Comparing Methods to Genetically Engineer Bacteriophage and Increase Host Range.

INTRODUCTION: Antibacterial resistance is an emerging problem in military medicine. Disruptions to the health care systems in war-torn countries that result from ongoing conflict can potentially exacerbate this problem and increase the risk to U.S. forces in the deployed environment. Therefore, novel therapies are needed to mitigate the impact of these potentially devastating infections on military operations. Bacteriophages are viruses that infect and kill bacteria. They can be delivered as therapeutic agents and offer a promising alternative to traditional antibiotic chemotherapy. There are several potential benefits to their use, including high specificity and comparative ease of use in the field setting. However, the process of engineering phages for military medical applications can be a laborious and time-consuming endeavor. This review examines available techniques and compares their efficacy. MATERIALS AND METHODS: This review evaluates the scientific literature on the development and application of four methods of bacteriophage genome engineering and their consideration in the context of military applications. Preffered Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed for a systematic review of available literature that met criteria for analysis and inclusion. The research completed for this review article originated from the United States Military Academy's library "Scout" search engine, which compiles results from 254 available databases (including PubMed, Google Scholar, and SciFinder). Particular attention was focused on identifying useful mechanistic insight into the nature of the engineering technique, the ease of use, and the applicability of the technique to countering the problem of antimicrobial resistance in the military setting. RESULTS: A total of 52 studies were identified that met inclusion criteria following PRISMA guidelines. The bioengineering techniques analyzed included homologous recombination (12 articles), in vivo recombineering (9 articles), bacteriophage recombineering of electroporated DNA (7 articles), and the CRISPR-Cas system (10 articles). Rates of success and fidelity varied across each platform, and comparative benefits and drawbacks are considered. CONCLUSIONS: Each of the phage engineering techniques addressed herein varies in amount of effort and overall success rate. CRISPR-Cas-facilitated modification of phage genomes presents a highly efficient method that does not require a lengthy purification and screening process. It therefore appears to be the method best suited for military medical applications.

Lessons Learned From the U.S. Military Experience With Hantavirus During the Korean War.

INTRODUCTION: The Korean War (1950-1953) consisted of two phases. The first was a rapid mobile phase, and the second was a slow and stationary phase. During the stationary phase, approximately 3,000 UN troops became infected with a then unknown agent. The resulting illness began with flu-like symptoms and often progressed to a severe hemorrhagic fever leading to kidney failure and death. However, the cause was not to be identified until well over 20 years following the conclusion of the war when Dr. Ho Wang Lee succeeded in isolating Hantavirus from field rodents. The U.S. Military experience with Hantavirus during the Korean War is a case study of the potential impact of war-related environmental change on disease transmission. The lessons learned from this experience should inform future military medical planning and serve as a reminder of the impact that an unknown agent can have on military operations. MATERIALS AND METHODS: A literature review of all available records with respect to the U.S. Military experience with Hantavirus was conducted. PubMed was the primary search engine used for this review. However, primary literature and historical accounts were also evaluated. All records were examined for environmental, epidemiological, and public health data regarding hemorrhagic fever outbreaks among U.S. forces during the Korean War. The quantitative and qualitative data from these sources were analyzed and evaluated within the context of military medical planning and force health protection to derive lessons learned that should be applied to the management and mitigation of viral disease in future wars. RESULTS: Widespread deforestation resulting from war-related efforts most likely played a significant role in the outbreaks of Hantavirus among UN forces during the war. A lack of cultural literacy and an overreliance on erroneous assumptions most likely delayed the identification of the true causative agent. It is conceivable that these delays led to an increased casualty rate and that they had a negative impact on military operations during the war. CONCLUSIONS: A basic understanding of the ecological mechanisms that maintain species diversity in the local environment coupled with an appreciation for the impact of environmental change on this diversity is of paramount importance for the prevention and mitigation of viral disease outbreaks in the deployed setting. Military medical planners should become familiar with the medical literature of the region in which they will be operating as this literature often describes the agents that will most likely be encountered by U.S. forces.

A Brief Primer on the Concept of the Neuroweapon for U.S. Military Medical Personnel.

The malevolent application of neuroscience is an emerging threat to the U.S. military. At present, U.S. military medical personnel are not capable of adequately diagnosing or treating the injuries and illnesses that may result from exposure to potential neuroweapons. This fact was illustrated in 2016 when U.S. diplomats serving in Havana, Cuba reported hearing strange noises accompanied by a constellation of unexplained health effects. Similar incidents have been reported in China and Russia. Although various hypotheses have been put forward to explain these symptoms, none of them have been verified. The reported symptoms were analogous to the physiological responses that have been produced in the laboratory by exposing volunteers to pulsed microwave energy. However, these incidents of undetermined origin demonstrate that widespread neurological illness can be disruptive to U.S. government operations and that it is currently not possible to identify the cause, determine the correct treatment, or ascribe attribution to potential neuroweapon use in an overseas setting. Since it is likely that Special Operations medical personnel will be among the first to respond to neuroweapon attacks in the deployed environment, it is essential that they be made aware of this emerging threat and that efforts be made to incorporate potential directed energy neuroweapons and other neuroweapon configurations into future Chemical, Biological, Radiological, Nuclear, and high yield Explosives (CBRN-E) training modules. The intention of this article is to introduce the concept of the neuroweapon to military medical personnel and to provide a brief review of the relevant literature.