D4 4 Month 12- Internal technical report

Progress in the first six months
For administrative reason there was a delay in completing the contractual arrangements between the institution of the project coordinator and the individual partners. It also took some time for all of the partners to fully understand the process and there respective responsibilities. As a consequence there was a delay in the start of the secondments.

In the first six month there were secondments between EIG and Cardiff and KAU and NCDC.

Dr Ekaterine Tevdoradze from EIG visited the group at CU for 1 month for training in how to develop a standardise soil microcosm with which to assess germinant efficacy (Task 2.1). Studies suggested that we need to optimise the ratio of phage to target bacteria to achieve efficacy killing of the pathogen in soil.

Dr Leila Kvachadze from EIG visited the group at CU for 1 month for training in how to work safely in a UK containment level 2 laboratory (task 3.5).

Dr Fatih Buyuk, Ms Aliye Gulmez and Ms Elif Tazegul from KAU visited the group at NCDC for 1 month to receive training in Characterization of the genetic diversity of B.anthracis isolates (task 3.2) and – Safe working with B.anthracis biosafety 2 and 3 (task 3.5). During the visit they spent two weeks in the Genome Laboratory were they were introduced to sequencing methods and the platform which is used at NCDC to genotype bacterial isolates. The final two weeks were spent visiting the biosafetly level 2 and 3 facilities for training in the various aspects of operating safely in such faciltities.

Progress by month 12

WP1- Development and characterisation of phage based decontamination agents

Isolation of B.anthracis specific phages using the Sterne vaccine strain of B.anthracis

1. The group at CU with help from colleagues from EIG and NCDC isolated novel B.anthracis specific phages from environmental samples and determine there biological activity. Electron microscopy images of the phages can be seen below.

micro 1 micro 2 micro 3
2. Ms Aleksandra Nakonieczna, from WiHe with support form the the group at EIG isolated and characterisation B.anthracis specific bacteriophages

Researchers from NCDC received training in the isolation and characterisation of B.anthracis specific bacteriophages (task 1.2) and as a consequence should be better equipped to isolate bacteriophages from there own B.anthracis contaminated sites.
WP2- Develop of methods to enhance the activity and stability of sporicidal agents

Researchers from Georgia have received training in how to set up and run soil microscosms. They were introduced to the method that shows the effect of germinants on spore reduction in soil and the principle of adding germinants to soil to trigger the conversion of spores to vegetative form. They observed that B. anthracis failed to persist in vegetative form after it had been introduced to germinate.
WP3- Field trials, efficacy testing and assessment of environmental impact
Dr Andriy Buzun from NSC IECVM visited the group at KAU for 1 month to receive training in the identification and characterisation of potential anthrax spore decontamination test sites with a view to identifying similar sites in Ukraine.

Researchers from KAU in Turkey visited the group at NCDC in Georgia for training in the use of molecular methods to characterize the genetic diversity of B.anthracis. Researchers from NSC IECVM in Ukriane underwent similar molecular characterization training at IZSPB Italy in the method of genotyping using 15 loci Multi Locus VNTR Analysis –MLVA.

Researchers from NCDC, EIG, KAU and NSC IECVM received training in how to operate safely 8in Biosafety level 2 and 3 facilities

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