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National Repository of Grey Literature 2 records found  Search took 0.01 seconds. 
Methods for the detection of Paenibacillus larvae and Melissococcus plutonius using real-time PCR
 Hrabák, Jaroslav ; Vlková, Kateřina ; Erban, Tomáš
The methodology is intended for diagnostic laboratories focused on bee infections. Detection of the causative agents of American foulbrood (Paenibacillus larvae) and European foulbrood (Melissococcus plutonius) is essential to prevent the spread of those highly contagious infections of a honey bee (Apis mellifera) diseases. In case of the suspicion, an examination of bee colonies in protective zones of the disease is indicated. The second possibility of use is the confirmation of the infection based on the detected clinical symptoms in the bee colony. The main target user in the Czech Republic are laboratories investigating animal diseases for the State Veterinary Administration (SVS) (accredited laboratories) in case of suspected disease and investigations in protective zones. Other target users are laboratories investigating bee colonies for reasons of preventive surveillance based on the request of beekeepers. The PCR method enables fast and reliable detection of P. larvae and M. plutonius from different samples taken from the bee colony. Compared to classic microbiological techniques, it allows to provide a result within a few hours of receiving the sample in the laboratory. In this way, it is possible to ensure, above all during the period of intense flight activity of bees, early intervention in bee colonies suspected of the occurrence of the mentioned diseases and to prevent the spread of the disease to nearby habitats of bee colonies.
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Molecular epidemiology of selected viral, bacterial and fungal disease of honeybees in the Czech Republic
 Ryba, Štěpán ; Stopka, Pavel (advisor) ; Voříšek, Karel (referee) ; Půža, Vladimír (referee)
4 Summary Altogether, the six most common bee viruses which infect the honey bee (Apis mellifera) were monitored in the territory of the Czech Republic between 2006 and 2009. Parallel infections of viruses (DWV, ABPV and BQCV) in bee adults and parallel co- infection of viruses with fungal diseases caused by Nosema apis and Nosema ceranae were confirmed by PCR tests. A new sensitive method of detection of the originator of the American foulbrood (Paenibacillus larvae) from bee debris was developed for the practical use of detection of AFB disease in bee populations. Various approaches for the extraction of spores from bee debris and lyses of spores were compared. The sensitivity of PCR tests for the presence of Paenibacillus larvae in debris was compared with the classic cultivation method. The PCR method for the detection American foulbrood was further studied and developed to be more efficient. A new method, based on a matrix-like sample re-arrangement and a use of pooled samples, has been developed for testing 1000 samples in 35 PCR reactions. Another goal was to develop a robust and fast screening method for American foulbrood based on the cultivation test using paper sheets RIDA®COUNT with a specific cultivation medium, specific selection conditions for Paenibacillus larvae and chromogen visualization...
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