P. Straat

In a solvent system of 10(-2)m phosphate buffer (pH 6.8)-ethanol (2:1, v/v) and in an iodine-induced reaction, the polycyclic hydrocarbons [(3)H]3,4-benzpyrene and [(3)H]3,4-BP/[(3)H]9, 10-dimethyl-1,2-benzanthracene (DMBA) can be covalently linked to deoxyribonucleic acid (DNA) at room temperature. By stepwise addition of the hydrocarbon and repeating the reaction two to three times after isolating the hydrocarbon DNA adduct, it was possible to introduce as many as one covalently bound hydrocarbon molecule per 100 nucleotide bases. When 3,4-BP and DMBA were linked in this way to biologically active transforming DNA of Bacillus subtilis, they caused (i) reduction of the transforming activity of the DNA accompanied by (ii) significant increases in the frequency of forward mutations. The majority of these hydrocarbon-induced mutations were not able to revert spontaneously. These samples of DNA covalently linked with hydrocarbons showed much lower levels of survival of biological activ...

One factor involved in the possibility that airborne microbes might contaminate the Jovian atmosphere, is whether microbes have the capacity to propagate in air. Prior to these studies, the evidence was that the airborne state was lethal to microbes. By mixing an aerosol of aerobic bacteria with another containing 14C glucose, we were able to detect the presence of 14CO2, showing that the airborne cells were metabolically active. In the same type of experiment, we were able to show that thymidine was incorporated into the acid-insoluble fraction of samples, indicating the formation of DNA. Finally, we were able to show, both by an increase in numbers of viable cells and a parallel increase in particle numbers, that at least two new generations of cells could occur. As of this writing, evidence for propagation of anaerobic bacteria has been negative.

As part of an effort to discover whether bacteria might propagate within airborne particles, we studied the incorporation of thymidine into the trichloroacetic acid-insoluble fraction of airborne cells of Serratia marcescens to seek evidence of the possible formation of new DNA. Two aerosols, one of S. marcescens and another of [3H]thymidine ([3H]dT) suspended in growth medium were caused to aggregate in air just prior to directing the aerosols into rotating-drum aerosol storage chambers. The age of the S. marcescens culture and other conditions for maximizing ([3H]dT) uptake were selected on the basis of prior in vitro trials. With 10-h cultures and addition of 2-deoxyadenosine to the [3H]dT, we showed that [3H]dT is incorporated into the trichloroacetic acid-insoluble fraction of cells recovered 6 h after aerosols were stored under the conditions of high humidity and 30 degrees C. Tests conducted in the same manner with Formalin-killed S. marcescens ruled out the possibility of ad...

A decade has passed since the first labeled-release (LR) Viking biology experiment produced an astonishing positive response on Mars. But that response was deemed unconvincing when no organic compounds was found. It is now contention that the survival of the LR data, together with other information not previously considered (including Viking lander image and spectral data that suggest the possible existence of martian lichen), justifies the conclusion that it is now more probable than not that the LR experiment did in fact detect life of Mars.

The first (and only) dedicated life detection experiments on another planet were performed by the Viking Landers of 1976. In the Viking Labeled Release (LR) experiment of Levin and Straat, injections of organic compounds into Martian soil samples caused radioactive gas to evolve approaching plateaus of 10,000 - 15,000 cpm over several sols (Martian days). These "actives" were run at lander sites 1 and 2 with similar results. In contrast, the LR response to the 160o C control sample soils was very low. In conjunction with the active experiment results this negative result from the controls satisfied the pre-mission criteria for life. However, a controversy immediately arose concerning a biologic interpretation of the data. In an attempt to resolve this issue in the current work, we have employed complexity analysis of the Viking LR data for the initial six sols, and of terrestrial LR pilot studies using bacteria-laden, active soil (Biol 5) and sterilized soil (Biol 6). . Me...

Three different types of biological experiments on samples of martian surface material ("soil") were conducted inside the Viking lander. In the carbon assimilation or pyrolytic release experiment, (14)CO(2) and (14)CO were exposed to soil in the presence of light. A small amount of gas was found to be converted into organic material. Heat treatment of a duplicate sample prevented such conversion. In the gas exchange experiment, soil was first humidified (exposed to water vapor) for 6 sols and then wet with a complex aqueous solution of metabolites. The gas above the soil was monitored by gas chromatography. A substantial amount of O(2) was detected in the first chromatogram taken 2.8 hours after humidification. Subsequent analyses revealed that significant increases in CO(2) and only small changes in N(2) had also occurred. In the labeled release experiment, soil was moistened with a solution containing several (14)C-labeled organic compounds. A substantial evolution of radioactive gas was registered but did not occur with a duplicate heat-treated sample. Alternative chemical and biological interpretations are possible for these preliminary data. The experiments are still in process, and these results so far do not allow a decision regarding the existence of life on the plonet Mars.

This report summarizes all results of the labeled release life detection experiment conducted on Mars prior to conjunction. Tests at both landing sites provide remarkably similar evolution of radioactive gas upon addition of a radioactive nutrient to the Mars sample. The "active" agent in the Mars sample is stable to 18 degrees C, but is substantially inactivated by heat treatment for 3 hours at 50 degrees C and completely inactivated at 160 degrees C, as would be anticipated if the active response were caused by microorganisms. Results from test and heat-sterilized control Mars samples are compared to those obtained from terrestrial soils and from a lunar sample. Possible nonbiological explanations of the Mars data are reviewed along with plans for resolution of the Mars data. Although such explanations of the labeled release data depend on ultraviolet irradiation, the labeled release response does not appear to depend on recent direct ultraviolet activation of surface material. Available facts do not yet permit a conclusion regarding the existence of life on Mars. Plans for conclusion of the experiment are discussed.

The Labeled Release extraterrestrial life detection experiment onboard the Viking spacecraft is described as it will be implemented on the surface of Mars in 1976. This experiment is designed to detect heterotrophic life by supplying a dilute solution of radioactive organic substrates to a sample of Martian soil and monitoring for evolution of radioactive gas. A significantly attenuated response by

Viking radiorespirometry ("Labeled Release" [LR]) experiments conducted on surface material obtained at two sites on Mars have produced results which on Earth would clearly establish the presence of microbial activity in the soil. However, two factors on Mars keep the question open. First, the intense UV flux striking Mars has given rise to several theories postulating the production of highly oxidative compounds. Such compounds might be responsible for the observed results. Second, the molecular analysis experiment has not found organic matter in the Mars surface material, and therefore, does not support the presence of roganisms. However, sensitivity limitations of the organic analysis instrument could permit as many as one million terrestrial type bacteria to go undetected. Terrestrial experiments with UV irradiation of Mars Analog Soil did not produce Mars type LR results. Gamma irradiation of silica gel did produce positive results, but not mimicking those on Mars. The life question remains open.