title: Bacterial Cooperation Causes Systematic Errors in Pathogen Risk Assessment due to the Failure of the Independent Action Hypothesis.
abstract: The Independent Action Hypothesis (IAH) states that pathogenic individuals (cells, spores, virus particles etc.) behave independently of each other, so that each has an independent probability of causing systemic infection or death. The IAH is not just of basic scientific interest; it forms the basis of our current estimates of infectious disease risk in humans. Despite the important role of the IAH in managing disease interventions for food and water-borne pathogens, experimental support for the IAH in bacterial pathogens is indirect at best. Moreover since the IAH was first proposed, cooperative behaviors have been discovered in a wide range of microorganisms, including many pathogens. A fundamental principle of cooperation is that the fitness of individuals is affected by the presence and behaviors of others, which is contrary to the assumption of independent action. In this paper, we test the IAH in Bacillus thuringiensis (B.t), a widely occurring insect pathogen that releases toxins that benefit others in the inoculum, infecting the diamondback moth, Plutella xylostella. By experimentally separating B.t. spores from their toxins, we demonstrate that the IAH fails because there is an interaction between toxin and spore effects on mortality, where the toxin effect is synergistic and cannot be accommodated by independence assumptions. Finally, we show that applying recommended IAH dose-response models to high dose data leads to systematic overestimation of mortality risks at low doses, due to the presence of synergistic pathogen interactions. Our results show that cooperative secretions can easily invalidate the IAH, and that such mechanistic details should be incorporated into pathogen risk analysis.
authors.affiliation: Department of Molecular Biosciences, The University of Texas, Austin, Austin, Texas, United States of America.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Cornforth
authors.affiliation: Department of Life Sciences, Imperial College London, Silwood Park, Ascot, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Matthews
authors.affiliation: Centre for Immunity, Infection and Immunity, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Sam P
authors.initials: SP
authors.lastname: Brown
authors.affiliation: Department of Life Sciences, Imperial College London, Silwood Park, Ascot, United Kingdom.
authors.forename: Ben
authors.initials: B
authors.lastname: Raymond
mesh_terms: D000465:Algorithms; D000818:Animals; D001413:Bacillus thuringiensis / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D003140:Communicable Disease Control / Q000379:methods*; D004731:Endotoxins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity*; D054884:Host-Pathogen Interactions; D006801:Humans; D007814:Larva / Q000187:drug effects / Q000382:microbiology; D056265:Microbial Interactions; D008954:Models, Biological; D009036:Moths / Q000187:drug effects / Q000382:microbiology*; D009154:Mutation; D011994:Recombinant Proteins / Q000378:metabolism / Q000633:toxicity; D018570:Risk Assessment; D013047:Specific Pathogen-Free Organisms; D013171:Spores, Bacterial / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology; D035501:Uncertainty
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis