The authors investigated the impact of the extension of the El Kantra Channel on the composition and structure of macrobenthic assemblages in Boughrara Lagoon (Gulf of Gabes, Tunisia along with the use of 4 biotic indices (AMBI, BENTIX, M-AMBI and TUBI). Thirteen stations were sampled seasonally in 2012–2013. Forty-one species were found in 2012–2013 not recorded in 2009–2010, including 20 species of polychaetes belonging to the trophic groups of deposit-feeders and carnivores which are expected to increase in areas disturbed by organic pollution. During the survey, we recorded a high fish mortality, essentially caused by the development of harmful algal blooms (HAB) which increased organic matter deposition, thus inducing polychaete development. This seems to weaken the bio-indicating power of biotic indices used here which, paradoxically, classified all sampled stations at a high ecological status. A review of these indices and their applicability to all marine environments is recommended.