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Bacterial defense against protozoan grazing has been shown to occur in many different bacteria. Predation resistance traits may however be plastic, making bacterial com munities resilient or resistant to predation perturbations. We studied the adaptation of pre dation resistance traits in bacteria isolated from a microcosm experiment. In the initial microcosm ex periment the predation pressure on bacteria varied markedly, while changes in the bacterial community composition could not be verified. Seven bacteria were isolated from the microcosm (Micrococcus sp., Rhodobacter sp., Paracoccus sp., Shewanella sp., Rhizobium sp. and 2 un identified species) and these were repeatedly exposed to high predation by the ciliate Tetrahymena pyriformis. High variations in edibility and rate of adaptation of predation resistance traits were observed among the strains. The initial mortality rate of the different bacterial taxa and the change over time varied by a factor of 7 and 24, respectively. Rhodobacter sp. was already predation resistant at the start of the experiment and did not change much over time, while Micrococcus sp., Paracoccus sp. and Shewanella sp. initially were relatively edible and later developed predation resistance. In conclusion, we show that rapid adaptation of predation resistance traits is common among bacteria in an aquatic microbial community, and that a single test of a bacterium’s edibility will in many cases not be enough to fully understand its ecological role, as it will not reveal the potential adaptive response. The results suggest the potential of rapid changes of predation resistance as a mechanism for bacterial communities to be resilient to variations in predation disturbances.

Predation-resistance has been suggested to be a key for persistence of pathogenic bacteria in aquatic environments. Little is known about driving factors for different types of protozoa resistant bacteria (PRB). We studied if presence of PRB is linked to specific plankton taxa, the aquatic nutrient state, or predation pressure on bacteria. Nineteen freshwater systems were sampled and analyzed for PRB, plankton composition and physicochemical variables. Three PRB genera were identified; Pseudomonas, Mycobacterium and Rickettsia. Use of minimum entropy decomposition algorithm and phylogenetic analysis showed that different nodes (representing OTUs of high taxonomic resolution) matched to environmental isolates of the three genera. Links between the PRB genera and specific plankton taxa were found, but showed different relationships depending on if 18S rRNA OTU or microscopy data were used in the analysis. Mycobacterium spp. was negatively correlated to aquatic nutrient state, while Pseudomonas showed the opposite pattern. Rickettsia spp. was positively related to predation pressure on bacteria. Both Mycobacterium and Rickettsia were more abundant in systems with high eukaryotic diversity, while Pseudomonas occurred abundantly in waters with low prokaryotic diversity. The different drivers may be explained by varying ecological strategies, where Mycobacterium and Rickettsia are slow growing and have an intracellular life style, while Pseudomonas is fast growing and opportunistic. Here we give an insight to the possibilities of newly advanced methods such as sequencing and oligotyping to link potential pathogens with biomarkers. This as a tool to assist predictions of the occurrence and persistence of environmental pathogens.

Pathogenic bacteria occur naturally in aquatic systems. Co-existence of bacteria and protozoa has led to development of predation resistance strategies, which is suggested to serve as a driver for evolution of pathogenic bacteria. However, the ecological mechanisms for selection for different types of predation resistant and pathogenic bacteria are poorly known. To disentangle effects from nutrient availability and protozoan predation pressure on the occurrence of varying predation resistant bacterial genera, an enrichment-dilution experiment was performed where an aquatic bacterial community was exposed to protozoa. Operational taxonomical units, specific for three predation resistant bacterial genera were identified; Pseudomonas, Rickettsia and Mycobacterium. These genera are also known to harbor species that are potentially pathogenic to mammals. Rickettsia and Mycobacterium were promoted where protozoa were abundant and the predation pressure high, while Pseudomonas dominated the bacterial community at the highest nutrient level where the predation pressure on bacteria were low. Our study thus indicates that waters of all nutrient states can harbor pathogenic bacteria, but that bacteria with different ecological strategies occur depending on nutrient level and perturbation. The generative model approach presented here provide a possibility to integrate environmental data in prediction models of pathogens in complex environments.

The long co-existence of bacteria and protozoa in natural ecosystems has led to the evolution of different bacterial predation-resistance mechanisms¹, which in turn may have triggered development of mammal pathogens², such as the tularemia bacterium Francisella tularensis³. We studied links between environmental persistence and pathogenicity of Francisella tularensis subsp. holarctica (F. t. holarctica), by comparing its growth in association with an aquatic amoeba and a murine macrophage. A virulent wild-type strain and four isogenic mutations with different functional protein deletions were compared; DsbA^{4, 5} a membrane lipoprotein with disulfide oxidoreductase activity important for proper folding in Francisella tularensis; Hfq⁶ a pleiotropic regulatory RNA binding protein; PilA^{7, 8} a type IV pilus subunit and PglA⁹ a protein involved in O-linked protein glycosylation. DsbA was found to be essential for bacterial growth in association with both amoeba and macrophage, while PglA did not affect bacterial persistence in any of the hosts. Absence of PilA and Hfq had marked negative effect on the bacterial cell counts in amoeba, while growth was only slightly impaired in the macrophage. Functional similarities for bacterial persistence in both hosts highlight eco-evolutionary links between persistence of intracellular pathogenic bacteria in aquatic systems and mammal hosts.