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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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Systemic symptoms. Systemic clinical score (a), intra-ruminal temperature (b) following intramammary challenge with E. coli P4 (0 h) and cumulative systemic score (c) over the first 80 hours following intramammary challenge in first lactation heifers injected with adjuvant only (black circles), in heifers immunized with intramuscular injections only (blue squares) or immunized with intramuscular and intramammary injections (red triangles). Means and error bars, corresponding to standard error of the mean (a and b) or standard deviation (c), are presented. Letters (a,b,c) indicate a difference (p value < 0.05) between control and IM groups, control and IMM groups, and IM and IMM groups respectively. Asterisk indicate a difference (P < 0.05) between immunization groups.