In order to explore the effects of long-term fertilization on the microbiological characters of red soil, soil samples were collected from a 19-year long-term experimental field in Qiyang of Hunan, with their microbial biomass carbon (MBC) and nitrogen (MBN) and microbial utilization ratio of carbon sources analyzed. The results showed that after 19-year fertilization, the soil MBC and MBN under the application of organic manure and of organic manure plus inorganic fertilizers were 231 and 81 mg x kg(-1) soil, and 148 and 73 mg x kg(-1) soil, respectively, being significantly higher than those under non-fertilization, inorganic fertilization, and inorganic fertilization plus straw incorporation. The ratio of soil MBN to total N under the application of organic manure and of organic manure plus inorganic fertilizers was averagely 6.0%, significantly higher than that under non-fertilization and inorganic fertilization. Biolog-ECO analysis showed that the average well color development (AWCD) value was in the order of applying organic manure plus inorganic fertilizers = applying organic manure > non-fertilization > inorganic fertilization = inorganic fertilization plus straw incorporation. Under the application of organic manure or of organic manure plus inorganic fertilizers, the microbial utilization rate of carbon sources, including carbohydrates, carboxylic acids, amino acids, polymers, phenols, and amines increased; while under inorganic fertilization plus straw incorporation, the utilization rate of polymers was the highest, and that of carbohydrates was the lowest. Our results suggested that long-term application of organic manure could increase the red soil MBC, MBN, and microbial utilization rate of carbon sources, improve soil fertility, and maintain a better crop productivity.