Predatory bacteria are a guild of heterotrophs that feed directly on other living bacteria. They belong to several bacterial lineages that evolved this mode of life independently and occur in many microbiomes and environments. Current knowledge of predatory bacteria is based on culture studies and simple detection in natural systems. The ecological consequences of their activity, unlike those of other populational loss factors like viral infection or grazing by protists, are yet to be assessed. During large-scale cultivation of biological soil crusts intended for arid soil rehabilitation, episodes of catastrophic failure were observed in cyanobacterial growth that could be ascribed to the action of an unknown predatory bacterium using bioassays. This predatory bacterium was also present in natural biocrust communities, where it formed clearings (plaques) up to 9 cm in diameter that were visible to the naked eye. Enrichment cultivation and purification by cell-sorting were used to obtain co-cultures of the predator with its cyanobacterial prey, as well as to identify and characterize it genomically, physiologically and ultrastructurally. A Bacteroidetes bacterium, unrelated to any known isolate at the family level, it is endobiotic, non-motile, obligately predatory, displays a complex life cycle and very unusual ultrastructure. Extracellular propagules are small (0.8-1.0 µm) Gram-negative cocci with internal two-membrane-bound compartmentalization. These gain entry to the prey likely using a suite of hydrolytic enzymes, localizing to the cyanobacterial cytoplasm, where growth begins into non-compartmentalized pseudofilaments that undergo secretion of vesicles and simultaneous multiple division to yield new propagules. I formally describe it as Candidatus Cyanoraptor togatus, hereafter Cyanoraptor. Its prey range is restricted to biocrust-forming, filamentous, non-heterocystous, gliding, bundle-making cyanobacteria. Molecular meta-analyses showed its worldwide distribution in biocrusts. Biogeochemical analyses of Cyanoraptor plaques revealed that it causes a complete loss of primary productivity, and significant decreases in other biocrusts properties such as water-retention and dust-trapping capacity. Extensive field surveys in the US Southwest revealed its ubiquity and its dispersal-limited, aggregated spatial distribution and incidence. Overall, its activity reduces biocrust productivity by 10% at the ecosystem scale. My research points to predatory bacteria as a significant, but overlooked, ecological force in shaping soil microbiomes.

There is an estimated five trillion pieces of plastic in the global ocean, with 4.8 to 12.7 million metric tons entering the ocean annually. Much of the plastic in the ocean is in the form of microplastics, or plastic particles <5mm in size. Microplastics enter the marine environment as primary or secondary microplastics; primary microplastics are pre-manufactured micro-sized particles, such as microbeads used in cosmetics, while secondary microplastics form from the degradation of larger plastic objects, such water bottles. Once in the ocean, plastics are readily colonized by a consortium of prokaryotic and eukaryotic organisms, which form dense biofilms on the plastic; this biofilm is termed the “plastisphere”. Despite growing concerns about the ecological impact of microplastics and their respective plastispheres on the marine environment, there is little consensus about the factors that shape the plastisphere on environmentally relevant secondary microplastics. The goal of my dissertation is to comprehensively analyze the role of plastic polymer type, incubation time, and geographic location on shaping plastisphere communities attached to secondary microplastics. I investigated the plastisphere of six chemically distinct plastic polymer types obtained from common household consumer products that were incubated in the coastal Caribbean (Bocas del Toro, Panama) and coastal Pacific (San Diego, CA) oceans. Genotyping using 16S and 18S rRNA gene amplification and next-generation Illumina sequencing was employed to identify bacterial and eukaryotic communities on the polymer surfaces. Statistical analyses show that there were no polymer-specific assemblages for prokaryotes or eukaryotes, but rather a microbial core community that was shared among plastic types. I also found that rare hydrocarbon degrading bacteria may be specific to certain chemical properties of the microplastics. Statistical comparisons of the communities across both sites showed that prokaryotic plastispheres were shaped primarily by incubation time and geographic location. Finally, I assessed the impact of biofilms on microplastic degradation and deposition and conclude that biofilms enhance microplastic sinking of negatively buoyant particles and reduce microplastic degradation. The results of my dissertation increases understanding of the factors that shape the plastisphere and how these communities ultimately determine the fate of microplastics in the marine environment.

Cyanobacteria and algae living inside carbonate rocks (endoliths) have long been considered major contributors to bioerosion. Some bore into carbonates actively (euendoliths); others simply inhabit pre-existing pore spaces (cryptoendoliths). While naturalistic descriptions based on morphological identification have traditionally driven the field, modern microbial ecology has shown that this approach is insufficient to assess microbial diversity or make functional inferences. I examined endolithic microbiomes using 16S rRNA genes and lipid-soluble photosynthetic pigments as biomarkers, with the goal of reassessing endolith diversity by contrasting traditional and molecular approaches. This led to the unexpected finding that in all 41 littoral carbonate microbiomes investigated around Isla de Mona (Puerto Rico) and Menorca (Spain) populations of anoxygenic phototrophic bacteria (APBs) in the phyla Chloroflexi and Proteobacteria, were abundant, even sometimes dominant over cyanobacteria. This was not only novel, but it suggested that APBs may have been previously misidentified as morphologically similar cyanobacteria, and opened questions about their potential role as euendoliths. To test the euendolithic role of photosynthetic microbes, I set a time-course experiment exposing virgin non-porous carbonate substrate in situ, under the hypothesis that only euendoliths would be able to initially colonize it. This revealed that endolithic microbiomes, similar in biomass to those of mature natural communities, developed within nine months of exposure. And yet, APB populations were still marginal after this period, suggesting that they are secondary colonizers and not euendolithic. However, elucidating colonization dynamics to a sufficiently accurate level of molecular identification among cyanobacteria required the development of a curated cyanobacterial 16S rRNA gene reference database and web tool, Cydrasil. I could then detect that the pioneer euendoliths were in a novel cyanobacterial clade (named UBC), immediately followed by cyanobacteria assignable to known euendoliths. However, as bioerosion proceeded, a diverse set of likely cryptoendolithic cyanobacteria colonized the resulting pore spaces, displacing euendoliths. Endolithic colonization dynamics are thus swift but complex, and involve functionally diverse agents, only some of which are euendoliths. My work contributes a phylogenetically sound, functionally more defined understanding of the carbonate endolithic microbiome, and more specifically, Cydrasil provides a user-friendly framework to routinely move beyond morphology-based cyanobacterial systematics.