m-positive Pleomorphic Bacilli (GPPB), especially the genus Actinomyces, are part of the oral microflora and they are generally associated with cement caries. They are also found in inactive sites in periodontal disease and in pulpar infections. The aim of the present work was to find an appropriate isolation culture medium for the recovery of the genus Actinomyces from oral samples, and to propose a minimum schema of biochemical tests for the identification and differentiation of Actinomyces species from other GPPB. Samples of saliva, subgingival plaque and post-extraction acute alveolar osteitis were cultivated in Starch Casein Agar (SCA), Yeast Extract Dextrose Agar (YDA), Columbia Blood Agar 5% (BA), both with cephadroxyl (10 μL/mL) and without antibiotic. The plates were incubated in aerobic conditions, in strict anaerobic conditions and in a candle jar for 5 days at 37°C. (1) The highest recovery of Actinomyces was obtained in BA without antibiotic in a candle jar for all oral samples. (2) In all the incubated in SCA, YDA and BA with antibiotic,Actinomyces , other GPPB, Gram-positive and -negative cocci and Gram-negative bacilli showed growth in all aerobic conditions. (3) In BA medium with antibiotic the Gram-negative microflora was inhibited, the Actinomyces recovery being lower than in the other media studied.