Detection of Paenibacillus larvae larvae spores in honey and diseased larvae Samples by culture and PCR

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Abstract

American foulbrood (AFB) is the most serious brood disease of the honey bee. Traditional methods are
reliable but rather slow simply because they are based on biochemical, morphological and physiological
identification of cultivated isolates. The aim of this study is the detection of Paenibacillus larvae larvae
spores in honey and diseased larvae samples by culture and PCR. Therefore 54 samples of diseased larvae
and 36 honey samples, were diluted with an equal volume of distilled water and centrifuged, then the pellet
was used for bacterial culture, DNA extraction and PCR. PCR products were electrophoresed on 0.8 %
agarose gel. Five of 54 (9.3 %) larvae samples and 5 of 36 (13.9 %) honey samples were positive for
Paenibacillus larvae larvae by culture and PCR. As a result, screening of honey and larvae samples by PCR
method proved to be a reliable, fast and useful method on regional and national scale for monitoring,
controlling, and using preventive measures before the occurrence of American foulbrood damages.

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