Abstract:[Objective] Biolog-FF system with 95 kinds of carbon source was used to exploit the metabolism feature of four fungi in chlortetracycline degradation. [Methods] The changes of absorbance values of the four strains in 95 kinds of carbon sources were determined in different time. The total organic carbon (TOC) contents were measured after the four strains were inoculated into the chlortetracycline solid waste in different time. [Results] The four strains digested different carbon sources, and the activities of them were quite distinct. 41 kinds, 39 kinds, 15 kinds and 14 kinds of carbon were used by Penicillium citrinum LJ318, Trichoderma harzianum LJ245, Penicillium spinulosum LJ236 and Penicillium oxalicum LJ302, respectively. The average activity digesting carbon of LJ245 and LJ318 were significantly higher than that of the strain LJ236 and LJ302. All the four strains could utilize sugars, amino acids, polymers and so on. [Conclusion] Carbon metabolism of LJ245 and LJ318 in residue were significantly faster than LJ236 and LJ302. This show the same results as what the Biolog system drew out. Biolog technology is an effect method in the study of carbon metabolic characteristics of fungi. This study would build a scientific foundation for investigating the characteristics of fungal carbon metabolism, and for the bio-degradation of chlortetracycline in environment.