ANÁLISIS DE RELACIONES

The definitive detection and identification of abiotic and biological lipids in the subsurface of an actively serpentinizing system would be an important advance in understanding the development of pre-biotic chemistry and life in extreme environments. Accessing these environments requires complex drilling and processing procedures with numerous opportunities for contamination to occur. Microbiologists working with these tools in low biomass environments have been particularly focused on minimizing and characterizing potential contamination. They have lead the development of chemical, microbiological, and particulate tracers in an attempt to quantify contamination throughout the entire drilling process (Smith et al., 2000; Lever et al., 2006; D’Hondt et al., 2007; Santelli et al., 2010; Frise et al., 2017; Orcutt et al., in prep). For example, methods involving PFC tracers and fluorescent microspheres have been extensively implemented to measure the intrusion of drilling fluids and nonindigenous microorganisms, respectively, into drilled cores (Lever et al., 2006; Orcutt et al., in prep). However, many significant microbiological and geochemical discoveries using ODP drilled

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rock or sediment cores have been claimed without explicitly discussing the possibility of contamination (Smith et al., 2000).

Similar issues face studies of lipid biomarkers and other organic compounds in subsurface environments. As highlighted in the study of French et al., (2015), it is difficult to ascertain whether previous conclusions made using subsurface lipid biomarkers accurately represent in situ processes without in depth characterization of contaminants. Such characterization is necessary to avoid ambiguity concerning any future conclusions made using deep biosphere organic compounds. However, as demonstrated in this study, the simple adoption of contamination techniques developed primarily for tracking cells is not sufficient when sample analyses focus on lipids.

Biologically derived compounds could not be definitively identified in the Atlantis Massif samples, indicating that any active biosphere in the serpentinite subsurface is present at levels below our detection limits. The cores recovered from

the subsurface contained a series of C18 to C30 n-alkanes with 13C signatures of

-30.9‰ to -28.8‰. These straight-chain hydrocarbons were only identified in samples that had their exteriors removed with a rock saw rather than being rinsed with Milli-Q water. The similarity in alkane distributions and isotope signatures between samples and swipes of the rock saw implicate the rock saw as the source of these compounds. While it is possible that abiotically-derived compounds could

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have been present, it was impossible to identify any due to contamination. This result highlights the critical importance of minimizing and thoroughly characterizing potential contamination at all steps along the sample processing pathway in order to prevent misinterpretation.

Despite contaminant-derived compounds being present in our cut samples, the remaining cores did not contain the same alkane contamination. This suggests that many of the steps taken to prevent contamination were effective and should be implemented in future endeavors. It provides some optimism for detecting compounds in future drilling projects despite the difficulties inherent in completely eliminating contamination during drilling. However, we advise caution in the use of a rock saw to remove core exteriors. Harsher cleaning methods than solvent cleaning are necessary to remove organics from the blade. The potential loss of material during Milli-Q rinsing also make this approach somewhat problematic. Alternative mechanisms could be more successful, such as chipping away the exterior of samples or implementing the cleaning procedures of French et al. (2015). These approaches should be thoroughly tested prior to and during implementation and, similar to the requirements for microbiological studies, future publications should include the results of these assessments.

Detecting lipid biomarkers in serpentinites will continue to be a goal for researchers interested in the development of pre-biotic chemistry and the survival

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of life in the subseafloor. As this research progresses, methodological improvements to the minimization and characterization of potential contaminants should also occur.

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