosol generation, maintenance in the aerosol phase, and sampling techniques. Potential tools for the future are examined at the interface between atmospheric chemistry, aerosol physics, and molecular microbiology where the heterogeneity and variability of aerosols can be explored at the single-droplet and single-microorganism levels within a bioaerosol. The review highlights the importance of method comparison and validation in bioaerosol research and the benefits that the application of novel techniques could bring to increasing the understanding of aerobiological phenomena in diverse research fields, particularly during the progression of atmospheric transport, where complex interdependent physicochemical and biological processes occur within bioaerosol particles.

Particle-size dependent effects in the Balb/c murine model of inhalational melioidosis.

Deposition of Burkholderia pseudomallei within either the lungs or nasal passages of the Balb/c murine model resulted in different infection kinetics. The infection resulting from the inhalation of B. pseudomallei within a 12 µm particle aerosol was prolonged compared to a 1 µm particle aerosol with a mean time-to-death (MTD) of 174.7 ± 14.9 h and 73.8 ± 11.3 h, respectively. Inhalation of B. pseudomallei within 1 µm or 12 µm particle aerosols resulted in a median lethal dose (MLD) of 4 and 12 cfu, respectively. The 12 µm particle inhalational infection was characterized by a marked involvement of the nasal mucosa and extension of bacterial colonization and inflammatory lesions from the olfactory epithelium through the olfactory nerves (or tracts) to the olfactory bulb (100%), culminating in abscessation of the brain (33%). Initial involvement of the upper respiratory tract lymphoid tissues (nasal-associated lymphoid tissue (NALT) and cervical lymph nodes) was observed in both the 1 and 12 µm particle inhalational infections (80-85%). Necrotising alveolitis and bronchiolitis were evident in both inhalational infections, however, lung pathology was greater after inhalation of the 1 µm particle aerosol with pronounced involvement of the mediastinal lymph node (50%). Terminal disease was characterized by bacteraemia in both inhalational infections with dissemination to the spleen, liver, kidneys, and thymus. Treatment with co-trimoxazole was more effective than treatment with doxycycline irrespective of the size of the particles inhaled. Doxycycline was more effective against the 12 µm particle inhalational infection as evidenced by increased time to death. However, both treatment regimes exhibited significant relapse when therapy was discontinued with massive enlargement and abscessation of the lungs, spleen, and cervical lymph nodes observed.

The cell membrane as a major site of damage during aerosolization of Escherichia coli.

This study aimed to provide data on the survival and site of damage of Escherichia coli cells following aerosolization using two different techniques, nebulization and flow focusing. Four metabolic stains were assessed for their ability to detect respiratory activities and membrane homeostasis in aerosolized E. coli cells. The degree of sublethal injury increased significantly over the 10-min period of aerosolization in E. coli cells aerosolized by using the Collison nebulizer, reaching up to 99.9% of the population. In contrast, a significantly lower proportion of the population was sublethally damaged during aerosolization using the flow-focusing aerosol generator (FFAG). Concomitantly, loss of membrane homeostasis increased at a higher rate in nebulized cells (68 to 71%) than in those aerosolized by using the FFAG (32 to 34%). The activities of respiratory enzymes decreased at increased rates in nebulized cells (27 to 37%) compared to the rates of decrease in cells aerosolized by using the FFAG (59 to 61%). The results indicate that the physiology of an aerosolized bacterium is linked to the method of aerosol generation and may affect the interpretation of a range of aerobiological phenomenon.

Systematic review of military motor vehicle crash-related injuries.

CONTEXT: Motor vehicle crashes account for nearly one third of U.S. military fatalities annually. The objective of this review is to summarize the published evidence on injuries due specifically to military motor vehicle (MMV) crashes. EVIDENCE ACQUISITION: A search of 18 electronic databases identified English language publications addressing MMV crash-related injuries between 1970 and 2006 that were available to the general public. Documents limited in distribution to military or government personnel were not evaluated. Relevant articles were categorized by study design. EVIDENCE SYNTHESIS: The search identified only 13 studies related specifically to MMV crashes. Most were case reports or case series (n=8); only one could be classified as an intervention study. Nine of the studies were based solely on data from service-specific military safety centers. CONCLUSIONS: Few studies exist on injuries resulting from crashes of military motor vehicles. Epidemiologic studies that assess injury rates, type, severity, and risk factors are needed, followed by studies to evaluate targeted interventions and prevention strategies. Interventions currently underway should be evaluated for effectiveness, and those proven effective in the civilian community, such as graduated driver licensing, should be considered for implementation and evaluation in military populations.

Receptor mimicry as novel therapeutic treatment for biothreat agents.

The specter of intentional release of pathogenic microbes and their toxins is a real threat. This article reviews the literature on adhesins of biothreat agents, their interactions with oligosaccharides and the potential for anti-adhesion compounds as an alternative to conventional therapeutics. The minimal binding structure of ricin has been well characterised and offers the best candidate for successful anti-adhesion therapy based on the Galß1-4GlcNAc structure. The botulinum toxin serotypes A-F bind to a low number of gangliosides (GT1b, GQ1b, GD1a and GD1b) hence it should be possible to determine the minimal structure for binding. The minimal disaccharide sequence of GalNAcß1-4Gal found in the gangliosides asialo-GM1 and asialo-GM2 is required for adhesion for many respiratory pathogens. Although a number of adhesins have been identified in bacterial biothreat agents such as Yersinia pestis, Bacillus anthracis, Francisella tularensis, Brucella species and Burkholderia pseudomallei, specific information regarding their in vivo expression during pneumonic infection is lacking. Limited oligosaccharide inhibition studies indicate the potential of GalNAcß1-4Gal, GalNAcß-3Gal and the hydrophobic compound, para-nitrophenol as starting points for the rational design of generic anti-adhesion compounds. A cocktail of multivalent oligosaccharides based on the minimal binding structures of identified adhesins would offer the best candidates for anti-adhesion therapy.