The aim of this work was to analyse the applicability of electrical conductivity sensors for on-line monitoring the start-up period of an anaerobic fixed-bed reactor. The evolution of bicarbonate concentration and methane production rate was analysed. Strong linear relationships between electrical conductivity and both bicarbonate concentration and methane production rate were observed. On-line estimations of the studied parameters were carried out in a new start-up period by applying simple linear regression models, which resulted in a good concordance between both observed and predicted values. Electrical conductivity sensors were therefore identified as an interesting method for monitoring the start-up period of anaerobic fixed-bed reactors due to its reliability, robustness, easy operation, low cost, and minimum maintenance compared with the currently used sensors.