David M. Klaus

Investigation of space flight effects on Escherichia coli and a proposed model of underlying physical mechanisms

Previous investigations have reported that space flight may produce a stimulating effect on microbial metabolism; however, the specific underlying mechanisms associated with the observed changes have not yet been identified. In an effort to systematically evaluate the effect of space flight on each phase of microbial growth (lag, exponential and stationary), a series of experiments was carried out using in vitro suspension cultures of Escherichia coli aboard seven US Space Shuttle missions. The results indicated that, as a result of space flight, the lag phase was shortened, the duration of exponential growth was increased, and the final cell population density was approximately doubled. A model was derived from these cumulative data in an attempt t associate gravity-dependent, extracellular transport phenomena with unique changes observed in each specific phase of growth. It is suggested that a cumulative effect of gravity may have a significant impact on suspended cells via their fluid environment, where an immediate, direct influence of gravity might otherwise be deemed negligible.

Bacterial growth in space flight : logistic growth curve parameters for Escherichia coli and Bacillus subtilis

Abstract Previous investigations have reported that bacterial suspension cultures grow to higher stationary concentrations in space flight than on Earth; however, none of these investigations included extensive ground controls under varied inertial conditions. This study includes extensive controls and cell-growth data taken at several times during lag phase, log phase, and stationary phase of Escherichia coli and Bacillus subtilis. The Marquardt-Levenberg, least-squares fitting algorithm was used to calculate kinetic growth parameters from the logistic bacterial growth equations for space-flight and control growth curves. Space-flight cultures grew to higher stationary-phase concentrations and had shorter lag-phase durations. Also, evidence was found for increased exponential growth rate in space.

Functional weightlessness during clinorotation of cell suspensions.

A clinostat is a device often used in gravitational biology studies. Selecting an appropriate speed of rotation, however, is a frequently debated topic, particularly for suspended cells. In an attempt to define the necessary criteria for determining an acceptable revolution speed, the primary forces governing particle behavior during clinorotation--gravity, diffusion and centrifugation--were mathematically assessed. In support of the theoretical exercise, bacterial growth experiments indicated that results obtained using a clinostat followed trends resembling previous space flight results. It is suspected that this is due, in part at least, to similarly altered external transport processes in each environment.

Effects of space flight, clinorotation, and centrifugation on the substrate utilization efficiency of E. coli.

Cultures of Escherichia coli grown in space reached a 25% higher average final cell population than those in comparably matched ground controls (p<0.05). However, both groups consumed the same quantity of glucose, which suggests that space flight not only stimulated bacterial growth as has been previously reported, but also resulted in a 25% more efficient utilization of the available nutrients. Supporting experiments performed in “simulated weightlessness” under clinorotation produced similar trends of increased growth and efficiency, but to a lesser extent in absolute values. These experiments resulted in increases of 12% and 9% in average final cell population (p<0.05), while the efficiency of substrate utilization improved by 6% and 9% relative to static controls (p=0.12 and p<0.05, respectively). In contrast, hypergravity, produced by centrifugation, predictably resulted in the opposite effect — a decrease of 33% to 40% in final cell numbers with corresponding 29% to 40% lower net growth efficiencies (p<0.01). Collectively, these findings support the hypothesis that the increased bacterial growth observed in weightlessness is a result of reduced extracellular mass transport that occurs in the absence of sedimentation and buoyancy-driven convection, which consequently also improves substrate utilization efficiency in suspended cultures.

Microgravity and its implications for fermentation biotechnology

Fermentation processes are highly dependent upon physical and chemical environmental parameters, many of which are influenced by gravity. Extending biotechnology into the realm of space flight provides researchers with an opportunity to investigate the role that gravity plays in natural growth processes. Physical factors governing cell sedimentation, nutrient mixing and byproduct dispersion are altered in the absence of the constant sedimenting force of gravity. In addition, space flight has also been shown to give rise to a wide variety of indirect consequences associated with the physiology of the organisms themselves.

The assessment of leaf water content using leaf reflectance ratios in the visible, near-, and short-wave-infrared

The common features of spectral reflectance from vegetation foliage upon leaf dehydration are decreasing water absorption troughs in the near‐infrared (NIR) and short‐wave‐infrared (SWIR). We studied which leaf water index in the NIR and SWIR is most suitable for the assessment of leaf water content and the detection of leaf dehydration from the laboratory standpoint. We also examined the influence of the thickness of leaves upon leaf water indices. All leaf water content indices examined exhibited basic correlations with the relative water content (RWC) of leaves, while the R 1300/R 1450 leaf water index also demonstrated a high signal strength and low variability (R 2>0.94). All examined leaf reflectance ratios could also be correlated with leaf thickness. The thickness of leaves, however, was not independent of leaf RWC but appeared to decrease substantially as a result of leaf dehydration.

The effects of space flight on the production of monorden by Humicola fuscoatra WC5157 in solid-state fermentation.

Abstract The effect of space flight on the production of the antibiotic monorden on two types of agar media, T8 and PG, by Humicola fuscoatra WC5157 was examined on board the US Space Shuttle mission STS-77 in May 1996. Paired space-flight and ground control samples were prepared using identical hardware, protocol, media, and inoculum. Inoculation occurred simultaneously for both groups 2.5 h after launch. The flight and ground samples were allowed to grow for the entire 10-day mission in a dark, thermally controlled (22 °C) environment. Post-flight HPLC analysis of the flight and ground sample extracts indicated that the production of monorden by H. fuscoatra WC5157 in the flight samples was higher than in the ground samples in both agar media. In the T8 medium, the production of monorden in the flight and ground samples was 11.6 ± 3.5 μg and 8.9 ± 1.1 μg respectively (30% increase). In the PG medium, the production of monorden in the flight and ground samples was 23.8 ± 3.3 μg and 8.2 ± 2.2 μg respectively (190% increase). The production of monorden in the flight and ground control samples was confirmed by HPLC-MS analysis.

The effect of space flight on the production of actinomycin D by Streptomyces plicatus

The effect of space flight on production of the antibiotic actinomycin D by Streptomyces plicatus WC56452 was examined onboard the US Space Shuttle mission STS-80. Paired space flight and ground control samples were similarly prepared using identical hardware, media, and inoculum. The cultures were grown in defined and complex media under dark, anaerobic, thermally controlled (20°C) conditions with samples fixed after 7 and 12 days in orbit, and viable residuals maintained through landing at 17 days, 15 h. Postflight analyses indicated that space flight had reduced the colony-forming unit (CFU) per milliliter count of S. plicatus and increased the specific productivity (pg CFU−1) of actinomycin D. The antibiotic compound itself was not affected, but its production time course was altered in space. Viable flight samples also maintained their sporulation ability when plated on agar medium postflight, while the residual ground controls did not sporulate.

Maintenance, reliability and policies for orbital space station life support systems

The performance of productive work on space missions is critical to sustaining a human presence on orbital space stations (OSS), the Moon, or Mars. Available time for productive work has potentially been impacted on past OSS missions by underestimating the crew time needed to maintain systems, such as the Environmental Control and Life Support System (ECLSS). To determine the cause of this apparent disconnect between the design and operation of an OSS, documented crew time for maintenance was collected from the three Skylab missions and Increments 4–8 on the International Space Station (ISS), and the data was contrasted to terrestrial facility maintenance norms. The results of the ISS analysis showed that for four operational and seven functional categories, the largest deviation of 60.4% over the design time was caused by three of the four operational categories not being quantitatively included in the design documents. In a cross category analysis, 35.3% of the crew time was found to have been used to repair air and waste handling systems. The air system required additional crew time for maintenance due to a greater than expected failure rate and resultant increased time needed for repairs. Therefore, it appears that the disconnect between the design time and actual operations for ECLSS maintenance on ISS was caused by excluding non-repair activities from the estimates and experiencing greater than expected technology maintenance requirements. Based on these ISS and Skylab analyses, future OSS designs (and possibly lunar and Martian missions as well) should consider 3.0–3.3h/day for crews of 2 to 3 as a baseline of crew time needed for ECLSS maintenance.

Microbial antibiotic production aboard the International Space Station.

Previous studies examining metabolic characteristics of bacterial cultures have mostly suggested that reduced gravity is advantageous for microbial growth. As a consequence, the question of whether space flight would similarly enhance secondary metabolite production was raised. Results from three prior space shuttle experiments indicated that antibiotic production was stimulated in space for two different microbial systems, albeit under suboptimal growth conditions. The goal of this latest experiment was to determine whether the enhanced productivity would also occur with better growth conditions and over longer durations of weightlessness. Microbial antibiotic production was examined onboard the International Space Station during the 72-day 8A increment. Findings of increased productivity of actinomycin D by Streptomyces plicatus in space corroborated with previous findings for the early sample points (days 8 and 12); however, the flight production levels were lower than the matched ground control samples for the remainder of the mission. The overall goal of this research program is to elucidate the specific mechanisms responsible for the initial stimulation of productivity in space and translate this knowledge into methods for improving efficiency of commercial production facilities on Earth.