Image j

SPSS 21.0, GraphPad Prism 7.0, R project and Image J were used for data processing and mapping. Data are expressed as the mean±standard deviation (SD), and count data are presented as the frequency. For variables that did not fit a normal distribution, Mann Whitney U-tests were performed for comparisons. Differences between two groups were evaluated using the Student’s
t-test when F-test validated the homogeneity of variance or the Welch
t-test when inhomogeneity of variance existed. All experiments were repeated at least three times. Differences are considered significant at
P ≤ 0.05.

Statistical Chi squared test for equality of proportions was applied in order to correlate urinary OspA outcome (detectable or non-detectable) to clinical LB diagnosis and serology. Power calculations were performed in order to estimate the power of the test, given the number of samples in each group, the proportions of urinary OspA outcome, and a significance level of 0.05. Calculations were performed using R software (http://www.r-project.com).
Western blotting band intensity was quantified with imagej software (imagej.nih.gov/ij/index.html">http://imagej.nih.gov/ij/index.html) by selecting the area of interest and calculating area, mean and standard deviation of selection peR software instructions. Blast analysis and protein alignment was performed using pBLAST [32 (link)]. Search parameters were as follows: query sequence: KTSTLTISVNSKKTTQLVFTKQDTITVQKYDSAGT, Database Name: non redundant, Program: BLASTP 2.2.31+.

All image analysis was done using ImageJ (http://rsb.info.nih.gov/ij), and data was further analyzed using self-developed Jupyter notebooks (https://jupyter.org/) and R-scripts (https://www.r-project.org/; available upon request).