Avian caeca contain a large and diverse population of bacteria. Certain genera, including Bifidobacterium, are thought to exert health-promoting effects. Two media were evaluated to determine their sensitivity and selectivity for bifidobacteria in the hen caecal samples: modified Wilkins-Chalgren agar (MW; Oxoid) with the addition of glacial acetic acid (1 ml/l) and mupirocin (100 mg/l) and modified TPY agar (MTPY; ADSA, Spain) with glacial acetic acid (1 ml/l) and mupirocin (100 mg/l). The colonies arising on the plates inoculated with caecal samples were Gram stained, screened for the presence of fructose-6-phosphate phosphoketolase activity, and tested for fermentation patterns using ANAEROtest (Lachema, Czech Republic) and API 50 CHL (BioMérieux, France) kits. Both agars were selective for bifidobacteria, however, MTPY agar showed higher cfu/g than MW agar. Bifidobacterial counts were higher than 10(10) cfu/g of caecal contents using MTPY agar. Most of strains isolated from this medium fermented melibiose, sucrose, and raffinose, but not glucose. Soya peptone (5 g/l; Oxoid) stimulated the growth of glucose non-fermenting strains in complex liquid media. The results suggest that the media for selective enumeration and isolation of bifidobacteria in poultry caecal samples should not contain glucose as the sole carbon source. It can be concluded that MTPY medium is highly selective and permits the growth of both glucose fermenting and glucose non-fermenting bifidobacteria.