In addition, 6 SP600125 classical reactions, i.e. nitrite, nitrate, pyrazinamidase, Voges-Proskauer medium, urease and H2S production, and three controls, i.e. peptidase control, pyrazinamidase control and assimilation control were included.
Figure 2 The Brucella specific Micronaut™ microtiter plate. Design of the newly developed Brucella specific Micronaut™ microtiter plate including 93 selected substances. Glu(pNA)-OH (ENAOH), Pyr-pNA (PYRNA) (constantly negative reaction), and H-hydroxyprolin-βNA (HP) (constantly strong positive reaction) turned out to be key substances useful for the identification of the genus Brucella and its differentiation from other bacteria [Additional file 7]. A stable negative buy PND-1186 reaction for D-threitol (D-TOL) and mostly positive reactions for L-alanine (L-Ala), D-alanine (D-Ala), propionic Selleckchem KPT-8602 acid (Propn), L-proline (L-Pro), D-proline (D-Pro), and D-serine (D-Ser) could be observed in B. melitensis. B. microti which also makes use of alanine and proline could be separated from B. melitensis by a constantly negative reactivity for Propn and D-Ser. A positive myo-inositol
(INOL) reaction seemed to be characteristic for most B. melitensis strains and B. inopinata. Bis-p-nitrophenyl phosphate pH 7.5 (BISPH7), p-nitrophenyl phosphate di(2-amino-2-ethyl-1,3-propanediol) pH 7.5 (PHOS7), and p-nitrophenyl-a-d-glucopyranoside pH 7.5 (aGLU7) were found positive frequently in B. suis and regularly
in B. microti strains, variable in B. melitensis and mostly negative in B. abortus. Glutarate (Gluta) and mesaconic acid (Mesac) which were almost exclusively metabolized by B. microti might be helpful for further differentiation. P-nitrophenyl-a-d-glucopyranoside Calpain pH 5.5 (aGLU5) and p-nitrophenyl-n-acetyl-β-d-glucosaminide pH 7.5 (CHIT7) showed weak positive reactions in B. suis and B. canis and strong positive reactions in B. microti and B. inopinata. B. microti and B. inopinata exhibited outstanding metabolic capabilities in comparison to all other brucellae, sharing a series of reactions with O. anthropi and O. intermedium. Most remarkably, both species were strongly positive in the Voges-Proskauer reaction. The slow growing strains of the B. ovis group did not metabolize any carbohydrates except for D-glucose-L-cysteine (GLUCY), L(+)-arabinose (L-ARA), D-TOL, and adonite (ADON) and only a few amino acids. In addition, B. ovis strains were usually not able to deoxidize nitrite (NTI, nitrite reduction) and nitrate (NTA, nitrate reduction). Ac-Gly-Lys-βNA (AcGK) tested strongly positive in B. ovis and B. canis whereas Trp-βNA (W) regularly tested negative in these species as compared to all other Brucella spp. In comparison with other species B.