Prism statistical software

Data were expressed as mean ± standard deviation (mean ± SD) of three experiments using monocytes from three different donors. Statistical significance was measured by Student’s t-test using GraphPad Prism Statistical Software (GraphPad Software, La Jolla, USA). *P < 0.05 and **P < 0.01 indicate statistic difference between compared groups.

All experiments were repeated three times and performed in triplicate. Data were analyzed with SPSS statistical analysis software (version 22.0, IBM Corp., Armonk, NY, USA) using the probit analysis statistical method. The LC₅₀ values (with 95% confidence limits) were calculated. Differences among the results were considered to be statistically significant when the p value was <0.05. MS Excel 2007 was used to determine the regression equation (Y = mortality; X = concentrations), and the LC₅₀ was derived from the obtained best-fit line. One-way ANOVA followed by post hoc Tukey test and two-way ANOVA followed by Duncan’s test and t test were applied to determine significant differences in teratogenicity, behavior, learning impairment assessment, and gene expression between exposed and control groups. Data are presented as mean values ± standard error of the mean (SEM), with significant differences relative to the control (p-values ≤ 0.05). GraphPad Prism statistical software (GraphPad Software, San Diego, CA, USA) was used for all graphs.

Growth curves were constructed by plotting the logarithm of the number of Salmonella vs. time at the different condition assays. Each point of the growth curve corresponds to the average value of all the samples analyzed (at least three replicates). The curves obtained were fitted with the Baranyi and Roberts model (Equations (1) and (2)) [22 (link)]: $Y_t=Y_0+{\mu }_{max}·A_t-\frac{Y_{max}-Y_0}{M}·ln\left[1-e^{-M}+\left(e^{-M}·\frac{Y_{max}-Y_0-{\mu }_{max}·A_t}{Y_{max}-Y_0}\right)\right]$
$A_t=t-\lambda ·\left[1-\frac{1}{h_0}·ln\left(1-e^{-h_0·\frac{t}{\lambda }}+e^{-h_0·\left(\frac{t}{\lambda }-1\right)}\right)\right]$
where Y_t is the Log₁₀ of cell concentration at time t (CFU/mL), Y₀ is the Log₁₀ of the initial cell concentration (CFU/mL), Y_max is the Log₁₀ of maximum cell concentration (CFU/mL), μ_max is the maximum growth rate (h⁻¹), λ is the lag phase (h), and M and h₀ are curvature parameters, taking them as constant values and with both set at a value of 10. For this purpose, GraphPad PRISM^® statistical software (GraphPad Prism version 8.00 for Windows, GraphPad Software, San Diego, CA, USA) was used. The same software was used for calculating the goodness of fit parameters (R² and RMSE) and to carry out the statistical analysis (Student’s t-tests and ANOVA). Differences were considered significant for p ≤ 0.05.

The normality of the variables was evaluated by the Kolmogorov-Smirnov test, and Student's t-test or Mann Whitney U test was used to compare the characteristics of the study population and DNA damage in relation to characteristics of the study population. The statistical differences of damage observed for groups by the comet assay and BMNCyt assay were determined by ANOVA test. Values of P < 0.05 were considered statistically significant. All analyses were performed using the GraphPad PRISM statistical software (GraphPad Inc., San Diego, CA, USA).