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. 2017 Jun 13;7:3441. doi: 10.1038/s41598-017-03724-7

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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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Humoral response to immunisation. Antibody titers were evaluated by ELISA, using HKEc P4 strain as antigen. Results obtained before immunisation (I0) and 2 weeks after the booster immunization (R14) are presented for the adjuvant only (Control), intramuscular (IM) and intramuscular and intramammary (IMM) groups. (a) antibody titers in the IgM isotype; (b) antibody titers in the IgG₂ sub-isotype. *p < 0.05, **p < 0.01, Friedman test and post hoc Dunn’s multiple comparison test.