The project aims to build competence at FOI to improve civil preparedness in laboratory capability to identify and track infectious microorganisms and to support the Armed Forces in a useful way in their future development within the B-analysis area for B-agents that are considered as potential antagonistic threats. The activities within the project during the past three years have been organized into five work packages: i) Collaboration in the European Biodefence Laboratory Network (EBLN), (ii) Development of metagenomics for identification of B-agents in environmental samples, (iii) Development of statistical frameworks for the calculation of evidence value in a microbial forensic investigation, (iv) Genotyping, geographic distribution and genetic diversity of tularemia in the environment, and (v) Updating and supplementing the existing microbial reference collections with new isolates and maintenance of the viability of the strains included in the FOI BSL-3 strain collections. The output of the cooperation in EBLN has resulted in access to reference strain collections for many different potential threat agents and the corresponding genome sequences, as well as the harmonization of methods for bioinformatic analysis of genomic data in order to be able to exchange raw data between laboratories. Reference databases of genomic data and reference strains collections of threat agents are invaluable tools for the development of modern methods for Banalysis. Two different variants of metagenomics, shotgun-sequencing and amplicon-sequencing have evaluated in work package (ii). The results show that shot-gun sequencing can be used to identify specific genes in a complex sample. However, further development is needed to definitively ascertain which bacteria are present in the sample. Amplicon-sequencing has been evaluated for the detection of fecal contamination in water. Overall, the results show that this is a useful method for detection of fecal contamination and for microbial source tracking. Frameworks for the calculation of evidentiary value in the event of a biological attack have been developed in work package (iii). We have shown in our examples that it is possible to calculate evidence values even for scenarios where near-identical strains have been used. The methodology developed is generic and can be used to evaluate the strength of different hypotheses in all types of microbial source tracking scenarios, not only in forensic contexts. Within the work package (iv) we have shown that knowledge of the local natural genotype distribution and genotype frequencies for infectious agents is crucial to be able to detect anomalies that may indicate deliberate release. We have also shown that there is a significant environmental background of closely related tularemia bacteria in the environment that is not harmful to humans but which cross-reacts with current genetic and immunological methods. Furthermore, we have identified a sharp boundary in the north-south direction through Germany that separates two different clones of tularemia. By mapping this distribution, we can now determine whether a tularemia sample comes from Western or Eastern Europe. We have in work package (v) continued the updating and supplementation of the existing strain collections with new isolates during the three year period. The project has, during the three-year period, followed the research plan with very small deviations and the objectives formulated at the beginning of 2012 has largely been achieved and in some areas even been surpassed.