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  • Author or Editor: Janusz Kocki x
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Authors: Katarzyna Dudek, Dariusz Bednarek, Roger D. Ayling, Maria Szczotka, Ewelina IWAN and Janusz Kocki

Mycoplasma bovis is a primary infectious agent of many disorders in cattle including bovine respiratory disease. No commercial vaccines against M. bovis are available in Europe. The immune response of calves to three saponin-based adjuvants combined with a field Polish M. bovis strain was evaluated. Four groups of six calves each were injected subcutaneously with the M. bovis strain combined with either saponin, saponin + Emulsigen®, saponin + Emulsigen® + alphatocopherol acetate, or with phosphate-buffered saline as control group. Blood and nasal swab samples were collected up to day 84 post injection. All formulations effectively stimulated the humoral and the cellular immune response of the calves, but the course of the response depended on the adjuvant formulation. These immunological data provide additional information supporting the findings of previous M. bovis saponin and Emulsigen® vaccine challenge studies to facilitate the development of successful M. bovis vaccines.

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Eleven Juniperus species and a total of 62 different varieties collected at different localities in Poland were extracted with methanol in order to determine the level of umbelliferone present in the extracts. Thin-layer chromatography (TLC) analysis was performed on glass silica gel 60 F254 plates first developed with n-hexane-ethyl acetate (70:30, 7p/7p) and redeveloped after drying with dichloromethane and diethyl ether (80:20, 7p/7p). Densitometric determinations were performed in fluorescence mode by using a deuterium lamp (D2) for excitation at 326 nm and a K400 filter for measurement of the emitted light using a CAMAG TLC Scanner 3. A linear regression equation was used to determine the amount of umbelliferone in the extracts. The mobile phases used for TLC separated well umbelliferone from other compounds present in the extracts. A good linear regression model was obtained in the range of 10-80 ng with a correlation coefficient (r) of 0.99844. Juniperus communis L. “Pendula” contained the highest (88.3 mg/100 g DW) UMB content. Umbelliferone quantitative screening in so many Juniperus species has been performed for the first time. There are no reports dealing with the quantitative assessment of umbelliferone in the Juniperus species. The described method has been validated and it is simple, fast, reliable, specific, precise, and useful for routine assays of Juniperus extracts containing umbelliferone.

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