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InSight System Acoustic Requirement Evolution

The InSight Mars lander is to be launched to Mars in 2018 on a rocket, contained within an aeroshell designed for Mars entry. The primary mission objective is to deliever an ultra-sensitive seismometer to the Martian surface for scientific investigations. The preliminary InSight PF acoustic requirement (the heritage Phoenix environment, 139.8 dB overall SPL maximum expected environment) was reduced significantly when the Launch vehicle was selected as the InSight launch vehicle (134.5 dB overall SPL maximum expected environment). This reduction would have resulted in a very low Lander Deck random vibration at SEIS, and other instruments inside the aeroshell, during the system Protoflight (PF) acoustic test with questionable workmanship value. To mitigate this concern JPL and Launch vehicle contractor agreed to increase the InSight PF acoustic test requirement to enhance workmanship testing for science instruments and assemblies inside the aeroshell. The actual InSight system acoustic test vibration responses at the SEIS instrument are compared to the predicted responses.

O'Connell, Michael↗

Overview of the Phoenix Entry, Descent and Landing System Architecture

NASA s Phoenix Mars Lander began its journey to Mars from Cape Canaveral, Florida in August 2007, but its journey to the launch pad began many years earlier in 1997 as NASA s Mars Surveyor Program 2001 Lander. In the intervening years, the entry, descent and landing (EDL) system architecture went through a series of changes, resulting in the system flown to the surface of Mars on May 25th, 2008. Some changes, such as entry velocity and landing site elevation, were the result of differences in mission design. Other changes, including the removal of hypersonic guidance, the reformulation of the parachute deployment algorithm, and the addition of the backshell avoidance maneuver, were driven by constant efforts to augment system robustness. An overview of the Phoenix EDL system architecture is presented along with rationales driving these architectural changes.

Grover, Myron R., III↗

NAIRAS Model transition to the CCMC: real-time dosimetric output and low-Earth orbit applications

NASA’s Nowcast of Atmospheric Ionizing Radiation for Aviation Safety (NAIRAS) model is areal-time, global, physics-based model originally developed to predict exposure to cosmic radiation to air travelers from both galactic and solar sources. A prototype operational NAIRAS model has provided tabular and graphical data products via its public web site for a number of years. We present the advances in the transition of NAIRAS to operations at the Community Coordinated Modeling Center (CCMC), located at NASA Goddard Space Flight Center, where global NAIRAS products are available for a real-time space weather version of the model. In addition, a runs-on-request version provides model output along user selected flight trajectories. We show the global dosimetric output from the model that are available in real-time as well as the new products developed to support commercial spaceflight missions to low-Earth orbit(LEO) and the International Space Station (ISS).

Guillaume Gronof↗

Phoenix Lander's Thermal Evolved Gas Analyzer: Differential Scanning Calorimeter and Mass Spectrometer Database Development

The Mars Scout Phoenix lander will land in the north polar region of Mars in May, 2008. One objective of the Phoenix lander is to search for evidence of past life in the form of molecular organics that may be preserved in the subsurface soil. The Thermal Evolved Gas Analyzer (TEGA) was developed to detect these organics by coupling a simultaneous differential thermal analyzer (SDTA) with a mass spectrometer. Martian soil will be heated to approx.1000 C and potential organic decomposition products such as CO2, CH4 etc. will be examined for with the MS. TEGA s SDTA will also assess the presence of endothermic and exothermic reactions that are characteristic of soil organics and minerals as the soil is heated. The MS in addition to detecting organic decompositon products, will also assess the levels of soil inorganic volatiles such as H2O, SO2, and CO2. Organic detection has a high priority for this mission; however, TEGA has the ability to provide valuable insight into the mineralogical composition of the soil. The overall goal of this work is to develop a TEGA database of minerals that will serve as a reference for the interpretation of Phoenix-TEGA. Previous databases for the ill-fated Mars Polar Lander (MPL)-TEGA instrument only went to 725 C. Furthermore, the MPL-TEGA could only detect CO2 and H2O while the Phoenix-TEGA MS can examine up to 144 atomic mass units. The higher temperature Phoenix-TEGA SDTA coupled with the more capable MS indicates that a higher temperature database is required for TEGA interpretation. The overall goal of this work is to develop a differential scanning calorimeter (DSC) database of minerals along with corresponding MS data of evolved gases that can used to interpret TEGA data during and after mission operations. While SDTA and DSC measurement techniques are slightly different (SDTA does not use a reference pan), the results are fundamentally similar and thus DSC is a useful technique in providing comparative data for the TEGA database. The objectives of this work is to conduct DSC and MS analysis up to 1000 C of select minerals that may be found in the martian soil.

Sutter, B.↗

Desert-Urban System Integrated Atmospheric Monsoon (DUSTIEAIM) in the Southwestern United States Science Plan

The Desert-Urban System Integrated Atmospheric Monsoon (DUSTIEAIM) campaign is a groundbreaking, high-impact scientific mission that will transform how we understand and respond to energy and water challenges in one of America’s fastest-growing and most heat-stressed urban regions: Phoenix, Arizona. Starting in April 2026, this 18-month field campaign harnesses the full power of the U.S. Department of Energy (DOE) Atmospheric Radiation Measurement (ARM) user facility and an interdisciplinary science team including national laboratories, universities, and agencies with a broad range of subject-matter expertise. With cutting-edge instruments, active and passive ground-based sensors, radars, and integrated modeling, DUSTIEAIM will deliver the most comprehensive environmental data set ever collected for a desert-urban-agricultural interface.

54 ENVIRONMENTAL SCIENCES↗

Participatory Technology Assessment for Mars Mission Planning: Public Values and Rationales

Public support and interest are needed to design an ambitious human spaceflight program. However, it is difficult to understand what the public values and would support. And it is even more challenging and rare to consider public views prior to actually developing a mission. Participatory technology assessment (pTA) is a method that aims to understand public preferences and values in order to inform upstream government decision-making. We assess a recently completed experiment in pTA, the "Informing NASA's Asteroid Initiative" project. Through a cooperative agreement with NASA, the Expert and Citizen Assessment of Science and Technology (ECAST) network conducted a pTA-based forum on NASA's Asteroid Initiative and the Journey to Mars. ECAST organized two citizen forums in Phoenix, Arizona and Boston, Massachusetts in November 2014, with a total of 183 citizens selected so as to minimize self-selection biases. This paper focuses on the "Journey to Mars" session, which had the primary goal of soliciting citizen perceptions about different Mars exploration scenarios and mission planning approaches. Citizens were given background information about three potential Mars exploration scenarios that NASA could carry out: 1) Crewed orbital mission to direct robots on the surface of Mars; 2) Short exploratory crewed mission to the surface of Mars; and 3) establishing a permanent settlement. Citizens then engaged in structured facilitated discussions about their preferences among the scenarios and NASA's mission planning approach. Using a grounded theory coding approach, we analyzed participants' written rationales and dialogue about Mars exploration. In general, participants did not show a strong preference for any particular mission profile, but there was a slight preference for the crewed orbital robotics scenario. Participants who supported this approach saw it as the quickest, safest, and least costly road to a successful mission. However, many participants were interested in seeing "boots on the ground," as they believed this would propel scientific advancement, increase excitement about space exploration, and make humans a "two-planet species."

Bertrand, Pierre↗

Phoenix: Preliminary design of a high speed civil transport

The goal of the Phoenix Design Project was to develop a second generation high speed civil transport (HSCT) that will meet the needs of the traveler and airline industry beginning in the 21st century. The primary emphasis of the HSCT is to take advantage of the growing needs of the Pacific Basin and the passengers who are involved in that growth. A passenger load of 150 persons, a mission range of 5150 nautical miles, and a cruise speed of Mach 2.5 constitutes the primary design points of this HSCT. The design concept is made possible with the use of a well designed double delta wing and four mixed flow engines. Passenger comfort, compatibility with existing airport infrastructure, and cost competitive with current subsonic aircraft make the Phoenix a viable aircraft for the future.

Aguilar, Joseph↗

Multibody Modeling and Simulation for the Mars Phoenix Lander Entry, Descent and Landing

A multi-body flight simulation for the Phoenix Mars Lander has been developed that includes high fidelity six degree-of-freedom rigid-body models for the parachute and lander system. The simulation provides attitude and rate history predictions of all bodies throughout the flight, as well as loads on each of the connecting lines. In so doing, a realistic behavior of the descending parachute/lander system dynamics can be simulated that allows assessment of the Phoenix descent performance and identification of potential sensitivities for landing. This simulation provides a complete end-to-end capability of modeling the entire entry, descent, and landing sequence for the mission. Time histories of the parachute and lander aerodynamic angles are presented. The response of the lander system to various wind models and wind shears is shown to be acceptable. Monte Carlo simulation results are also presented.

Queen, Eric M.↗

AutoGen Version 5.0

Version 5.0 of the AutoGen software has been released. Previous versions, variously denoted Autogen and autogen, were reported in two articles: Automated Sequence Generation Process and Software (NPO-30746), Software Tech Briefs (Special Supplement to NASA Tech Briefs), September 2007, page 30, and Autogen Version 2.0 (NPO- 41501), NASA Tech Briefs, Vol. 31, No. 10 (October 2007), page 58. To recapitulate: AutoGen (now signifying automatic sequence generation ) automates the generation of sequences of commands in a standard format for uplink to spacecraft. AutoGen requires fewer workers than are needed for older manual sequence-generation processes, and greatly reduces sequence-generation times. The sequences are embodied in spacecraft activity sequence files (SASFs). AutoGen automates generation of SASFs by use of another previously reported program called APGEN. AutoGen encodes knowledge of different mission phases and of how the resultant commands must differ among the phases. AutoGen also provides means for customizing sequences through use of configuration files. The approach followed in developing AutoGen has involved encoding the behaviors of a system into a model and encoding algorithms for context-sensitive customizations of the modeled behaviors. This version of AutoGen addressed the MRO (Mars Reconnaissance Orbiter) primary science phase (PSP) mission phase. On previous Mars missions this phase has more commonly been referred to as mapping phase. This version addressed the unique aspects of sequencing orbital operations and specifically the mission specific adaptation of orbital operations for MRO. This version also includes capabilities for MRO s role in Mars relay support for UHF relay communications with the MER rovers and the Phoenix lander.

Gladden, Roy E.↗

The Rapid Response Radiation Survey (R3S) Mission Using the HiSat Conformal Satellite Architecture

The Rapid Response Radiation Survey (R3S) experiment, designed as a quick turnaround mission to make radiation measurements in Low Earth Orbit (LEO), will fly as a hosted payload in partnership with NovaWurks using their Hyper-integrated Satlet (HISat) architecture. The need for the mission arises as the Nowcast of Atmospheric Ionization Radiation for Aviation Safety (NAIRAS) model moves from a research effort into an operational radiation assessment tool. Currently, airline professionals are the second largest demographic of radiation workers and to date their radiation exposure is undocumented in the USA. The NAIRAS model seeks to fill this information gap. The data collected by R3S, in addition to the complementary data from a NASA Langley Research Center (LaRC) atmospheric balloon mission entitled Radiation Dosimetry Experiment (RaD-X), will validate exposure prediction capabilities of NAIRAS. The R3S mission collects total dose and radiation spectrum measurements using a Teledyne μDosimeter and a Liulin-6SA2 LED spectrometer. These two radiation sensors provide a cross correlated radiometric measurement in combination with the Honeywell HMR2300 Smart Digital Magnetometer. The magnetometer assesses the Earth's magnetic field in the LEO environment and allows radiation dose to be mapped as a function of the Earth's magnetic shielding. R3S is also unique in that the radiation sensors will be exposed on the outer surface of the spacecraft, possibly making this the first measurements of the LEO radiation environment with bare sensors. Viability of R3S as an extremely fast turnaround mission is due, in part, to the nature of the robust, well-defined interfaces of the conformal satellite HiSat Architecture. The HiSat architecture, which was developed with the support of the Defense Advanced Research Projects Agency's (DARPA's) Phoenix Program, enabled the R3S system to advance from the first concept to delivery of preliminary design review (PDR) level documents in 29 calendar days. The architecture allows for interface complexities between the specific devices and the satellite bus to be resolved in a standardized interface control document (ICD). The ICD provided a readymade framework to interface to the modular satellite bus. This modularity allowed for approximately 90% of the R3S system to be designed and fabricated in two months without constraint of the hosting satellite's development cycle. This paper discusses the development of the R3S experiment as made possible by use of the HiSat architecture. The system design and operational modes of the experiment are described, as well as the experiment interfaces to the HiSat satellite via the user defined adapter (UDA) provided by NovaWurks. This paper outlines the steps taken by the project to execute the R3S mission in the 4 months of design, build, and test. Additionally portrayed is the ground work done at LaRC to posture the organization for a fast response and the process by which the opportunity was identified as aligning with key strategic goals. Finally, a description of the engineering process is provided, including the use of facilitated rapid/concurrent engineering sessions, the associated documentation, and the review process employed.

Miller, Nathanael A.↗

Mars in Situ Resource Utilization Technology Evaluation

We have examined the technologies required to enable Mars In-Situ Resource Utilization (ISRU) because our understanding of Mars resources has changed significantly in the last five years as a result of recent robotic missions to the red planet. Two major developments, (1) confirmation of the presence of near-surface water in the form of ice in very large amounts at high latitudes by the Phoenix Lander and (2) the likely existence of water at lower latitudes in the form of hydrates or ice in the top one meter of the regolith, have the potential to change ISRU technology selection. A brief technology assessment was performed for the most promising Mars atmospheric gas processing techniques: Reverse Water Gas Shift (RWGS) and Methanation (aka Sabatier), as well as an overview of soil processing technology to extract water from Martian soil.

Muscatello, Anthony C.↗

Electrochemical Life Detection Methods for Ocean World Exploration

Ubiquitous across terrestrial life is cellular machinery that allows chemical energy flow by facilitating and regulating electron-transfer and chemical modification pathways. Key classes of energy transport molecules enable this movement of electrons for a variety of biological purposes. Additionally, biological enzymes function to add or remove functional groups such asphosphate moieties to redox biomolecules. Presumably, extraterrestrial life is likely to rely on similar energy transport mechanisms. With the search for life in our solar system focused on the icy satellites of Jupiter and Saturn, Europa and Enceladus, developing instrumentation capable of measuring electrochemical redox signatures representative of biomolecules or enzymatic activity in seawater appears a promising and novel means of life detection. Here, we report our adaptation of the Mars Phoenix Wet Chemistry Laboratory (WCL) electroanalytical voltammetry capabilities to assay life-critical redox molecules in synthetic seawater representative of a saline alkaline solution similar to what has been predicted from the Cassini mission data of Enceladus’ sub-surface ocean. In addition, we employ a well-established electrochemical assay that indicates phosphatase activity by comparing substrate and product redox signatures. Our study demonstrated a 10 nM limit of detection for biological redox molecules and a 3 aM limit of detection for alkaline phosphatase in seawater. Incorporation of these methods into next generation WCL payloads aimed at ocean world life detection will enable the search for biological redox-active species and enzymatic activity as indicators of life.

planetary instruments↗

The ARADS Project as an Astrobiological Survey Mission Prototype

An astrobiological survey prior to human arrival will be important for preservation of signs of any existing or past evidence of Martian life, to reduce the risk of unintentional forward contamination of Martian environments in which terrestrial microbes might be able to gain a foothold (e.g., [1]), and to potentially protect the health of any human crews and enable their safe return to Earth. Evaluating for the presence of life and biosignatures may become a critical-path Mars exploration precursor in the not-so-far future, circa 2030-35. The near-subsurface, with its potential combination of volatiles, vapor exchange with the surface, and shelter from surface radiation, will be a primary initial target for in-situ astrobiological investigation. Sample acquisition, drilling and sample handling are key elements of robotic missions that target the subsurface [2], as well as near-surface ground ice deposits. In the mid 2000s, the Mars Astrobiology Field Laboratory (AFL) was proposed as a follow-on mission concept to the Mars Science Laboratory (MSL)/Curiosity rover mission [3]. The AFL would have included precision drilling to 2m, automated sample handling and remote processing of cores. However, the AFL concept needed further development and maturation. The Atacama Rover Astrobiology Drilling Studies (ARADS) project concept [4] was in part AFL-inspired. The 2014 ARADS concept included a medium sized rover, with habitability instruments, including the Phoenix Wet Chemistry Laboratory (WCL) [5], and an enhanced version of the ExoMars Mars Organic Molecule Analyzer (MOMA) Laser Desorption-Linear Ion Trap Mass Spectrometer (LITMS) [6]. ARADS likewise included two biosignature detection instruments -- the Signs of Life Detector (SOLID) immunoassay instrument [7,8] and the Chemical Laptop-derived [9] Planetary In-Situ Capillary Electrophoresis System (PISCES), while mitigating Planetary Protection-relevant contamination issues [10]. ARADS project objectives were to demonstrate the potential science return for such a mission in the context of the search for evidence of life, and technically to demonstrate the feasibility of roving with drilling missions to Mars. ARADS technology goals targeted analog-site demonstrations of automated drilling and sampling, integrated with its mobile habitability and biosignature detection instruments. And ARADS science operations goals developed remote science, contamination mitigation, and communications methods to enable simulating AFL-like science operations with time delays, directed by a remote science team[11]. Five subsequent Mars analog site field demonstrations (in Chile’s hyperarid Atacama Desert, 2016-19) provided a step-by-step development pathway for the ARADS platform’s science capabilities and systems integration, culminating in an integrated remote roving mission simulation in the final ARADS year (2019).

biosignatures↗

Mars In-Situ Resource Utilization Technology Evaluation

We have examined the technologies required to enable Mars Tn-Situ Resource Utilization (ISRU) because our understanding of Mars resources has changed significantly in the last five years as a result of recent robotic missions to the red planet [1-4]. Two major developments, (1) confirmation of the presence of near-surface water in the form of ice in very large amounts at high latitudes by the Phoenix Lander and (2) the likely existence of water at lower latitudes in the form of hydrates or ice in the top one meter of the regolith, have the potential to change ISRU technology selection. A brief technology assessment was performed for the most promising Mars atmospheric gas processing techniques: Reverse Water Gas Shift (RWGS) and Methanation (aka Sabatier), as well as an overview of soil.processing technology to extract water from Martian soil.

Santiago-Maldonado, Edgardo↗

Relay Telecommunications for the Coming Decade of Mars Exploration

Over the past decade, an evolving network of relay-equipped orbiters has advanced our capabilities for Mars exploration. NASA's Mars Global Surveyor, 2001 Mars Odyssey, and Mars Reconnaissance Orbiter (MRO), as well as ESA's Mars Express Orbiter, have provided telecommunications relay services to the 2003 Mars Exploration Rovers, Spirit and Opportunity, and to the 2007 Phoenix Lander. Based on these successes, a roadmap for continued Mars relay services is in place for the coming decade. MRO and Odyssey will provide key relay support to the 2011 Mars Science Laboratory (MSL) mission, including capture of critical event telemetry during entry, descent, and landing, as well as support for command and telemetry during surface operations, utilizing new capabilities of the Electra relay payload on MRO and the Electra-Lite payload on MSL to allow significant increase in data return relative to earlier missions. Over the remainder of the decade a number of additional orbiter and lander missions are planned, representing new orbital relay service providers and new landed relay users. In this paper we will outline this Mars relay roadmap, quantifying relay performance over time, illustrating planned support scenarios, and identifying key challenges and technology infusion opportunities.

Edwards, C.↗

Science in Exploration: From the Moon to Mars and Back Home to Earth

NASA is embarking on a grand journey of exploration that naturally integrates the past successes of the Apollo missions to the Moon, as well as robotic science missions to Mars, to Planet Earth, and to the broader Universe. The US Vision for Space Exporation (VSE) boldly lays out a plan for human and robotic reconnaissance of the accessible Universe, starting with the surface of the Moon, and later embracing the surface of Mars. Sustained human and robotic access to the Moon and Mars will enable a new era of scientific investigation of our planetary neighbors, tied to driving scientific questions that pertain to the evolution and destiny of our home planet, but which also can be related to the search habitable worlds across the nearby Universe. The Apollo missions provide a vital legacy for what can be learned from the Moon, and NASA is now poised to recapture the lunar frontier starting with the flight of the Lunar Reconnaissance Orbiter (LRO) in late 2008. LRO will provide a new scientific context from which joint human and robotic exploration will ensue, guided by objectives some of which are focused on the grandest scientific challenges imaginable : Where did we come from? Are we alone? and Where are we going? The Moon will serve as an essential stepping stone for sustained human access and exploration of deep space and as a training ground while robotic missions with ever increasing complexity probe the wonders of Mars. As we speak, an armada of spacecraft are actively investigating the red planet both from orbit (NASA's Mars Reconnaissance Orbiter and Mars Odyssey Orbiter, plus ESA's Mars Express) and from the surface (NASA's twin Mars Exploration Rovers, and in 2008 NASA's Phoenix polar lander). The dramatically changing views of Mars as a potentially habitable world, with its own flavor of global climate change and unique climate records, provides a new vantage point from which to observe and question the workings of our own planet Earth. By 2010 NASA will have its first mobile analytical laboratory operating on the surface of Mars (Mars Science Laboratory) in search of potentially subtle expressions of past life or at least of life-hospitable environments. Meanwhile back here on Planet Earth, NASA will be continuing to implement an increasingly comprehensive program of robotic missions that address major issues associated with global climate variability, and the "state variables" that affect the quality of human life on our home planet. Ultimately, the fmits of NASA's emergent program of Exploration (VSE) will provide never-beforepossible opportunities for scientific leadership and advancement, culminating in a new state of awareness from which to better plan for the sustainability of life on Earth and for extending Earth life to the Moon and eventually to Mars. As NASA nears its 50th anniversary, the unimaginable and unexpected wealth of strategic knowledge its missions have generated about Earth, the Universe, and our local Solar System boggles the mind and serves as a legacy of knowledge for Educators to inspire future generations.

Garvin, James B.↗

Autonomous Measurement of Electrochemical Redox Couples for In-Situ Characterization of Enceladus Plume Samples

A 3D-printed microfluidic electrochemical cell with integrated electrodes was developed to demonstrate autonomous in-situ electrochemical characterization of terrestrial seawater samples. The system is designed to extend the capabilities of the 2008 Phoenix Lander’s Wet Chemistry Laboratory (WCL) that analyzed the soluble chemistry of the Martian soil, by miniaturizing its fluidic architecture for analysis of µL-volume samples as expected from an Enceladus fly-by mission. The microfluidic device has a total fluid volume of 48 µL and includes dual channels each populated with 7 electrodes. Measurement of pH was measured using an iridium oxide electrode. Cyclic voltammetry with Au, Pt, and glassy carbon working electrodes was used to measure redox couples in synthetic seawater that contained inorganic and bio-organic molecules. Glassy carbon was found to have minimal risk of interference from constituents of synthetic seawater when measuring molecular organic redox species; however, care must be used to prevent interference due to potential oxidation or dissolution in the case of Au working electrodes. A completely autonomous end-to-end run including sample delivery was performed, to demonstrate the feasibility of microfluidic-based cyclic voltammetry measurements for in-situ characterization of redox couples in Ocean World samples.

Planetary Instruments↗

Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled from the coupon to recover the spores. One hundred μl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

cleanroom↗