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. 2021 Mar 17;11:6095. doi: 10.1038/s41598-021-85624-5

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Immunization with EGF domains affects tick feeding and interferes with tick transmission of B. burgdorferi. B. burgdorferi-infected ticks were allowed to feed on mice immunized with Is86 EGF-1, -2, or -3. Ticks that fed on mice immunized with adjuvant only were used as controls. The parameters of tick feeding were recorded. (A) Mean number of replete ticks, with error bars showing standard error of mean (± SEM). Fully replete ticks were collected and counted (n = 30, 24, 29, 27/group, respectively). The amount of EGF-1 ticks collected on day 4 was significantly lower (**p < 0.01). (B) Median tick engorgement weight, with error bars showing 95% confidence intervals (CI). The weights of individual fully replete ticks (n = 30, 27, 30, 30/group, respectively) were measured. Data points with zero values indicate unattached live or dead ticks collected 24 h post infestation. Fully replete EGF-1 ticks weighed significantly less (**p < 0.01). (C) Mean tick molting rate, with error bars showing ± SEM. The collected ticks were allowed to molt (n = 21, 15, 20, 17/group, respectively) in the incubator, and the percentage of molted ticks was calculated. The asterisks indicate statistically significant differences at p < 0.05. (D) Mean B. burgdorferi burden in mouse skin. This experiment represents the transmission of spirochetes from infected ticks to naïve mice. Murine skin samples were collected 10 days after tick feeding. Spirochete levels were evaluated by measuring copies of flaB using qPCR and normalizing against mouse β-actin. The B. burgdorferi burden was significantly reduced in mice immunized with EGF-1, as compared to controls (*p < 0.05). Bars represent the mean ± SEM of four qPCR analyses of B. burgdorferi levels from two independent animal experiments. (E) Median spirochete burden in ticks (n = 3/group), with error bars showing 95% CI. Spirochete burdens in fully replete ticks were assessed by measuring copies of flaB using qPCR and normalizing against tick β-actin. There was no difference among all EGF domain-immunized and control groups.