B. melitensis attenuated strain B115 was provided by the Veterinary Laboratories Agency (VLA) of Weybridge (U.K.) and cultured to prepare the bacterial extract according to previously described protocols [21 (link), 22 (link)].
Briefly, the bacteria were cultured in 1 l of Brucella broth (Becton Dickinson, France) at 37 °C in aerobic conditions under stirring for 3 days. When the culture reached an optical density (OD) of 2.080, the broth was centrifuged at 5000 g (ALC PK131R centrifuge, Milan, Italy) for 20 min. The pellet was washed with saline solution, inactivated at 100 °C for 10 min, sonicated, centrifuged and the supernatant was dialysed against distilled water before measurement of the protein content as previously described by Corrente et al. 2015.
Briefly, the bacteria were cultured in 1 l of Brucella broth (Becton Dickinson, France) at 37 °C in aerobic conditions under stirring for 3 days. When the culture reached an optical density (OD) of 2.080, the broth was centrifuged at 5000 g (ALC PK131R centrifuge, Milan, Italy) for 20 min. The pellet was washed with saline solution, inactivated at 100 °C for 10 min, sonicated, centrifuged and the supernatant was dialysed against distilled water before measurement of the protein content as previously described by Corrente et al. 2015.
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