Discriminative canonical analysis of 87 biometric parameters in the marine and lagoon atherinids of 'Atherina boyeri' complex from the Mediterranean Sea helps defining three distinct atherinid groups: marine punctuated, marine unpunctuated and lagoon atherinids. Each atherinid group constitutes a clearly independent original entity. Besides, each one is a more or less heterogeneous group with geographical disparities, characterising specimen collected from France and Tunisia. Concordance of biometric, biochemical and genetic results as well help define two new species of atherinids: Atherina punctata = punctuated marine atherinids, and Atherina lagunae = atherinids living in lagoon environments.