Nematodes infections are responsible for debilitating conditions and economic losses in domestic animals as well as livestock and are considered an important public health problem due to the high prevalence in humans. The nematode resistance for drugs has been reported for livestock, highlighting the importance for development of new anthelmintic compounds. The aim of the current study was to apply and compare fluorimetric techniques using Sytox and propidium iodide for evaluating the viability of C. elegans larvae after treatment with anthelmintic drugs. These fluorescent markers were efficient to stain larvae treated with ivermectin and albendazole sulfoxide. We observed that densitometric values were proportional to the concentration of dead larvae stained with both markers. Furthermore, data on motility test presented an inverse correlation with fluorimetric data when ivermectin was used. Our results showed that lower concentrations of drugs were effective to interfere in the processes of cellular transport while higher drugs concentrations were necessary in order to result in any damage to cell integrity. The methodology described in this work might be useful for studies that aim to evaluate the viability of nematodes, particularly for testing of new anthelminthic compounds using an easy, economic, reproducible, and no time-consuming technique.