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43 records · Page 3

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NASA NTRSDate not supplied

Microbiological Analysis of Mizuna Grown in the Veggie Hardware to Define Critical Control Points and Ensure the Safety of Space Grown Crops

The Veggie facility on the International Space Station has been utilized as a “pick and eat” plant growth system to provide fresh produce for crew consumption. The VEG-04 experiments completed in 2019 examined the effect of red-rich and blue-rich light treatments on the growth of mizuna as well as harvest method and resulting yield. Analysis was performed on plant tissues and associated hardware to evaluate the influence of these experimental variables on the microbial population. VEG-04A plant pillows with pre-planted Mizuna mustard seeds were launched on SpaceX-16 in December 2018. The pillows were initiated, and a single 35-day harvest was performed. Veg 04B pillows were pre-planted with Mizuna seeds and launched on SpaceX-18 in July 2019, initiated and subsequently harvested at days 29, 43 and 58. The crew consumed approximately half of the produce, and the remainder was frozen and returned for analysis. Leaves, swabs, wicking material, substrate, and roots were processed and plated on media for the enumeration and isolation of bacteria and fungi. Isolated bacterial colonies were identified using Biolog Micro ID system or MicroSEQ16S rDNA sequencing technique. Fungal colonies were identified using the MicroSEQ D2 rDNA kit. Sample extracts were plated onto specialized media to identify Escherichia coli/coliforms, Staphylococcus aureus and Salmonella sp. Results indicate that bacterial and fungal counts were higher in plants grown in red-rich lightin VEG-04A, while the opposite was true in the Veg-04B third harvest. Microbial counts increased with the repeated harvest method used in Veg-04B. These data support the understanding of environmental and horticultural practices that can affect the microbiological quality of space-grown produce grown and aid in identification of critical control points for the development of a hazard analysis critical control point plan for ISS-grown crops. This research was co-funded by the NASA’s Human Research Program and Space Biology.

Mary E Hummerick↗

NASA NTRSDate not supplied

Effects of Spaceflight Relevant Carbon Dioxide Levels on Pathogenesis Related Microbial Characteristics

BACKGROUND Multiple stressors in the spaceflight environment have the potential to alter microbial pathogenesis and virulence characteristics, including microgravity, radiation, and hostile/confined conditions. This combination of factors may lead to a stacking of risks, potentially creating unexpected increase in the risk of infectious disease. One understudied variable is the higher CO2 atmospheric concentration. The current Spaceflight Maximum Allowance Concentration (SMAC) for 24-hour average CO2 is 0.4% (3 mm Hg). These elevated ambient CO2 levels aboard the ISS could potentially influence the diversity and phenotypic responses of the resident microbial communities from both the spacecraft environment (air, surface, water) and crewmembers (gut, nasal, skin microbiomes). METHODS For these studies, we will evaluate the response of Staphylococcus aureus, Streptococcus pneumoniae and Enterobacter aerogenes, which are opportunistic pathogens that are of medical significance and have been or are likely to be found aboard spacecraft. These microorganisms will be cultured in both spaceflight analog and control conditions at CO2levels representing terrestrial atmospheric concentration of 0.04% (0.3 mmHg), ISS elevated concentration of 0.4% (3 mm Hg), and an unexpected elevated concentration of 1 % (7.6 mmHg) to represent a worst case scenario. DISCUSSION Overwhelming evidence from studies performed in food microbiology, marine biology and terrestrial environmental biology suggest altered gene expression, selective bacterial inhibition, increased growth and diversity, and increased antibiotic resistance of bacterial communities individually and in biofilm formation when exposed to increased levels of CO2. While many of these findings are based on levels of CO2 higher than would be found during spaceflight exploration missions, the potential for subtle changes in CO2 levels to exacerbate infectious disease risks warrants an evaluation of pathogen responses to spaceflight CO2 conditions. Our study will provide answers to spaceflight relevant atmospheric conditions, as described in the Risk of Adverse Health Effects Due to Host-Microorganism Interaction (GapMicro 103).

A A Medina-Colorado↗

NASA NTRSDate not supplied

Effects of Spaceflight Relevant Carbon Dioxide Levels on Pathogenesis Related Microbial Characteristics

Multiple stressors in the spaceflight environment, including microgravity and radiation, have the potential to alter microbial pathogenesis and virulence characteristics. Understanding how these and other factors may exacerbate the risk of infectious disease for astronauts is important to ensure the success of the mission. One understudied question in our assessment of infectious disease risk is the contribution carbon dioxide (CO2) at atmospheric concentrations higher than would be observed in terrestrial settings. Previous terrestrial studies have shown altered gene expression, selective bacterial inhibition, increased growth and diversity, and increased antibiotic resistance of bacterial communities individually and in biofilms when exposed to increased levels of CO2. While many of these findings are based on levels of CO2 higher than would be found during spaceflight exploration missions, the potential for subtle changes in CO2 levels to exacerbate infectious disease risks warrants an evaluation of pathogen responses to spaceflight CO2 conditions. To address this question, we are evaluating the effects of elevated atmospheric CO2 on microbial concentration, diversity, and pathogenesis-related characteristics using medically significant microorganisms cultured at CO2 levels representing terrestrial atmospheric concentration (≈0.04 %), ISS elevated concentration (0.4 %), and an unexpectedly elevated concentration (1 %) to represent a worst-case scenario. To determine if microbial responses to these CO2 concentrations may be exacerbated in response to the spaceflight environment, microorganisms are being cultured under static conditions and both spaceflight analogue and control conditions using the Rotating Wall Vessel (RWV) bioreactor (Synthecon, Houston, TX). For these studies, we will evaluate the response of Staphylococcus aureus, Streptococcus pneumoniae, and Enterobacter aerogenes, which are opportunistic bacterial pathogens that are of medical significance and have been or are likely to be found aboard human-inhabited spacecraft. Bacteria will be evaluated individually and together as a consortium. The following Specific Aims are be investigated: Aim 1. Characterize the growth, diversity, and pathogenesis-related stress responses of medically significant microorganisms grown under multiple CO2 concentrations under static growth in a liquid medium. Stress responses include acid stress, oxidative stress, and thermal stress. Aim 2. Characterize the growth, diversity, and pathogenesis-related stress responses of medically significant microorganisms grown under multiple CO2 concentrations in both spaceflight analogue and control conditions. Stress responses include acid stress, oxidative stress, and thermal stress.

S G Thornhill↗

NASA NTRS2022

Plasma Activated Water for Crewed Transit and Planetary Habitation: A Study of Gas Type, Electrode Material, and Power Supply Selection and the Impact on the Final Frontier - FY21 CIF

An in-depth study of plasma activated water (PAW) generation was conducted to link changes in power supply, electrode material, input gas, and treatment time to the resulting reaction chemistry. These changes in chemistry can help tailor PAW for different space applications. An AC, DC, and nanosecond (ns) pulsed power supply were each used to generate PAW with stainless steel, copper, tungsten, or platinum (Pt) electrodes while utilizing air, nitrogen (N2), carbon dioxide (CO2), helium (He), or argon (Ar) as the feed gas. Tap or deionized (DI) water was treated for 1 to 15 minutes, and the generated PAW was tested for changes in pH, conductivity, oxidation reduction potential, nitrates (NO3-), ammonium (NH4+), and peroxide. Calculations showed that the production of reactive nitrogen species was the leading cause of pH and conductivity changes. The DC generated air plasma was able to reduce the pH of DI water and generate NO3‑. The pulsed supply, operating at 20% of the input power of the DC supply, lowered the pH generated NO3‑. When a simulated Martian gas mixture of 95% CO2 and 5% N2 was used as the feed gas, NO3‑ was generated with the DC and pulsed supplies, respectively. Mixing PAW with plasma generated ash from inedible biomass allowed pH control, thus enhancing PAW’s potential use for sanitation applications. The large shift in pH was used to study sanitation effects of Escherichia coli (E. coli) reduction and Staphylococcus aureus (S. aureus), in which log reductions were found to be negligible. Additionally, the plasma generated ash in combination with PAW was also implemented in 10-day microgreen growth trials, in which PAW and ash resulted in quicker emergence of the microgreens compared to the standard growth conditions and comparable dry masses to Hoagland’s nutrient solution treated samples.

plasma activated water (PAW)↗

NASA NTRSDate not supplied

Strategies for Determining Safety of Fermented Foods Produced in Space

The BioNutrients experiments examine the feasibility of using microbially produced foods – yogurt, kefir, fermented food products– to supplement the current prepackaged food system to provide freshly synthesized nutrients and genetically engineered protein therapeutics to support crew health for sustainable and long duration crewed exploration. Safety of the products must be established as direct consumption of such products is a risk due to potential contamination with pathogenic organisms. Identifying pathogen contamination is particularly challenging as the fermenting cultures can mask the presence of contaminating organisms. Here we consider current methods of detecting coliforms, aerobic colonies, non-lactic acid bacteria, molds (yeasts), and specific pathogens including Staphylococcus aureus and salmonella. We also consider alternate and additional testing including sequencing and quantitative PCR. We investigate the use of the E-Nose developed by NASA Ames Research Center. This portable device senses volatile organic compounds in real time and can be trained to recognize “good” uncontaminated products from contaminated ones as a means of predicting food safety. Finally, we consider methods of in-flight pasteurization to increase safety and decrease overall microbial load of these products. This presentation will summarize testing status and outline the strategy that will be employed in the BioNutrients-3 ISS fermented foods experiment.

Natalie Ball↗

NASA NTRSDate not supplied

Strategies for Determining Safety of Fermented Foods Produced in Space

Food↗

NASA NTRS1995

Staphylococcal enterotoxins bind H-2Db molecules on macrophages

We screened a panel of monoclonal antibodies against selected macrophage cell surface molecules for their ability to inhibit enterotoxin binding to major histocompatibility complex class II-negative C2D (H-2b) macrophages. Two monoclonal antibodies, HB36 and TIB126, that are specific for the alpha 2 domain of major histocompatibility complex class I, blocked staphylococcal enterotoxins A and B (SEA and SEB, respectively) binding to C2D macrophages in a specific and concentration-dependent manner. Inhibitory activities were haplotype-specific in that SEA and SEB binding to H-2k or H-2d macrophages was not inhibited by either monoclonal antibody. HB36, but not TIB126, inhibited enterotoxin-induced secretion of cytokines by H-2b macrophages. Lastly, passive protection of D-galactosamine-sensitized C2D mice by injection with HB36 antibody prevented SEB-induced death. Therefore, SEA and SEB binding to the alpha 2 domain of the H-2Db molecule induces biological activity and has physiological consequences.

Non-NASA Center↗