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NASA NTRSDate not supplied

Nutrient Stability in NASA Spaceflight Experiment Rodent Food Bars

Nutrient-upgraded Rodent Food Bars are the standard diet for mice involved in spaceflight experiments aboard the International Space Station as part of NASA’s Rodent Research Project. These Food Bars are a twin-screw extruded AIN-93G based product with nutrient additives to preserve a healthy and complete diet for the animals, and the bar form factor is necessary for use in the microgravity environment. Due to the nature of NASA flight projects and the lengthy production process of these Food Bars, a shelf-life assessment was undertaken to determine nutrient stability over an extended storage time. To assess palatability and stability over an extended duration, a panel of lipid oxidation markers and both fat- and water-soluble vitamins were assessed under various time and temperature conditions via AOAC International standardized methodology. Thiamine was the least stable vitamin at room temperature. Bars that had been continually kept at 4C (t=6, 12, 27 months) showed significantly higher levels of thiamine (via Student’s t-test) compared to all other bars that underwent some degree of ambient temperature storage. This suggests that refrigeration is a key component in maintaining adequate nutrient levels in these relatively high moisture (average 29.3%) food bars. Also of note was the level of Vitamin A in the form of retinol. There was no correlation to storage temperature, but rather the only factor having an effect on retinol levels was time. As low, yet consistent decrease in retinol was observed over the course of the experiment, though at no time did the retinol levels drop below the rodent Nutrient Requirements for Laboratory Animals (NRC). In conclusion, over the 27-month time course, all analyzed vitamins and lipid oxidation factors remained relatively stable and at levels which provide fully sufficient nutrition to rodents during spaceflight. Some groups however, displayed higher-than-expected aerobic and anaerobic bacteria colony forming units, which is believed to stem from a since-revised handling technique.

Food↗

DOE OSTI2023

Concurrent colonization of rodent kidneys with multiple species and serogroups of pathogenic Leptospira

ABSTRACT Rodents are important reservoir hosts of pathogenic leptospires in the US Virgin Islands. Our previous work determined that trapped rodents were colonized withLeptospira borgpeterseniiserogroup Ballum (n= 48) and/orLeptospira kirschneriserogroup Icterohaemorrhagiae (n= 3). In addition, nine rodents appeared to be colonized with a mixed population comprising more than one species/serogroup. The aim of this study was to validate this finding by characterizing clonal isolates derived from cultures of mixed species. Cultures of presumptive mixed species (designated LR1, LR5, LR37, LR57, LR60, LR61, LR68, LR70, and LR72) were propagated in different media including Hornsby-Alt-Nally (HAN) media, incubated at both 29℃ and 37℃, and T80/40/LH incubated at 29℃. Polyclonal reference antisera specific for serogroup Ballum and Icterohaemorrhagiae were used to enrich for different serogroups followed by subculture on agar plates. Individual colonies were then selected for genotyping and serotyping. Of the nine cultures of mixed species/serogroups, a single clonal isolate was separated in five of them:L. borgpeterseniiserogroup Ballum in LR1, LR5, and LR37, andL. kirschneriserogroup Icterohaemorrhagiae in LR60 and LR72. In four of the cultures with mixed species (LR57, LR61, LR68, and LR70), clonal isolates of bothL. borgpeterseniiserogroup Ballum andL. kirschneriserogroup Icterohaemorrhagiae were recovered. Our results definitively establish that rodents can be colonized with more than one species/serogroup ofLeptospiraconcurrently. The identification and characterization of multiple species/serogroups ofLeptospirafrom individual reservoir hosts of infection are essential to understand the epidemiology and transmission of disease to both human and domestic animal populations. IMPORTANCE PathogenicLeptospira, the causative agent of human and animal leptospirosis, comprise a diverse genus of species/serogroups which are inherently difficult to isolate from mammalian hosts due to fastidious growth requirements. Molecular evidence has indicated that reservoir hosts ofLeptospiramay shed multiple species concurrently. However, evidence of this phenomena by culture has been lacking. Culture is definitive and is essential for comprehensive characterization of recovered isolates by high-resolution genome sequencing and serotyping. In this work, a protocol using recently developed novel media formulations, in conjunction with reference antisera, was developed and validated to demonstrate the recovery of multiple species/serogroups of pathogenicLeptospirafrom the same host. The identification and characterization of multiple species/serogroups ofLeptospirafrom individual reservoir hosts of infection are essential to understand the epidemiology and transmission of disease to both human and domestic animal populations.

Biotechnology & Applied Microbiology↗

NASA NTRS1989

The rodent research animal holding facility as a barrier to environmental contamination

The rodent Research Animal Holding Facility (RAHF), developed by NASA Ames Research Center (ARC) to separately house rodents in a Spacelab, was verified as a barrier to environmental contaminants during a 12-day biocompatibility test. Environmental contaminants considered were solid particulates, microorganisms, ammonia, and typical animal odors. The 12-day test conducted in August 1988 was designed to verify that the rodent RAHF system would adequately support and maintain animal specimens during normal system operations. Additional objectives of this test were to demonstrate that: (1) the system would capture typical particulate debris produced by the animal; (2) microorganisms would be contained; and (3) the passage of animal odors was adequately controlled. In addition, the amount of carbon dioxide exhausted by the RAHF system was to be quantified. Of primary importance during the test was the demonstration that the RAHF would contain particles greater than 150 micrometers. This was verified after analyzing collection plates placed under exhaust air ducts and rodent cages during cage maintenance operations, e.g., waste tray and feeder changeouts. Microbiological testing identified no additional organisms in the test environment that could be traced to the RAHF. Odor containment was demonstrated to be less than barely detectable. Ammonia could not be detected in the exhaust air from the RAHF system. Carbon dioxide levels were verified to be less than 0.35 percent.

Savage, P. D., Jr.↗

NASA NTRS1989

The rodent Research Animal Holding Facility as a barrier to environmental contamination

The rodent Research Animal Holding Facility (RAHF), developed by NASA Ames Research Center (ARC) to separately house rodents in a Spacelab, was verified as a barrier to environmental contaminants during a 12-day biocompatibility test. Environmental contaminants considered were solid particulates, microorganisms, ammonia, and typical animal odors. The 12-day test conducted in August 1988 was designed to verify that the rodent RAHF system would adequately support and maintain animal specimens during normal system operations. Additional objectives of this test were to demonstrate that: (1) the system would capture typical particulate debris produced by the animal; (2) microorganisms would be contained; and (3) the passage of animal odors was adequately controlled. In addition, the amount of carbon dioxide exhausted by the RAHF system was to be quantified. Of primary importance during the test was the demonstration that the RAHF would contain particles greater than 150 micrometers. This was verified after analyzing collection plates placed under exhaust air ducts and and rodent cages during cage maintenance operations, e.g., waste tray and feeder changeouts. Microbiological testing identified no additional organisms in the test environment that could be traced to the RAHF. Odor containment was demonstrated to be less than barely detectable. Ammonia could not be detected in the exhaust air from the RAHF system. Carbon dioxide levels were verified to be less than 0.35 percent.

Savage, P. D., Jr.↗

NASA NTRS1997

Use of the Aromascan(TM) Instrument for Nonsubjective Evaluation of Rodent Spaceflight Hardware

This report describes the verification and utilization of the AromaScan(TM) (Hollis, NH) instrument for the ground-based evaluation of odor containment by various spaceflight habitats developed at NASA's Ames Research Center (ARC). The AromaScan(TM) instrument is an electronic odor detection system consisting of 32 polymer sensors that respond differentially to 10 different chemical groups present in an air sample. The AromaScan(TM) system also includes neural network software for constructing a database of known odors, against which an unknown odor can be compared. At present, the standard method for characterizing rodent odor containment during the development and testing of spaceflight hardware is the use of a human odor assessment panel. However, this can be a very time consuming and costly process, and the results are inherently subjective. The AromaScan(TM) system should produce more consistent and objective results, as well as a cost savings in the long term. To test and verify the AromaScan(TM) instrument, daily air samples will be collected from the exhaust port of rodent habitats, during experiment development tests, then injected into the instrument and used to create a database of recognizable odors. Human sniff tests will be performed in conjunction with the AromaScan(TM) analysis, and the results will be correlated. We will then teach the neural network to differentiate between an acceptable and an unacceptable odor profile, as defined by the human sniff test, and to be able to accurately identify an odor that would not pass a sniff panel. The results of our efforts will be to verify that the AromaScan(TM) system is a valuable alternative to human sniff panel assessments for the early iterative process of designing and testing rodent waste filters for spaceflight. Acceptance by a human panel will remain one of the final criteria for successful rodent habitat development.

Scribner, K. A.↗

NASA NTRS2008

Pulmonary Toxicity Studies of Lunar Dusts in Rodents

NASA will build an outpost on the lunar surface for long-duration human habitation and research. The surface of the Moon is covered by a layer of fine, reactive dust, and the living quarters in the lunar outpost are expected to be contaminated by lunar dust. NASA established the Lunar Airborne Dust Toxicity Advisory Group (LADTAG) to evaluate the risk of exposure to the dust and to establish safe exposure limits for astronauts working in the lunar habitat. Because the toxicity of lunar dust is not known, LADTAG has recommended investigating its toxicity in the lungs of laboratory animals. After receiving this recommendation, NASA directed the JSC Toxicology Laboratory to determine the pulmonary toxicity of lunar dust in exposed rodents. The rodent pulmonary toxicity studies proposed here are the same as those proposed by the LADTAG. Studies of the pulmonary toxicity of a dust are generally done first in rodents by intratracheal instillation (ITI). This toxicity screening test is then followed by an inhalation study, which requires much more of the test dust and is labor intensive. We succeeded in completing an ITI study on JSC-1 lunar dust simulant in mice (Lam et al., Inhalation Toxicology 14:901-916, 2002, and Inhalation Toxicology 14: 917-928, 2002), and have conducted a pilot ITI study to examine the acute toxicity of an Apollo lunar (highland) dust sample. Preliminary results obtained by examining lung lavage fluid from dust-treated mice show that lunar dust was somewhat toxic (more toxic than TiO2, but less than quartz dust). More extensive studies have been planned to further examine lung lavage fluid for biomarkers of toxicity and lung tissues for histopathological lesions in rodents exposed to aged and activated lunar dust samples. In these studies, reference dusts (TiO2 and quartz) of known toxicities and have industrial exposure limits will be studied in parallel so the relative toxicity of lunar dust can be determined. The ITI results will also be useful for choosing an exposure concentration for the animal inhalation study on a selected lunar dust sample, which is included as a part of this proposal. The animal inhalation exposure will be conducted with lunar dust simulant prior to the study with the lunar dust. The simulant exposure will ensure that the study techniques used with actual lunar dust will be successful. The results of ITI and inhalation studies will reveal the toxicological risk of exposures and are essential for setting exposure limits on lunar dust for astronauts living in the lunar habitat.

Lam, Chiu-wing↗

NASA NTRS2016

Rodent Habitat On ISS: Spaceflight Effects On Mouse Behavior

The NASA Decadal Survey (2011), Recapturing a Future for Space Exploration: Life and Physical Sciences Research for a New Era, emphasized the importance of expanding NASA life sciences research to long duration, rodent experiments on the International Space Station (ISS). To accomplish this objective, flight hardware, operations, and science capabilities supporting mouse studies in space were developed at NASA Ames Research Center. The first flight experiment carrying mice, Rodent Research Hardware and Operations Validation (Rodent Research-1), was launched on Sept 21, 2014 in an unmanned Dragon Capsule, SpaceX4, exposing the mice to a total of 37 days in space. Ground control groups were maintained in environmental chambers at Kennedy Space Center. Mouse health and behavior were monitored for the duration of the experiment via video streaming. Here we present behavioral analysis of two groups of five C57BL/6 female adult mice viewed via fixed camera views compared with identically housed Ground Controls. Flight (Flt) and Ground Control (GC) mice exhibited the same range of behaviors, including eating, drinking, exploratory behavior, self- and allo-grooming, and social interactions at similar or greater levels of occurrence. Mice propelled themselves freely and actively throughout the Habitat using their forelimbs to push off or by floating from one cage area to another, and they quickly learned to anchor themselves using tails and/or paws. Overall activity was greater in Flt as compared to GC mice, with spontaneous ambulatory behavior including the development of organized ‘circling’ or ‘race-tracking’ behavior that emerged within the first few days of flight and encompassed the primary dark cycle activity for the remainder of the experiment. We quantified the bout frequency, duration and rate of circling with respect to characteristic behaviors observed in the varying stages of the progressive development of circling: flipping utilizing two sides of the habitat, circling, multi-lap circling and group-circling. Once begun, mice did not regress to flipping behavior or other previous behavioral milestones for the remainder of flight. An overall upward trend in circling frequency, rate, duration, participation, and organization was observed over the course of the 37-day spaceflight experiment. In this presentation, we will summarize qualitative observations and quantitative comparisons of mice in microgravity and 1g conditions. Behavioral analyses provide important insights into the overall health and adaptation of mice to the space environment, and identify unique behaviors and social interactions to guide future habitat development and research on rodents in space.

Mouse↗

NASA NTRS2015

NASA's Rodent Research Project on ISS: Validation of a New Platform for Conducting Biomedical and Basic Research into the Consequences of Long Duration Habitation in Space

Rodent research has played a key role in advancing biomedical discoveries both on Earth and in space. The National Research Counsel’s Decadal survey(1) emphasized the importance of expanding NASAs life sciences research to perform long duration, rodent experiments on the International Space Station (ISS). To accomplish this objective, flight hardware, operations, and science capabilities were developed at NASA ARC to support both commercial and government-sponsored rodent research.Rodent Research-1 (RR-1) was the first mission in which animals were delivered and maintained in the ISS for a long duration mission in modified Animal Enclosure Module hardware. Both RR validation and commercial science objectives were pursued on the RR-1 mission. Adult female mice (20 total Flight, FLT) were launched Sept 21, 2014 in RR hardware within a Dragon Capsule (SpaceX4), then after 4 days in transit, were transferred for habitation on the ISS for 17 days (commercial) or 33 days (validation), when animals were euthanized and select tissues recovered on orbit. Various controls groups consisted of: 1) Basal mice from the same cohorts as FLT mice, but tissues were recovered at time of launch, 2) Vivarium (VIV) were housed in standard cages 3) Ground Controls (GC) were housed in flight hardware within an environmental chamber at Kennedy Space Center. The health and behavior of all mice on the ISS were monitored by video feed on a daily basis. Mice were euthanized by injection of Euthasol, then either fast frozen intact or dissected to preserve livers (fast frozen) and spleens (RNAlater). Samples were stored at ≤ -80˚C until their return to Earth for later analyses.Hardware performed nominally throughout the mission and the planned in-flight science operations were completed successfully. FLT mice appeared generally more physically active on orbit than respective GC groups. After 33 days on the ISS, mean body weights of FLT mice did not differ from GC, with both groups showing a 6% rise compared to time of launch, while VIV mice showed an 8% rise over the same period. Importantly, there were no significant differences in body weights between groups at the end of 33 days on the ISS, providing an indication that the RR hardware supported the health of the mice both on Earth and in space. Based on the preliminary data obtained from the livers and spleens of mice after 17 days on the ISS, purified RNA was of high quality (RIN values of spleen: FLT=9.48 +0.40, GC=9.28 +0.44, n=5/group); therefore, RNA quality from samples retrieved on orbit was acceptable for even the most demanding transcriptomic analyses. In addition, liver enzyme activity levels (units/mg protein) of FLT mice (after 17d on ISS) and all control mice were similar in magnitude to samples that were optimally prepared by freezing in liquid nitrogen in the laboratory (enzymes analyzed included catalase, glutathione reductase and glyceraldehyde-3-phosphate dehydrogenase). Validation analyses still in progress include behavior and tissue biochemistries, as well as optimization of science return by post-flight recovery of tissues for biospecimen sharing and global expression analyses.Together, these preliminary findings demonstrate new capability for supporting long duration rodent research on the ISS to achieve both basic science and biomedical objectives.

Globus, R. K.↗

NASA NTRSDate not supplied

Reanalysis of Rodent Data from Spacelab Life Sciences-1

The space bioscience field has long been plagued by the challenge of spaceflight with effects of radiation and microgravity. Having multiple and repeated spaceflight experiments for model organisms to solve these space stressors is costly and time consuming. Therefore, reusing and reanalyzing legacy experiments is one way that scientists can draw new conclusions in a timely manner and without using too many resources. Moreover, advances in general biological knowledge allows legacy experiments to be placed into more complete context.Here we aim to analyze all data and metadata taken from rats flown on the SLS-1 mission to create a comprehensive biological model that can be supplemented with current data to allow new discoveries in how space flown organisms adapt to the space environment. Our approach begins with the identification of all the data and metadata, including graphs and tables, for SLS-1 in NASA archives and other sources. Then, each piece of data and metadata will be digitized, reformatted and analyzed. Lastly, a previously developed astronaut model will be used to create the data framework and a comprehensive biological rodent model. The datasets we are using is from the 1991 SpaceLab Life Science 1 (SLS-1) NASA Mission. This was the first designated spacelab mission flown. All 29 rodents were tested for nine days in two different habitats: Research Animal Holding Facility (RAHF) and Animal Enclosure Module (AEM). The rodents were prepared for a live return and compared to a ground control. A total of 30 rodent experiments were accepted as flight studies on the mission. By digitization and reorganizing SLS-1 rat data we will both directly generate new insights and indirectly enable other scientists to by providing the data and metadata in a digitized form.

Space Biology↗

NASA NTRSDate not supplied

Reanalysis of Rodent Data from Spacelab Life Science-1

Space Biology↗

NASA NTRS2014

Rodent Research Development for Long Duration Studies on the International Space Station

Rodent research in space is needed to advance our understanding of the health risks,consequences and possible countermeasures to protect crew during future, long duration missions. TheAnimal Enclosure Module (AEM) was designed originally to support habitation of rats and mice onrelatively short duration, Shuttle missions (<19 days). The AEM was flown previously on 27 SpaceShuttle missions, and recently was modified extensively to support future long duration space biology andbiomedical research on the International Space Station (ISS). In consultation with a Science WorkingGroup comprised of veterinarians and investigators experienced in rodent spaceflight experimentation inspace, the Rodent Habitat project team at Ames Research Center modified existing hardware, developednew hardware, operations, and science activities, and performed a series of ground-based operational andscience habitat verification tests in preparation for the first validation flight.

long duration studies↗

DOE OSTI2022

Assessing rodents as carriers of pathogenic Leptospira species in the U.S. Virgin Islands and their risk to animal and public health

Leptospirosis is a global zoonotic disease caused by pathogenic bacteria of the genus Leptospira. We sought to determine if rodents in U.S. Virgin Islands (USVI) are carriers of Leptospira. In total, 140 rodents were sampled, including 112 Mus musculus and 28 Rattus rattus. A positive carrier status was identified for 64/140 (45.7%); 49 (35.0%) were positive by dark-field microscopy, 60 (42.9%) by culture, 63 (45.0%) by fluorescent antibody testing, and 61 (43.6%) by real-time polymerase chain reaction (rtPCR). Molecular typing indicated that 48 isolates were L. borgpetersenii and 3 were L. kirschneri; the remaining nine comprised mixed species. In the single culture-negative sample that was rtPCR positive, genotyping directly from the kidney identified L. interrogans. Serotyping of L. borgpetersenii isolates identified serogroup Ballum and L. kirschneri isolates as serogroup Icterohaemorrhagiae. These results demonstrate that rodents are significant Leptospira carriers and adds to understanding the ecoepidemiology of leptospirosis in USVI.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1990

Revision of the Wind River faunas, early Eocene of central Wyoming. IX - The oldest known hystricomorphous rodent (Mammalia: Rodentia)

The rostral portion of the skull of a new genus and species of rodent, Armintomys tullbergi, from the earliest middle Eocene of the Wind River Basin (Wyoming) provides the geologically oldest known record of the hystricomorphous zygomasseteric structure. Armintomys also preserves the oldest known occurrence of incisor enamel that is transitional from pauciserial to uniserial. Other dental characters include: anteriorly grooved incisor, small premolars, and relatively primitive sciuravidlike molars. Analysis of this unique combination of characters implies that Armintomys is the oldest known myomorph rodent and the only known representative of a new family. Armintomyidae, which is referred, with question, to the myomorph superfamily Dipodoidea. Armintomys is more primitive, especially in premolar retention and structure, than the Bridgerian zapodid Elymys from Nevada, but adds to evidence from the latter for an early origin and radiation of dipodoid rodents.

Dawson, Mary R.↗

NASA NTRS2015

Multigenerational Independent Colony for Extraterrestrial Habitation, Autonomy, and Behavior Health (MICEHAB): An Investigation of a Long Duration, Partial Gravity, Autonomous Rodent Colony

The path from Earth to Mars requires exploration missions to be increasingly Earth-independent as the foundation is laid for a sustained human presence in the following decades. NASA pioneering of Mars will expand the boundaries of human exploration, as a sustainable presence on the surface requires humans to successfully reproduce in a partial gravity environment independent from Earth intervention. Before significant investment is made in capabilities leading to such pioneering efforts, the challenges of multigenerational mammalian reproduction in a partial gravity environment need be investigated. The Multi-generational Independent Colony for Extraterrestrial Habitation, Autonomy, and Behavior health is designed to study these challenges. The proposed concept is a conceptual, long duration, autonomous habitat designed to house rodents in a partial gravity environment with the goal of understanding the effects of partial gravity on mammalian reproduction over multiple generations and how to effectively design such a facility to operate autonomously while keeping the rodents healthy in order to achieve multiple generations. All systems are designed to feed forward directly to full-scale human missions to Mars. This paper presents the baseline design concept formulated after considering challenges in the mission and vehicle architectures such as: vehicle automation, automated crew health management/medical care, unique automated waste disposal and hygiene, handling of deceased crew members, reliable long-duration crew support systems, and radiation protection. This concept was selected from an architectural trade space considering the balance between mission science return and robotic and autonomy capabilities. The baseline design is described in detail including: transportation and facility operation constraints, artificial gravity system design, habitat design, and a full-scale mock-up demonstration of autonomous rodent care facilities. The proposed concept has the potential to integrate into existing mission architectures in order to achieve exploration objectives, and to demonstrate and mature common capabilities that enable a range of destinations and missions.

Rodgers, Erica M.↗

NASA NTRS2017

Vascular Patterning Analysis by VESGEN 2D/3D with Bioinformatics: Updates for Rodent Tissues

Fractally branching vascular systems are a complex physiological requirement shared by humans with all higher terrestrial life forms, including other vertebrates, insects, and higher land plants. Vascular trees, networks, and tree-network composites are therefore mapped and quantified by the VESsel GENeration Analysis (VESGEN) software according to weighted physiological vascular rules that include vessel connectivity, tapering and bifurcational branching. According to fluid dynamics, successful vascular transport depends upon a complex distributed system of highly regulated laminar flow. VESGEN has elucidated changes in vascular patterning resulting from inflammatory, developmental and other signaling pathways within numerous tissues of major model organisms important for Space Biology, especially for rodents. Important early stage regenerative opportunities have been identified by VESGEN vascular analysis for visual impairments in the human retina, and is currently being used for research into astronaut visual and ocular disorders associated with long duration missions. The VESGEN 2D software is a mature, automated, widely published capability for which beta testing and public release by NASA is planned for the upcoming year. Early-stage capabilities for VESGEN 3D analysis are under development for the rodent retina and intestine as prototype tissues. A prototype VESGEN 2D Bioinformatics software capability has also been developed to associate phenotypic changes in molecular expression with vascular structure and function. By new VESGEN bioinformatic innovations, expression patterns of the genetic, transcriptional, protein and other markers for regulatory molecules such as vascular endothelial growth factor (VEGF) and their receptors, often indicators of tissue oxygenation status, are co-localized with alterations in vascular pattern. Biomarkers are therefore mapped and quantified as information dimensions directly correlated with the spatial dimensions of a vascular pattern. Further important technology innovations by NASA include substantial image segmentation advances for more automated binary extraction of the grayscale vascular patterns, together with informative associated image quality assessments. Vascular mapping and quantification capabilities for the rodent retina and intestine are illustrated for VESGEN 2D, along with technology status reports on VESGEN 3D and Bioinformatic capabilities. Research partially supported by Ames Center Innovation Awards.

Parsons-Wingerter, P.↗

NASA NTRS2017

Environmental Enrichment in the ISS Rodent Habitat Hardware System

Responses of animals exposed to microgravity during in-space experiments were observed via available video recording stored in the NASA Ames Life Sciences Data Archive. These documented observations of animal behavior, as well as the range and level of activities during spaceflight, demonstrate that weightlessness conditions and the extreme novelty of the surroundings may exert damaging psychological stresses on the inhabitants. In response to a recognized need for in-flight animals to improve their wellbeing we propose to reduce such stresses by shaping and interrelating structures and surroundings to satisfying vital physiological needs of inhabitants. A Rodent Habitat Hardware System (RHHS) based housing facility incorporating a tubing network system, to maintain and monitor rodent health environment with advanced accessories has been proposed. Placing mice in a tubing-configured environment creates more natural space-restricted nesting environment for rodents, thereby facilitating a more comfortable transition to living in microgravity. A sectional tubing structure of the RHHS environment will be more beneficial under microgravity conditions than the provision of a larger space area that is currently utilized. The new tubing configuration was found suitable for further incorporation of innovative monitoring technology and accessories in the animal holding habitat unit which allow to monitor in real-time monitoring of valuable health related biological parameters under weightlessness environment of spaceflight.

modifications↗

NASA NTRS2015

NASA's Rodent Research Project: Validation of Flight Hardware, Operations and Science Capabilities for Conducting Long Duration Experiments in Space

Research using rodents is an essential tool for advancing biomedical research on Earth and in space. Rodent Research (RR)-1 was conducted to validate flight hardware, operations, and science capabilities that were developed at the NASA Ames Research Center. Twenty C57BL/6J adult female mice were launched on Sept 21, 2014 in a Dragon Capsule (SpaceX-4), then transferred to the ISS for a total time of 21-22 days (10 commercial mice) or 37 (10 validation mice). Tissues collected on-orbit were either rapidly frozen or preserved in RNA later at less than or equal to -80 C (n=2/group) until their return to Earth. Remaining carcasses were rapidly frozen for dissection post-flight. The three controls groups at Kennedy Space Center consisted of: Basal mice euthanized at the time of launch, Vivarium controls, housed in standard cages, and Ground Controls (GC), housed in flight hardware within an environmental chamber. FLT mice appeared more physically active on-orbit than GC, and behavior analysis are in progress. Upon return to Earth, there were no differences in body weights between FLT and GC at the end of the 37 days in space. RNA was of high quality (RIN greater than 8.5). Liver enzyme activity levels of FLT mice and all control mice were similar in magnitude to those of the samples that were optimally processed in the laboratory. Liver samples collected from the intact frozen FLT carcasses had RNA RIN of 7.27 +/- 0.52, which was lower than that of the samples processed on-orbit, but similar to those obtained from the control group intact carcasses. Nonetheless, the RNA samples from the intact carcasses were acceptable for the most demanding transcriptomic analyses. Adrenal glands, thymus and spleen (organs associated with stress response) showed no significant difference in weights between FLT and GC. Enzymatic activity was also not significantly different. Over 3,000 tissues collected from the four groups of mice have become available for the Biospecimen Sharing Program. Together, these validation flight findings demonstrate the capability to support long-duration RR on the ISS to achieve both basic science and biomedical objectives.

rodents↗

NASA NTRS2016

Biospecimen Retrieval from NASA's Rodent Research-1: Maximizing Science Return from Flight Missions

Rodent Research (RR)-1 was conducted to validate flight hardware, operations, and science capabilities that were developed to support long duration missions on the International Space Station. After 37 days in microgravity twenty mice were euthanized and frozen on orbit. Upon return to Earth the carcasses were dissected and yielded 32 different types of tissues from each mouse and over 3200 tissue aliquots. Many tissues were distributed to the Space Life and Physical Sciences (SLPS) Biospecimen Sharing Program (BSP) Principal Investigators (PIs) through the Ames Life Science Data Archive (ALSDA). A second round of dissections was performed to collect additional tissues from the remaining carcasses thawed for a second time for additional BSP PIs. Tissues retrieved included vaginal walls, aorta, pelvis, brown adipose tissue, tail, spine and forearms. Although the analyses are still in progress, some of the PIs have reported that the quality of the tissues was acceptable for their study. In a separate experiment we tested the RNA quality of the tissues that were dissected from frozen carcasses that were subjected to euthanasia, freezing, first and second thaw dissections. Timelines simulated the on-orbit RR-1 procedures to assess the quality of the tissues retrieved from the second thaw dissections. We analyzed the RIN values of select tissues including kidney, brain, white adipose tissue (WAT) and brown adipose tissue (BAT). Overall the RIN values from the second thaw were lower compared to those from the first by about a half unit; however, the tissues yielded RNA that are acceptable quality for some quantitative gene expression assays. Interestingly, RIN values of brain tissues were 8.4+/-0.6 and 7.9+/-0.7 from first and second round dissections, respectively (n=5). Kidney and WAT yielded RIN values less than 8 but they can still be used for qPCR. BAT yielded higher quality RNA (8.2+/-0.5) than WAT (5.22+/-0.9), possibly due to the high fat content. Together, these data show that select tissues can be utilized for gene expression studies even if they are retrieved from carcasses that were subjected to at least two freezing and thawing processes; this further expands science return from valuable and infrequent rodent experiments in space.

rodent↗