Detection of Paenibacillus larvae spores in the debris and wax of honey bee by the Tween 80 method
2007
Bzdil, J.,Statni Veterinarni Ustav, Olomouc (Czech Republic)
The aim of the present study was to validate a new method of detection of Paenibacillus larvae spores in the debris and wax of honey bee and compare it with the method commonly used in the Czech Republic, i.e. the method based on dissolving wax components of samples in toluene or benzene and releasing them in a liquid medium. The new method uses homogenization of the material at 70+/-2 deg C with distilled water and the agent Tween 80. The spores are transferred from the suspension thus created to a liquid medium diluted and treated by heat (90+/-2 deg C) in order to destroy vegetative saprophytic microflora. This medium is then inoculated on solid culture media (blood agar and MYPGP with nalidixic acid) at individual doses of 0.2 mL. After five to eight-day incubation at 37 deg C, the suspected P. larvae colonies are counted. The new method was tested using control samples and 1,509 field samples of winter debris collected in protection areas around American foulbrood outbreaks. Fourty-six field samples were P. larvae positive. The comparison of the number of spores found concurrently by both methods in 21 control samples and their benchmarking by the paired t-test method showed that the Tween method was significantly more effective in detection of P. larvae spores than the commonly used Toluene method.
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