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. 2017 Dec 1;7:16719. doi: 10.1038/s41598-017-16749-9

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Feeding of uninfected Ixodes ricinus nymphs on chronically infected mice does not affect the Borrelia spirochete load in the mouse skin surrounding the tick feeding lesion. Mice were infected with Borrelia burgdorferi ss strain N40 via (A) intradermal or (B) intraperitoneal inoculation. At 40 days post-infection, the mice were infested with 5 to 10 uninfected I. ricinus nymphs. At different time points during nymphal attachment (5 h, 24 h, 3d, 5d, 7d), the mice were sacrificed and the mouse skin surrounding the tick feeding lesions was excised. The Borrelia load in the mouse skin was estimated using qPCR. Different mouse organs were tested by culture for seven days to determine the presence of Borrelia. Tissue biopsies that yield spirochete-negative cultures were retested using qPCR. The blood remained spirochete-negative over the duration of the tick blood meal, with the exception of one mouse at day 3 that had been inoculated intradermally.