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. 2019 May 21;11(5):462.
doi: 10.3390/v11050462.

First Isolation and Phylogenetic Analyses of Tick-Borne Encephalitis Virus in Lower Saxony, Germany

Mathias Boelke  1   2 , Malena Bestehorn  3   4 , Birgit Marchwald  5 , Mareike Kubinski  6   7 , Katrin Liebig  8   9 , Julien Glanz  10   11 , Claudia Schulz  12   13 , Gerhard Dobler  14   15 , Masyar Monazahian  16 , Stefanie C Becker  17   18
Affiliations

First Isolation and Phylogenetic Analyses of Tick-Borne Encephalitis Virus in Lower Saxony, Germany

Mathias Boelke et al. Viruses. .

Abstract

Tick-borne encephalitis (TBE) is the most important tick-borne arboviral disease in Europe. Presently, the main endemic regions in Germany are located in the southern half of the country. Although recently, sporadic human TBE cases were reported outside of these known endemic regions. The detection and characterization of invading TBE virus (TBEV) strains will considerably facilitate the surveillance and assessment of this important disease. In 2018, ticks were collected by flagging in several locations of the German federal state of Lower Saxony where TBEV-infections in humans (diagnosed clinical TBE disease or detection of TBEV antibodies) were reported previously. Ticks were pooled according to their developmental stage and tested for TBEV-RNA by RT-qPCR. Five of 730 (0.68%) pools from Ixodes spp. ticks collected in the areas of "Rauher Busch" and "Barsinghausen/Mooshuette" were found positive for TBEV-RNA. Phylogenetic analysis of the whole genomes and E gene sequences revealed a close relationship between the two TBEV isolates, which cluster with a TBEV strain from Poland isolated in 1971. This study provides first data on the phylogeny of TBEV in the German federal state of Lower Saxony, outside of the known TBE endemic areas of Germany. Our results support the hypothesis of an east-west invasion of TBEV strains in Western Europe.

Keywords: field study; phylogenetic analysis; surveillance; tick; tick-borne encephalitis virus.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Phylogenetic tree of whole genome sequences available in GenBank and the two TBEV-LS-Barsinghausen/Mooshuette isolates HB IF06 8033 (MK922615) and 8040 (MK922616) and TBEV-LS-Rauher Busch P19 (MK922617) obtained in the present study (marked red). Virus isolate names are given as follows: TBEV-isolate name or number-three letter country code according to UN standard (accession number for the sequences as available from NCBI). UN country codes are: AUT: Austria; CHE: Switzerland; CZE: Czech Republic; EST: Estonia; FIN: Finland; GER: Germany; KOR: Korea; NLD: Netherlands; RUS: Russian Federation; SVN: Slovenia; SVK: Slovakia; SWE: Sweden. All sequences obtained from German isolates are marked bold, the new sequences obtained during this study are additionally marked in red. As an outgroup the Louping ill virus (KF056331.1) and TBEV-Vasilchenko (AF069066) were used.
Figure 2
Figure 2
Phylogenetic tree (Maximum likelihood Tree, MEGA6.0) of the five “TBEV-LS” isolates collected in “Barsinghausen/Mooshuette” and “Rauher Busch” (TBEV-LS-Rauher Busch-P16 (MK903683), -P19 (MK903681) and TBEV-LS-Barsinghausen/Mooshuette-P51 (MK903682), -HB IF06 8040 (MK903679) and –HB IF06 8033 (MK903680) in the German federal state of Lower Saxony, 2018, using E gene sequences. Virus isolate names are given as follows: TBEV-isolate name or number-three letter country code according to UN standard (accession number of the sequence as available from NCBI). UN country codes are: AUT: Austria; CHE: Switzerland; CZE: Czech Republic; EST: Estonia; FIN: Finland; GER: Germany; KOR: Korea; NLD: Netherlands; POL: Poland; RUS: Russian Federation; SVN: Slovenia; SVK: Slovakia; SWE: Sweden. As an outgroup strain Louping ill virus (KF056331.1), TBEV-NL (LC171402) and TBEV-Vasilchenko (AF069066) were used. All sequences obtained from German isolates are marked bold, the new sequences obtained during this study are additionally marked in red.
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