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A RANDOM GRID BASED MOLECULAR EPIDEMIOLOGICAL STUDY ON EBLV ISOLATES FROM GERMANY

A RANDOM GRID BASED MOLECULAR EPIDEMIOLOGICAL STUDY ON EBLV ISOLATES FROM GERMANY. C. Freuling 1 , N. Johnson 2 , D. Marston 2 , T. Selhorst 1 , L. Geue 1 , A. Fooks 2 , N.Tordo 3 and T. Müller 1

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A RANDOM GRID BASED MOLECULAR EPIDEMIOLOGICAL STUDY ON EBLV ISOLATES FROM GERMANY

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  1. A RANDOM GRID BASED MOLECULAR EPIDEMIOLOGICAL STUDY ON EBLV ISOLATES FROM GERMANY C. Freuling1, N. Johnson2, D. Marston2, T. Selhorst 1, L. Geue1, A. Fooks2, N.Tordo3 and T. Müller 1 Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, D-16868 Wusterhausen, Germany Veterinary Laboratories Agency (Weybridge), Surrey, United Kingdom Unité de la Rage and Laboratoire des Lyssavirus, Institut Pasteur, Paris, France

  2. Objective • Molecular characterization of a representive panel of German EBLV isolates • Germany has one of the highest numbers • Geographic distribution as a key determination for selection in contrast to random sampling • Spatiotemporal correlation to phylogenetic information

  3. Bat rabies in Europe 1977-2006 N= 831

  4. Materials and Methods • Application of a random grid with 30 km length • Selection of 1 isolate per grid cell • 48 out of 120 isolates selected • Phylogenetic analysis based on complete N-gene sequences

  5. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results • Identification of EBLV1b isolates from the southwest of Germany 5776N.seq EBLV1a EBLV1b

  6. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results 5776N.seq

  7. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results 5776N.seq

  8. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results 5776N.seq

  9. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results 5776N.seq

  10. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results 5776N.seq

  11. 4895N.seq 5778N.seq 67 5782N.seq 96 3132N.seq 13450N.seq 80 13452N.seq 59 6795N.seq 15548N.seq 57 13394N.seq 36 38 915N.seq 11647N.seq 13323N.seq Bat-Hamburg-N.seq 50 76 12865N.seq 13415N.seq 68 4644N.seq 74 64 13416N.seq 13414N.seq 13445N.seq 5226N.seq 15 13408N.seq 13406N.seq 13409N.seq 8624N.seq 65 12871N.seq 13407N.seq 5254N.seq 2976-N.seq 19 64 992.seq 65 3131N.seq 13418N.seq 5250N.seq 63 59 976N.seq 30 13401N.seq 60 13428N.seq 33 70 13438N.seq 41 13453N.seq 13439N.seq 15730N.seq 933N.seq 99 52 5248N.seq 65 10927N.seq 58 77 10924N.seq 13422N.seq 13403N.seq 15571N.seq 99 5006N.seq 0.005 Results 5776N.seq

  12. Results • short genomic insertions in the 3‘ UTR of the N – gene • 6 bp insertion in EBLV-1b (Johnson et al, 2007) • 1 bp insertion in EBLV-1a M N G L P 3‘ 5‘ Is. No 976 AATTGGAAAGAAAAA------AA-CTAACACCACT Is. No 5006 AATTGAAAAGAAAAAGAAAAAAA-CTAACACCACT Is. No 15571 AATTGAAAAGAAAAAGAAAAAAA-CTAACACCACT Is. No 5776 AATTGGAAAGAAAAA------AAACTAACACCACT Is. No 3132 AATTGGAAAGAAAAA------AAACTAACACCACT

  13. Conclusions/Discussion • Further confirmation of EBLV1b in southern Germany • High sequence homology in N-Gene • Indication for geographical clustering of EBLV1a • No species differences • Identification of short genomic insertions in the 3‘ UTR of the N-gene • Further research nessesary • Other gene segments (e.g. G, ψ – pseudogene) • More isolates from different time periods

  14. Thanks for attention!!

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