Bioaerosol transport in the atmosphere disperses microbial species between continents, affects human and plant health, and may influence hydrologic cycling. However, there have been few quantitative observations of bioaerosols at altitudes more than a few kilometers above the surface. Lack of data on bioaerosol distributions in the atmosphere has impeded efforts to assess the aerial dissemination of microbes and their vertical extent in the biosphere. In this study, a helium balloon payload system was used to sample microbial cells and dust particles in air masses as high as 38 km above sea level over three locations in the southwestern United States. The cell concentrations at altitudes between 3 and 29 km were highly similar (2–5 × 10⁵ cells m⁻³) and approximately threefold lower than those observed in the convective boundary layer (CBL; 1 × 10⁶ cells m⁻³), decreasing to 8 × 10⁴ cells m⁻³ at 35–38 km. The detection of adenosine triphosphate (ATP) and recovery of bacteria possessing extreme tolerance to desiccation and shortwave ultraviolet radiation confirmed that certain microorganisms have the capacity to persist at lower altitudes of the stratosphere. Our data and related calculations provide constraints on the upper altitudinal boundary for microbial habitability in the biosphere.

The study of the forces which govern the geographical distributions of life is known as biogeography, a subject which has fascinated zoologists, botanists and ecologists for centuries. Advances in our understanding of community ecology and biogeography—supported by rapid improvements in next generation sequencing technology—have now made it possible to identify and explain where and why life exists as it does, including within the microbial world. In this review, we highlight how a unified model of microbial biogeography, one which incorporates the classic ecological principles of selection, diversification, dispersion and ecological drift, can be used to explain community dynamics in the settings of both health and disease. These concepts operate on a multiplicity of temporal and spatial scales, and together form a powerful lens through which to study microbial population structures even at the finest anatomical resolutions. When applied specifically to curious strains of conjunctivitis-causing, nonencapsulated Streptococcus pneumoniae, we show how this conceptual framework can be used to explain the possible evolutionary and disease-causing mechanisms which allowed these lineages to colonize and invade a separate biogeography. An intimate knowledge of this radical bifurcation in phylogeny, still the only known niche subspecialization for S. pneumoniae to date, is critical to understanding the pathogenesis of ocular surface infections, nature of host-pathogen interactions, and developing strategies to curb disease transmission.

When Carl Sagan and Ed Salpeter envisioned potential sinkers, floaters, and hunters living in Jupiter’s clouds in 1976, the nature of life in Earth’s atmosphere remained widely unknown. Decades later, research has revealed a remarkable variety of microorganisms in our atmosphere. However, the spectral features of airborne microbes as biomarkers for detecting atmospheric life remained a mystery. Here, we present the first reflectance spectra of biopigments of atmospheric microorganisms based on laboratory cultivars of seven microbial strains isolated from Earth’s atmosphere. We show their distinct UV-resistant biosignatures and their impacts on models of diverse planetary scenarios, using Habitable Worlds Observatory parameters. The reflectance of these biopigments from aerial bacteria creates the means to detect them on other Earth-like planets. It provides a paradigm shift that moves the search for life beyond the surface of a planet to ecosystems in atmospheres and clouds.

Nanopore sequencing, as represented by Oxford Nanopore Technologies’ MinION, is a promising technology forin situlife detection and for microbial monitoring including in support of human space exploration, due to its small size, low mass (∼100 g) and low power (∼1W). Now ubiquitous on Earth and previously demonstrated on the International Space Station (ISS), nanopore sequencing involves translocation of DNA through a biological nanopore on timescales of milliseconds per base. Nanopore sequencing is now being done in both controlled lab settings as well as in diverse environments that include ground, air and space vehicles. Future space missions may also utilize nanopore sequencing in reduced gravity environments, such as in the search for life on Mars (Earth-relative gravito-inertial acceleration (GIA)g= 0.378), or at icy moons such as Europa (g= 0.134) or Enceladus (g= 0.012). We confirm the ability to sequence at Mars as well as near Europa or Lunar (g= 0.166) and lowerglevels, demonstrate the functionality of updated chemistry and sequencing protocols under parabolic flight, and reveal consistent performance acrossglevel, during dynamic accelerations, and despite vibrations with significant power at translocation-relevant frequencies. Our work strengthens the use case for nanopore sequencing in dynamic environments on Earth and in space, including as part of the search for nucleic-acid based life beyond Earth.