Dispersant application is used as a response technique to minimize the environmental risk of an oil spill. In nearshore areas, dispersant application is a controversial countermeasure: environmental benefits are counteracted by the toxicity of dispersant use. The effects of the use of chemical dispersants on meiobenthic organisms and nematodes were investigated in a mesocosm experiment. A 20 day experiment was performed in four experimental sets of mesocosms. In three of them, sediments were contaminated, respectively by oil (500 mg kg−1), dispersed oil (oil + 5% dispersant), and dispersant alone, whereas in the last set sediments were kept undisturbed and used as a reference (Re). Our results showed that the meiobenthic response to oil contamination was rapid, for copepods and nematodes. One-way ANOVA showed a significant decrease of the abundance of copepods. In the case of nematodes, univariate and multivariate analyses indicated a clear decrease of the abundance of the species after only 20 days of pollutant exposure and thus reducing Shannon–Wiener diversity and Pielou's evenness. In contrast, Sphaerolaimus gracilis and Sabateria sp. became more frequent within disturbed assemblages and appeared to be resistant and/or opportunistic species in the presence of these kinds of toxicants. Moreover, responses of copepods and nematodes to the treatment seemed to be the same irrespective of whether only oil or oil + dispersant was performed. The main toxicities of dispersed oil come not from the “composition of a newly formed oil and oil spill dispersant mixture” but from the “quantities of increased dispersed oil droplets”.