Identification and characterization of the Francisella sp.strain 08HL01032 isolated in air condition systems

Abstract:

Abstract: [Objective] To identify and characterize the strain 08Hl01032 was isolated from air condition systems in the routine investigations of Legionella in Guangzhou, China, in 2008. [Methods] We adopted several phenotypic and genotypical methods, such as the growth status on various media, morphological, physical and biochemical characteristics, animal test, antibiotic susceptivities, PCR identification, sequence analysis of 16S RNA and RNA polymerase β-subunit (ropB) gene etc, to determinate the phylogenetic position and outline the basic biological characteristics. [Results] Strain 08Hl01032 was Gram-negative with polymorphic short rods or coccobacillus; with no flagella; devoid of spores; well growth on buffered charcoal yeast extraction (BCYE) agar and BCYE supplemented with glycine (3 g/L), polymyxin B sulfate (80,000 iu/L), vancomycin (1 mg/L) and cycloheximide (80 mg/L) (GVPC medium) within 2 days, but delayed growth on ordinary sheep blood agar untill 5~7 days; catalase positive; oxidase negative; no reduction of nitrate; no hydrolysis of urea; delayed fermention of glucose to produce acid; which was primarily considered as Legeionella. It was lastly identified to the genus Fransicella, characterized by a variety of biochemical and molecular phylogenetic tests, which shared the highest similarities to F. Philomiragia with 95.3 % to 16S rRNA gene of 1377 oligo nucleotides and 87.3 % to ropB gene of 367 oligo nucleotides (GenBank accession number: FJ591095, FJ939309). Growth were observed after a treatment for 10 minutes with the KCl-HCl buffer of pH 2.2, 20oC, and at 25℃, 37 oC (optimum 25~28 oC ), but not at 42 oC. The cells had capsule-like construction by transmission electron microscopy, however no virulence found to mice. [Conculsions] Strain 08Hl01032 was a potential new species of the genus Fransicella with a typical characteristic of L-cysteine growth stimulating activity, distinguishingly to Legionella with L-cysteine growth dependent activity.