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A continuation of base-line studies for environmentally monitoring space transportation systems at John F. Kennedy Space Center. Volume 3, part 2: Ichthyological studies, sailfin molly reproduction study

The applicability of monitoring populations of Poccilia latipinna (sailfin molly) and its reproductive efforts as reliable indicators of environmental effects of aerospace activities in the Kennedy Space Center area was investigated. Results show that the sailfin molly experiences drastic fluctuations in population and reproductive success and is not an appropriate factor for monitoring to establish perturbations of the environment due to space transportation system related activities.

Snelson, F. F., Jr.

The Role of Gravity on the Reproduction of Arabidopsis Plants

The presence of gravity as a necessary environmental factor for higher plants to complete their life cycle was examined. Arabidopsis thalliana (L.) Heynh. Columbia strain plants were grown continuously for three generations in a simulated micro-g environment as induced by horizontal clinostats. Growth, development and reproduction were followed. The Arabidopsis plants were selected for three generations on clinostats because: (1) a short life cycle of around 35 days; (2) the cells of third generation plants would in theory be free of gravity imprint; and (3) a third generation plant would therefore more than likely grow and respond like a plant growing in a micro-g environment. It is found that gravity is not a required environmental factor for higher plants to complete their life cycle, at least as tested by a horizontal clinostat. Clinostatting does not prevent the completion of the plant life cycle. However, clinostatting does appear to slow down the reproductive process of Arabidopsis plants. Whether higher plants can continue to reproduce for many generations in a true micro-g environment of space can only be determined by long duration experiments in space.

Hoshizaki, T.

The pineal gland - Its possible roles in human reproduction

The paper discusses the role of the pineal gland in controlling mammalian reproduction, with particular attention given to the role of melatonin in polyestrus mammals, like humans and laboratory rodents. Evidence is cited indicating the influence of melatonin production and blood content on the age of puberty, the timing of the ovulatory cycle, gonadal steriodogenesis, and patterns of reproductive behavior. It is suggested that abnormal patterns of melatonin might be associated with amenorrhea, anovulation, unexplained infertility, premature menopause, and habitual abortions.

Brzezinski, Amnon

The Effect of Hindlimb Suspension on the Reproductive System of Young Male Rats

Colonization of space requires the ability to reproduce in reduced gravity. Following spaceflight, astronauts and male rats exhibit decreased testosterone (T). This has important implications as T effects the testes and accessory sex glands. To our knowledge no studies have examined the effects of spaceflight on accessory sex glands. Due to the rarity of spaceflight opportunities, ground models have been used to simulate weightlessness. The objective of this study was to determine the effect of long-term (21 d) weightlessness on the reproductive system of male rats. Weightlessness was simulated using the Morey-Holton hindlimb suspension (HLS) model. Age 10 week old, male Sprague-Dawley rats weighing (209.0 +9.7g) were randomly assigned (n=10/group) to either HLS or ambulatory control. In HLS rats, testes mass was 33% lower (p<0.05) than ambulatory controls. However, HLS had no effect on prostate (0.65 +0.09g vs 0.69 +0.12g) or seminal vesicles (1.01 +0.35g vs 0.75 +0.22g) weights compared to controls. The absence of effects on plasma T in this study contrasts previous reports of reduced plasma T in HLS male rats. This discrepancy may have been due to the age of animal and timing of sampling. T levels vary dramatically during testes development as well as within normal diurnal cycles. In young HLS rats, testes weight was reduced but not plasma T. Subsequently there was no effect on accessory sex glands. However, this may not be the case in older rats. More studies using standardized methods are needed to gain a better understanding of male reproduction function and capability in weightlessness. Funding provided by NASA.

Tou, Janet

The effects of feedback lighting on the circadian rhythm of locomotor activity and the reproductive maturation of the male Djungarian hamster (Phodopus sungorus)

The non-parametric model of entrainment suggests that brief pulses of light, delivered between dusk and dawn can simulate the phasing effects of full photoperiods or even constant light (LL). Feedback lighting (LDFB) is a lighting condition where individual animals, otherwise in constant darkness (DD), are exposed to light in response to a monitored behavior. The specific purpose of this type of illumination is to expose the circadian cycle to light only during the subjective night. LDFB has been used to support this hypothesis in several species of nocturnal rodents and one species of diurnal primate by producing similar free-running periods in LDFB as in LL. This lighting condition has also been used to test the hypothesis that exposing the subjective night to even short duration light pulses will maintain reproductive function in long day breeders. In the Syrian hamster (Mesocricetus auratus), however, LDFB is not as photostimulatory as LL despite extensive light exposure during the subjective night. In the experiments presented here, a group of immature male Djungarian hamsters (Phodopus sungorus) were placed in individual light-tight sound attenuated chambers where they had free access to food, water and an activity wheel. The animals were exposed to one of four lighting conditions [DD, LL, LDFB or a neighbor control of feedback lighting (LDFB NC)] for approximately 30 days shortly after weaning. LDFB NC is a lighting condition where a neighbor control hamster receives the identical lighting regime as a paired animal exposing itself to LDFB, yet the neighbor has no control over it. A fifth group was exposed to a light-dark cycle of 16 hours of light and 8 hours of dark (LD16:8). This group was housed in cages in a colony room and did not have access to a running wheel. The free-running periods of the locomotor activity rhythms for hamsters exposed to LDFB and LL were not similar, unlike the results for rats, Syrian hamsters, mice, monkeys and even mature Djungarian hamsters. Immature hamsters exposed to DD and LDFB NC developed more slowly than animals exposed to LL or LD16:8, while hamsters in LDFB developed at an intermediate rate. Thus, it appears that LDFB, although capable of inducing reproductive function in immature Djungarian hamsters, is not as photostimulatory as may have been expected from current photoperiodic models, despite substantial light exposure during the subjective night. Furthermore, this data may suggest that the circadian system of 18-48 day old Djungarian hamsters are still undergoing organizational maturation.

NASA Program Space Biology

Onset of and recovery from nitrogen stress during reproductive growth of soybean

Photosynthetic rates and allocation of dry matter, nitrogen, and nonstructural carbohydrates were determined during onset of and recovery from a nitrogen stress for reproductive soybean (Glycine max [L.] Merrill cv Ransom) plants. Until the beginning of seed fill, non-nodulated plants were grown in flowing solution culture with 1.0 mM NO3- in a complete nutrient solution. One set of plants then was transferred to minus-nitrogen solution for 24 d of seed fill; a second set was transferred to a minus-nitrogen solution for 14 d followed by return to the complete solution with 1.0 mM NO3- for the remaining 10 d of seed fill; and a third set was continued on the complete solution. Net CO2 exchange rates of individual leaves, which remained nearly constant during seed fill for nonstressed plants, declined at an accelerated rate during onset of nitrogen stress as the specific content of reduced nitrogen in the leaves was decreased by remobilization of nitrogen to support pod growth. The rate of nitrogen remobilization out of leaves initially was relatively greater than the decrease in photosynthetic rate. While rate of pod growth declined in response to the developing nitrogen stress, photosynthetic assimilation of carbon exceeded reproductive demand and nonstructural carbohydrates accumulated within tissues. Following resupply of exogenous NO3-, specific rate of NO3- uptake by roots was enhanced relative to nonstressed plants. While there was little increase in content of reduced nitrogen in leaves, net remobilization of nitrogen out of leaves ceased, and the decline in photosynthetic rate stabilized at about 51% of that for nonstressed plants. This level of photosynthesis, combined with the availability of elevated pools of carbohydrates accumulated during stress, was sufficient to support the increases in both the specific rates of NO3- uptake and the rate of pod growth during recovery.

Non-NASA Center

Reproductive Ontogeny of Wheat Grown on the MIR Space Station

The reproductive ontogeny of 'Super-Dwarf' wheat grown on the space station Mir is chronicled from the vegetative phase through flower development. Changes in the apical meristem associated with transition From the vegetative phase to floral initiation and development of the reproductive spike were all typical of 'Super Dwarf' wheat up to the point of anthesis. Filament elongation, which characteristically occurs just prior to anthesis and moves the anthers through the stigmatic branches thus facilitating pollination, did no1 xcur in the flowers of spikes grown on Mir. While development of spikes on tillers typically occurs later :han that of spikes on the main stem, all flowers appear to be arrested at the same developmental point.

Bubenheim, David L.

Quantifying the basic reproduction number and underestimated fraction of Mpox cases worldwide at the onset of the outbreak

In 2022, there was a global resurgence of mpox, with different clinical-epidemiological features compared with previous outbreaks. Sexual contact was hypothesized as the primary transmission route, and the community of men having sex with men (MSM) was disproportionately affected. Because of the stigma associated with sexually transmitted infections, the real burden of mpox could be masked. We quantified the basic reproduction number (R 0 ) and the underestimated fraction of mpox cases in 16 countries, from the onset of the outbreak until early September 2022, using Bayesian inference and a compartmentalized, risk-structured (high-/low-risk populations) and two-route (sexual/non-sexual transmission) mathematical model. Machine learning (ML) was harnessed to identify underestimation determinants. Estimated R 0 ranged between 1.37 (Canada) and 3.68 (Germany). The underestimation rates for the high- and low-risk populations varied between 25–93% and 65–85%, respectively. The estimated total number of mpox cases, relative to the reported cases, is highest in Colombia (3.60) and lowest in Canada (1.08). In the ML analysis, two clusters of countries could be identified, differing in terms of attitudes towards the 2SLGBTQIAP+ community and the importance of religion. Given the substantial mpox underestimation, surveillance should be enhanced, and country-specific campaigns against the stigmatization of MSM should be organized, leveraging community-based interventions.

60 APPLIED LIFE SCIENCES

Reproduction during spaceflight by plants in the family Brassicaceae

Researchers report on studies of reproduction in Arabidopsis thaliana in space during during the Chromex-03 on STS-54, Chromex-04 on STS-51, and Chromex-05 on STS-68 missions. The obstacles to seed formation were related to carbon dioxide levels. Other experiments examined in flight pollination and seed production in Brassica rapa during parabolic flight, a 4-1/2 month stay on Mir, and on STS-87. During the Mir experiment, Brassica seeds were harvested from seeds sown in flight. The second generation seeds grew to produce new seeds that contained more starch and less protein and lipid when compared to ground control seeds.

manned

Successful post-translocation reproduction and genetic integration of eastern box turtles

Translocation is a conservation tool increasingly used in the recovery of at-risk species, including turtles, which are one of the world's most imperiled taxa. Post-release monitoring is essential to determine the outcomes of a given intervention and inform future efforts. However, monitoring typically focuses on post-release survival and spatial ecology whereas few studies assess the genetic and demographic outcomes. The eastern box turtle (Terrapene carolina carolina) is in decline throughout its range and is increasingly likely to be subject to translocations, including efforts to repatriate animals confiscated from the illegal wildlife trade. In 2019–2021, we translocated two groups of box turtles to the Savannah River Site in South Carolina, USA, including confiscated turtles (n = 208) and surrendered long-term captive turtles (LTC; n = 35). In 2022, we monitored a subset of confiscated (n = 12), LTC (n = 15), and sympatric resident (n = 8) females for reproductive output and genotyped their offspring and candidate sires to assign parentage. We found that all groups of females produced eggs at a similar rate and produced viable offspring but that the most recently translocated group (LTCs) displayed lower hatching success. Parentage assignment revealed that all groups sired offspring and mated with each other. Furthermore, our results broadly indicate that confiscated and LTC box turtles can successfully reproduce and genetically integrate following their release into wild populations, and that translocation may serve as a valuable tool for local population recovery.

59 BASIC BIOLOGICAL SCIENCES

The efficacy and ecological impact of mercury sorbents on the physiology and reproductive fitness of Daphnia magna

Environmental mercury (Hg) contamination threatens ecosystems due to its bioaccumulation and toxicity. Effective and sustainable remediation strategies are essential, with carbon-based sorbents like biochar (BC) and activated carbon (AC) gaining attention for their high adsorption capacity and cost-effectiveness. However, their ecological impacts remain poorly understood. This study evaluates the Hg removal efficacy and ecological effects of three sorbents—BC, granular AC (GAC), and AC fiber (ACF)—in a flow-through column system with and without Hg additions. We tested Hg removal at two concentrations (1 and 4 µg/L Hg) and assessed chronic exposure effects on a sentinel and sensitive freshwater organism, Daphnia magna, over 30 days of exposure to the columns’ effluent. GAC removed 100 % of Hg, while BC and ACF achieved 99 % removal. The ACF + 4 µg/L Hg effluent induced 70 % lethality in D. magna after nearly three weeks of exposure, while the GAC effluent significantly reduced fecundity, even in the absence of Hg exposure. In contrast, the BC effluent enhanced reproductive output, likely due to inputs of carbon and nitrogen in the effluent. These findings highlight the need to balance Hg removal efficacy with ecological safety, emphasizing the importance of considering both remediation effectiveness and potential environmental impacts.

Activated carbon

Vaccine Efficacy of a Replication-Competent Interferon-Expressing Porcine Reproductive and Respiratory Syndrome (PRRS) Virus Against NADC-34 Challenge

Background/Objectives: Porcine reproductive and respiratory syndrome virus (PRRSV) significantly impedes swine production due to rapid genetic variation and suppression of antiviral interferon (IFN) responses, leading to ineffective immunity. To address this, we developed IFNmix, a replication-competent PRRSV modified live vaccine (MLV) candidate co-expressing three Type I IFN subclasses (IFNα, IFNβ, IFNδ) to enhance antiviral immunity. Methods: In two independent in vivo experiments, we compared the protection of IFNmix and a commercial PRRSV MLV vaccine during challenge with a virulent PRRSV strain. Clinical signs, antibody and cytokine production, viral replication, and lung pathology in IFNmix-vaccinated pigs were compared to those of commercial PRRSV vaccines and controls. Results: Pigs vaccinated with IFNmix exhibited similar anti-PRRSV antibody development, serum viral loads, lung lesions, and cytokine responses post-challenge with the virulent NADC34 strain, with comparable or lower body temperatures and weight gain, to pigs vaccinated with the commercial vaccines. While IFNmix showed early viral load reduction compared to the commercial vaccine (Days 7–14 post-challenge), it demonstrated similar efficacy in controlling PRRSV replication and lung pathology. Conclusions: These findings suggest that IFNmix, by expressing multiple IFNs, can potentially enhance innate and adaptive immune responses, offering a promising approach to improving PRRSV vaccine efficacy. Further studies are needed to evaluate IFNmix against a broader range of PRRSV strains and to optimize its attenuation and immunogenicity.

Miller, Laura C. (ORCID:0000000289469416)