Biological Experiments in Space: 30 Years Investigating Life in Space Orbit

By Galina Nechitailo and Alexey Kondyurin

Biological Experiments in Space: 30 Years Investigating Life in Space Orbit covers investigations of plant, algae, animals, fish, microorganisms and tissue cultures on space flights, beginning with the first orbital space station on Salyut 1. The book includes results on the influence of the entire complex of physical factors associated with spaceflight on biological systems, including analysis of the impact of microgravity on organisms, as well as the effects of electric and magnetic fields. This book offers important insights for researchers of space biology and astrobiology, as well as space agency and industry specialists developing future space stations and missions.

Lack of gravity, temperature and chemical gradients, magnetic and electrical fields, spectral composition and intensity of light, and high-energy cosmic radiation influence many important metabolic and physiological processes in animals, plants, and microorganisms, as well as transfer phenomena in and around them. Success of future space exploration depends on understanding the effects of these factors on biological organisms and developing appropriate countermeasures, aimed at improving growth, development, and reproduction in microgravity.

• Includes results on the influence of the entire complex of physical factors associated with spaceflight on a range of biological systems
• Analyzes the impacts of microgravity, as well as electric and magnetic fields, on organisms
• Covers pioneering investigations of plants, algae, animals, fish, microorganisms and tissue culture in space flights

• Language: English
• Publisher: Elsevier Science
• Release date: Jul 10, 2021
• ISBN: 9780128205013

Book preview

Chapter 1

First long-term station Salyut 1: first steps to establish a space garden

Abstract

First biological experiments on Salyut space station. First growing plants and other biological objects in space: potato, flax, Crepis, bony fish, Chlorella, yeast, frog spawn and tulips. First flower in spaceflight. Our success and failure. Can we grow plant in space or not?

Keywords

Space station; Salyut 1; potato; flax; Crepis; bony fish; Chlorella; yeast; frog spawn; tulips

Back in 1962 Sergey Korolev determined the priority of fundamental problems for the scientific organizations of the USSR to provide manned astronautics, including a program of botanical and agriculture research in space with the aim to create the biological systems for life support systems in spaceflight. These research activities on crop production for biological life support systems in spaceflight, which were carried out by different scientific organizations, progressed very slowly. Vasily Mishin, the head of the Central Design Bureau of Experimental Mechanical Engineering (TsKBEM) after the death of Sergey Korolev, decided to start a laboratory for the biomedical support of spaceflights in 1970. Specialist biologists, medics, and engineers were engaged for the laboratory. The aim of the laboratory was the implementation of studies focused on creating a scientifically substantiated background for the biological life support systems in spaceflights. The best scientists and engineers from Moscow, Leningrad (now St. Petersburg), Krasnoyarsk, and Novosibirsk and from the republics of Lithuania, Latvia, Belorussia, Moldova, Ukraine, Azerbadgan, Georgia, and Uzbekistan collaborated on the project. The laboratory provided biological experiments on spaceships and space stations from Salyut 1, followed by Salyut 4, 6, and 7, and Mir space station over more than 30 years. The problem of space crops during spaceflight, which was overviewed by Sergey Korolev, was found to be quite difficult. Only after 20 years of intensive investigations and experiments in spaceflight did the first wheat plants pass the entire life cycle from seeds to seeds in spaceflight, in 1991. During 30 years about 1000 spaceflight experiments on genetics, embryology, biochemical, and biotechnological studies had been established in space stations.

From an interview with Galina Nechitailo:

When I graduated with my PhD under supervision of Professor Nikolai Dubinin, the question arose of what to do next? Where to go to work? My friend, a very famous designer in the field of rocketry, Dmitry Knyazev (Fig. 1.1) told me: If you want, you can go to our company. The company was TsKBEM, which was started by Sergey Korolev. We will open a unit there and you will be engaged in our biology. He came to Professor Nikolai Dubinin and said: Nikolai Petrovich. There is such an idea: refer your favorite graduate student to our company. Nikolai Dubinin always said that he had several favorite graduate students. I was among them. Nikolai Dubinin replied: Good, but how to do it? A letter from the Institute is not enough. Then Dmitry Knyazev went to Mstislav Vsevolodovich Keldysh. Dmitry Knyazev was friends with Mstislav Keldysh, who at that time was President of the USSR Academy of Sciences. Dmitry Knyazev and Nikolai Dubinin came and agreed with Mstislav Keldysh, and Mstislav Keldysh wrote to the company that there was such an idea to send a biologist to them to carry out the genetic research that Nikolai Dubinin had begun on unmanned ships. When this letter came to the company and they started applying for work for me, there were no biologists there except Vladimir Pravetskiy, who had previously been the head of the third main department of the Ministry of Health.

I was introduced to Vladimir Pravetskiy, but he was terribly unhappy and did not want any development in this field. However, since this was a decision from above him, he had to agree with this and told me: Do whatever you want. TsKBEM at that time was a large industrial plant, many industrial buildings, towers. What is it possible to do there for a biologist? I was attached to a group that worked on food, spacesuits, and linen for astronauts: T-shirts, underpants, overalls. I was assigned to this group at first, and then I was made the leader of the group. I tried to figure out how science can be done here. It was all quite difficult. Then I asked Dmitry Knyazev: I want to see what is the company as a whole doing. He said: Well, there are workshops that are completely off the topic of biology and it’s hard to go there. They won’t let you in because of secrecy. But I said: Well, what can be done? I have to understand where and what? See what spaceships are being built and what equipment can be placed in them. Finally, he made me different birds (signs) in my pass to get access to all the workshops and I began to look at them. What is there? Where is that? I looked at all these spaceships. Everything was amazingly interesting. Soon after another biologist was engaged in TsKBEM–Alexander Lvovich Mashinsky, whom I already knew. He worked at the Institute of Biomedical Problems and supervised the plant program for the planned annual lunar experiment. When he came, I was happy: Another biologist. And then together we began to think, what experiments can be installed? We had a look the program of the next spaceflights. Dmitry Knyazev was familiar with all the key leaders and helped us a lot. He was a very close friend of Boris Viktorovich Rauschenbakh, a great man, designer, and scientist. I must say that all people of that generation were very versatile; they were fond of science, art, music, and even religion. Boris Rauschenbach collected the Christian icons, and Victor Pavlovich Legostaev did too. It was a very intelligent team. I worked together with so many talented people. Vladimir Sergeyevich Syromyatnikov, designer of docking ports, is also an amazing person. He had so many incredible ideas! It was he who created the androgenic docking unit, which was used during the flight of the Soyuz–Apollo and is even being used now. Alexander Mashinsky and I had decided to prepare the first program of biological experiments for the next spaceflight. The next flight was Soyuz 9 with Andrian Nikolaev and Vitaly Sevastyanov. We had overviewed all the literature. What's to do? What is done? What were the experiments? But there were few experiments with living organisms. In addition to experiments with dogs Belka and Strelka, dry seeds, pollen of plants, and microorganisms, that is, not actively functioning organisms, were sent to space. Therefore, we very quickly, in 3 months, wrote a program for living plants. Then there was a commission on space science under the supervision of Mstislav Keldysh, where scientists and designers gathered, discussed future space programs, and decided what would be installed on board of the spaceship and what would not. I invited geneticists to our program. At the Paton Institute in Ukraine, we were made very quickly some devices for biological experiments on the board, which we later called the Biofixator and Biocontainer. And in such containers, on Soyuz 9, we sent potato tubers, Guppy fish fry, and seeds of higher plants. Drosophila flies were sent in a small container. Vitaly Sevastyanov wanted them to fly out and called them Nyurka fly. But they were in a closed container and could not fly out.

Figure 1.1 Dmitry Knyazev and Galina Nechitailo on holiday.

1.1 Soyuz 9

The first experiments during a long flight were carried out on a Soyuz 9 spaceship. The spaceship was piloted by cosmonauts Andriyan Nikolaev and Vitaly Sevastyanov. The flight duration was 17 days and 16 hours. The launch was on June 1, 1970 and the landing was June 19. The main goal of biomedical research was to study the effect of a multiday spaceflight on the functionality of the crew and its performance. After the flight Andrian Nikolaev suggested that long flights, more than 18 days, are impossible for a human.

From interview with Galina Nechitailo:

This 18-day spaceflight, of course, was critical for the cosmonauts. Nikolaev after the flight said: Well, that’s it. We cannot fly longer, because this is the limit for a human. But the fact is that when they flew, they only had a spaceship and a crew compartment. Very limited volume for crew. It was impossible to do any physical exercise. The most that they had was a small expander with which they could stretch a little. Therefore, at the stations under construction, and at that time the Salyut station was already under construction, various fitness equipment had already been designed so that the astronauts could run, stretch, that is, maintain physical fitness.

In this flight, a number of biological experiments were installed. For the first time, living plants were sent to the orbit. Biological studies were conducted with seeds, potato tubers, and Drosophila in flight conditions. Chlorella algae grew in one of the containers.

The first experiments over a long period in orbit were carried out in the BB-1 device (biological block). The BB-1 device consisted of a number of plates; seeds were fixed on some of them, and other plates were used as radiation detectors. The BB-1 device was placed on board the spaceship. The seeds of Crepis, Arabidopsis, wheat, and some garden plants were sent.

The problem of exposing seeds on board spaceships was raised by the main designer Sergey Korolev. How can cosmonauts be provided with food on a long flight? If enough seeds for a long spaceflight are to be taken to the spaceship and plants are to be grown in spaceflight, therefore the seeds must maintain the ability to germinate for a long time, for example, several years. This was the main goal of these studies.

In this first long spaceflight on Soyuz 9, the seeds were exposed in orbit for 18 days. After landing, the seeds were analyzed in a laboratory on Earth. Germination energy of the seeds was analyzed and genetic analysis of the seeds was done.

The major characteristics of seeds are germinating capacity and energy of germination. These characteristics were measured for flight samples and for ground samples cultivated in the ground after the flight. The germinating capacity was calculated by the following formula:

(1.1)

where A is the total number of seeds and A′ is the number of sprouts from these seeds.

The energy of germination was calculated by the following formula:

(1.2)

where A is the total number of seeds and A′ is the number of sprouts from these seeds at t days after sowing.

Wheat of type Saratovskaya-29 was used in the experiment. The wheat seeds were selected before the experiment: empty and deformed seeds were excluded from the experiment. The germination results of wheat seeds are presented in Table 1.1.

Table 1.1

After spaceflight the wheat plants from the ground and spaceflight seeds were grown in a phytotron for 114 days until full ripeness. Wheat was cultivated using aquatic culture in the Knop solution in 3 L vegetation vessels, which were automatically aerated for 5 minutes every hour. Each vessel contained 14 plants. Water and mineral metabolism was continuously monitored. A morphological analysis of the plants at the early stage showed no morphological difference between ground and spaceflight plants. The analysis of water consumption showed that the flight plants consumed slightly less water than the ground plants. The microelement analysis showed that the utilization of phosphorus, potassium, calcium, and magnesium by ground and spaceflight plants are similar. The nitrogen and major minerals content in harvest is similar for ground and spaceflight plants. These results showed that 18 days spaceflight does not influence the harvest of wheat grown on the ground after the flight.

The potato was considered as an important source of food in future space missions. Soyuz 9 carried the potato tubers, germinating them during the 18-day flight mission. The Priekulsky Ranny type was selected for the experiment. Three tubers of 8–10 g each were placed in a perforated container with a mineral enriched substrate. The substrate was moistened with 2:1 water/dry weight ratio before the launch. Ground samples were in a similar container and moisture substrate. After the mission the container was returned to the laboratory within 24 hours after landing of the spaceship (Fig. 1.2). The flight tubers had prominent shoots and small roots (Fig. 1.3). The ground tubers had more developed shoots and roots in comparison with the flight samples. The ground tubers lost less mass (Table 1.2) and the ground shoots and roots were longer. The shoots of the flight samples had some branches, while the ground shoots did not have branches. Two of the three main shoots of the flight tubers had become brown at the apices. They were lost in following days and lateral shoots appeared.

Figure 1.2 First potato tubers germinated in spaceflight. Ground control and flight vessels with the tubers after landing of the Soyuz 9 spaceship. Taking the flight samples from the vessel after the landing. The ground vessel with the control ground tubers is near.

Figure 1.3 The potato tubers germinated in the Soyuz 9 spaceflight over 18 days (right, marked o that means "opit or experiment") and ground control samples (left, marked k that means "kontrol or control sample"). Shoots of the flight tubers are branched, while each ground sample shows one long shoot.

Table 1.2

The potato tubers from the flight and ground were grown in a phytotron on ceramist substrate for 93 days. Carbon dioxide content in the air was 0.1%. Luminescent lamps with water filters provided 125 W/m² of illumination power for 16 hours per day.

The cultivated potato showed some morphological difference between the flight and ground samples, when the first five to eight leaves appeared (Fig. 1.4). One plant had larger and rounded terminal leaves, prominent stalk pubescence, with longer hairs in comparison with the ground plants. Another plant also had rounded leaves. Leaf vein counts varied in all flight samples. The height of the flight plants was lower than the ground samples for the first 2 weeks of growth (Fig. 1.5).

Figure 1.4 The cultivated potato from the tubers germinated in Soyuz 9 spaceflight (left, marked o that means "opit or experiment") and on ground (right, marked k that means "kontrol or control sample") at the same time. The photo was taken 14 days after landing. The terminated leaves of the flight sample are large and rounded, while the new top leaves are usual, like the ground plant.

Figure 1.5 Growing height of potato shoots from tubers germinated in Soyuz 9 flight and ground tubers. The value of STD is the same as the size of signs.

The harvest was analyzed after 93 days of cultivation (Fig. 1.6). The tubers of ground and flight samples were viewed as similar (Fig. 1.7). After that the height of the flight and ground plants become equal. Table 1.2 shows the harvest parameters. The flight plants have bigger total biomass including tubers than the ground plants.

Figure 1.6 Potato harvest from ground and Soyuz 9 spaceflight tubers in the phytotron on keramzit substrate, lightweight expanded clay aggregate, under the luminescence lamps.

Figure 1.7 Harvest of potato grown from tubers germinated in Soyuz 9 spaceflight (1–3) and germinated on ground (4–6).

The general conclusion is that the 18 days vegetation of potato tubers during spaceflight had some effect on the growing plants, while the following cultivation on the ground made this difference neglectable.

Thus for the first time potato tubers were germinated in spaceflight.

Chlorella is the first candidate for a self-sufficient ecological system for future long-term spaceflight. This is why Chlorella was at the center of investigations from the first spaceflights in Russia and the United States. The first experiments with Chlorella cells were done in the second spaceflight of Vostok 2 with cosmonaut German Titov during 25 hours on August 6–7, 1961 and the Discoverer 17 unmanned mission during 72 hours on November 12, 1960. The postflight analysis showed the viability of Chlorella pyrenoidosa culture and normal activity with regard to photosynthesis, growth, and proliferation, while immediately after the spaceflight the culture showed lower photosynthesis activity and a large number of dead cells. The first experiments with Chlorella were installed on Soyuz 9 too. The Chlorella vulgaris cells were exposed during this flight in active and resting states. It was the start of widespread Chlorella investigations in subsequent space station flights.

For the experiments, special equipment was designed and developed that met the requirements of spaceflight safety, especially with regard to the crew, and the requirements for living organisms. In particular, already in the first flights, proposals were made to set up the experiment directly on the launch pad of the spacecraft, literally the hours before launch (from 2.5 to 6 hours), and in some cases, even directly at the time the crew boarded the ship, when the astronauts took the sample cassette with them into the elevator in a suit pocket. The experimental equipment included not only ensuring the vital activity of organisms during the flight, but also providing conditions for the delivery of samples from the laboratory to the cosmodrome, during launch and landing, and for the delivery of samples to the laboratory for research. The entire chain of sample preparation and transfer was calculated and tightly controlled to eliminate all possible risks of sample loss due to transportation. The effects of transportation and storage on biological objects were also taken into account when comparing flight and ground samples.

Compliance with these conditions was facilitated by the direct presence and participation of space biologists in the development and production of spaceships, close cooperation with the designers of spaceships, and the close attention and interest of the chief designers in the formulation and conducting of biological experiments. As a result of such attention, the biological program on spacecraft over more than 30 years has been the longest and most successful of all scientific space programs carried out on spaceships, given the difficulty of the experiments and the success of the pioneering results.

From interview with Galina Nechitailo:

The first time I arrived at the Baikonur cosmodrome at the start of Soyuz 9 and it was necessary to prepare biological samples for the spaceflight, there was no room and equipment at the cosmodrome for the preparation of our experiments. We turned to the sanitary and epidemiological station, where there was a sterile box. There we prepared our biological samples for the first flight on Soyuz 9. And since we were puzzled that there was no laboratory and equipment to prepare the biological samples, we wrote proposals. For the next flight on the Salyut we already had a biological laboratory in MIK (Installation and Testing Complex), which was established about 250 m from the spaceship. The biological samples could be taken straight out of the laboratory and delivered to the spaceship. Later biological equipment was usually installed 4 hours before the start. That was practically at the crew arrival time. Unfortunately, now, as a rule, biological samples are installed significantly earlier than the launch time, which can affect the experimental requirements for living biological samples.

It is important to say that we had the opportunity to receive biological material immediately after landing. Alexander Mashinsky was always at the landing site (Fig. 1.8). Sometimes the rescue team was still taking the astronauts out of the landing capsule, but the biological samples were taken first. Since the landing capsule of the spaceship does not have a particularly large capacity, the total returned weight from the orbit did not exceed 5–6 kg. It included different samples of blood, urine, and so on, and usually 3–4 kg of biological samples. Our biological equipment was on almost every spaceship. Our staff immediately removed the biological equipment from the landing capsule at the landing site, processed the material, when it was required, and transferred it to transportation containers (Fig. 1.9). Our specialists had portable thermostats in which biological samples were immediately placed and the biological samples were delivered to the laboratory under controlled conditions. Thus, quite quickly, after the landing of the astronauts, biologists in Moscow got the samples under maintained storage and transportation conditions. At this point, all our collaborators from different Institutes came to us. We had collaborations with the best scientists from Russia, Ukraine, the Baltic republics, Belarus, and Moldova. And then the final steps were taken with the samples to distribute them to the laboratories for detailed and sophisticated investigations.

Figure 1.8 Alexander Mashinski gets set with biological samples (potato) at the landing site.

Figure 1.9 Alexander Mashinski treats the biological samples after a flight in an airplane from the landing site.

1.2 Kosmos 368

Together with manned flights we took part in biological experiments on unmanned satellites. It was a good experience for us and we saw what results our colleagues had got from the flights. Thus Kosmos 368 with payload Nauka 3 was launched October 8, 1970 with the Zenit 2M rocket and landed October 14 (6 days). The orbit at the apogee was 411 km, at the perigee 211 km. The orbital inclination was 65 degrees. The temperature on board was maintained at 20°C–22°C.

The satellite Kosmos 368 carried various kinds of units and containers with microorganisms, insects, seeds, plants, and cultures of animal and vegetable cells. The purpose of the biotechnical tests was to establish the possibility of using new equipment to conduct biological tests in prolonged spaceflights. The biological tests in the AES Kosmos 368 were the natural extension of experiments conducted earlier in other spacecraft instruments. The biological samples were delivered to the laboratory within 24 hours after