Research Journal of Microbiology1816-4935xxxx-xxxxScience International10.3923/jm.2008.1.8Brucella abortus Fermenter Cultural Conditions and LPS Extraction Method for Antigen Production]]>ShapouriReza MobarezAshraf MohabatiAhmadiHojat TabaraieBahman DoustReza Hosseini NorozianDariush HosseiniAhmad Zavaran SiadatDavar 1200831We modified Brucella fermentation medium from FAO/WHO for enhancing of Brucella abortus S99 biomass. The modified media composed of 15 g L-1 peptic digest animal tissue, 15 g L-1 pancreatic digest of casein, 10 g L-1 yeast extract and 0.10 g L-1 sodium bisulphate. Glucose was added during the incubation by fed-batch method (1-30 g L-1). Agitation speed and air flow rates were controlled at 300-500 rpm and 4-8 L min-1, respectively. Cell density was 9-10% and viable count was 3-3.3x1011 mL-1. The modified conditions enhance the biomass production to more 2 times than the FAO/WHO method. Three methods were accomplished for LPS production: Extraction by butanol with enzymatic digestion, hot phenol extraction with enzymatic digestion, modified hot phenol with trichloroacetic acid (TCA) procedure. Yield of LPS extraction was 0.2, 0.8 and 1.3%, respectively. Method III results in a greater yield of LPS which is 6 and 1.5 times the yields of methods I and II, respectively. Protein contamination of LPS was <2, <2 and <2.9% and nucleic acid contamination of LPS was <1, <1 and <1.4%, respectively. The ketodeoxyoctonate content of LPS (in each of the three methods) was in agreement with ketodeoxyoctonate values obtained previously for highly purified LPS of B. abortus. According to present study, hot phenol with trichloroacetic acid (TCA) procedure is the most suitable procedure for large-scale LPS production from Brucellae, which can be employed for the production of Brucella biomass for vaccine and antigen preparations.]]>Alton, G.G., L.M. Jones, R.D. Angus and J.M. Verger,19881st Edn.,Pages: 174Pages: 174Bhattacharjee, A.K., L.L. Van De Verg, M.J. Izadjoo, L. Yuan, T.L. Hadfield, W.D. Zollinger and D.L. Hoover,2002Brucella melitensis lipopolysaccharide as a noncovalent complex with Neisseria meningitides group B outer membrane protein.]]>7033243329Campos, M.A., G.M.S. Rosinha, I.C. Almeida, X.S. Salgueiro and B.W. Jarvis et al.,2004Brucella abortus infection in mice.]]>72176186Celik, E. and P. Calik,2004Bacillus species.]]>20491499Cheema, J.J.S., N.V. Sankpal, S.S. Tambe and B.D. Kulkarni,20021813561365Corbel, M.J., C.D. Bracewell, E.L. Thomas and K.P.W. Gill,1979Brucella Species.]]>1979pp: 86-89pp: 86-89Corbel, M.J. and D.F.D. Hendry,19831983Dodge, T.C. and J.M. Gerstner,2002E. coli.]]>7712381245Fernandez-Prada, C.M., E.B. Zelazowska, M. Nikolich, T.L. Hadfield, R.M. Roop, G.L. Robertson and D.L. Hoover,2003Brucella melitensis and human phagocytes: Bacterial surface O-polysaccharide inhibits phagocytosis, bacterial killing and subsequent host cell apoptosis.]]>7121102119Freer, E., E. Moreno, I. Moriyon, J. Pizarro-Cerda, A. Weintraub and J.P. Gorvel,1996Brucella sp. counterparts.]]>17858675876Godfroid, J. and A. Kasbohrer,200290135145Goldstein, J., T. Hoffman, C. Frasch, E.F. Lizzio and P.R. Beining et al.,1992Brucella abortus is less toxic than that from Escherichia coli, suggesting the possible use of B. abortus or LPS from B. abortus as a carrier in vaccines.]]>6013851389Gorvel, J.P. and E. Moreno,2002Brucella intracellular life: From invasion to intracellular replication.]]>90281297Hendry, D.F.D., M.J. Corbel, R.A. Bell and J.A. Stack,1985Brucella antigen production and standardization.]]>1985Huang, L.Y., A.M. Krieg, N. Eller and D.E. Scott,19996762576263Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall,1951193265275Moreno, E.,2002903138Morrison, D.C. and L. Leive,1978E. coli O111:B4 prepared by two extraction procedures.]]>25029112919Nielsen, K.,200290447459Phillips, M., G. Pugh and B. Deyoe,198950311317Porte, F., A. Naroeni, S. Ouahrani-Bettache and J.P. Liautard,2003Brucella suis lipopolysaccharide O antigen in phagosomal genesis and in inhibition of phagosome-lysosome fusion in murine macrophages.]]>7114811490Pulendran, B., P. Kumar, C.W. Cutler, M.V. Mohamadzadeh and D.J. Banchereau,2001in vivo.]]>16750675076Refai, M.,20029081110Rittig, G., A. Kaufmann, A. Robins, B. Shaw and H. Sprenger et al.,2003Brucella induce different intracellular trafficking and cytokine/chemokine release in human monocytes.]]>7410451055Schurig, G.G., N. Sriranganathan and M.J. Corbel,200290479496Tsai, C.M. and C.E. Frasch,1982119115119Weissbach, A.,1959Escherichia coli B.]]>1234705709Weynants, V., D. Gilson, A. Cloeckaert, A. Tibor and P.A. Denoel et al.,1997Brucella species by competition binding assays with monoclonal antibodies.]]>6519391943