Cellular Fatty Acids as Chemical Markers for Differentiation of Acinetobacter baumannii and Acinetobacter calcoaceticus

Objective Gas chromatography （GC） was used to investigate the cellular fatty acid （CFA） composition of 141 Acinetobacter boumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria. Methods Whole cell fatty acid methyl esters （FAMEs） were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System （MIS） analysis. Results All A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18：1 co9c, 16：0, Sum In Feature 3, 12：0, 17：1co8c, 3-OH-12：0, 17：0, Sum In Feature 2, 2-OH-12：0, and 18：0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18：1 co9c versus 16：0/18：1 co9c and Sum In Feature 3/18：1 co9c versus unknown 12.484/18：1 co9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16：0 versus 17：0 and Sum In Feature 3/2-OH-12：0 versus17：0 fatty acids. Conclusion The ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. colcoaceticus.

Use of Rich BHI Medium Instead of Synthetic TMH Medium for Gene Regulation Study in Yersinia pestis

Objective This study is to verify the use of rich BHI medium to substitute synthetic media for gene regulation studies in Yersinia pestis. Methods The transcriptional regulation of rovA by PhoP or via temperature upshift, and that of pla by CRP were investigated when Y. pestis was cultured in BHI. After cultivation under 26 ~C, and with temperature shifting from 26 to 37 ~C, the wild-type （WT） strain or its phoP or crp null mutant （AphoP or Acrp, respectively） was subject to RNA isolation, and then the promoter activity of rovA or plo in the above strains was detected by the primer extension assay. The rovA promoter-proximal region was cloned into the pRW50 containing a promoterless lacZ gene. The recombinant LacZ reporter plasmid was transformed into WT and AphoP to measure the promoter activity of rovA in these two strains with the ^-Galactosidase enzyme assay system. Results When Y. pestis was cultured in BHI, the transcription of rovA was inhibited by PhoP and upon temperature upshift while that ofpla was stimulated by CRP. Conclusion The rich BHI medium without the need for modification to be introduced into the relevant stimulating conditions （which are essential to triggering relevant gene regulatory cascades）, can be used in lieu of synthetic TMH media to cultivate Y. pestis for gene regulation studies.