Transposon mutagenesis in bifidobacterium breve: construction and characterization of a Tn5 transposon mutant library for bifidobacterium breve UCC2003

Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities.

Baseline analysis of the existing capacities and needs for capacity building for Marine Strategy Framework Directive implementation in the Celtic Seas sub-region.

This report evaluates the existing situation in the Celtic Seas sub-region and determines the current state of preparedness for transboundary management of marine ecosystems and MSFD implementation. Recommendations for capacity building are provided through the analysis of the existing conflicts and potential synergies between relevant policies, institutions and information resources for MSFD implementation across the region. This report strives to empower stakeholders through the provision of a sound baseline with accurate and up-to-date information on the current status of MSFD implementation, potential opportunities and suggested approaches for building capacities in their region and across the Celtic Seas. It is evident that there are a number of national marine planning processes currently underway and at different stages throughout the United Kingdom and the pre-planning context for MSP in Ireland. On a similar note, this evaluation of MSFD implementation progress to-date in the United Kingdom, Ireland and France highlights that each Member State has implemented the legal and procedural requirements of preparatory steps in differing manners and using different time scales. This variance across the sub-region has the potential to impact the achievement of GES by 2020 across the Celtic Seas.