Application of a polymerase chain reaction (PCR) method as a complementary tool to microscopic analysis for the detection of bones and other animal tissues in home-made animal meals

A polymerase chain reaction (PCR) method was compared with a variation of the official microscopic technique (Directive 98/88/EC) for the detection in animal meals of cereals (wheat and corn) and animal parts (bone, feathers, meat, liver, fat and blood). Microscopy successfully detected animal bones in raw feeds with a sensitivity of 1 g kg^-1, while the sensitivity of the PCR method was in the range of 5-10 g kg^-1. Microscopy also allowed the detection of animal bones and feathers in feeds processed at 115 and 133 °C but failed to detect other animal materials. The PCR method successfully detected cereals (wheat and corn) as well as meat, bone, liver, fat and feathers after processing at 115 °C for 20 min. Heating at 133 °C under overpressure (autoclave) conditions resulted in more intense DNA fragmentation and lower DNA extractability. Nevertheless, bone and liver, as well as wheat and corn in home-made animal meals, were successfully detected even after heating at 133 °C for 20 min.