Complexity Analysis of the Viking Labeled Release Experiments
heating the soil to 160° C). However, controversy has reignedever since over these ndings. Until recently, chemicalinterpretation o the LR results has dominated but discoverieso Martian atmospheric methane [5, 6], sub-surace water iceon Mars , drops o liquid water at the Phoenix landing site, and the incredible hardiness o terrestrial extremophiles have all led to the re-examination o the possibility o extant Martian microbial lie.In past work , we have shown that the “active” (gas-evolving) Viking LR experiments exhibited strong circadianrhythms in radiolabeled gas release. Tese oscillations rapidly grew in amplitude and regularity in the rst two sols (onesol=24.66 hr, a Martian solar day) o the active experiments toreach a near steady state o constant amplitude and period.Perhaps, this reects the synchronization o a population o microbes to the temperature cycle imposed by the Vikinglanders. When tested, heat-treated (control) samples o thesame soil showed a greatly attenuated rhythm, or no rhythm whatsoever. In the two experiments in which the active soilsamples were stored or several months beore administeringthe nutrient solution, rhythmicity was almost completely absent.
2. New Approach
We now report a new methodological approach to thesedata, complexity analysis. Due to the high order present inbiological systems  , time series o biological variables, withtheir short- and long-range correlations, scale-invariance,complex periodic cycles, quasi-periodicities, positive andinverse “memory” and the like, exhibit behaviours thatare dierent rom the complete unpredictability o purerandom physical processes (white noise). Moreover, they are also distinguishable rom the trivially smooth landscapeo a completely predictable deterministic process, otenmaniesting themselves with icker (pink) noise (temporalscale statistical invariance) [12, 13]. We have now ound thata set o complexity measures (appendix#1 or denition)unambiguously distinguishes the active LR experiments, orportions thereo, rom various abiotic controls (p<0.001).Tese measures very strongly suggest, in agreement withterrestrial analyses, that the active LR experiments in alllikelihood detected microbial lie on Mars.
3. LR Results on Mars
Summary o initial analysesIn the thousands o tests that were conducted on a wide variety o terrestrial microorganism-laden soils in 20 years o testing beore and ater the Viking mission, radiolabeled gas,presumably CO
(or possibly CO
plus some other carbon-containing gas such as CH
) was produced by cellularmetabolism, always evolving immediately ater the injectiono the radiolabeled LR nutrient (e.g. Biol 5, see Methods).Heat-treated control soils produced insignicant responses(e.g. Biol 6). Tere was never a alse positive or ambiguousresult in the terrestrial experiments. In the current study,terrestrial LR pilot experiments using bacteria-laden active(Biol 5) and sterilized (Biol 6) soil samples were analyzed,using the same nonlinear approaches that were employedor analysis o the Martian data.On Mars, injected soil samples evolved radioactive gas [3,14] rapidly, subsequently approaching plateaus o 10,000 –15,000 cpm ater several sols (Fig 3, top panel). Tese “actives”(VL1c1, VL1c3, VL2c1, VL2c3), were run at Viking Landersites 1 and 2, with similar results. In contrast, the LR responsein VL1c2, the 160° C control, was very low, essentially nil,thereby, in conjunction with the active experiment results,satisying the pre-mission criteria or lie (see appendix #2or a brie description o the Viking LR results).Martian soil heated or three hours at 51° C produced anerratic succession o declining low-amplitude oscillations,each rising or about a sol, then precipitously alling tobaseline (VL2c2). Soil treated or three hours at 46° Cresponded with typical “active” kinetics, but 70% reducedin amplitude (VL2c4). Further, ormerly “active” soils storedat 10° C or three and ve months, at Lander 2 (VL2c5), andLander 1 (VL1c4), respectively, ailed to respond to thenutrient . A second nutrient injection was made to each “active” soilater seven sols (VL1c1, VL2c1, VL2c3) or 16 sols (VL1c3).Each time, the gas briey spiked, ollowed immediately by a24% mean decrease in the accumulated
C gas. Laboratory simulations  showed absorption o CO
by wetted Marsanalog soils (pH 7.2) indicating that the Viking LR gas was,at least in part, CO
. In a terrestrial experiment, upon secondinjection  to an Antarctic soil with known bacterialcontent (pH 8.1) a brie spike also occurred, ollowed by adecrease in the accumulated gas. CH
, now known to be acomponent o the Martian atmosphere (6) and a possiblebiological metabolite, is virtually insoluble in aqueousmedia at temperatures and pressures recorded in the VikingLanders. I produced in such experiments, then it must haveremained in the non-reabsorbed
C-labeled gas raction.Tese results indicate that a signicant raction o the
C-labeled gas evolved on Mars was CO
, at least a part o which (~24%) was reabsorbed on wetting o what was likely an alkaline soil , while the unabsorbed raction could