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This project uses fast, high-throughput in vitro approach to study new nanomaterials applicable to Chemical Biological Defense (CBD) for their environmental and human related toxicity. Four CBD nanomaterials were evaluated for their aquatic toxicity by Microtox test and human cytotoxicity by NRU assay. These studies fill the data gaps and provide toxicity information useful in risk assessment when the environmental and human exposure to these new CBD nanomaterials. In addition, Transepithelial Electrical Resistance (TER) and Transepithelial Permeability (TEP) assays were developed and tested on EpiAirway, a 3-D human tracheal/bronchial epithelial equivalent. Further evaluation of the effects of these CBD nanomaterials on tissue tight junction and barrier function is the on-going study. This project supports the efforts of USAPHC/OTSG in CBRN incident response for protecting the work force, civilian and military, from the unintended consequences of nanotechnology processes and materials exposure.

Sustainable use of training ranges requires the development of compounds that have a minimal impact on the environment when used in a weapon system. Triaminoguanidinium-1-methyl-5- nitriminotetrazolate (TAG-MNT) is a novel, explosive, military compound of interest for application in some weapon systems. Little is known of its toxicologic properties. To ensure the health of potentially exposed personnel and the environment, several initial toxicity investigations were conducted and the results compared with another widely used energetic (hexahydro-1,3,5-trinitro-1,3,5-triazine; RDX). In a novel microplate Ames assay, TAG-MNT was a weak mutagen only at the limit concentration of 2 g/L. However, TAG-MNT was cytotoxic to bacteria and a human liver cell line at 250 mg/L and greater. Unlike RDX, TAG-MNT did not have an affinity for the GABAa receptor convulsant site, and was predicted not to induce seizure. After acute oral dosing in female rats, TAG-MNT had no apparent adverse effect up to the limit dose of 2 g/kg. However, daily oral dosing for 14 days at exposures of 1000 mg/kg-d and above caused reduction in food intake, weight loss, increased kidney weight, leucopenia, and elevated blood urea nitrogen and creatinine levels. Leucopenia, increased liver mass, evidence of liver hepatocyte necrosis and centrilobular hypertrophy were observed at 500 mg/kg-d and above. TAG-MNT was negative in the rat micronucleus assay of blood samples. Based on these data, the 14-day oral No observed adverse effect level (NOAEL) and the lowest observed adverse effect level (LOAEL) is 250 and 500 mg/kg/day, respectively., Published in Journal of Toxicology and Environmental Health Sciences, v3 n3 p80-94, March 2011.

Environmental consequences of using specific compounds in military applications have lead to undesirable outcomes. Examples include expensive clean up operations, off-site groundwater migrations, and closing of operational ranges. Additionally, the use of specific weapon systems containing compounds with unknown or limited toxicity data may lead to adverse health consequences to soldiers and civilians. Often incomplete health information has led to inaccurate full life cycle cost estimates. The Army is currently exploring replacement substances for compounds identified as hazardous to health from an environmental and/or occupational (ESOH) perspective. To evaluate the environmental and occupational health consequences of new replacement compounds, a tiered approach has been developed and used within the program. Early in the research stage models are primarily relied upon (e.g. QSAR approaches) and as the technology progresses, a greater reliance is placed on experimental data beginning with in vitro techniques. As greater investment is devoted to system development, less uncertain in vivo data are collected. Together, these data and weighted lines of evidence are used to help guide life cycle decisions in the development of new systems. Examples will be provided., Presented at the Annual Partners in Environmental Technology Technical Symposium & Workshop (15th), Washington, DC, 30 Nov - 2 Dec 2010. Sponsored by SERDP and ESTCP. U.S. Government or Federal Rights License

Polymerase chain reaction (PCR) is an exquisitely sensitive method for the amplification and detection of genetic material (DNA and RNA sequences). We have developed several assays using a variation of PCR that proceeds very quickly and allows the monitoring of the progress of the reaction in real time. We describe here assays for the detection of the gene encoding staphylococcal enterotoxin A (SEA), a toxin produced by the bacterium Staphylococcus aureus and an agent responsible for a significant fraction of food poisoning incidents worldwide. We also describe several assays for the detection of a simulant of viral pathogens, the bacteriophage MS2., See also ADM001523.

Antibodies are essential components in biosensors and enable the detection of biological warfare agents with both sensitivity and selectivity. A biomanufacturing process was developed for the production of a monoclonal antibody (mAb) specific for detecting the Frencisella tularensls organism in the environment. This report describes a process for the production of 12 g of monocional antibody from an in vitro call culture system. Hybridoma calls were grown in 2-L gas permeable call culture bags. The numbers of both total and live calls in culture media were measured throughout the process and correlated with antibody concentration over time. Antibody concentrations in samples collected during culture were monitored and during the purification process were analyzed by analytical high-performance liquid chromatography. Antibodies were purified from spent culture media, using a single-step Protein-A affinity chromatography procedure. The overall yield for the downstream purification process is approximately 78%. The purifies of the purified mAb lots were around 94 - 99%, as determined by an analytical gel permeation chromatographic method. The purified mAb was active as reference lots of the antibody produced elsewhere, as determined by direct enzyme-linked immunosorbent assays.