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Introduction to JavaScript Programming Test Bank Chapter 6
with XML and PHP
Test Bank for Chapter 6
MULTIPLE CHOICE
1. Where are <form></form> tags placed?
a. Directly after the opening <body> tag b. Anywhere in the HTML document
and must be closed right before the
closing </body> tag
c. Anywhere in the body of a web page d. In the <head> section
ANS: C
2. Buttons that can be automatically created using the type attribute are:
a. submit b. reset
c. hidden d. (a) and (b) only e. (a), (b), and (c)
ANS: E
3. Which of the following are ways to submit form data?
a. By email b. to a JavaScript program
c. to a server d. (b) and (c) only e. (a), (b), and (c)
ANS: E
4. When using a set of radio buttons, which attribute must be the same for all buttons in the set
a. name b. value
c. id d. selected
ANS: A
5. Which line of code should be used to make the following code snippet work?
var longString = "Great day, isn't it?";
var shortString = _____???_______
document.write("It's going ton rain to" + shortString);
a. shortString = longString.substr(20, 7);
b. shortString = longString.substr(6, 3);
c. shortString = substr(longString, 6, 3);
d. shortString = longString.substr(7, 3);
ANS: B
© 2014 Pearson Education 1
Introduction to JavaScript Programming Test Bank Chapter 6
with XML and PHP
6. Which of the following will send form results from a form named “importantInfo” to the email
address [email protected] with the subject “Read this!”
a. <form name=“info” method=“post” id=“important info” action =
[email protected]?subject = Read this!” enctype=“plain/text”>
b. <form name=“importantInfo” method=“post” id=“info” action=
“mailto:[email protected]?subject=Read this!” enctype=“plain/text”>
c. <form name=“importantInfo” method=“email” id=“info” action =
mailto:[email protected] “subject=Read this!” enctype=“plain/text”>
d. <form name=“importantInfo” method=post id=info action =
“mailto:[email protected] ? subject=Read this!” enctype=text/plain>
ANS: B
7. Which line of code will check if any character in the string variable pword is the letter X and
return true for the variable check?
var check = false;
for (i = 0; i < pword.length; i++)
{
_____???_______
check = true;
}
a. if (var charX == "X");
b. if (var charX == charX.charAt[i]);
c. if (var charX.charAt[i] == "X");
d. if (var charX.charCodeAt[i] == "X");
ANS: C
8. Given the following line of code, what does the this keyword refer to?
<input type = "button" name = "murgatroyd" id = "Mortimer"
onclick = "doSomething(this.id)" />
© 2014 Pearson Education 2
Introduction to JavaScript Programming Test Bank Chapter 6
with XML and PHP
a. the button type b. the name attribute
c. the id attribute d. the onclick() event
ANS: C
9. Which of the following will check to see if a password contains a # sign, given that the
character code for "#" is 37? The password is 8 characters long and is stored in a variable named
pword.
a. var check = false;
for (j = 1; j < 7; j++)
{
if (pword.charCodeAt[j] == 37);
check = true;
}
b. var check = false;
for (j = 0; j < 8; j++)
{
if (pword.charCodeAt(j) == 37)
check = true;
c. var check = false;
for (j = 0; j <= 8; j++)
{
if (pword.charCodeAt()== 37)
check = true;
}
d. var check = true;
for (j = 1; j < 9; j++)
{
if (pword.charCodeAt(37)== "#")
check = true;
}
ANS: B
10. Which of the following sets or changes the tab order of form controls?
a. tab b. tab = 0; c. tab index d. index
ANS: C
© 2014 Pearson Education 3
Introduction to JavaScript Programming Test Bank Chapter 6
with XML and PHP
11. Which of the following will substitute an image named redButton.jpg that is stored in the same
place as the web page for a generic button? The doStuff() function is called when the button is
clicked.
a. button type="image" src=redButton.jpg onclick="doStuff()" /></button>
b. <a href="Javascript:doStuff()"> <img src = "redButton.jpg"> </a>
c. <button type="image" onclick="doStuff()"><img src="redButton.jpg" />
d. <a href=:Javascript:doStuff()" src ="redButton.jpg" type="button" />
ANS: B
12. Which of the following will call a function named setBlue() when a text box with id =
"blue" gets the focus?
a. onfocus = "blue" onclick = ("setBlue()");
b. onfocus.blue = setBlue()
c. id = "blue" onclick = setBlue()
d. onfocus = "setBlue('blue')"
ANS: D
13. Which of the following is the correct way to set a background color of blue to an HTML element with
id = "color_change"?
a. document.getElementById("color_change").style.background = "blue";
b. document.getElementById("color_change").innerHTML = "blue";
c. document.getElementById("color_change").style = blue(this.id);
d. document.getElementById("color_change").this.id = background("blue");
ANS: A
14. Which of the following checks if the sixth character of a string variable named myMail is the @ sign
using a Boolean variable named atSign set to true if this is true?
a. for (k = 0; k < myMail.length; k++)
{
if(myMail.charAt(k) == @)
atSign = true;
}
b. atSign = "@";
for (k = 0; k < 7; k++)
{
if(myMail.charAt(k) == atSign)
flag = true;
}
c. if(myMail.substr(5, 1) == "@")
atSign = true;
d. if(myMail.substr(1, 5) == "@")
© 2014 Pearson Education 4
Introduction to JavaScript Programming Test Bank Chapter 6
with XML and PHP
atSign = true;
ANS: C
15. Which of the following checks to see if the number of characters in a given string named myName is
greater than 2 and less than 11?
a. if(myName.length > 2 && myName.length < 11)
b. if(myName.length > 2 || myName.length < 11)
c. if(myName.length > 1 && myName.length <= 10)
d. if(myName.length >= 2 || myName.length <= 11)
ANS: A
TRUE/FALSE
1. True/False: Radio buttons are used to allow users to select several options at one time.
ANS: F
2. True/False: A form using the <form></form> tag pair can be placed anywhere within a web page.
ANS: T
3. True/False: When a form is enhanced with JavaScript, an event handler must be used to evoke the
JavaScript code.
ANS: T
4. True/False: The Common Gateway Interface (CGI) allows web pages to be generated as executable
files.
ANS: T
5. True/False: CGI scripts are normally saved in a folder named cgi-bin that exists on every client's
hard drive.
ANS: F
6. True/False: The property of each radio button in a group of radio buttons that must be the same for
each button is the id property.
ANS: F
7. True/False: The checked property can be used to return the state of a checkbox to a JavaScript
function.
ANS: T
8. True/False: The properties that determine the size of a text box are cols and rows.
ANS: F
© 2014 Pearson Education 5
Introduction to JavaScript Programming Test Bank Chapter 6
with XML and PHP
9. True/False: If the information entered into a textarea box exceeds the number of rows originally set,
a scroll bar is created.
ANS: T
10. True/False: The two types of buttons that display masked text (such as *'s or #'s) to hide what a user
enters are "hidden" and "password".
ANS: F
© 2014 Pearson Education 6
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with Unrelated Content
number the bacterial population of the Earth. This potential could
result not only in competition with any Martian life, but in drastic
changes in the geochemical and atmospheric characteristics of the
planet. To avoid such a disaster, certainly the first, and probably
many succeeding landers on Mars, must be sterile—devoid of
terrestrial life ([ref.55]). Since the space environment will not in itself
kill all life aboard, the lander must leave the Earth in a sterile
condition.
The sterility of an object implies the complete absence of life. The
presence of life or the lack of sterility may be proven; but the
absence of life or sterility cannot be proven, for the one viable
organism that negates sterility may remain undetected. Many
industrial products which must be guaranteed as sterile cannot be
tested for sterility in a nondestructive manner. A similar situation
exists in determining the sterility of a spacecraft. Certification of
sterility—based on experience with the sterilizing process used,
knowledge of the kinetics of the death of micro-organisms, and
computation of the probability of a survivor from assays for sterility
—is the only accurate approach to defining the sterility of such
treated items.
Macroscopic life can be readily detected and kept from or removed
from the spacecraft, but the detection and removal of microscopic
and submicroscopic life is an extremely difficult task. The destruction
of micro-organisms can be achieved by various chemical and
physical procedures. Sterilizing agents have been evaluated not only
for their ability to kill microbial life on surfaces and sealed inside
components, but also for the agents' effects on spacecraft reliability
as well (refs. [ref.56]-[ref.59]). Of the available agents, only heat
and radiation will penetrate solid materials. Radiation is expensive,
hazardous, difficult to control, and apparently damages more
materials than does heat. Heat, therefore, has been selected as the
primary method of spacecraft sterilization and will be used, except in
specific instances where radiation may prove to be less detrimental
to the reliability of critical parts ([ref.60]).
The sterilization of spacecraft is a difficult problem if flight reliability
is not to be impaired. The development of heat-resistant parts will
enable the design and manufacture of a heat-sterilizable spacecraft.
Without careful microbiological monitoring of manufacture and
assembly procedures, many bacteria could be trapped in parts and
subassemblies. To permit sterilization at the lowest temperature-time
regimen that will insure kill of all organisms, the microbiological load
inside all parts and subassemblies must be held to a minimum.
The role of industrial clean rooms in reducing the biological load on
spacecraft is currently being defined. NASA-supported studies
indicate that biological contamination in industrial clean rooms for
extended time periods is about 1 logarithm less (tenfold reduction),
compared with conditions in a well-operated microbiological
laboratory ([ref.61]). With the use of clean-room techniques and
periodic decontamination by low heat cycles or ethylene oxide
treatment, it should be possible to bring a spacecraft to the point of
sterilization with about 106 organisms on board ([ref.60]).
The sterilization goal established for Mars landers is a probability of
less than 1 in 10 000 (10-4) that a single viable organism will be
present on the spacecraft. Laboratory studies of the kinetics of dry-
heat kill of resistant organisms show that at 135° C the number of
bacterial spores can be reduced 1 logarithm (90 percent) for every 2
hours of exposure (refs. [ref.58] and [ref.62]). The reduction in
microbial count needed is the logarithm of the maximum number on
the spacecraft (106) plus the logarithm of the reciprocal of the
probability of a survivor (104), or a total of 10 logarithms of
reduction in microbial count. Thus, with an additional 2 logarithms
added as a safety factor, a total of 12 logarithms of reduction in
count has been accepted as a safe value which can be achieved by a
dry-heat treatment of 135° C for 24 hours. This is the heat cycle that
is currently under study and being developed for use in spacecraft
sterilization ([ref.60]). However, other heat treatments at
temperatures as low as 105° C for periods of 300 hours or longer
are under study ([ref.63]).
Based on results to date, it is reasonable to believe that a full
complement of heat-sterilizable hardware will be available when
needed for planetary exploration. Every effort is being made to
improve the state of the art to a point where spacecraft can not only
withstand sterilization temperatures, but will be even more reliable
than the present state-of-the-art hardware that is not heated.
chapter 3
Environmental Biology
BIOLOGICAL EFFECTS OF WEIGHTLESSNESS AND ZERO GRAVITY
H
igh priority has been given to studies of weightlessness.
Gravity is one of the most fundamental forces
that acts on living organisms, and all life on Earth except the smallest appears to be oriented with
respect to gravity, although certain organisms are more responsive to it than others. The gravity force
on Earth is 1 g, but this force may be experimentally varied from zero g, or weightlessness, to many
thousands of g's.
Zero gravity or decreased gravity occurs during freefall, in parabolic trajectory, or during orbit around
the Earth. Gravitational force decreases by the square of the distance away from the Earth's center. It is
reduced about 5 percent at about 200 nautical miles' altitude. Gravitational force greater than 1 g can
be obtained by acceleration, deceleration, or impact. It also can be increased by using a centrifuge
which adds a radial acceleration vector to the 1 g of Earth.
On the ground, the biological effects of gravity have been studied at 1 g, and experimentally, forces of
many g have been produced. In addition, modifications of the effects of the 1-g force have been
induced by suspension of the organism in water or by horizontal immobilization of an erect animal such
as man. The biological effects of such modification have been of significant value in understanding some
of the possible consequences of human exposure to the zero-g environment of space.
Weightlessness in an Earth-orbiting satellite occurs when the continuous acceleration of Earth's gravity
is exactly counterbalanced by the continuous radial acceleration of the satellite. In such a weightless
state, organisms are liberated from their natural and continuous exertion against 1 g, but this liberation
may carry with it certain serious physical penalties.
Some of the physical processes which probably have the greatest biological effects are (1) convective
flow of fluid, e.g., protoplasmic streaming, transport of nutrient materials, oxygen, waste products, and
CO2 from the immediate environment of the cell, and (2) sedimentation occurring within cells;
substances of higher density sediment in a gravitational field, and those of lighter density rise. A
separation of particles of different densities probably occurs. The removal of gravity would change a
distribution of particles like mitochondria by 10 percent ([ref.64]).
Gravity has effects on the physical processes involved in mitosis and meiosis. Study under
weightlessness might contribute to our understanding of the general cellular information-relay process.
A gravitational effect is known in the embryonic development of the frog Rana sylvatica. After
fertilization, the eggs rotate in the gravitational field so that the black animal hemisphere is uppermost.
Development becomes abnormal if this position is disturbed. If the egg is inverted following the first
cleavage and held in this position, two abnormal animals result, united like Siamese twins. This
phenomenon appears to be related to the gravitational separation of low- and high-density components
of the egg. The size of the egg is about 1 to 2 mm and is suspended in water of about the same
density. This system is very sensitive to gravity; and, under weightlessness, the separation of different
density components might be irregular, leading to aberrant development. When certain aquatic insect
eggs are inverted, subsequent development results in shortened abnormal larvae.
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
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) Env
Atmospheric
Survival, Temperature, N2
Species Moisture pressure,
months °C perce
mm Hg
85, 25±15,
Conditions on Mars: 14µ±7µ -70 to +30 3 to
11
Anaerobic sporeformers
Low,
Clostridia, Bacillus 6 -60 to +20 76 95
(CaSO4)
planosarcina
Atmospheric
Survival, Temperature, N2
Species Moisture pressure,
months °C perce
mm Hg
Anaerobic
nonsporeformers Low,
6 -60 to +20 76 95
Pseudomonas, (CaSO4)
Rhodopseudomonas
Anaerobes
Aerobacter
Growth Very wet -75 to +25 760 100
aerogenes,
Pseudomonas sp.
Clostridium,
1 percent
Corynebacteria 10 -25 to +25 65 100
or less
"Thin short rod"
0.5
Bacillus cereus 2 percent -25 to +25 65 94
soil
1 8.4
Clostridium sporogenes -25 to +25 65 94
(growth) percent
Clostridium botulinum 10 Lyophilized -25 to +25 65 95
Klebsiella pneumoniae 6 Lyophilized -25 to +25 65 95
Bacillus subtilis var.
4 2 percent -25 to +25 85 95
globigii
0.5
Sarcina aurantiaca 4 -25 to +25 85 95
percent
Clostridium tetani 2 or less 1 percent -60 to +25 85 95
Aspergillus niger Over 6 hr Very dry -60 to +25 76 95.
Aspergillus oryzae Over 6 hr Very dry -60 to +25 76 95.
Mucor plumbeus Over 6 hr Very dry -60 to +25 76 95.
Atmospheric
Survival, Temperature, N2
Species Moisture pressure,
months °C perce
mm Hg
Rhodotorula rubra Over 6 hr Very dry -60 to +25 76 95.
Pea, bean, tomato, rye,
0.3 Moist +25 75 100
sorghum, rice.
Winter rye 0.6 Moist -10 to +23 76 98
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 gauss) Human
Gravity 0g 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 desulfuricans Grows and reduces sulfate
atm)
Magnetism 167 000 gauss Neurospora 1 hr—no effect, Arbacia
Arbacia development delayed
Drosophila
Gravity 400 000 g Ascaris eggs 1 hr—eggs hatch, 40 days'
growth
110 000 g Escherichia coli
Pressure 1400 atm Marine organisms Growth
Microwave 2450 Mc/sec 0.3 Drosophila 68 hr, growth not affected
to 1 W/cm2
Visible 50 000 ft-c Chlorella, Seconds, recurrently
17 000 ft-c higher plants 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 6500 Man Threshold of pain
2
dyne/cm at 0.02
to 4.8 kcs/sec
Table IV.—Extreme Low and High Temperature Effects Permitting Life Processes
Minimum
temperature, Organism Activity or condition
°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 glaucus Growth (in glycerol)
-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
temperature, Organism Activity or condition
°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 stearothermophilus Cultured in laboratory
83 Sulfate-reducing bacteria Found in a well
89 Sulfate-reducing a bacteria Found in oil waters
65 to 85 Sulfate-reducing a bacteria Cultured in laboratory
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 desulfuricans Grow and reduce sulfate at 1000 atm
Table V.—Extreme Temperature Limits of Survival
Minimum
temperature Organism
°C
-190 Yeast bacteria, 10 species
-197 Trebouxia erici from lichens
-197 Protozoa, Anguillula
Minimum
temperature Organism
°C
-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
temperature Organism Time of exposure
°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. actinomycetes 5 days at 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
Maximum
temperature Organism Time of exposure
°C
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%
H2 O Aw 0.48 Pleurococcus vulgaris
Aw 0.5 Xenopsylla cheopis (prepupae)
H 2 O2 0%
He 0%
CO 0%
CO2 0%
CH4 0%
CH2O 0%
CH3OH 0%
Chemical Minimum Organism
factor
N2 0%
NO 0%
NO2 0%
N2 O 0%
Ar 0%
NaCl,
Na2SO4,
NaHCO3
H2 S 0%
H2SO4 0%
Cu++
Zn++
pH 0 Acontium velatum
Thiobacillus thioodixans
Eh -450 mV at Sulfate-reducing bacteria
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, animals Growth
O3 100 ppm 5 Armillaria mellea Growth
(ozone)
500 ppm 5 Light emission
H2 100% Various plants Germination
Chemical Maximum Pressure, Time, Organism Activity
factor atm days
H2 O Aw 1.0 1 Various aquatic Growth
organisms
H 2 O2 0.34% Rye Germination
enhanced
He 100% Wheat, rye, rice Germination
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
N2 O 100% 1.2 4 Rye Germination
96.5% 1.7 Rye Germination
Tenebrio molitor Survival
Ar 100% 1.2 2 Rye Germination
NaCl, 67% Photosynthetic Growth
Na2SO4, bacteria
NaHCO3
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