ABSTRACT
OBJECTIVE: To characterize a Bartonella strain M9HN-SHQ from a blood culture of cat from Henan Province,China. METHODS: The organisms were subcultured in 5% CO2 at 37 degrees C on trypticase soy agar containing 5% sheep blood for 6 to 7 days. We analyzed the isolate using whole-cell fatty acid analysis,Etest for susceptibility testing, random amplified polymorphic DNA (RAPD), pulsed-field gel electrophoresis(PFGE) and sequence analysis of 16S rRNA, gltA, groEL, ftsZ, rpoB, ribC and 16S-23S rRNA intergenic spacer region. RESULTS: Isolate M9HN-SHQ stained faintly as a gram-negative rod but was easier to visualize when stained by the Gimenez technique. Most of the biochemical and cellular fatty acid properties of strain M9HN-SHQ were typical for bacteria of the Bartonella genus. The strain was susceptible to Cefotaxime sodium, Rifampin, Ciprofloxacin and other four antibiotics. Genotypic characterization of strain M9HN-SHQ, including RAPD, PFGE was distinguishable from the reference strains of B. henselae, B. elizabethae, B. vinsonii subsp. berkhoffii and B. grahamii. Sequence analysis of the genes from the seven chromosomal regions identified the strain M9HN-SHQ as B. clarridgeiae. CONCLUSION: To our knowledge,this is the first report that documents Bartonella clarridgeiae infections of domestic cats in China.
Subject(s)
Bartonella/genetics , Cats/microbiology , Animals , Anti-Infective Agents/pharmacology , Bacterial Proteins/genetics , Bartonella/classification , Bartonella/drug effects , Cefotaxime/pharmacology , Ciprofloxacin/pharmacology , Electrophoresis, Gel, Pulsed-Field , Microbial Sensitivity Tests , Phylogeny , RNA, Ribosomal, 16S/genetics , Random Amplified Polymorphic DNA Technique , Rifampin/pharmacologyABSTRACT
Bartonella is a significant human pathogen and is the world's most common bacterial zoonosis acquired from companion animals. However, there is no uniform method for Pulse-Field Gel Electrophoresis (PFGE) for Bartonella population genetics studies. Further, some genes of Bartonella can mutate frequently and may affect the use of PFGE for Bartonella. Here we designed methods to solve these problems. We standardized the bacterial concentration, selected the appropriate digestion enzyme, optimized the electrophoretic parameters and characterized reproducibly two Bartonella species strains. Thus we optimized the PFGE procedure and determined how often Bartonella mutated. Our data shows a practical protocol for inter- and intra-species identification of Bartonella and was reproducible using two species strains that showed no mutation occurred after two passages for B. elizabethae; but mutation did occur in B. henselae.
