Fluorescence microscopy as a tool for in situ life detection

The identification of extant and, in some cases, extinct bacterial life is most convincingly and efficiently performed with modern high-resolution microscopy. Epifluorescence microscopy of microbial autofluorescence or in conjunction with fluorescent dyes is among the most useful of these techniques. We explored fluorescent labeling and imaging of bacteria in rock and soil in the context of in situ life detection for planetary exploration. The goals were two-fold: to target non-Earth-centric biosignatures with the greatest possible sensitivity and to develop labeling procedures amenable to robotic implementation with technologies that are currently space qualified. A wide panel of commercially available dyes that target specific biosignature molecules was screened, and those with desirable properties (i.e., minimal binding to minerals, strong autofluorescence contrast, no need for wash steps) were identified. We also explored the potential of semiconductor quantum dots (QDs) as bacterial and space probes. A specific instrument for space implementation is suggested and discussed.     

Planetary Protection and the astrobiological exploration of Mars: Proactive steps in moving forward

Future efforts towards Mars exploration should include a discussion about the effects that the strict application of Planetary Protection policies is having on the astrobiological exploration of Mars, which is resulting in a continued delay in the search for Martian life. As proactive steps in the path forward, here we propose advances in three areas. First, we suggest that a redefinition of Planetary Protection and Special Regions is required for the case of Mars. Particularly, we propose a definition for special places on Mars that we can get to in the next 10–20?years with rovers and landers, where try to address questions regarding whether there is present-day near-surface life on Mars or not, and crucially doing so before the arrival of manned missions. We propose to call those special places “Astrobiology Priority Exploration” regions (APEX regions). Second, we stress the need for the development of robotic tools for the characterization of complex organic compounds as unequivocal signs of life, and particularly new generations of complex organic chemistry and biosignature detection instruments, including advances in DNA sequencing. And third, we advocate for a change from the present generation of SUV-sized landers and rovers to new robotic assets that are much easier to decontaminate such as microlanders: they would be very small with limited sensing capabilities, but there would be many of them available for launch and coordination from an orbiting platform. Implementing these changes will help to move forward with an exploration approach that is much less risky to the potential Mars biosphere, while also being much more scientifically rigorous about the exploration of the “life on Mars” question – a question that needs to be answered both for astrobiological discovery and for learning more definitive lessons on Planetary Protection.     

Life at the wedge: the activity and diversity of arctic ice wedge microbial communities

The discovery of polygonal terrain on Mars underlain by ice heightens interest in the possibility that this water-bearing habitat may be, or may have been, a suitable habitat for extant life. The possibility is supported by the recurring detection of terrestrial microorganisms in subsurface ice environments, such as ice wedges found beneath tundra polygon features. A characterization of the microbial community of ice wedges from the high Arctic was performed to determine whether this ice environment can sustain actively respiring microorganisms and to assess the ecology of this extreme niche. We found that ice wedge samples contained a relatively abundant number of culturable cells compared to other ice habitats (～10(5) CFU·mL(-1)). Respiration assays in which radio-labeled acetate and in situ measurement of CO(2) flux were used suggested low levels of microbial activity, though more sensitive techniques are required to confirm these findings. Based on 16S rRNA gene pyrosequencing, bacterial and archaeal ice wedge communities appeared to reflect surrounding soil communities. Two Pseudomonas sp. were the most abundant taxa in the ice wedge bacterial library (～50%), while taxa related to ammonia-oxidizing Thaumarchaeota occupied 90% of the archaeal library. The tolerance of a variety of isolates to salinity and temperature revealed characteristics of a psychrotolerant, halotolerant community. Our findings support the hypothesis that ice wedges are capable of sustaining a diverse, plausibly active microbial community. As such, ice wedges, compared to other forms of less habitable ground ice, could serve as a reservoir for life on permanently cold, water-scarce, ice-rich extraterrestrial bodies and are therefore of interest to astrobiologists and ecologists alike. .