Acta Astronautica Vol. 33, pp.

167-177, 1994
Copyright © 1994 Elsevier Science Ltd
Pergamon 0094-5765(94)00107-3 Printed in Great Britain. All rights reserved
0094-5765/94 $7.00+0.00

C.E.B.A.S.-AQUARACK PROJECT: THE MINI-MODULE AS

TOOL IN ARTIFICIAL ECOSYSTEM RESEARCH

V. Blfim (M3), E. Stretzke and K. Kreuzberg* (CM3)

C.E.B.A.S. Center of Excellence, Faculty of Biology, Ruhr-University Bochum, FRG

and *German Aerospace Establishment,Ececntive Department, Cologne Porz, FRG

ABSTRACT

The evolution of the C.E.B.A.S-AQUARACK project including results of the scientific frame program
was frequently presented at the IAA Man in Space Symposia 1989 and 1991 and the IAF/IAA congres-
ses since 1990. C.E.B.A.S. (Closed Equilibrated Biological Aquatic System) is a combined
animal/plant system for long-term multi-generation experiments with aquatic organisms in ground la-
boratories and in a space station. For short-term missions a miniaturized version was developed which
fits into a spaceiab middeck locker together with all surrounding equipment. The latest development is
an optimized prototype with a total volume of about 11 liters which consists of a main animal tank
(Zoological Component) with integrated bacteria filter, a semibiological coarse filter, an illuminated
higher plant container (Botanical Component) and combined small animal and electrode compartment.
A silastic tubing gas exchanger in a closed side-loop serves as an emergency unit in case of the mal-
function of the Botanical Component and the water is driven through the system by rotatory pumps. It
is operative for several weeks in closed state. This C.E.B.A.S. Mini-Module also represents an aquatic
artificial ecosystem in which basic scientific problems of component interactions and system theory can
be solved with the side aspects of combined production of animal and plant food in bioregenerative life
support systems. The paper presents details of the current statuts of the hardware development and data
about the function of the fully biological life support of the system, e. g. mid-term registrations of
water parameters. Moreover, morphological and pysiological data of the experimental animals (-the te-
leost fish Xiphophorus hel/erJ-) and plants (-a tropical CeratophyUum species-) demonstrate the biologi-
cal stability of the system. These are used to elaborate first details of population interactions and inter-
dependencies as a basis of a disposed comprehensive system analysis which is the essential pre-
condition for the design of possible aquatic modules for bioregenerative life support systems.

INTRODUCTION

There is a certain tradition of the utilization of aquatic animah in German space biology. The first suc-
cessful experiment was the BIOSONDE containing a leecha. It was followed by the STATEX system
in the Spacelab-Dl mission 2 where the behaviour and the morphology of the vestibularis system in
clawed toad tadpoles was studied and an improved version of this was flown with fish fry and tadpoles
in the Spacelab-D2 mission in 1993. The STATEX life support system is simple and does not involve a
water recycling component so that its time of utilization is limited. The scientific demand for long-term
space experiments with aquatic organi.~ms lasting over several subsequent generations3 led to the deve-
lopment of a multi-purpose "space aquarium" for aquatic animals including a highly effective water re-
cycling system which then was combined with a module for aquatic plants. This basic scheme was
further improved and resulted in the development of a so-called "Closed Equilibrated Aquatic System"
(C.E.B.A.S.) which was paralleled by the formation of a wide-spread and multidisciplinary internation-
al zoological research program covering all space-relevant areas like genetics, embryology/teratology,
skeleton research, reproductive biology and vestibular research, e. g.. The common experimental ani-
mals are the ovoviviparous teleost fish X/phophorus hel/er/and the pulmonate water snail B/ompha/ar/a
glabrata. The development of C.E.B.A.S. led until today to two subsequent laboratory prototypes
which were all successfully tested4 - 7. Currently the last version (HI) leading to the spaceflight hard-
ware is under construction. The C.E.B.A.S.-system still is disposed as a a national German option for
the COLUMBUS space station.

167
 168 lOth IAA Man in Space Symposium

The original C.E.B.A.S. consists of 3 main components. The first is the animal tank, the Zoological
Component, with a water recycling system consisting of a semibiological coarse filter (for physical fil-
tering and degradation of oragnic material), a bacteria filter (converting NH + excreted by the fishes
. . . . 4. ° o

into NO, and N O by Nitrobacter and Nitrosomonas bacteria), a nitrogen compound elmunator, a gas
exchanger (silastic~tubing spiral or array), a heat/cold exch_an~erand an UV-sterilizer. The second is
the Botanical Component. It was primarily disposed to be a microalgal bioreactor containing the photo-
heterotrophous and motile species Chlamydomonas reinhardtii. The construction of this device was de-
layed by several reasons and it was substituted by a higher plant growth chamber. A tropical
CeratophyUum species turned out to he very effective for the elimination of NO -ions from the water
so that the Botanical Component is able to take over the tasks of the nitrogen compound eliminator.
Moreover, in a properly sized and illuminated plant growth chamber the plants produce enough oxygen
to fulfil the demands of the animals and micoorganisms in the filter subcomponents. So, the gas ex-
changer of the water recycling system may be placed into a side loop of the system which is only open
in cases of emergency . The third, the Electronical Component, controls, monitors and stabilizes the
biological equilibrium between the Zoological and the Botanical Component. In the initial concept it re-
presents a computerized process control system of high complexity. In the normal operation of
C.E.B.A.S. the system, however, this is stable enough that the latter only serves as a data acquisition.
In emergency cases it activates a tele-alert to the person in duty and compensates within certain narrow
limits system disturbances. In practical operation, however, the function of the Electronical Component
is more equilibrating than stabilizing.

HARDWARE DESIGN OF THE C.E.B.A.S. MINI MODULES AND METHODOLOGICAL TEST

CONCEPT

The development of C.E.B.A.S. includes 3 time levels: the ground laboratory phase, the space test
phase and the scientific space phase. From an economical and practical point of view a hardware test of
the original system in space is not very senseful and therefore it was decided to develop a miniaturized
version of the system for the utilization in a spacelab middeck locker or a biosattelite. The basic
disposition was to scale down the volume of the original C.E.B.A.S. system (90 to 150 liters in the
different versions) to about 10 liters but to maintain all components and s u b c o ~ t s . Because the
C.E.B.A.S. MINI MODULE is dedicated only to short and mid-term missions the water recycling
system has not to be redundant in this case.

Two different basic designs of the C.E.B.A.S. MINI MODULE are existing. The first is the modular
one including separate subcomponents which are easily detachable with the exception of one filter com-
ponent integrated into the animal tank and which may be used as bacteria filter or coarse filter. This
type is dedicated to laboratory experiments where manipulation of subcomponent is essential. The
second design is the monolithic one. In this type all subcomponents are integrated into a single block.
It represents the design of the flight hardware which fits into a middeck locker tray together with video
equipment, control systems and data acquisition. Currently this MINI MODULE exists only on paper
but it went into industrial construction as real flight hardware in July 1993.

In contrast to the latter the laboratory protopyes are operative and exist in 3 versions. The first has an
animal tank of 6.7 liters volume and the integrated filter has 1.33 liters. The external components are
cylindrical. Two chambers with 0.95 liters volume each serve as filters (one of them may be used as a
second animal chamber or for measuring purposes). The plant chamber has a volume of 1.45 liters and
contains a centered 4 W-fluorescent lamp. The gas exchanger is a spiral of silastic tubing attached to
the plant chamber in a side loop. The water is driven by a low-power rotatory pump. Together with
with all auxiliary equitnnent this results in a total volume of about 11 liters. The material of all
subcomponents is PLEXIGLASR. The animRi tank is illuminated by a second 4 W-fluorescent lamp.
This system, "version I", was tested with 4 adult X/phophorus he//en and Cerat~t~y//um spec.
successfully for 3 weeks already in early 1991 and the results were published elsewhere ".

The device described above device brought nearly optimal results in the laboratory test but it had to be
redesigned by reasons of energy saving. This resulted in the C.E.B.A.S. MINI MODULE "version II"
which has the same components and dimensions as version I with the exception of the plant chamber.
To eliminate the separate illumination of the Aninu13chamber the plant chamber is constructed as a flat
rectangular block with a volume of 0.95 liters. It is positioned in parallel with the animal tank and both
are illuminated by a 4 W-fluorescent lamp in between. The amount of light is high enough for the sup-
ply of the latter. So 4 W of energy are saved. In this connection it has to be pointed out, that the
photoperiod and light intensity is not only of high importance for the photosynthesis of the plants but
 10th IAA Man in Space Symposium 169

= ball valve 11

1o 0

20

7
14~

000 t--13

18 17 16 15
\j/ l/~

Figure 1: Construction principle of the C.E.B.A.S. MINI M O D U L E version III: 1 = animal tank and
bacteria filter, 2 = lamps, 3 = plant chamber, 4 = coarse filter, 5 = fine grain filter/ani-
mal/measuring chamber, 6 = flowmeter, 7 = rotatory pumps, 8 = electrode chamber, 9 =
water sample collecting module, 10 = oxygen electrode, 11 = emergency gas exchanger, 12
= thermocryostate, 13 = heat exchanger, 14 = coolant loop, 15 = luxmeter plate, 16 =
standard measuring plate, 17 = stainless steel grid, 18 = neutral gray filter foils, 19 = light
shielding, 20 = sample ports.
170 lOth IAA Man in Space Symposium

also for the fish reproduction. Light is an essential external stimulus for the maintenace of the function
of the reproductive system of teleosts. In addition, a certain light intensity (min. 800 Ix at the water
surface) is essential and own experiments showed that also the spectral composition is of high impor-
lance 9. The initial construction principle of the C.E.B.A.S. MINI MODULE was further optimized
and adapted to the special laboratory conditions in 3 further steps. The system alterations are not very
spectacular and involve the implementation of a simple temperature control (the hot summer 1992 over-
rode the laboratory air condition) and some redesigned tube connections and valves for easier handling.
This "version II" of the C.E.B.A.S.-MINI Module was also successfully tested and was biologically
stable for more than 7 weeks in 1992. The results of this experiment were also already published else-
where 1°. The next target, however, was a test run for more than 8 weeks. Therefore further modifica-
tions of the system had to implemented resulting in the "version HI". The "common illumination
principle" for animal and plant containers was revised and separate lamps for each were installed be-
cause the latter had to be set to permanent illumination to maintain an increaseded oxygen production.
Resulting from this the animal tank had to be optically separated from the plant illuminantion and the
intensity of the latter bacame variable by removable neutral gray foils and a stainless steel grid. The
plant chamber was fitted with two sample ports and the oxygen electrode chamber was redesigned. The
inplementation of a second pump was necessary and a special "water sample module" was added to
allow a bubble-free and compensated removal of water samples for chemical analysis. Together with all
sucomponents like heat exchanger, sample module, tube pumps and filter substrates the total volume
was about 12,4 1 of which 11 l were water. The construction principle of the tested MINI MODULE is
shown in figure 1 . The water temperature was adjusted to 25 +/- 0.5 °C and the flow rate set to 2.5
l/vain. The light intensity in the animal tank was about 800 Ix with a 12:12 hours photoperiod. After
the essential adaptation period of 5 days the plants were illuminated permantly with an intensity of
about 5000 lx. Temperature, pressure, oxygen concentration, flow rate, pH value, redox potential and
conductivity were on-line monitored by electrodes connected to the data acquisition/control system.
The concentrations of the following ions and total hardness were dally determined with conventional
chemical test kits: NH +, NO , NO -, PO 3- and C1-. CO concentrations were taken in the same
• • 3 2 4
way occasionally. In a~diUon, water samples were taken for 6(] days in 3 to 15 day intervals to deter-
mine number of colony-forming ammonia and nitrite oxidizing bacteria in the filter systems by inocula-
tion, incubation and evaluation of suitable selective culture media.

C.E.B.A.S. MINI MODULE V.III TEST RESULTS

In general the laboratory experiments with the C.E.B.A.S. MINI MODULE have two different pur-
poses. On the one hand, they are simply hardware tests to investigate the biological stability of the
systems for a certain time span as a preparation for the spaceflight hardware construction. On the other
there are a series of different scientific aspects which go far over the initial concept of C.E.B.A.S.. It
is important to realize that C•E.B.A.S. is a partially closed artificial or man-made aquatic ecosystem
which can be utilized as a scientific tool in the field of biospherics. In this context it is of high interest
to investigate the interdependences and interactions between the single biological subcomponents of the
system which represents a highly complex and time-consuming task. In contrast to the preliminary one
this expe"rtment was disposed as a "whole sytem test"• It has to be pointed out that this is only the very
beginning of a disposed comprehensive research program analyzing the C.E.B.A.S. as an aquatic eco-
system instead of its primary purpose as a device for mnlti-generation experiments in space biology.
Nevertheless, this new reserach program is also space-related because it may lead to utilizations of the
system as a aquaculture module in bioregenerative life support systems of lunar or planetary bases.

The C.E.B.A.S. MINI MODULE version HI was filled with water at november 16, 1992 and at
december 2 the filter substrates were added. At december 8 the plant chamber was inoculated with 20 g
(wet weight) Ceratophyllum spec. In contrast to all former experiments the bacteria filter was not
inoculated to observe the development of microrganisms from the water. The coarse filter contained
lava grains (7-10 mm diameter) which also served as the substrate in the bacteria filter. The second
filter chamber was partially also filled with lava grain but with a diameter of 4-7 ram. The rest volume
was PERLONR wool. At december 14, 1993, 3 adult Xiphophorus helleri (strain H3) were inserted.
The fishes derived from a controlled breeding. Two of them were mature virgin females, one a mature
male. The total fish biomass was 7260 rag. They were fed irregularily with 30/~g food per nag fish
weight. A dry standard diet used in minimal quantities to avoid water pollution and to guarantee the
survival of the animals. At december 15 some Biomphalaria glabrata were added to run the system
under realistic conditions. From december 17 the plant chamber was permanently illuminated. In the
beginning in the water-filled system the oxygen saturation quickly decreased from about 80 to 37 %
and increased again after opening the system. The developing microorganisms in the filter substrates
 10th IAA Man in Space Symposium 171

induced a sharp decrease down to about 15% which demanded again oxigenization. The plants in the
system kept under low light intemity and 12:12 hour photoperiod ind~ed a day/night rythm of
40:20%. After increasing light intensity up to 5000 Ix and permanent illumination the oxygen salarati-

8,20 ,, Fil P l 24h ~ . .- e l l +, +_~~ +

,.oo .................
!.................
i..-..I
...........
~J...............
I.......
J.........................
+....-i, ..........
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7,80 .................
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- t'"'"~'I" i~................

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..............i..............!+
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i
+::o::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::::
6,40 .................
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6,20 ! '. i + '~ i i i ~ +
0 1 2 3 4 5 6 7 8 9 10 ~,veeks

500 .... i ....... !. . . . . . . . . . . . . . i.................

4oo ................. i .................. i ................ + ........... ' ...... i ................. i .................. i ................. i ............... ~ ................. i .............

2OO + ! i i i i ! ! ! !
0 1 2 3 4 5 6 7 8 9 10 weeks

Figure 2: oxygen saturation, pH values and redox potential in the C.E.B.A.S. MINI MODULE V.III
 172 lOth IAA Man in Space Symposium

on varied between 90 and 55 %. The only exception was a single decrease to 30% which was induced
by overfeeding and which the fishes survived. Switching off the plant chamber after week 9 and repla-
cing it by the gas exchanger for about 2 weeks resulted in oxygen levels between 55 and 75% saturati-
on which is more than sufficient for the survival of the fishes. This clearly demonstrates that as well
the plants as the gas exchanger are able to provide the system with enough oxygen for the survival of
the fishes also for longer time spans. The pH value varied between 6.8 and 7.9 for more than 9 weeks
of the test which is optimal for Xiphophorus helleri. In this range the pH curve parallels nearly that of
the oxygen saturation. After switching off the plant chamber and inserting the gas exchanger the pH
value decreased quickly to 6.4 to 6.8 which is tolerable. The redox potential increased from about 250
to 500 mV in 8 weeks. After that the elctrode had to be changed which resulted in a sharp decrease to
300 to 400 mV which is possibly due to an electrode shift. Oxygen saturation, pH and redox potential
values during the test period are shown in figure 2. The ammonia ion concentration was about 0.3 mg/l
before the filter substrates were in the system. After that it varied between 0.2 and 0.05 mg/l with
decreasing tendency. The NO2 concentration at first increased up to 55 #g/l and went down after filter
activation to near zero in the course of the test period. In comparison to former experiments the NO -
concentrations were low with 8 to 16 mg/l until the fith week. Thereafter it went down to 4 to 12 mg~.
Figure 3 gives an overview of the nitrogen compound ions during the test period. The PO4.3-
concetration was initially low with 0.15 to 0.2 mg/l and increased with the insertion of the lava gram
filter substrates up to 0,5 mg/l. In the further course of the experiment it decreased 0.2 to 0.4 rag/1 and
then to 0.02 to 0.2 mg/l. The CI concerntration went down from about 21 rag/1 at the beginning to 12
to 14 mg/l at the ent of the test. The total hardness started with 5°dH and decreased to 2 °dH until the
plant chamber was removed, after that it increased to 3 °dH. The CO concentrations varied between 3
and 20 mg/l; during the period in which fishes and plants were integrated they were between 3 and 13
mg/l. In the gas exchanger phase of the experiment the CO concentrations were slightly elevated and
ranged between 8 and 20 mg/ml. The conductivity was around 250/zS at the beginning and decreased
from the point of permanent plant illumination on continuously to 160/tS. In the gas exchanger phase
it slightly increased again to 175 #S.

After an adaptation period of 3 days the fish fed normally and showed an undisturbed reproductive be-
haviour, i. e. that the male tried to copulate in the normal frequency and that the females responded in
the adequate way. After the experiment the individual body weights of the fishes were determined. It
turned out that the total weight had decreased to 6850 mg which corresponds to a weight loss of 5.8 %.
Adult X. heUeri are not able to increase the normal bodyweight in the given time of the experiment
with a qualitatively and quantitatively specified diet so that it makes no big sense to calculate fish bio-
mass production at all. The weight loss is a result of the minimal feeding. The water snails reproduced
in the system in a normal way. When the plant chamber was harvested after about 6.5 weeks test dura-
tion the biomass was 82 g which represents a biomass production of 310%. The results of the
microbiological microbiological investigations are currently in evaluation. A first view indicates that
the ammonia and nitrite oxidizing bacteria developed well also without inoculation of the filters and
that they exhibit a nearly identical population dynamics.

CONCLUSIONS AND FUTURE ASPECTS

The most important result of the test reported here is that the C.E.B.A.S. MINI MODULE maintains a
biological stability for more than 50 days with an adequate load of animals, plants and filter microor-
ganisms without any process control by the Electronlcal Component which can be replaced by a simple
data acquisition with the exception of temperature and photopefiod control. It is of further importance
that in contrast to the preliminary experiments nitrate was not longer a "deadlock substance" increasing
permanently up to toxic levels thus limiting the biological stability of the system. All ion concentra-
tions -and with them the conductivity- showed a decreasing tendency during the experiment. We sup-
pose that the increased fight intensity and the permanent illumination stimulated the plant's metabolism
to high nitrate elimination levels thus resulting in a very ion-poor water. For longer experiment dura-
tions this could turn out to be a limiting factor so that "ion feeding" -possibly combined with an in-
creased food supply for the animals- might be essential from time to time. As already mentioned above,
the newly defined operation conditions of the plant chamber led to a highly increased production of
biomass and a more than sufficient net oxygen supply for the fishes and snails in the system. On the
other hand, this increased plant biomass production is another limiting factor for the operation time
of the C.E.B.A.S. MINI MODULE because after a maximum of about 9 weeks it has to be harvested
to avoid a breakdown. In practice this problem migt be solved by the implementation of a second plant
chamber which has to be illuminated with the minimal light intensity of S00 Ix in a 12:12 hour photo-
period (together with the animal tank) which is set to 5000 Ix and permanent illumination after about 7
 10th 1AA Man in Space Symposium 173

weeks when the first is nearly filled and then set down to minimal light conditions. On the other hand
the present experiment showed also clearly that the emergency gas exchanger works very well and is
able to guarantee the oxygen supply for all organisms in the system for a minimum of two weeks. In
i I

I Fil IPl IAn124h i o t.. Pl-,Ox+

I °'351- i I il [ I i AmmomamWl|i i' '~ I
'o,3o L..~..-...i .................. i...l ............ i.[ .............. H ......... i ................. !.................. !.-.".............. i ............ ......i......l ......... i ................. '

0,25 ...... !..................

i..........;..............................
i.................
i..................
~.................
i..................
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o,~o ................
~........i...............................................
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...............
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o,15 ...........
:,...... ! ....................
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0,011 ; i ~ i i
2 4 6 8 10 Weeks

[Fil IPl IAn124h . l Pl-/Ox+

6°1 ~ ~ I I II ~ ~Nitrite~--/li [ i I
i i i i i
50 ................. ~............. i ................ i ............................... i ................. i .................. i .................. i ................. ~ ............... !..................

4o ................. i ............. i................ i ............................... i ................. i.................. i.................. i .................. i ............... i . . . . . . . . . . . . . . . . . .

,o ............... ................. ................ ............................... i ................. i ......................

,o ............. i ................. . . . . . . . . . . . i .....

0 . . . . . . .

t 2 4 6 8 10 Weeks

IFil 24h F" Pl-/Ox+

.................,....I
.................. li' l ......... f........ [.Nitrat_e. m....~_. t : ........... .................

. . . . . . . . . . . .

2 4 6 8 lo Weeks

Figure 3: NH 4 + • NO 2 - and NO - concentrations in the C.E.B.A.S. MINI MODULE V.III -

174 lOth IAA Man in Space Symposium

that case, however, C.E.B.A.S. would not longer fulfil the demands to a fully biological system. For
the initially disposed long-duration experiments this is without any importance but it would be a heavy-
to-tolerate disadvantage and handicap for its role as man-made aquatic ecosystem and tool for bio-
spheries research.

The results of present experiment can be discussed generally under two different aspects. The first is
primary target of the C.E.B.A.S. project and the role of the MINI MODULE in this connection,
namely its function as a spaceflight hardware test in the space test phase for the development of the
space station version. The test of the complete system showed clearly that it maintains biological sta-
bility for the more than the fivefold time span of a spacelab mission and that -provided that preliminary
mathematical modelling results are verified in another experiment series- it is also suitable for bio-
sattelite flights or a mid-term test in the MIR space station. This was the reason that it was decided to
start the industrial construction of the spaceflight hardware immediately. Moreover, there is a fully de-
veloped disposition of a series of scientific experiments which go far over the necessary but superficial
hardware test aspect and which can be performed within the time span of a spacelab mission with best
chances of success. In the foreground there is a skeleton research experiment with neonate X. hel/er/
which on Earth mineralize their cell-free axial skeleton in the first week after birth and which can be
maintained in numbrers over 100 in the C.E.B.A.S. MINI MODULE. This central experiment is ac-
companied by several "backup experiments" dealing with immunology, embryology/teratology and re-
productive biology. In addition, in the small second animal chamber several biomineralization and
embryological studies are disposed with the puimonate water snail Biomphalaria glabrata the embryo-
nic development of which lasts only 7 days. So the C.E.B.A.S. MINI MODULE will also serve as a
scientific tool in a cooperative German/NASA scientific spacelab middeck locker experiment.

The second aspect is -as already mentioned above- that C.E.B.A.S. is a partially closed artificial or
man-made aquatic ecosystem which may be optimized and serve as a valuable tool in biospehrics rese-
arch. Like the biosphere Earth C.E.B.A.S. is energetically open, i. e. that in any case energy has to be
permanently introduced into the system. In addition, the current C.E.B.A.S. systems are still "nutri-
tionally" open, i. e. that the animal food must be procvided from outside. The C.E.B.A.S.-MINI
MODULE is a very suitable and easy-to-handle tool to establish a self-maintaining food chain which
closes the system. To perform this task a comprehensive system analysis is a conditio sine qua non for
further development. This is only possible with the knowledge of the interdependeces and interactions
of the single system components including the determination of limiting "deadlock substances". The
"fishes only" and "plants only" experiments 1° were first steps on this laborious and time-consuming
trail. The next task is the development of "Nutritional Components" which are suitable to close the
food chain. Adult X. hel/er/can be fed in an optimal way with small live crustacea, like Daphnia spe-
cies. Redfem and Lock 11 developed a closed microalgae/Daphn/a culture system which easily could be
combined with the C.E.B.A.S. and the only negligible technical problem is the implementation of 2
magnetic valves and a timer. A similar system is operative at the Institute of Biophysics of the Russian
Academy of Sciences (Kokova, personal communication) at Krasnoyarsk, Russia, which is open and
has much smaller dimensions. In a cooperative disposition this shall be closed and connected with the
C.E.B.A.S. MINI MODULE. Crustacea in the size of the genus Daphn/a are not suitable, however,
for the nurtrition of X. hel/er/neonates. Therefore it is disposed to add a microalgae-rotatoria link to
the system which was also developed at Krasnoyarsk by Kokova 12" 13. In the final stage this may re-
sult in a nutritioanally closed "5-Component C.E.B.A.S" which is schematically shown in figure 4.

Although the development of this man-made cloded ecosystem will last several years it opens innovati-
ve aspects for the development of bioregenerative life support systems for space exploration. So, e. g.,
the low gravity in a lunar or martian station is the most serious problem for breeding tetrapods. The
disturbed reproductive behaviour easily could be replaced by artificial insemination or blastocyst im-
plantation but the influence of low gravity conditions on the musculo- sceletal system will disturb pro-
tein production within economical limits. In addition, urine and feces will cause serious waste
management problems which can only be solved by highly complex and sophisticated (and herewith
extremely expensive) technology. So aquaculture seems to be the only suitable way. The theoretical
possibility mentioned above to drive forward C.E.B.A.S. system into a development of an aquatic
CELSS does not mean that this is practicable or suitable. The general question is why plant food has to
be converted at all into animal protein and vice versa. PlRntS are "producers" and Animals are "consu-
mers" in natural ecosystems. So basically there is no need to produce food by raising heterotrophic
organisms if plant protein is available in a consumable form. So, e. g., cell wall-free mutants of uni-
cellular algae might serve as a sufficient protein source for human nutrition. Recycling of human waste
products can be used for nutrient solutions or fertilizing higher plants and are useless for animal feed-
 10th 1AA Man in Space Symposium 175

ing. With respect to C.E.B.A.S. that means that there is primarily a production of algal biomass which
has to be removed frequently from the system without delivering food for the animals. These, however,
have to be fed by a special diet which must be stored and supplemented from time to time. Provided
the the amount of food given is exactly adequate to the animal biomass present the feces may deliver
only a minimum amount of nutrient even for photoheterotrophous algae. In the experimental system
only occasionally an animal will die in a natural way because the removal of animal samples simulates
this process. So the aspects basically are not so positive to develop an aquatic CELSS. On the other

Algae ROTATORIA Rotatoria

SYSTEM

Algae ~ | Algae ~ | Fishes

BACTERI- -
AL SYSTEM
II CRUSTACE-
Alq SYSTEM
II CRUSTACE-
FISH-
AN SYSTEM

Bacteria ~ I Crustacea ~ I Crustecea

Figure 4: Scheme of the disposed "5-Component-C.E.B.A.S."

hand there is the possibility to try the utilization of plant-eating fish species to convert possibly bad
tasting algal biomass or plant pans inedible for humans into animal protein. Grass carp could be a
suitable species as it is explained later. In this case it is doubtful, however, if a succesful reproduction
of free spawning fish which release their eggs and sperm into the water is possible at all under
conditions of near weightlessness. In free spawning fish, however, there might be a certain amount of
"StatiStiCal fertilizations" because as a rule these animgls produce tremendous amounts of gametes.
Artificial fertilization could be a solution of this problem. Embryonic development of the young then is
another problem. In fish species performing parental care as the genus T/kspia which are suitable for
aquaculture the prediction for successful reproduction under space conditions is near zero.

Another aspect is the capacity of the system at all. C.E.B.A.S. simulates in a ZOOLOGICAL COM-
PONENT a natural ecosystem with a stable population density of about 5 g of biom_~s per 1. It would
be an extremely difficult but not impossible task to adapt the animal holding tank to the conditions of
intensive aquaculture on ground. Large amounts of fish in a limited water volume result in extreme
conditions which have to be stabilized by additional oxygen supply, extremely effective and manpower-
consuming filter systems and large amounts of food. In this case the animal system could change to a
"simulated producer" because excess food and large amounts of feces could deliver nutrients for algae
or higher plants. As already mentioned above on ground there are already systems which use
aquaculture water for growing lettuce, e.g.. The economy and efficiency of these types of hydroponics
are questionary on ground as well as in a space station. Own preliminary experiments with higher wa-
ter plants show that these might be extremely useful in a water recycling system but until today there
are no indications for an utilization for human or animal nutrition. In this field a tremendous amount of
work has to be done if there are serious intentions to develop aquatic CELSS. There are some
possibilities to hit this target, e.g. by utilizing the edible rootless tropical duckweed Wo~a arlsiza in
suspension culture as it is performed already in Southeast Asia. Nevertheless, it will be a long and
laborious way which possibly is not worth the efforts from an economical point of view.
176 lOth IAA Man in Space Symposium

Bearing in mind that higher plants can be utilized for bioregenerative life support with a high efficiency
a different concept for a aquaculture system may be developed. A considerable part of the higher plants
is inedible for man and must be recycled in any way. These plant parts theoretically could be fed to
herbivorous fishes suitable for aquaculture. In a first view, e. g. the cichlid fish Oreochromis mos-
sambicus (formerly gIlapia mossambica or Sarotherodon mossambicus) could be a prospective candid-
ate. This species can be bred and maintained easily in terrestrial intensive aquaculture with best results
but there are two seroius reasons for exspected difficulties. On the one hand it is not solely herbivorous
(-it is in parallel a very active predator-) and on the other it exhibits a very complex courtship and
spawning behaviour in combination with an intensive parental care in the course of the reproductive
cycle which without any doubt will be heavily disturbed under lowered gravity. Although they are a
minority amongst the fish feeding types there are several herbivorous fresh water species of teleosts but
only a few are used in practical aquaculture. Of special interest in this connection is the Chinese grass
carp (Ctenopharyngodon idellus) which has a considerable importance in aquaculture and can be bred
easily with the help of hormonal treatment. There are a lot of imponderabilities to verify such a system
but this is considered to investigate in connection with the Russian BIOS project at Krasnoyarsk. In any
case, the basic idea is worth to be further investigated and possibly a especially suitable freshwater
teleost species may be selected to consume at least a part of the man-inedible plant biomass thus
transforming it into animal protein for human nutrition.

There is, however, another aspect which illuminates the problem from a totally different direction: the
human factor. One has to realize that human life is essentially correlated with special demands the scale
of which goes from simple and basic ones up to the general field of culture. One of these demands is
the taste of food, its smell and its sight. The ritual of "having a meal" with tasty food at a specially
prepared table in a cultivated athmosphere together with other people is a very important aspect in hu-
man personal and social life. An astronaut may be able to consume freeze-dried or pasted food for a
certain time without physical and psychical disturbances but this would be impossible in long-term
missions lasting, e. g., several years. Provided that there is no transported supply with fresh food the
autonomous production of both plants and animals for human nutdtuion are necessary (except the
astronauts are selected vegetarians). Accepting this problem in general the production of fish protein
indeed could be a suitable way for human food supply in a space station or a planetary base. In this
case however, it would be much more easy to try to adapt methods of modern intensive aquaculture to
space conditions and to develop special devices for water recycling than to use a complex and very dif-
ficult-to-stabilize combined animal/plant system like an "advanced C.E.B.A.S." for this purpose. The
same is valid in general for plant production with the exception that indeed waste water from an inten-
sive aquaculture system could be used in hydroponics in any way. The problem for a reproductive
biologist is in this connection the development of an efficient hatchery which is the essential precondi-
tion for a intensive aquaculture system. This creates questions of fertilization, embryonic development,
growth and differentiation in space which only can be solved in long-term multi-generation space expe-
riments. This closes the loop to the experimental C.E.B.A.S. system which is a suitable tool to invest-
igate all these problems. So all dispositions to create an aquatic CELSS must remain sole speculations
until a tremendous amount of basic and space research is performed. On the other hand: innovative
ideas demand for creativity and phantasy and may lead to results in combination with well disposed and
carefully performed basic and applied research.

The C.E.B.A.S. Center of Excellence which was newly founded at the beginning of 1993 at the Facul-
ty of Biology of the Ruhr-University of Bochum, FRG, is responsible for the further develpment of the
C.E.B.A.S. system as well for space as for terrestrial research. With the flight hardware construction
by the industry now a first step into the space test phase is done and the first spaceiab flight is disposed
for 1995. Moreover, it integrates and combines the the initial international multidisciplinary zoological
C.E.B.A.S.-AQUARACK research program with new disciplines as gravireception research,
microbiology, plant physiology, system theory and biospherics covering all relevant areas of
C.E.B.A.S.. Together with the possibility to expand the international cooperations thus optimizing the
already existing worldwide scientific network this institution now offers the intellectual, methodologi-
cal and financial power to verify most of the innovative and challenging ideas mentioned above.

ACKNOWLEDGEMENTS

The C.E.B.A.S. project and the C.E.B.A.S. Center of Excellence are funded by the German Federal
Ministry of Research and Technology v/a the German Space Agency (DARA, grant # 50 WB 9319-3)
and the Ministry of Science and Research of the Country of of Nordrhein-Westfalen (grant # IV A 1-
216-002 92).
 lOth IAA Man in Space Symposium 177

REFERENCES

1. Lotz, R. G. A., Baum, P., Bowman, G. H., Klein, K. D., yon Lohr, R. and L. Schroetter: Test of
a life support system with Hirudo medicinalis in a sounding rocket. Life Sci. Space
Res. 10, 133-143 (1972)
2. Neubert, J., Briegleb, W., Schatz, A., Hertwig, I. and B. Ktuse: Observations on structure and
function of the gravireceptor in a vertebrate (Xenopus laevis D.) exposed to near
weightlessness. In: Sabra: P R__ Ian.*,~.n_ R. and M_ H. K~il~r (~1~ V Scientific r~-
gnit~ of the German .~paeo~lah mi~icm DI (lOg6 Norderney) W P F K61n, 423-430
0987)
3. Blfim, V. und R. Fr6se: Experimente mit aquatischen Tieren im Weltraum. I~MFT Ah.q~hlul~h~rlcht
znm Fnr.~chnn~nfia'n~ fll Qv g~ 474 (1989)

4. Blfnn, V., and Krenzberg, K.: The GERMAN CELSS Program with Special Respect of the
C.E.B.A.S.-AQUARACK Project. Acta Astronautica 23, 245-252 (1991)

5. Blfan, V.: C.E.B.A.S., a Closed Equilibrated Biological Aquatic System as a Possible Precursor for
a Long-Term Life Support System. Adv. Space Res. 12, 193-204 (1992)

6. Blfim, V. and K. Kreuzberg: The C.E.B.A.S.-AQUARACK project: the laboratory prototype and
first results of the scientific frame program. Microgravity Quarterly 2, 1-15 (1992)

7 Blfim, V. and K. Kreuzberg: C.E.B.A.S.-AQUARACK: second generation hardware and latest

scientific results. Acta Astronautica 27, 197-204 (1992)

8. Blfim, V., Kreuzberg, K. and E. Stretzke: C.E.B.A.S.-AQUARACK-project: middeck locker mini-

module test results. ASGSB Bull. 5, 51 (1991)

9. Blfun, V: Current status and first scientific results of the C.E.B.A.S.-AQUARACK project. ASGSB
Bull. 3, 52 (1989)

10. Blfim, V., Kreuzberg, K. and E. Stretzke : C.E.B.A.S.-MINI MODULE: test remits of an
artificial aquatic ecosystem. Adv. Space Res. (in press)

11. Redfern, P. R. and M. A. Lock: An integrated study on nutrient cycling and population biology in
closed aquatic ecosystems. Proc. ~nd Int_ Svmn l J ~ g n p p Rin~ru~h~ricg,
Huntsville, Febr. 1992 (1992)

12. Kokova, V. E.: Continuous cultivation in invertebrates. Nauka, Novosibirsk, 168 pp. (1982)

13. Prolulmikov, A. M. and Kokova, V. E.: Influence of different livin food on growth and develop-
ment of carp larvae (Cyprinus carpio U , Cyprinidae. J. Woprocy Ichthyology 24,
495-499 (1984)