While the phenomenon of increased population tolerance to environmental contaminants has been widely reported for aquatic animal species living in contaminated ecosystems, the question of the nature of the tolerance, i.e., genetic adaptation or phenotypic plasticity, has not yet been studied to the same degree and remains under debate when explaining the variability of toxicological sensitivity among field populations. The occurrence of both processes challenges the ecological risk assessment (ERA) of chemicals, questioning the relevance of toxicity assessment procedures based on bioassays employing organisms from naive reference populations. Nonetheless, given that genetic adaptation and phenotypic acclimation are based on different mechanisms, with different persistence periods over time when exposure is stopped, gaining insight into the mechanism of tolerance to contaminants of field populations is a major concern for ERA. In this context, this study aimed to test whether genetic adaptation underlies the tolerance to cadmium (Cd) in a field Gammarus fossarum population identified in a previous study. Breeding and exposure experiments conducted in laboratory conditions revealed that: (1) first and second cohorts of neonates produced from field-collected parents present an increased Cd tolerance, which is explained by a reduction of between-brood variability of Cd sensitivity with an absence of sensitive broods compared to two reference populations rather than by the existence of more tolerant individuals; and (2) Cd tolerance was no longer evident for broods of the third cohort produced by these breeders when maintained under Cd-free conditions in the laboratory. Hence, this study drew the striking conclusion that the observation of Cd tolerance inheritance within the field population studied does not demonstrate genetic adaptation, but it results from an effect of parental exposure inducing transgenerational plasticity, which deeply influences the variability of population sensitivity. To our knowledge, this study is only the second report of increased tolerance to contaminants in a field population explicitly attributed to the phenomenon of transgenerational acclimation.