Chapter II: Part 2

The directional growth of plant shoots and plant roots is probably due to this sedimentation phenomenon, particularly the effect on movement of auxins ([ref.65]).

Free convection flow is a major transport process, and under its influence the mixing of substances is much more effective than when diffusion operates alone. Free convection flow is a macroscopic phenomenon which increases not only with g, but varies also approximately with the five-fourths power of the bulk concentration involved. Whether or not convection is important at the microscopic level remains an experimentally unsolved question. The Grashoff number limits free convection to the macroscopic domain. It would appear in weightlessness that the contribution of free convective flow would be small and that only diffusion should occur. This phenomenon would cause equilibration to occur much more slowly than that occurring with free convection and diffusion. The absence of convective transfer raises a problem as to how nutrients may be obtained and waste products removed in living cells during weightlessness. In a liquid substrate, nutrients and oxygen would be depleted, and waste products would accumulate around the cell.

Absence of gravity may have far-reaching consequences in the homeostatic aspects of cell physiology. The outstanding characteristics of living cells which are most likely to be influenced by the absence of gravity are the ability of the cell to maintain its cytoplasmic membrane in a functional state, the capacity of the cell to perform its normal functions during the mitotic cycle, and the capacity of the cytoplasm to maintain the constant reversibility of its sol-gel system ([ref.66]).

Two-phase systems, e.g., air-in-water and air-in-oil, possess entirely different characteristics at zero g than at 1 g. These physical differences in phase interaction could well be suspected of interfering with the orientation and flow pattern of cell constituents, thus hindering the cellular processes involved in the movement, metabolism, and storage of nutrients and waste.

On the basis of theoretical calculations, weightlessness can be expected to have some effect even on one individual cell if its size exceeds 10 microns in diameter ([ref.64]). Cell colonies might be affected. In larger cells there may be a redistribution of enzyme-forming systems which give rise to polarization. The low surface tension of the cell membrane lends itself to hydrostatic stress distortion, implying an alteration in permeability and thus an almost certain alteration of cell properties under low gravity conditions.

Another aspect of gravity that affects the growth and development of living organisms is the directionality of the gravitational field. In fact, some plants are so sensitive that they are able to direct their growth with as little stimulus as a 1×10⁻⁶ gravitational field. Investigations of plant growth in altered gravitational fields are underway at Argonne National Laboratory and Dartmouth College.

The Argonne Laboratory has designed and developed a 4-pi, or omnidirectional, clinostat. By rotating a plant so that the force of gravity is distributed evenly over all possible directions, the directional effects of gravity are eliminated, simulating some aspects of the zero-g state. It was shown that certain plants grew more slowly and had fewer and smaller leaves, while others had about 25 percent greater replication of fronds and had greater elongation of certain plant parts. It will be extremely interesting to compare these effects under zero-g conditions in orbiting spacecraft.

The effect of gravity in transporting growth hormones in plants has been demonstrated at Dartmouth College using radiocarbon-labeled growth hormones. Plant geotropisms and growth movements have been studied and biosatellite experiments developed.

Anatomy is considered a derivative adaptation to gravity ([ref.67]). A large background of plant research exists on the effect of orientation on plant responses. Information from clinostat experiments is considered susceptible of extrapolation to low gravity conditions because the threshold period for gravitational triggering is relatively long.

Once over critical minimum dimensions, the major effects of low gravity would be assumed to occur in those heterocellular organisms that develop in more or less fixed orientation with respect to terrestrial gravity and which respond to changes in orientation with relatively long induction periods; these are the higher plant orders. On the other extreme are the complex primates which respond rapidly, but whose multiplicity of organs and correlative mechanisms are susceptible to malfunction and disorganization. It may be suggested that the heterocellular lower plants and invertebrates will be less affected. Perturbations of the environment to which the experimental organism is exposed must be limited or controlled to reduce uncertainties in interpretation of the results. At the same time, the introduction of known perturbations may assist in isolating the effects due solely to gravity. Study of _de novo_ differentiation and other phenomena immediately after syngamy may be of particular importance. Study of anatomical changes after exposure of the organism to low gravity is important.

BIOLOGICAL EFFECTS OF SPACE RADIATION¹

¹ This section includes part of the Summary of the Panel on Radiation
Biology of the Environmental Biology Committee Space Science Board,
NAS/NRC (1963), and results of research by the Bioscience Programs,
NASA.

Radiation sources in space are of three types: galactic cosmic radiation, Van Allen belts, and solar flares with an intense proton flux. Cosmic radiation has higher energy levels than radiation produced by manmade accelerators.

The Panel on Radiation Biology, while recognizing the need for radiobiological studies of an applied nature with reference to manned flight programs, stated that it would be shortsighted for the United States to confine its efforts to the solution of immediate problems since, in the long run, successful exploration of space will be aided by the contributions of basic research. Both the immediate biological research program and the continuing program for basic studies should be built upon the large body of existing knowledge of radiation effects. The attitude that all radiobiological experiments need be repeated in the space environment should be resolutely rejected. Since fundamental radiobiology cannot be performed easily in space, it has been recommended that, wherever possible, these investigations be carried out in ground laboratories in preference to flying laboratories.

Space environment does vary from the terrestrial environment, but the variations are not so great as to lead to the expectation of strikingly different biological effects of radiation in space. However, it is conceivable that radiations whose effects are well known under terrestrial conditions may have some unsuspected biological effects when combined with unusual features of the space environment: e.g., zero g. Previous space radiobiological studies have depended solely on very low and inaccurately measured doses of ambient space radiation. It has been difficult to distinguish between the observed response levels and the random noise; thus, experiments have been inconclusive.

Biological Effects of Heavy Ions and Mesons

The biological effects of heavy ions (especially Z>2) and mesons are of specific interest to space radiobiology.

Controlled Radiobiological Experiments in Space

There is the remote possibility that the radiobiological response may be modified by factors as yet unknown and perhaps not susceptible to terrestrial study. Experiments have been designed to settle this matter including the exposure of biological materials during space flight which meet the following criteria of reliability: (1) the use of well-known biological systems, e.g., mutation induction or chromosome breakage; (2) the use of a sufficient number of individuals in the experiment to guarantee statistical precision on the results; (3) the exposure of the system to known quantities and qualities of radiation; (4) the use of adequate controls.

High-altitude balloon ascents of the 1930’s initiated study of the biological effects of cosmic rays. They were limited to the exploration of secondary cosmic radiation effects. After World War II, the research extended to the use of V-2 rockets fired from the White Sands Proving Ground. Interest returned to balloons and a significant program was underway by 1950, first using mice and then hamsters, fruit flies, cats, and dogs. These flights gave no evidence of radiation damage. However, it was realized that the flights were too far south to obtain a significant exposure, and more northerly flights began in 1953. Mice and guinea pigs were flown on these later flights. Chase ([ref.68]) showed the most unequivocal results to that time, a statistically significant increase in light hairs on black animals and the streaks of white hair up to 10 times wider than expected. Brain lesions were detected in the guinea pigs flown on Man High in 1957. Many other types of biological material were sent aloft in an effort to further corroborate existing information and to investigate genetic and developmental effects of cosmic radiation.

From the earlier V-2 rocket flights to the Jupiter missile launchings of the monkeys Able and Baker, cosmic-ray research was continued, but the short flight durations of these vehicles did not provide substantial information. The USAF Discoverer satellite program has given impetus to cosmic-ray research and provided for longer "staytimes."

It has been difficult to separate radiation effects from other space-flight factors: therefore, some of the alterations observed are still subject to debate. Vibration, acceleration, and weightlessness appear to be the three most important additional parameters. Measurements of radiation dosage have been made by chemical and photographic dosimetry, ion chambers, and biological dosimetry. All evidence to date indicates that radiation exposure levels are not hazardous to man at present orbital altitudes up to 200 nautical miles. Most biological materials flown so far have been for the express purpose of investigating space-radiation levels and effects. The biological materials have ranged from tissue cultures to entire organisms and from phage and bacterial cells to man. The studies have required much of the space and weight resources allotted biology by the U.S.S.R. and the United States. They have been accompanied by ground-based controls.

The Vostok series provided the following data:

(1) A small, but statistically significant, increase was observed in
the percentage of chromosome aberrations in the rootlet cells of
air-dried wheat and pea seeds after germination. In this case
only, the increase did not depend on flight duration.
(2) Lysogenic bacteria exhibited an increase of genetic alterations
and increased phage production. Length of flight was associated
with increased bacteriophage production by the lysogenic bacteria.
There was an increase of recessive lethals coupled with
nonconvergence of chromosomes (sex linked) in the fruit fly. A
stimulation of cell division in wheat and pea seeds was observed.
Cultures of human cells exposed to space-flight factors did not
differ significantly from terrestrial controls with respect to
such indicators as proliferation rate, percentage of mortality and
morphological, antigenic, and cultural properties. Repeated
flights of the identical HeLa cells revealed that there was a
longer latent period for restoration of growth capacity than in
cells carried into space once or not flown at all.
(3) The most definite radiation effects observed were only revealed in
genetic tests. No harmful influence on those characteristics
affecting the viability of the organism has been discovered.

The Air Force Discoverer series launched from the west coast had a few successful flights incorporating organisms. With severe environmental stress and long recovery times, data on radiation exposure were equivocal up to Discoverer XVII and XVIII when cultures of human tissue were flown, recovered, and assessed for radiation exposure effects. Comparison with ground-based controls revealed no measurable differences.

Radiation dosimetry from the Mercury series established that minimal exposures were encountered at those orbital altitudes. A typical example is the MA-8 flight of W. M. Schirra, Jr., during which the body surface dosage was less than 30 millirads.

NASA has supported fundamental radiation studies at the Oak Ridge National Laboratory and the Lawrence Radiation Laboratory. Emphasis has been placed on the biological effects of high-energy proton radiation and particulate radiation from accelerators.

At the NASA Ames Research Center extensive fundamental studies are being carried out on the effects of radiation, especially in the nervous system. It has been demonstrated that deposits accumulate in the brain following exposure to large doses of ionizing particle radiation as well as after X-irradiation. These deposits, referred to as a "chemical lesion," result from an accumulation of glycogen. The formation of these deposits during exposure to large doses of X-irradiation was not increased in environments of 99.5 percent oxygen and increased atmospheric pressure.

SIMULATION OF PLANETARY (MARTIAN) ENVIRONMENTS

Attempts have been made to simulate to some degree the various parameters of the Martian environment, such as atmospheric composition, pressure, radiation flux, temperatures, and the day-night as well as seasonal cycles. Certain factors for Mars cannot yet be simulated, such as soil composition, gravitational field, magnetic field, and electrical field.

Caution is required in interpreting all simulation experiments. How Earth organisms respond to simulated Martian environments probably has nothing to do with life on Mars, but these experiments may show whether or not anything in the environment of Mars makes life as we know it impossible. We must expect that on Mars, life will have evolved and have adapted over long periods of time under conditions which are quite different from conditions on Earth. The simulation experiments also provide some information about the possibility of contaminating the planet Mars, or any planet, with organisms from Earth. In addition, they give us some clues about the possibilities of adaptation and evolution of life under these conditions.

From an evolutionary point of view, if life has developed on Mars, we expect it to have evolved at least to a microbial stage. On Earth, micro-organisms are the most ubiquitous and numerous forms of life. This fact should be considered in studying extraterrestrial bodies.

Micro-organisms have been selected as the best test organisms, and bacteria and fungi have been used because they are durable and easy to grow. Also, because of their rapid growth, many generations can be studied in a relatively short period of time. The organisms include chemoautotrophic bacteria, which are able to synthesize their cell constituents from carbon dioxide by energy derived from inorganic reactions; anaerobic bacteria, which grow only in the absence of molecular oxygen; photoautotrophic plants such as algae, lichens, and more complex seed plants; and small terrestrial animals.

Organisms have been collected from tundra, desert, hot springs, alpine, and saline habitats to obtain species with specialized capabilities to conserve water, balance osmotic discrepancies, store gases, accommodate to temperature extremes, and otherwise meet stresses. An attempt is made in these simulation experiments to extend these processes across the possible overlapping microenvironments which Earth and Mars may share.

Scientists have developed various special environmental simulators, including "Mars jars" and "Marsariums." These have made possible controlled temperatures, atmospheres, pressures, water activities, and soil conditions for duplicating assumed Martian surface. A complex simulator, developed by Young et al. ([ref.52]), reproduces the formation of a permafrost layer with some water tied up in the form of ice beneath the soil surface. This simulator serves as a model to study the wave of darkening, thus supporting the hypothesis that the pole-to-equator wave of darkening is correlated with the availability of subsurface water. The simulator is a heavily insulated 2-cu-ft capacity chamber with an internal pressure of 0.1 atm. The chamber contains a soil mixture of limonite and sand and an atmosphere of carbon dioxide and nitrogen. With the use of a liquid nitrogen heat exchanger at one end and an external battery of infrared lamps at the other end, the temperature simulates that of Mars from pole to equator. Thermocouples throughout the soil monitor the temperatures in the chamber.

Zhukova and Kondratyev ([ref.69]) designed a structure measuring 100×150×180 cm. Micro-organisms were placed at the surface of a copper bar made in a special groove separated by glass cloth. Copper was selected as one of the best heat-conduction materials permitting a rapid change of temperature. The lower end of the bar was immersed into a mixture of dry ice and ethyl alcohol, which made it possible to create a temperature of -60° C. Heating was performed by an incandescent spiral.

As the knowledge concerning the Martian environment becomes more refined, scientists can more accurately simulate this environment under controlled conditions in the laboratory. Determination of the effects of the Martian environment on Earth organisms will permit better theorization on the forms of life we might find on Mars and will permit us to estimate the potential survival of Earth contaminants on Mars.

However, until the environmental conditions of Mars are defined more accurately, the experiments must be changed continually to fit newly determined conditions. Therefore, existing simulation data are made less valid for comparison. The data resulting from the simulation experiments for Mars have been compiled in table II, and the experiments are summarized below.

The earliest simulation studies were carried out by the Air Force, and the studies during the past 6 years have been supported by NASA. Recently, these studies have received less support or have been terminated in favor of critical studies on the effects of biologically important environmental extreme factors on Earth organisms. These critical studies permit establishing the extreme environmental factor parameters in which Earth life can grow or survive. These data will have valuable application to the consideration of life on any planet, to the design of life-detection instruments, to the sterilization of space vehicles, and to the problem of contamination of planets.

Some exploratory experimental studies are in progress to study the capabilities of organisms to grow under the assumed conditions on Jupiter. These include studies at high pressure with liquid ammonia, methane, and other reducing compounds.

Early experiments simulating Martian conditions using soil bacteria were carried out by Davis and Fulton ([ref.70]) at the Air Force School of Aviation Medicine, San Antonio, Tex. Mixed populations of soil bacteria were put in "Mars jars" with the following conditions: 65-mm Hg pressure, 1 percent water or less, nitrogen atmosphere, sandstone-lava soil, and a temperature day-night cycle of +25° to -25° C. The moisture was controlled by desiccating the soil and adding a given amount of water. Experiments, conducted up to 10 months, demonstrated that obligate aerobes died quickly. The anaerobes and sporeformers survived. Although a small increase in the total number of organisms indicated growth, the increases in the number of bacteria may have been due to breaking up clumps of dirt.

Roberts and Irvine ([ref.71]) reported that, in a simulated Martian environment, colony counts of a sporeforming bacterium, _Bacillus cereus_, increased when 8 percent moisture was added. Moisture was considered more important than temperature or atmospheric gases inasmuch as a simulated Martian microenvironment containing 8 percent moisture permitted germination and growth of endospores of _Clostridium sporogenes_. Increases in colony counts of _Bacillus cereus_ appeared to be influenced by temperature cycling ([ref.72]).

Table II.—_Survival and Growth of Organisms in Simulated Planetary
(Martian) Environments_

------------------------------------------------------------------
Species Survival, Moisture Temperature,
months °C

------------------------------------------------------------------
Conditions on Mars: 14µ±7µ -70 to +30
------------------------------------------------------------------
Anaerobic 6 Low, -60 to +20
sporeformers (CaSO₄)
_Clostridia_,
_Bacillus
planosarcina_
------------------------------------------------------------------
Anaerobic 6 Low, -60 to +20
nonsporeformers (CaSO₄)
_Pseudomonas_,
_Rhodopseudomonas_
------------------------------------------------------------------
Anaerobes Growth Very wet -75 to +25
_Aerobacter
aerogenes_,
_Pseudomonas sp._
------------------------------------------------------------------
_Clostridium_, 10 1 -25 to +25
_Corynebacteria_ percent
"Thin short rod" or less
------------------------------------------------------------------
_Bacillus cereus_ 2 0.5 -25 to +25
percent
soil
------------------------------------------------------------------
_Clostridium sporogenes_ 1 8.4 -25 to +25
(growth) percent
------------------------------------------------------------------
_Clostridium botulinum_ 10 Lyophilized -25 to +25

------------------------------------------------------------------
_Klebsiella pneumoniae_ 6 Lyophilized -25 to +25

------------------------------------------------------------------
_Bacillus subtilis_ var. 4 2 percent -25 to +25
_globigii_
------------------------------------------------------------------
_Sarcina aurantiaca_ 4 0.5 percent -25 to +25

------------------------------------------------------------------
_Clostridium tetani_ 2 or less 1 percent -60 to +25
------------------------------------------------------------------
_Aspergillus niger_ Over 6 hr Very dry -60 to +25

------------------------------------------------------------------
_Aspergillus oryzae_ Over 6 hr Very dry -60 to +25
------------------------------------------------------------------
_Mucor plumbeus_ Over 6 hr Very dry -60 to +25
------------------------------------------------------------------
_Rhodotorula rubra_ Over 6 hr Very dry -60 to +25
------------------------------------------------------------------
Pea, bean, tomato, rye, 0.3 Moist +25
sorghum, rice.
------------------------------------------------------------------
Winter rye 0.6 Moist -10 to +23
------------------------------------------------------------------

Table II.—_Survival and Growth of Organisms in Simulated Planetary
(Martian) Environments_

----------------------------------------------------------------------
Species Atmospheric N₂, CO₂, Substrate
pressure, percent percent
mm Hg
----------------------------------------------------------------------
Conditions on Mars: 85, 3 to 30
25±15, 11
----------------------------------------------------------------------
Anaerobic 76 95 5 Air-dried
sporeformers soil
_Clostridia_,
_Bacillus
planosarcina_
----------------------------------------------------------------------
Anaerobic 76 95 5 Air-dried
nonsporeformers soil
_Pseudomonas_,
_Rhodopseudomonas_
----------------------------------------------------------------------
Anaerobes 760 100 (?) Difco
_Aerobacter infusion
aerogenes_, broth
_Pseudomonas sp._
----------------------------------------------------------------------
_Clostridium_, 65 100 (?) Soil
_Corynebacteria_
"Thin short rod"
----------------------------------------------------------------------
_Bacillus cereus_ 65 94 2.21 Sandstone
soil

----------------------------------------------------------------------
_Clostridium sporogenes_ 65 94 2 Enriched
soil
----------------------------------------------------------------------
_Clostridium botulinum_ 65 95 0 to Lava soil
0.5
----------------------------------------------------------------------
_Klebsiella pneumoniae_ 65 95 0 to Lava soil
0.5
----------------------------------------------------------------------
_Bacillus subtilis_ var. 85 95 0.3 Media
_globigii_
----------------------------------------------------------------------
_Sarcina aurantiaca_ 85 95 0.3 Desert
soil
----------------------------------------------------------------------
_Clostridium tetani_ 85 95 0.3 Soil
----------------------------------------------------------------------
_Aspergillus niger_ 76 95.5 0.25 Glass
cloth on
copper
bar
----------------------------------------------------------------------
_Aspergillus oryzae_ 76 95.5 0.25 Do.
----------------------------------------------------------------------
_Mucor plumbeus_ 76 95.5 0.25 Do.
----------------------------------------------------------------------
_Rhodotorula rubra_ 76 95.5 0.25 Do.
----------------------------------------------------------------------
Pea, bean, tomato, rye, 75 100 0 Filter
sorghum, rice. paper
----------------------------------------------------------------------
Winter rye 76 98 0.24 Soil
----------------------------------------------------------------------

Studies of the effects of simulated Martian environments on sporeforming anaerobic bacteria were carried out by Hawrylewicz et al. ([ref.49]). They showed that the encapsulated facultative anaerobe, _Klebsiella pneumoniae_, survived under simulated Martian atmosphere for 6 to 8 months, but were less virulent than the freshly isolated organisms. Spores of the anaerobe _Clostridium botulinum_ survived 10 months in the simulator. Hagen et al. ([ref.53]) found that the addition of moisture to dry-simulated Martian soil did not improve the survival of _Bacillus subtilis_ or _Pseudomonas aeruginosa_. _Bacillus cereus_ spores survived, with added organic medium plus moisture, but no germination of the spores resulted.

Hawrylewicz et al. ([ref.49]) put rocks from Antarctica bearing various lichens in simulated Martian conditions in a large desiccator. They found that the algal portion of a lichen, _Trebouxia erici_, showed only slight resistance to the Martian environment. They also pointed out the effect moisture had on the physical condition of lichens. The undersurface of a lichen has great water-absorbing capability, and the slightest amount of moisture on a rock surface is absorbed by the lichen which can turn green in 15 minutes.

Scher et al. ([ref.51]) exposed desert soils to simulated environmental conditions and diurnal cycles of Mars. The atmosphere consisted of 95 percent nitrogen and 5 percent carbon dioxide (no oxygen) and was dried, using calcium sulfate as a desiccant. The total atmospheric pressure was 0.1 atm. The temperature ranged from -60° to +20° C in 24-hour cycles. One hour was spent at the maximum and at the minimum temperatures. The chambers were irradiated with ultraviolet, 2537 Å, with a dose of 10⁹ ergs/cm², which is comparable to a daily dose found on Mars, and easily exceeds the mean lethal dose for unprotected bacteria. Soil aliquots were removed weekly and incubated at 30° C. The scoring was done both aerobically and anaerobically. Sporeforming obligate and facultative anaerobes, including _Clostridium_, _Bacillus_, and _Planosarcina_, and nonsporeforming facultative anaerobes, including _Pseudomonas_ and _Rhodopseudomonas_, were found. The experimental chambers were frozen and thawed cyclically up to 6 months. Organisms that were able to survive the first freeze-thaw cycle were able to survive the entire experiment. The ultraviolet irradiation did not kill subsurface organisms, and a thin layer of soil served as an ultraviolet shield. All of the samples showed survivors.

Young et al. ([ref.52]) assumed that water is present on Mars, at least in microenvironments, and that nutrients would be available. The primary objective of their experiments was to determine the likelihood of contaminating Mars with Earth organisms should a space probe from Earth encounter an optimum microenvironment in terms of water and nutrients. The experiments used bacteria in liquid nutrient media. The environment consisted of a carbon dioxide-nitrogen atmosphere, and the temperature cycling was -70° to +25° C, with a maximum time above freezing of 4½ hours. _Aerobacter aerogenes_ and _Pseudomonas sp._ grew in nutrient medium under Martian freezing and thawing cycles. Atmospheric pressure was not a significant factor in the growth of bacteria under these conditions.

Silverman et al. ([ref.47]) studied bacteria and a fungus under extreme—but not "Martian"—conditions. Spores of five test organisms (_B. subtilis_ var. _niger_, _B. megaterium_, _B. stearothermophilus_, _Clostridium sporogenes,_ and _Aspergillus niger_) and soils were exposed while under ultrahigh vacuum to temperatures of from -190° to +170° C for 4 to 5 days. Up to 25° C there was no loss in viability; at higher temperatures, differences in resistivity were observed. At 88° C, only _B. subtilis_ and _A. niger_ survived in appreciable numbers; at 107° C, only _A. niger_ spores survived; none were recoverable after exposure to 120° C. _B. subtilis_ survived at atmospheric pressure and 90° C for 5 days, but none of the other spores were viable alter 2 days. Four groups of soil organisms (mesophilic, aerobic, and anaerobic bacteria, molds, and actinomycetes) were similarly tested in the vacuum chamber. From one sample only actinomycetes survived 120° C, while one other soil sample yielded viable bacteria after exposure to 170° C. Several organisms resisted 120° C in ultrahigh vacuum for 4 to 5 days. When irradiated with gamma rays from a cobalt 60 source, differences were observed between vacuum-dried spores irradiated while under vacuum and those exposed to air immediately before irradiation. A reduction of from one-third to one-ninth of the viability of spores irradiated in vacuum occurred with vacuum-treated spores irradiated in air.

Siegel et al. ([ref.73]), in approximate simulations of Martian environments, studied tolerances of certain seed plants, such as cucumbers, corn, and winter rye, to low temperatures and lowered oxygen tensions. Lowered oxygen tensions enhanced the resistance of seedlings, particularly cucumber and rye to freezing, and lowered the minimum temperature required for germination. Germination of seeds in the absence of liquid water has also been studied. In this case, seeds of xerophytes have been suspended in air at 75-mm Hg pressure above water. The air was thus saturated. Germination was slow but did occur.

Siegel et al. (refs. [ref.73] and [ref.74]) found that the growth rate of several higher plants was enhanced by certain gases usually thought to be toxic, such as N₂O. This finding is significant inasmuch as the presence of nitrogen oxides in the Martian atmosphere has been cited as evidence for the nonexistence of plants on that planet by Kiess et al. ([ref.75]). Exploratory survival tests showed that various mature plants, as well as the larvae, pupae, and adult specimens of a coleopteran insect, were undamaged when exposed to at least 40 hours of an atmosphere containing 96.5 percent N₂O, 0.7 percent O₂, and 2.8 percent N₂.

Lichens are of interest because of their ability to survive and thrive under extreme environmental conditions on Earth. Biological activity of slow-growing lichens was detected by metabolic gas exchange, CO₂ detection being especially convenient. Siegel points out that this method is sensitive and nondestructive, to be preferred to staining techniques, which at present are limited because they are only semiquantitative, subjective, and destructive of the lichen.

A Russian study of simulated planetary environments has been performed with good simulation but for periods of only 2 to 6 hours. Comments on simulation experiments made by Zhukova and Kondratyev ([ref.69]) are presented as follows:

On the basis of modern conceptions on Martian conditions it is
difficult to imagine that higher forms of animals or plants
exist on the planet. A Martian change of seasons similar to that
of our planet empowers us to think that there is a circulation
of an organic substance on Mars, which cannot exist without
participation of microbic forms of life. Microorganisms are the
most probable inhabitants of Mars although the possibility is
not excluded that their physiological features will be very
specific. That is why the solution of the problem concerning the
character of life on Mars is of exceptional interest. But still
the answer to this question can be verified only by simulating
Martian conditions, taking into account the information obtained
from astrophysicists.

Experiments aimed at creating artificial Martian climatic
conditions have been started quite recently; their number is not
large since they cannot be combined with the results of numerous
experiments investigating the effect of extreme factors on
microorganisms. The result of the effect of such physicochemical
parameters of the medium as pressure, sharp temperature changes,
the absence of oxygen and insolation, depends on their
combination and simultaneity. These examples convincingly show
that while simulating Martian conditions one should strive to
the most comprehensive complex of simultaneously acting factors.
The creation of individual climatic parameters acting
successively leads to absolutely different, often opposite
results. It should be mentioned also that refusal to imitate
insolation and the performance of experiments with specimens of
soil which itself has protective effect on cells of
microorganisms, but not with pure culture of bacteria, are usual
shortcomings in the bulk of studies on this problem.

It appears that organisms from Earth might survive in large numbers when introduced to Martian environment. Whether these organisms will be capable of growth and explosive contamination of the planet in a biological sense or not is highly questionable. The likelihood of an organism from Earth finding ideal conditions for growth on Mars seems extremely low. However, the likelihood of an organism from Earth serving as a contaminant for any life-detection device flown to Mars for the purpose of searching out carbon-based life is considerably higher. The chance that life has originated and evolved on Mars is a completely separate question and much more difficult to answer.

It would be interesting to attempt to determine possible evolutionary trends which might occur on a planet by means of selection of organisms in a simulated planetary environment. Rapid genetic selection combined with radiation and chemicals to speed up mutation rate under these conditions should reveal possible evolutionary trends under the planetary environmental conditions. This could be attempted after the planetary environments are more accurately defined.

EXTREME AND LIMITING ENVIRONMENTAL PARAMETERS OF LIFE

The question of the existence of extraterrestrial life is one of the most important and interesting biological questions facing mankind and has been the subject of much controversial discussion and conjecture. Many of the quantitative, and even qualitative, environmental constituents of the planets also are still subjects of controversy and speculation. Best guesses about a relatively unknown planetary environment, combined with lack of information about the capabilities of Earth life to grow in extreme environments, do not provide the basis for making informed scientific estimates.

Life on Earth is usually considered to be relatively limited in its ability to grow, reproduce, or survive in extreme environmental conditions. While many common plants and animals (including man) are quite sensitive to, or incapable of, surviving severe chemical and physical changes or extremes of environment, a large number of micro-organisms are highly adapted and flourish in environments usually considered lethal. Certain chemoautotrophic bacteria require high concentrations of ammonia, methane, or other chemicals to grow. Anaerobic bacteria grow only in the absence of oxygen.

Besides adapting to the extremes of environments on Earth, life is also capable of growing and reproducing under extreme environmental conditions not normally encountered: e.g., from a few rad of radiation in normal habitats to 10⁶ or more rad from artificial sources, from 0.5 gauss of Earth magnetism to 167 000 gauss in manmade magnetic fields, and from 1-g force of gravity to 110 000 g. The extreme ranges of physical and chemical environmental factors for growth, reproduction, and survival for Earth micro-organisms are phenomenally large.

Life is ubiquitous on Earth and is found in almost every possible environment, including the most severe habitats, from the bottom of the ocean to the highest mountain tops and from cold Arctic habitats to hot springs, as well as in volcanic craters, deep wells, salt flats, and mountain snowfields. Earth life has become adapted to, and has invaded, nearly every habitat, no matter how severe. The physiological and morphological adaptations of life are exceedingly diverse and complex.

Surprisingly, the extreme parameters or ranges of the physical and chemical environmental factors permitting growth, reproduction, and other physiological processes of Earth organisms have not been critically compiled. A partial compilation of certain selected environmental factors has been made by Vallentyne ([ref.76]). A compilation of available published data on certain environmental extremes, particularly from recent NASA-supported research (compiled by Dale W. Jenkins, in press), is presented in tables III to VI. These data can serve as a starting point for a more intensive literature review by specialists, critical evaluation, standardization of end points, and especially to point out areas where critical experimentation is urgently needed.

This critical compilation involves a review of a very broad and complex range of subjects involved in many different disciplines with widely scattered literature. Since the effects of many of the specific environmental factors are harmful, it is difficult to select a point on a scale from no effect to death and use some criteria to say that normal or even minimal growth and reproduction are occurring. The effects of environmental factors are dependent on (1) the specific factor, times, (2) the concentration or energy, times, (3) the time of exposure or application of the factor. Many reports, especially older ones, do not give all of the necessary data to permit proper evaluation. A complicating factor is that the effect of each factor depends on the other factors before, during, and after its application. The condition of the organism itself is a great variable. Proper evaluation requires the critical review by a variety of biological specialists, physicists, and chemists.

To determine the potential of Earth organisms to survive or grow under other planetary environmental conditions, a number of experiments have been carried out attempting to simulate planetary environments, especially of Mars, as reviewed previously. While the results are of real interest, they do not provide much basic information. Further, as the Martian environment is more accurately defined, the experimental conditions are changed. In addition, some experimenters have altered certain factors, such as water content, to allow for potential microhabitats or for areas which might contain more water at certain times.

Table III.—_Extreme Physical Environmental Factors_

-----------------------------------------------------------------
Physical Minimum Organism
factors
-----------------------------------------------------------------
Temperature -30° C Algae (photosynthesis),
pink yeast (growth)
-----------------------------------------------------------------
Magnetism 0-50 gamma (=×10⁻⁵ Human
gauss)

-----------------------------------------------------------------
Gravity 0 g Human, plants, animals

-----------------------------------------------------------------
Pressure 10⁻⁹ mm Hg (5 days) _Mycobacterium_
_smegmatis_
-----------------------------------------------------------------
Microwave 0 W/cm²

-----------------------------------------------------------------
Visible 0 ft-c Animals, fungi,
bacteria

-----------------------------------------------------------------
Ultraviolet 0 erg/cm²

-----------------------------------------------------------------
X-ray 0 rad
-----------------------------------------------------------------
Gamma ray 0 rad

-----------------------------------------------------------------
Acoustic 0 dyne/cm²

-----------------------------------------------------------------

Table III.—_Extreme Physical Environmental Factors_

----------------------------------------------------------------------
Physical Maximum Organism Activity
factors
----------------------------------------------------------------------
Temperature 104° C (1000 _Desulfovibrio Grows and reduces
atm) desulfuricans_ sulfate
----------------------------------------------------------------------
Magnetism 167 000 _Neurospora_ 1 hr—no effect,
gauss _Arbacia_ _Arbacia_
_Drosophila_ development delayed
----------------------------------------------------------------------
Gravity 400 000 g _Ascaris_ eggs 1 hr—eggs hatch, 40
110 000 g _Escherichia coli_ days’ growth
----------------------------------------------------------------------
Pressure 1400 atm Marine organisms Growth

----------------------------------------------------------------------
Microwave 2450 Mc/sec _Drosophila_ 68 hr, growth not
0.3 to 1 affected
W/cm²
----------------------------------------------------------------------
Visible 50 000 ft-c _Chlorella_, Seconds,
17 000 ft-c higher plants recurrently
continuous
----------------------------------------------------------------------
Ultraviolet 10⁸ erg/cm², Bean embryos Suppressed growth
2537 Å
----------------------------------------------------------------------
X-ray 2×10⁶ rad Bacteria Growth
----------------------------------------------------------------------
Gamma ray 2.45×10⁶ rad _Microcoleus_ Continued growth
_Phormidium_
_Synechococcus_
----------------------------------------------------------------------
Acoustic 140 db or Man Threshold of pain
6500
dyne/cm² at
0.02 to 4.8
kcs/sec
----------------------------------------------------------------------

Table IV.—_Extreme Low and High Temperature Effects Permitting
Life Processes_

-----------------------------------------------------------------
Minimum Organism Activity or condition
temperature,
°C
-----------------------------------------------------------------
-11 Bacteria Growth (on fish)
-----------------------------------------------------------------
-12 Bacteria Growth
-----------------------------------------------------------------
-12 Molds Growth
-----------------------------------------------------------------
-15 _Pyramidomonas_ Swimming
-----------------------------------------------------------------
-15 _Dunaliella salina_ Swimming
-----------------------------------------------------------------
-18 Mold Growth
-----------------------------------------------------------------
-18 Yeast Growth
-----------------------------------------------------------------
-18 _Aspergillus Growth (in glycerol)
glaucus_
-----------------------------------------------------------------
-18 to -20 Mold Growth (in fruit juice)
-----------------------------------------------------------------
-18 to -20 _Pseudomonads_ Growth (in fruit juice)
-----------------------------------------------------------------
-20 Bacteria Growth
-----------------------------------------------------------------
-20 Bacteria Growth
-----------------------------------------------------------------
-20 Bacteria Luminescence development
accelerated
-----------------------------------------------------------------
-20 to -24 Insect eggs
(diapause)
-----------------------------------------------------------------
-30 Algae Photosynthesis
-----------------------------------------------------------------
-30 Pink yeast Growth (on oysters)
-----------------------------------------------------------------
-30 Lichens Photosynthesis
-----------------------------------------------------------------
-20 to -40 Lichens and conifers Photosynthesis
-----------------------------------------------------------------
-44 Mold spores Sporulation and germination
-----------------------------------------------------------------

Table IV.—_Extreme Low and High Temperature Effects Permitting
Life Processes_

------------------------------------------------------------------
Maximum Organism Activity or condition
temperature,
°C
------------------------------------------------------------------
73 Thermophilic organisms Growth (P³² metabolism)
------------------------------------------------------------------
73 _Phormidium_ (alga) Acclimatized
------------------------------------------------------------------
70 to 73 _Bacillus calidus_ Growth and spore
germination
------------------------------------------------------------------
70 to 74 _Bacillus cylindricus_ Growth and spore
germination
------------------------------------------------------------------
70 to 75 _Bacillus tostatus_ Growth and spore
germination
------------------------------------------------------------------
80 _Bacillus Cultured in laboratory
stearothermophilus_
------------------------------------------------------------------
83 Sulfate-reducing Found in a well
bacteria
------------------------------------------------------------------
89 Sulfate-reducing Found in oil waters
bacteria
------------------------------------------------------------------
65 to 85 Sulfate-reducing Cultured in laboratory
bacteria
------------------------------------------------------------------
89 Micro-organisms Found in hot springs
------------------------------------------------------------------
95 _Bacillus coagulans_ In 80 min. sporulation
activation
------------------------------------------------------------------
110 _Bacillus coagulans_ In 6 min, sporulation
activation
------------------------------------------------------------------
104 _Desulfovibrio Grow and reduce sulfate
desulfuricans_ at 1000 atm
------------------------------------------------------------------

Table V.—_Extreme Temperature Limits of Survival_

--------------------------------------------------
Minimum Organism
temperature
°C
--------------------------------------------------
-190 Yeast bacteria, 10 species
--------------------------------------------------
-197 _Trebouxia erici_ from lichens
--------------------------------------------------
-197 Protozoa, _Anguillula_
--------------------------------------------------
-252 Yeasts, molds, bacteria, 10 species
--------------------------------------------------
-253 Black currant, birch
--------------------------------------------------
-273 Bacteria, many species
--------------------------------------------------
-273 Bacteria, many species
--------------------------------------------------
-272 Desiccated rotifers
--------------------------------------------------
-269 Human spermatozoa
--------------------------------------------------

Table V.—_Extreme Temperature Limits of Survival_

------------------------------------------------------------------
Maximum Organism Time of exposure
temperature
°C
------------------------------------------------------------------
140 Bacterial spores 5-hr immersion
------------------------------------------------------------------
170-200 Desiccated rotifers 5 min
------------------------------------------------------------------
151 Desiccated rotifers 35 min
------------------------------------------------------------------
150 _Clostridium tetani_ 180 min
------------------------------------------------------------------
170 Aerobic bacteria, molds. 5 days at
actinomycetes 6×10⁻⁹mm Hg
------------------------------------------------------------------
127 (dry) Bacteria (in activated charcoal) 60 min
------------------------------------------------------------------
110 (wet) _Bacillus subtilis_ var. _niger_ 400 min
------------------------------------------------------------------
120 _Bacillus subtilis_ var. _niger_ 400 min
------------------------------------------------------------------
141 _Bacillus subtilis_ var. _niger_ 70 min
------------------------------------------------------------------
160 _Bacillus subtilis_ var. _niger_ 15 min
------------------------------------------------------------------
180 _Bacillus subtilis_ var. _niger_ 2 min
------------------------------------------------------------------
188 _Bacillus subtilis_ var. _niger_ 1 min
------------------------------------------------------------------
120 (wet) _Bacillus stearothermophilus_ 25 min
------------------------------------------------------------------
120 (dry) _Bacillus stearothermophilus_ 100 min
------------------------------------------------------------------
141 _Bacillus stearothermophilus_ 12 min
------------------------------------------------------------------
160 _Bacillus stearothermophilus_ 2 min
------------------------------------------------------------------
166 _Bacillus stearothermophilus_ 1 min
------------------------------------------------------------------

Table VI.—_Extremes of Chemical Environmental Factors
Permitting Growth or Activity_

--------------------------------------------------------
Chemical Minimum Organism
factor
--------------------------------------------------------
O₂ 0% HeLa cells, _Cephalobus_,
anaerobic bacteria
--------------------------------------------------------
O₃ (ozone) 0%

--------------------------------------------------------
H₂ 0%

--------------------------------------------------------
H₂O Aw 0.48 _Pleurococcus vulgaris_
------------------------------------------
Aw 0.5 _Xenopsylla cheopis_
(prepupae)
--------------------------------------------------------
H₂O₂ 0%

--------------------------------------------------------
He 0%

--------------------------------------------------------
CO 0%

--------------------------------------------------------
CO₂ 0%

--------------------------------------------------------
CH₄ 0%
--------------------------------------------------------
CH₂O 0%
--------------------------------------------------------
CH₃OH 0%
--------------------------------------------------------
N₂ 0%

--------------------------------------------------------
NO 0%

--------------------------------------------------------
NO₂ 0%

--------------------------------------------------------
N₂O 0%

--------------------------------------------------------
Ar 0%
--------------------------------------------------------
NaCl,
Na₂SO₄,
NaHCO₃
--------------------------------------------------------
H₂S 0%

--------------------------------------------------------
H₂SO₄ 0%

--------------------------------------------------------
Cu⁺⁺

--------------------------------------------------------
Zn⁺⁺

--------------------------------------------------------
pH 0 _Acontium velatum_
_Thiobacillus thioodixans_

--------------------------------------------------------
Eh -450 mV Sulfate-reducing bacteria
at pH 9.5
--------------------------------------------------------

Table VI.—_Extremes of Chemical Environmental Factors
Permitting Growth or Activity_

----------------------------------------------------------------------
Chemical Maximum Pressure, Time, Organism Activity
factor atm days
----------------------------------------------------------------------
O₂ 100% 1 Plants, Growth
animals
----------------------------------------------------------------------
O₃ 100 ppm 5 _Armillaria Growth
(ozone) --------------------------- mellea_ -----------------
500 ppm 5 Light emission
----------------------------------------------------------------------
H₂ 100% Various Germination
plants
----------------------------------------------------------------------
H₂O Aw 1.0 1 Various Growth
aquatic
organisms

----------------------------------------------------------------------
H₂O₂ 0.34% Rye Germination
enhanced
----------------------------------------------------------------------
He 100% Wheat, rye, Germination
rice
----------------------------------------------------------------------
CO 100% Rye Germination
--------------------------------------------------------------
80% 1.1 4 _Hydrogenomonas_ Growth
----------------------------------------------------------------------
CO₂ 100% 1.1 4 Rye Growth and
germination
----------------------------------------------------------------------
CH₄ 100% 1.1 4 Rye Germination
----------------------------------------------------------------------
CH₂O 50% Rye Germination
----------------------------------------------------------------------
CH₃OH 50% Rye Germination
----------------------------------------------------------------------
N₂ 100% .1 10 Various plants Germination and
root growth
----------------------------------------------------------------------
NO 18% .018 10 Sorghum, rice Germination and
root growth
----------------------------------------------------------------------
NO₂ 18% .018 10 Rye, rice Germination and
root growth
----------------------------------------------------------------------
N₂O 100% 1.2 4 Rye Germination
--------------------------------------------------------------
96.5% 1.7 Rye Germination
------------------------------------
_Tenebrio Survival
molitor_
----------------------------------------------------------------------
Ar 100% 1.2 2 Rye Germination
----------------------------------------------------------------------
NaCl, 67% Photosynthetic Growth
Na₂SO₄, bacteria
NaHCO₃
----------------------------------------------------------------------
H₂S 0.96 _Desulfovibrio Growth
g/liter desulfuricans_
----------------------------------------------------------------------
H₂SO₄ 7% _Acontium Growth
velatum_
-------------------------------
Thiobacilli Growth,
reproduction
----------------------------------------------------------------------
Cu⁺⁺ 12 _Thiobacillus Growth
g/liter ferrooxidans_
----------------------------------------------------------------------
Zn⁺⁺ 17 _Thiobacillus Growth
g/liter ferrooxidans_
----------------------------------------------------------------------
pH 13 _Plectonema Growth
nostocorum_
-------------------------------
_Nitrobacter_ Growth
-------------------------------
_Nitrosomonas_ Growth
----------------------------------------------------------------------
Eh 850 mV Iron bacteria Growth
at pH 3
----------------------------------------------------------------------

chapter 4

_Behavioral Biology_

EFFECTS OF THE SPACE ENVIRONMENT ON BEHAVIOR

NASA was established in 1958, shortly after the Russian launching of the second Earth satellite Sputnik II, the first vehicle to carry life into orbit around the Earth. This accomplishment was preceded by the pioneering work of Henry et al. ([ref.77]), in which animals were exposed briefly to low-gravity states in Aerobee rockets. A motion-picture camera photographed the behavior of two white mice in rotating drums during this series of flights, which marked the first time that simple psychological tests were made on animals in the weightless condition. While this behavioral experiment was relatively simple, it provided the basic concepts for recent studies which involved rotation of animals during the weightless state. Subsequent flights such as Project MIA (Mouse-in-Able) reflected a preoccupation with physiologic measures (refs. [ref.78] and [ref.79]), although the flights of Baker and Able included preflight and postflight performance studies ([ref.80]). Able’s behavior was recorded in detail on in-flight film, but none of the behavior was programed or under experimental control.

The first flights in which behavior or performance was explicitly programed were those of Sam and Miss Sam in flights of the Little Joe rocket with the Mercury capsule, launched from Wallops Island in 1959 and 1960 ([ref.81]). The first major space achievement in the behavioral sciences was the successful in-flight measurement of the behavior of the chimpanzee Ham in early 1961, in which the pretrained animal performed throughout the flight. The second achievement along these lines was in 1962 when the chimpanzee Enos made several orbits around Earth and performed continuously on a complex behavioral task. The tasks which the animals performed during these flights have been described in detail by Belleville et al. ([ref.82]), and the results of the in-flight performance have been presented by Henry and Mosely ([ref.83]). These early flights provided much of the technological framework on which current biological experiments on organisms during flights of extended duration are based. Due largely to the efforts of Grunzke (refs. [ref.84] and [ref.85]), the apparatus needed to sustain animals during space flight, such as zero-g watering and feeding devices, are now commonplace ([ref.86]). Advanced systems of programing stimulus presentations and recording responses, developed for Project Mercury, may now be seen in many basic research laboratories throughout the country.

Several other noteworthy advances have been made as an outgrowth of the Mercury animal flights. Immediately before the orbital flight MA-5, in which the chimpanzee Enos was employed, it was unexpectedly found that this 5-year-old animal was hypertensive. Subsequent centrifuge studies showed that its vascular responses exceeded those of a control group. Consideration of the animal’s preflight experience led to speculation concerning the origin of this hypertension. An explanation of the high-blood-pressure responses detected in Enos has been pursued by Meehan et al. ([ref.87]). Persistent hypertension has been produced in other laboratory chimpanzees restrained in the same manner as those participating in space flight and exposed to demanding performance tasks, a demonstration which has important implications for prolonged manned space flight and for cardiovascular medicine in general.

Studies more directly concerned with behavior and performance have been extended from those of Project Mercury. These extensions have been in the following directions: (1) the establishment and maintenance of complex behavioral repertoires under conditions of full environmental control, (2) the refinement of behavioral techniques for assessing sensory and motor processes, and (3) the maintenance of sustained performance under conditions of long-term isolation and confinement and preliminary extension of such experimental analysis to man.

Numerous studies with primate subjects, including several at Ames Research Center, have been devoted to developing methods for maintaining optimum performance in environments with limited sources of stimulation. Monkeys, baboons, and chimpanzees, for example, have been isolated for periods of longer than 2 years with no decrement in performance on complicated behavioral tasks ([ref.88]). The behavioral techniques used in these studies are closely related to those employed on human subjects under NASA sponsorship at the University of Maryland ([ref.89]). The essence of these techniques is in the proper programing of environmental stimuli ([ref.90]). It is not sufficient to provide the subject with his physiological requirements for survival, but he must be given the psychological motivation for using these provisions. This statement, of course, is an oversimplification of the problem, but it serves to illustrate the essence of these experimental programs.

Gravity has long been known as one of the major factors influencing various life processes and the orientation of both plants and animals. One of the most challenging problems of space research has been to define this influence more precisely. Related to the effect of gravity on living processes is the problem of the effects of weightlessness. Of particular interest to psychologists are the possible modifications an altered gravitational environment might produce in behavioral patterns basic to the animal’s maintenance and survival, such as eating, sensory and discriminative processes, development and maturation, and learning capacity ([ref.91]).

One prominent method of studying gravitational effects is to simulate an increase in gravity by centrifugation. Smith et al. ([ref.92]) and Winget et al. ([ref.93]) have investigated the effects of long-term acceleration on birds, primarily chickens, while Wunder (refs. [ref.94] and [ref.95]) and his coworkers (refs. [ref.96]-[ref.99]) have used fruit flies, mice, rats, hamsters, and turtles. The general findings are that, when animals are subjected to a prolonged period of acceleration of moderate intensity, they exhibit decreased growth, delayed maturation, and an increase in the size of certain muscles and organs, dependent on the species. With regard to the decreased growth effect, the data of these investigators show some exceptions. When the gravitational increase is kept below a certain limit, growth was greater than that of controls in the fruit fly, turtle, mouse, and chicken. The limit below which enhancement of growth was observed varied with the species studied.

The data on food intake do not present a consistent picture. Wunder ([ref.94]) found that food intake in accelerated mice was markedly reduced from that of nonaccelerated control animals. Smith, however, found that in chickens, food intake increased up to 36 percent over controls and has derived an exponential relation between food intake and acceleration. After six generations of selective breeding, Smith has produced a strain of chickens better adapted to prolonged exposure to high g.

A very relevant finding of their research with birds was that exposure to chronic acceleration in some way appears to interfere with habituation to rotatory stimulation. Chickens who were being subjected to chronic acceleration were given repeated rotatory stimulation tests to estimate their labyrinthine sensitivity. This study revealed that centrifuged animals showed a marked reduction in labyrinthine sensitivity. This result appeared to persist after the acceleration was terminated. In animals who developed gait or postural difficulties as a result of acceleration, there was no evidence of a postnystagmus in response to the rotatory stimulation test, which the investigators point out may be evidence of a lesion in the labyrinth or its neural pathways.

Smith has implicated social factors as interfering with acceleration effects. His subjects were typically accelerated four or six to a cage. When groups were mixed midway through the experiment, they exhibited a higher mortality rate and incidence of acceleration symptoms than did groups whose constituency remained unchanged.

At the U.S. Naval School of Aerospace Medicine, numerous studies have been conducted on the effects of slow rotation on the behavior and physiology of humans and animals ([ref.100]). Rotation initially produces decrements in performance, but adaptation to a rotating environment ensues quite rapidly (refs. [ref.101]-[ref.103]). Perceptual distortion, nystagmus, nausea, and other signs of discomfort are common responses to slow rotation. These symptoms are generally reduced with continued exposure (adaptation). Interestingly, however, adaptation is delayed when the subjects are exposed to a fixed reference outside their rotating environment.

At NASA-Ames, rodents have been used in experiments by Weissman and Seldeen to delimit the stimulus effects of rotation. In these experiments the subjects must discriminate between different speeds of rotation in order to obtain food reinforcement. The results thus far provide evidence that these animals are capable of discriminating between the different speeds at which they are being rotated. The range of speeds studied was 0-25 rpm, with tests of discrimination being made at intervals of less than 5 rpm. Experiments such as these will lead to the development of techniques for measuring rotational sensitivity in many species, including man.

The optimum configuration of manned spacecraft will depend, in part, upon biomedical considerations. A voluminous literature now exists on the possible hazards to man of prolonged exposure to zero-g conditions. Should prolonged weightlessness prove to be a serious detriment to health, consideration must be given to design concepts which provide artificial gravity.

No data exist on the minimum gravity requirements necessary to sustain basic biological functions for extended periods. A limit of 0.2 g has been given as the lower level at which man can walk unaided ([ref.104]). It has also been recommended that angular velocity be maintained at the lowest possible level in order to minimize the occurrence of vestibular disturbances. These recommendations are based on human-factor requirements, rather than upon biological considerations, which may significantly modify these values. In recent studies, a technique has been devised which promises to provide reliable criteria for biological acceptability, since it is based on fundamental biological and behavioral principles.

As animals progress up the evolutionary stale, their survival depends less and less upon stereotyped physiological reactions which occur in reflex fashion, in response to environmental stimulation. In higher organisms, survival depends more upon the capacity of organisms to modify their behavior. At the highest levels of functional efficiency, the ultimate form of adaptation is seen—the manipulation of the environment by the organism. Developments in behavioral science now permit us to utilize the adaptive behavior of animals to investigate many problems of biological interest. Recent studies on the self-selection of gravity levels represent a further attempt to exploit the adaptive capacities of animals, in order to provide information relevant to problems of space exploration.

One such project allows animals to select their own gravity environment in an apparatus designed to create g-forces through centrifugal action by rotation at 60 rpm ([ref.105]). The surface of this centrifuge is parabolic, so that the resultant of the centrifugal g and the Earth’s gravity is always normal to the surface. When the animal moves away from the center, increasing the radius of rotation, it is exposed to increasing gravity. Motion toward the center reduces the gravity level. By this means, an animal is free to select its own gravity environment.

When the animal moves toward or away from the center, he is moving from one tangential velocity to another. He is therefore acted upon by a third force—due to Coriolis acceleration. The effects of Coriolis forces are a major problem difficult to eliminate in studies such as these, but they must be taken into account in the design of spacecraft which produce artificial gravity by rotation. Motion of the head in any direction not parallel to the centrifugal force vector would result in bizarre stimulation of the semicircular canals and consequent motion sickness. This effect is likely to become even more pronounced if the sensitivity of these organs is increased by prolonged exposure to reduced gravity. Methods such as these are currently being developed for conducting a refined psychophysical analysis of gravity, including studies by Lange and Broderson on the perception of angular, linear, and Coriolis acceleration.

The results of animal studies such as these will be of great value in arriving at a decisive judgment concerning the need for artificial gravity in a manned orbiting space station, or other vehicles designed for long-term occupancy.

To aid in the interpretation of in-flight data, other studies are underway to determine the functions of the vestibular system, as a principal brain center related to orientation in space and to the physiology of posture and movement, as well as with the influences of acceleration, rotation, and weightlessness. Experiments are presently being conducted on monkeys and cats in order to trace these complex neurological connections and to determine their functional organization.

BIOLOGICAL INFORMATION SYSTEMS

The nature of memory has been the subject of considerable speculation in the past. It has long been felt intuitively that retention of information in the central nervous system involves either an alteration of preexisting material or structure, or, alternatively, synthesis of materials not present previously. The cellular site of operational alteration was unknown but, again intuitively, was felt to be closely associated with the synapses. The problems faced by early investigators were great; but nevertheless much information relevant to the question of biological information storage was obtained. With the relatively recent advent of more refined tools and methodologies, there has been rapid progress.

A significant amount of the work which has been conducted in the area of biological information and communication systems is easily classified as "basic research" (refs. [ref.106]-[ref.109]). This discussion will be limited to those aspects closely related to the fields of molecular biology and experimental psychology, which seem to have universal application to all known animal life forms. Studies involving the basic principles of acquisition, processing, storage, and retrieval of information in living systems are emphasized.

Early Work

Early speculations on the operational nature of memory have been based upon relatively little experimental evidence. Charles Darwin observed that domestic rabbits had smaller brains than their wild counterparts, and attributed this to lack of exercise of their intellect, senses, and voluntary movements. Unfortunately, subsequent studies of the brains of men with greatly differing intellectual capability did not substantiate the hypothesis. Idiots sometimes had larger brains than geniuses. Later, an idea proposed by Ramon y Cajal came into favor. Since brain cells did not increase in number after birth, he proposed that memory involved the establishment of new and more extended intercortical connections. Unfortunately, methods were not available to test this hypothesis adequately and it has remained until quite recently in the realm of conjecture.