Diagnosis of honeybees' American foulbrood's causative agent Paenibacillus larvae from suspicious brood and honey by molecular and conventional methods

In Turkey, diagnosis of American Foulbrood Disease (AFB) is carried out with conventional bacteriological methods according to the "Laboratory Method Unification Manual" of Ministry of Food, Agriculture and Livestock. Diagnosis with conventional culture method is time consuming and expensive. In some countries and laboratories, molecular methods recommended by OIE could be used for faster results. This research was designed as a method comparison work where conventional methods, PCR and Real Time PCR methods will be applied parallel regarding to application on the diagnosis process. For this purpose, 96 samples consisting of larval broods, honey broods and phenomenal amounts of harvested broods and adult bees were taken from different sources. Samples were initially examined by culture methods, and then by conventional PCR and real time PCR methods. In molecular studies optimization was achieved with different primers for conventional PCR and real time PCR methods. As a result, 10 samples were found to be positive with conventional culture methods, 16 were positive with PCR and 25 were positive with Real-Time PCR. All samples found positive with culture method were also found to be positive with molecular methods. According to the findings of our work, an AFB suspicious sample can be considered as negative if it is also was found negative with molecular methods. If it is found to be positive with molecular methods, it would be appropriate that the sample should be examined and results should be evaluated with culture methods. Keywords: Honeybee, American Foulbrood, Diagnosis, PCR, Real-Time PCR