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Moreover, this study is the first to highlight some biofilm development in in vitro gastrointestinal modelling systems.Directed acyclic graphs (DAGs) have had a major impact on the field of epidemiology by providing straightforward graphical rules for determining when estimates are expected to lack causally interpretable internal validity. Much less attention has been paid, however, to what DAGs can tell researchers about effect measure modification and external validity. In this work, we describe two rules based on DAGs related to effec