Growth comparison of several Escherichia coli strains exposed to various concentrations of lactoferrin using linear spline regression

摘要

BackgroundWe wanted to compare growth differences between 13 Escherichia coli strains exposed to various concentrations of the growth inhibitor lactoferrin in two different types of broth (Syncase and Luria-Bertani (LB)). To carry this out, we present a simple statistical procedure that separates microbial growth curves that are due to natural random perturbations and growth curves that are more likely caused by biological differences.Bacterial growth was determined using optical density data (OD) recorded for triplicates at 620 nm for 18 hours for each strain. Each resulting growth curve was divided into three equally spaced intervals. We propose a procedure using linear spline regression with two knots to compute the slopes of each interval in the bacterial growth curves. These slopes are subsequently used to estimate a 95% confidence interval based on an appropriate statistical distribution. Slopes outside the confidence interval were considered as significantly different from slopes within. We also demonstrate the use of related, but more advanced methods known collectively as generalized additive models (GAMs) to model growth. In addition to impressive curve fitting capabilities with corresponding confidence intervals, GAM’s allow for the computation of derivatives, i.e. growth rate estimation, with respect to each time point.