Date of Award

December 2025

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Space Studies

First Advisor

Keith Crisman

Abstract

This study examines culture-dependent microbial monitoring in a closed lunar/Martian analog habitat, focusing on viable community dynamics relevant to long-duration missions. A standardized sampling and cultivation pipeline was implemented in ILMAH to track microbial succession and persistence under realistic operational conditions. Surface samples were collected across eight habitat zones at five mission timepoints (Days 0, 5, 10, 15, 20) during a 21-day crewed mission. Samples were concentrated post-collection and plated on chromogenic film media to quantify total aerobic bacteria (AC-R), fungi (YM-R), E. coli (EC), and S. aureus (SA). Plates were imaged and enumerated digitally; representative colonies were isolated and identified via PacBio long-read sequencing to resolve strain-level diversity and spatial-temporal patterns of viable taxa within the habitat. This research addresses significant gaps in confined-habitat biosurveillance by pairing serial, site-resolved sampling with reproducible, viability-focused workflows. The approach evaluates how material type, moisture, and human activity shape viable loads and persistence, establishing a comparable baseline for culture-based monitoring in analogs. Findings from this study aim to contribute to unified, resource-efficient microbial surveillance strategies that support contamination control and risk assessment for future exploration missions to the moon and Mars, as well as here on Earth in low-biomass settings such as cleanrooms and clinical facilities.

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