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At least 127 records · Page 7

Science and payload options for animal and plant research accommodations aboard the early Space Station

The resources to be allocated for the development of the Initial Operational Capability (IOC) Space Station Animal and Plant Research Facility and the Growth Station Animal and Plant Vivarium and Laboratory may be limited; also, IOC accommodations for animal and plant research may be limited. An approach is presented for the development of Initial Research Capability Minilabs for animal and plant studies, which in appropriate combination and sequence can meet requirements for an evolving program of research within available accommodations and anticipated budget constraints.

Hilchey, John D.↗

Biosafety and biosecurity in veterinary laboratories

Here, with recent outbreaks of MERS-Cov, Anthrax, Nipah, and Highly Pathogenic Avian Influenza, much emphasis has been placed on rapid identification of infectious agents globally. As a result, laboratories are building capacity, conducting more advanced and sophisticated research, increasing laboratory staff, and establishing collections of dangerous pathogens in an attempt to reduce the impact of infectious disease outbreaks and characterize disease causing agents. With this expansion, the global laboratory community has started to focus on laboratory biosafety and biosecurity to prevent the accidental and/or intent ional release o f these agents. Laboratory biosafety and biosecurity systems are used around the world to help mit igate the risks posed by dangerous pathogens in the laboratory. Veterinary laboratories carry unique responsibilities to workers and communities to safely and securely handle disease causing microorganisms. Many microorganisms studied in veterinary laboratories not only infect animals, but also have the potential to infect humans. This paper will discuss the fundamentals of laboratory biosafety and biosecurity.

59 BASIC BIOLOGICAL SCIENCES↗

Animated software training via the internet: lessons learned

The Mission Execution and Automation Section, Information Technologies and Software Systems Division at the Jet Propulsion Laboratory, recently delivered an animated software training module for the TMOD UPLINK Consolidation Task for operator training at the Deep Space Network.

simulattions training animations flash↗

Gravitational considerations with animal rhythms

As established in the laboratory and largely confirmed by others, simulated high-g environments influence growth and development of animals as small as or smaller than baby turtles, sometimes accelerating and sometimes decelerating these processes. High-g environments result in many functional changes or adjustments in feeding, metabolism, circulation, fluid balances, and structures for support, and influence life expectancy. An assembly of equipment suitable for measuring oxygen consumption of small mammals as influenced by chronic centrifugation and/or by day-night rhythms is discussed.

Wunder, C. C.↗

Past and Future Work on Radiobiology Mega-Studies: A Case Study At Argonne National Laboratory

Between 1952 and 1992, more than 200 large radiobiology studies were conducted in research institutes throughout Europe, North America, and Japan to determine the effects of external irradiation and internal emitters on the lifespan and tissue toxicity development in animals. At Argonne National Laboratory, 22 external beam studies were conducted on nearly 700 beagle dogs and 50,000 mice between 1969 and 1992. These studies helped to characterize the effects of neutron and gamma irradiation on lifespan, tumorigenesis, and mutagenesis across a range of doses and dosing patterns. The records and tissues collected at Argonne during that time period have been carefully preserved and redisseminated. Using these archived data, ongoing statistical work has been done and continues to characterize quality of radiation, dose, dose rate, tissue, and gender-specific differences in the radiation responses of exposed animals. The ongoing application of newly-developed molecular biology techniques to the archived tissues has revealed gene-specific mutation rates following exposure to ionizing irradiation. The original and ongoing work with this tissue archive is presented here as a case study of a more general trend in the radiobiology megastudies. These experiments helped form the modern understanding of radiation responses in animals and continue to inform development of new radiation models. Recent archival efforts have facilitated open access to the data and materials produced by these studies, and so a unique opportunity exists to expand this continued research.

59 BASIC BIOLOGICAL SCIENCES↗

Applicability of the Nasa Galactic Cosmic Ray Simulator for Mice, Rats, and Minipigs

Space radiation poses multiple health risks to astronauts including cancer, cardiovascular disease, and damage to the central nervous system. Radiation from galactic cosmic rays (GCR) is the main source of these risks for missions beyond low Earth orbit to the moon and beyond. A galactic cosmic ray simulator (GCRsim) was developed at the NASA Space Radiation Laboratory to better understand and mitigate these risks by simulating the complex mixed field of the GCR environment at a ground-based facility. The GCRsim delivers a radiation field to cell and animal models in a laboratory setting that is comparable to the shielded radiation environment within internal organs of astronauts in deep space missions. Previous verification studies using Monte Carlo simulations with mouse (Digimouse) and rat (Digirat) digital phantoms showed that the GCRsim yields acceptable dose homogeneity within the internal organs of these animals. Spectral characteristics of the intended space radiation environment are also accurately reproduced at radiosensitive sites in both phantoms. In this work, similar Monte Carlo simulations were performed in a minipig model (Digipig) to show the applicability and limitations of the current GCRsim for larger animal model systems. The results showed dose homogeneity in internal organs of the three animal models with GCRsim irradiation. While slightly lower average doses were seen in the minipig compared to the external beam dose, the individual voxel doses in slices of the phantoms were found to be within 6%–8% of the average voxel dose, establishing that the GCRsim can still provide a relatively homogeneous irradiation within larger animals. Furthermore, simulated dose and fluence spectral results across the relevant linear energy transfer (LET) range agreed well with the reference field in the most relevant regions of the spectra, further verifying that the GCRsim beam represents the reference field in larger animals.

Galactic cosmic rays↗

Large Game Distribution Camera Study at Los Alamos National Laboratory

The unique habitats at the Laboratory support a diverse community of wildlife. The habitat types that include Pinon Juniper, Ponderosa Pine, and Mixed-conifer forests each provide different resources important for a variety of species. Wildlife monitoring for large game animals can be challenging because they avoid interactions with humans, and prefer undeveloped areas. This study was conducted to analyze the distribution of predators and other game animals seasonally across the Laboratory. This research informs decisions regarding wildlife management recommendations for conservation and protection. This study utilized game cameras to remotely monitor wildlife. As camera technology has advanced, game cameras have become adequate for documenting individuals, and are commonly used in wildlife studies on predation, abundance, occupancy, diversity, and endangered species detection. Twenty cameras were placed across the LANL landscape using a modified systematic sampling design. They were deployed in February 2018 and retrieved in January 2019, thus each camera sampled for one full year. All images were reviewed and the species, number, age, and sex of all animals in the photo were recorded. Here we present data for the most abundant species encountered: deer, elk, coyote, bear, bobcat, mountain lion, and fox respectively. The data were summarized by species and season. Future work will include occupancy modelling for each species. This will allow the development of heat maps showing species distributions across the Laboratory.

59 BASIC BIOLOGICAL SCIENCES↗

Large Game Distribution Camera Study at Los Alamos National Laboratory [Poster]

The unique habitats at the Laboratory support a diverse community of wildlife. The habitat types that include Pinon Juniper, Ponderosa Pine, and Mixed-conifer forests each provide different resources important for a variety of species. Wildlife monitoring for large game animals can be challenging because they avoid interactions with humans, and prefer undeveloped areas. This study was conducted to analyze the distribution of predators and other game animals seasonally across the Laboratory. The data gathered will be used to make recommendations regarding wildlife management for conservation and protection. Utilizing game cameras to remotely monitor wildlife was determined to be the most effective, noninvasive method to gather the necessary data. As camera technology has advanced, game cameras have become adequate for documenting individuals, and are commonly used in wildlife studies on predation, abundance, occupancy, diversity, and endangered species detection. Twenty cameras were placed across the LANL landscape using a modified systematic sampling design. They were deployed in February 2018 and retrieved in January 2019, thus each camera sampled for one full year. All images were reviewed and the species, number, age, and sex of all animals in the photo were recorded. Here we present data for the most abundant species encountered: deer, elk, coyote, bear, bobcat, mountain lion, and fox respectively. The data were summarized by species and season. Future work will include occupancy modelling for each species. This will allow the development of heat maps showing species distributions across the Laboratory.

54 ENVIRONMENTAL SCIENCES↗

Effect of 16-Day Spaceflight on the Morphology of Thick-Toed Geckos (Pachydactylus Bibronii Smith, 1846)

There are grounds to believe that space flown experiments on thick-toed geckos may help solve the problem of floatation of vertebrates in microgravity. Geckos of this species carry on the lower surface of their toes numerous setae, which allow them to remain attached to any surfaces regardless of the gravitational effects. Experiments were performed on 5 animals in each of the following groups: flight, basal, synchronous and laboratory controls. 32 hours after a 16- day flight the animals were euthanazed and examined using traditional histology and X-ray microtomography. Body mass losses were 10% in the flight animals, 7.4% in the synchronous controls, and 12.3% in the laboratory controls. Since the flight and synchronous animals were kept at 15-19 C, whereas the laboratory controls - at 26-28 C, it can be inferred that environmental temperatures impacted animal metabolism no less than flight induced stress. Blood tests of the flown animals showed a 12% decrease of erythrocytes and a 40% decrease of dark-nuclear granulocytes, with the number of light-nuclear granulocytes remaining unchanged. In the small intestine the number of goblet cells increased allowing them to occupy a large portion of the cyptal surface. Enhanced secretion was accompanied by the appearance of dead intestinal cells in the lumen. Clusters of degraded hepatocytes were found at the liver edges of flight animals. Signs of liver involution were similar to the changes produced by alcohol consumption but did not spread to its central part. In the heart, insignificant hypertrophy and excessive blood supply that still remained within the physiological norm were detected. No significant changes were found in the pancreas, lungs, nervous systems or the snouts of the flown animals, but the volume of their gallbladders was greater than in controls. The epithelium of toe pads of the flight animals became thinner. Histological examination of the humerus did not demonstrate significant mineral losses. However, X-ray microtomography showed changes in the trabecular structure in the subepyphyseal zone of bones in flight animals compared to the controls. In summary, all the changes detected in the flight animals were adaptive. Therefore, geckos of this species can be used as an animal model for morphological studies in longer-duration space flights.

Gulimova, V. I.↗

SU-E-U-02: The Development of a Practical Ultrasonic System for Cross-Sectional Imaging of Small Animals

Purpose: To test the feasibility of developing a practical medium frequency ultrasound tomography method for small animal imaging. The ability to produce cross-sectional or full body images of a live small animal using a low-cost tabletop ultrasound scanner without any special license would be very beneficial to long term biological studies, where repeated scanning is often required over an extended period of time. Methods: The cross sectional images were produced by compounding multiple B-scans of a laboratory phantom or an animal acquired at different projection angles. Two imaging systems were used to test the concept. The first system included a programmable 64-channel phased array controller driving a 128-channel, 5–10 MHz linear probe to produce 143 B-Mode projections of the spinning object. The second system designed and manufactured in house, produced 64 or 128 B-Mode projections with a single unfocused 8 MHz transducer scanning with a 0.116 mm step size. Results: The phased array system provided good penetration through the phantoms/mice (with the exception of the lungs) and allowed to acquire data in a very short time. The cross-sectional images have enough resolution and dynamic range to detect both high- and low-contrast organs. The single transducer system takes longer to scan, and the data require more sophisticated processing. To date, our images allow seeing details as small as 1–2 mm in the phantoms and in small animals, with the contrast mostly due to highly reflecting bones and air inclusions. Conclusion: The work indicates that very detailed and anatomically correct images can be created by relatively simple and inexpensive means. With more advanced algorithms and improved system design, scan time can be reduced considerably, enabling high-resolution full 3D imaging. This will allow for quick and easy scans that can help monitor tumor growth and/or regression without contributing any dose to the animal. The authors would like to acknowledge the financial and engineering support from Tessonics.

60 APPLIED LIFE SCIENCES↗

Proposed mechanistic description of dose-dependent BDE-47 urinary elimination in mice using a physiologically based pharmacokinetic model

Polybrominated diphenyl ethers (PBDEs) have been used in a wide variety of consumer applications as additive flame retardants. In North America, scientists have noted continuing increases in the levels of PBDE congeners measured in human serum. Some recent studies have found that PBDEs are associated with adverse health effects in humans, in experimental animals, and wildlife. This laboratory previously demonstrated that urinary elimination of 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) is saturable at high doses in mice; however, this dose-dependent urinary elimination has not been observed in adult rats or immature mice. Thus, the primary objective of this study was to examine the mechanism of urinary elimination of BDE-47 in adult mice using a physiologically based pharmacokinetic (PBPK) model. To support this objective, additional laboratory data were collected to evaluate the predictions of the PBPK model using novel information from adult multi-drug resistance 1a/b knockout mice. Using the PBPK model, the roles of mouse major urinary protein (a blood protein carrier) and P-glycoprotein (an apical membrane transporter in proximal tubule cells in the kidneys, brain, intestines, and liver) were investigated in BDE-47 elimination. The resulting model and new data supported the major role of m-MUP in excretion of BDE-47 in the urine of adult mice, and a lesser role of P-gp as a transporter of BDE-47 in mice. This work expands the knowledge of BDE-47 kinetics between species and provides information for determining the relevancy of these data for human risk assessment purposes. - Highlights: • We report the first study on PBPK model on flame retardant in mice for BDE-47. • We examine mechanism of urinary elimination of BDE-47 in mice using a PBPK model. • We investigated roles of m-MUP and P-gp as transporters in urinary elimination.

60 APPLIED LIFE SCIENCES↗

Molecular Evolution of the H5 and H7 Highly Pathogenic Avian Influenza Virus Haemagglutinin Cleavage Site Motif

ABSTRACT Avian influenza viruses are ubiquitous in the Anatinae subfamily of aquatic birds and occasionally spill over to poultry. Infection with low pathogenicity avian influenza viruses generally leads to subclinical or mild clinical disease. In contrast, highly pathogenic avian influenza viruses emerge from low pathogenic forms and can cause severe disease associated with extraordinarily high mortality rates. Here, we describe the natural history of avian influenza virus, with a focus on H5Nx and H7Nx subtypes, and the emergence of highly pathogenic forms; we review the biology of AIV; we examine cleavage of haemagglutinin by host cell enzymes with a particular emphasis on the biochemical properties of the proprotein convertases, and trypsin and trypsin‐like proteases; we describe mechanisms implicated in the functional evolution of the haemagglutinin cleavage site motif that leads to emergence of HPAIVs; and finally, we discuss the diversity of H5 and H7 haemagglutinin cleavage site sequence motifs. It is crucial to understand the molecular attributes that drive the emergence and evolution of HPAIVs with pandemic potential to inform risk assessments and mitigate the threat of HPAIVs to poultry and human populations.

Luczo, Jasmina M. [Australian Animal Health Labora↗

Detection of Apparent Early Rabies Infection by LN34 Pan-Lyssavirus Real-Time RT-PCR Assay in Pennsylvania

The Pennsylvania Department of Health Bureau of Laboratories (PABOL) tested 6855 animal samples for rabies using both the direct fluorescent antibody test (DFA) and LN34 pan-lyssavirus reverse transcriptase quantitative PCR (RT-qPCR) during 2017–2019. Only two samples (0.03%) were initially DFA negative but positive by LN34 RT-qPCR. Both cases were confirmed positive upon re-testing at PABOL and confirmatory testing at the Centers for Disease Control and Prevention by LN34 RT-qPCR and DFA. Rabies virus sequences from one sample were distinct from all positive samples processed at PABOL within two weeks, ruling out cross-contamination. Levels of rabies virus antigen and RNA were low in all brain structures tested, but were higher in brain stem and rostral spinal cord than in cerebellum, hippocampus or cortex. Taken together, the low level of rabies virus combined with higher abundance in more caudal brain structures suggest early infection. These cases highlight the increased sensitivity and ease of interpretation of LN34 RT-qPCR for low positive cases.

59 BASIC BIOLOGICAL SCIENCES↗

Skeletal and body composition evaluation

Research on radiation detectors for absorptiometry; analysis of errors affective single photon absorptiometry and development of instrumentation; analysis of errors affecting dual photon absorptiometry and development of instrumentation; comparison of skeletal measurements with other techniques; cooperation with NASA projects for skeletal evaluation in spaceflight (Experiment MO-78) and in laboratory studies with immobilized animals; studies of postmenopausal osteoporosis; organization of scientific meetings and workshops on absorptiometric measurement; and development of instrumentation for measurement of fluid shifts in the human body were performed. Instrumentation was developed that allows accurate and precise (2% error) measurements of mineral content in compact and trabecular bone and of the total skeleton. Instrumentation was also developed to measure fluid shifts in the extremities. Radiation exposure with those procedures is low (2-10 MREM). One hundred seventy three technical reports and one hundred and four published papers of studies from the University of Wisconsin Bone Mineral Lab are listed.

Mazess, R. B.↗

Experiment K-6-13. Morphological and biochemical examination of heart tissue. Part 1: Effects of microgravity on the myocardial fine structure of rats flown on Cosmos 1887. Ultrastructure studies. Part 2: Cellular distribution of cyclic ampdependent protein kinase regulatory subunits in heart muscle of rats flown on Cosmos 1887

The left ventricle of hearts from rats flown on the Cosmos 1887 biosatellite for 12.5 days was compared to the same tissue of synchronous and vivarium control animals maintained in a ground based laboratory. The volume density of the mitochondria in the myocardium of the space-flown animals was statistically less (p equal less than 0.01) than that of the synchronous or vivarium control rats. Exposure to microgravity resulted in a certain degree of myocardial degeneration manifested in mitochondrial changes and accumulation of myeloid bodies. Generalized myofibrillar edema was also observed.

Philpott, D. E.↗

Real-time PCR assays that detect genes for botulinum neurotoxin A–G subtypes

The role of Real-Time PCR assays for surveillance and rapid screening for pathogens is garnering more and more attention because of its versatility and ease of adoption. The goal of this study was to design, test, and evaluate Real-Time TaqMan PCR assays for the detection of botulinum neurotoxin (bont/A-G) genes from currently recognized BoNT subtypes. Assays were computationally designed and then laboratory tested for sensitivity and specificity using DNA preparations containing bont genes from 82 target toxin subtypes, including nine bivalent toxin types; 31 strains representing other clostridial species; and an extensive panel that consisted of DNA from a diverse set of prokaryotic (bacterial) and eukaryotic (fungal, protozoan, plant, and animal) species. In addition to laboratory testing, the assays were computationally evaluated using in silico analysis for their ability to detect bont gene sequences from recently identified toxin subtypes. Seventeen specific assays (two for each of the bont/C, bont/D, bont/E, and bont/G subtypes and three for each of the bont/A, bont/B, and bont/F subtypes) were designed and evaluated for their ability to detect bont genes encoding multiple subtypes from all seven serotypes. These assays could provide an additional tool for the detection of botulinum neurotoxins in clinical, environmental and food samples that can complement other existing methods used in clinical diagnostics, regulatory, public health, and research laboratories.

59 BASIC BIOLOGICAL SCIENCES↗