Widespread and largely unknown prophage activity, diversity, and function in two genera of wheat phyllosphere bacteria.

Environmental bacteria host an enormous number of prophages, but their diversity and natural functions remain largely elusive. Here, we investigate prophage activity and diversity in 63 Erwinia and Pseudomonas strains isolated from flag leaves of wheat grown in a single field. Introducing and validating Virion Induction Profiling Sequencing (VIP-Seq), we identify and quantify the activity of 120 spontaneously induced prophages, discovering that some phyllosphere bacteria produce more than 108 virions/mL in overnight cultures, with significant induction also observed in planta. Sequence analyses and plaque assays reveal E. aphidicola prophages contribute a majority of intraspecies genetic diversity and divide their bacterial hosts into antagonistic factions engaged in widespread microbial warfare, revealing the importance of prophage-mediated microdiversity. When comparing spontaneously active prophages with predicted prophages we also find insertion sequences are strongly correlated with non-active prophages. In conclusion, we discover widespread and largely unknown prophage diversity and function in phyllosphere bacteria.

Pheno- and Genotyping of Three Novel Bacteriophage Genera That Target a Wheat Phyllosphere Sphingomonas Genus.

Bacteriophages are viral agents that infect and replicate within bacterial cells. Despite the increasing importance of phage ecology, environmental phages-particularly those targeting phyllosphere-associated bacteria-remain underexplored, and current genomic databases lack high-quality phage genome sequences linked to specific environmentally important bacteria, such as the ubiquitous sphingomonads. Here, we isolated three novel phages from a Danish wastewater treatment facility. Notably, these phages are among the first discovered to target and regulate a Sphingomonas genus within the wheat phyllosphere microbiome. Two of the phages displayed a non-prolate Siphovirus morphotype and demonstrated a narrow host range when tested against additional Sphingomonas strains. Intergenomic studies revealed limited nucleotide sequence similarity within the isolated phage genomes and to publicly available metagenome data of their closest relatives. Particularly intriguing was the limited homology observed between the DNA polymerase encoding genes of the isolated phages and their closest relatives. Based on these findings, we propose three newly identified genera of viruses: Longusvirus carli, Vexovirus birtae, and Molestusvirus kimi, following the latest ICTV binomial nomenclature for virus species. These results contribute to our current understanding of phage genetic diversity in natural environments and hold promising implications for phage applications in phyllosphere microbiome manipulation strategies.

Simulation and identification of foodborne outbreaks in a large supermarket consumer purchase dataset.

Foodborne outbreaks represent a significant public health burden. Outbreak investigations are often challenging and time-consuming, and most outbreak vehicles remain unidentified. The development of alternative investigative strategies is therefore needed. Automated analysis of Consumer Purchase Data (CPD) gathered by retailers represents one such alternative strategy. CPD-aided investigations do not require trawling questionnaires to create a hypothesis and can provide analytical measures of association by direct data analysis. Here, we used anonymized CPD from 920,384 customers enrolled in Norway's largest supermarket loyalty program to simulate foodborne outbreaks across a range of different parameters and scenarios. We then applied a logistic regression model to calculate an odds ratio for each of the different possible food vehicles. By this method, we were able to identify outbreak vehicles with a 90% accuracy within a median of 6 recorded case-patients. The outbreak vehicle identification rate declined significantly when using data from only one of two retailers involved in a simulated outbreak. Performance was also reduced in simulations that restricted analysis from product ID to the product group levels accessible by trawling questionnaires. Our results show that-assuming agreements are in place with major retailers-CPD collection and analysis can solve foodborne outbreaks originating from supermarkets both more rapidly and accurately than than questionnaire-based methods and might provide a significant enhancement to current outbreak investigation methods.