Chitin deacetylases including CDA1 and CDA2, containing a chitin deacetylase domain and an LDL domain, have been reported to be essential for cuticle structure differentiation in different insect species. However, it is yet unexplored whether CDA1 and CDA2 activity is needed for the function of the cuticle as a barrier against pathogen and xenobiotics penetration. In this study, we studied the efficiency of fungal infection in the migratory locust Locusta migratoria in dependence of LmCDA1 and LmCDA2 function. Second instar nymphs injected with dsRNA against LmCDA1 and LmCDA2 transcripts were less resistant against the infection by the fungus Metarhizium anisopliae than control nymphs. At the same time, permeability to organophosphorus pesticides was increased in these nymphs. Interestingly, the CHC amounts at the cuticle surface were unaffected upon LmCDA1 and LmCDA2 reduction. These results suggest that the barrier function of the locust cuticle not only depends on surface CHCs, but also on an intact procuticle.