[Research on the procedure for recovery and species identification of Legionella from surface environmental water].
OBJECTIVE
To establish a set of procedure for recovery and species identification of Legionella from the surface environmental water.


METHODS
Forty-four water samples were collected in eight parks of Guangzhou city from August to November in 2006. The bacteriologic examination was performed by cultivation on BCYEalpha plate, and 108 presumptive Legionella colonies were picked and their homogeneous relationship was analyzed by using an amplified fragment length polymorphism (AFLP) method. Species identification was carried out by latex agglutination test, biochemical characterization, analysis of cellular fatty acids composition, 16 S rRNA gene and mip gene sequencing.


RESULTS
Legionella was recovered among 27 (61.36%) samples of all eight parks, and 31 different strains were identified from those 108 presumptive Legionella isolates by AFLP method, including 20 strains of L. pneumophila, five strains of L. feeleii, four strains of L. longbeachae, one strain of L. oakridgensis and one strain of L. sainthelensi, and L. pneumophila could be easily differentiated by phenotypic and biochemical characteristics, latex agglutination test or analysis of the cellular fatty acids composition . However, uncertain factors were existing in those phenotypic identification methods as compared to the sequence analysis.


CONCLUSIONS
The taxonomic analysis of the Legionellae family should be dependent on the 16 S rRNA gene or mip gene.
Qu
Ping-Hua
PH
Department of Laboratory Medicine, Chongqing University of Medical Science, Chongqing 400016, China.
Yin
Yi-Bing
YB
Hu
Zhao-Hui
ZH
Zhu
Qing-Yi
QY
Song
Ya-Jun
YJ
Yang
Rui-Fu
RF
Liu
Yuan-Li
YL
Li
Pu
P
D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D007875:Legionella / Q000235:genetics / Q000302:isolation & purification*; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D014871:Water Microbiology
D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S