The present work aimed to study the cellular, biochemical and molecular biomarkers in the digestive glands and hemocytes of Modiolus barbatus and whether there is a hierarchy in their response to thermal stress. We determined a) the neutral red retention assay (NRR) in heamotocytes and b) the lysosomal membrane stability (LMS), the levels of second messenger cAMP, the activity of acetylcholinesterase (AChE) in the digestive glands of Modiolus barbatus after acclimation to 18°C, 24°C, 28°C or 30°C for 30 days. Moreover, in order to estimate the threshold of temperature inducing expression of stress proteins we determined the levels of Hsp70 and Hsp90 in the digestive glands. Hsps are expressed at lower temperature than those causing reduction in the LMS and NNR times. The reduction in the LMS and NNR times at high temperatures of acclimation might be related to inability of Modiolus barbatus to gain energy from the ingested food. © 2011 Elsevier Ltd.