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Volume 2, Number 3—July 1996

Dispatch

Legionella-Like Amebal Pathogens––Phylogenetic Status and Possible Role in Respiratory Disease

Adenike Adeleke*, Janet Pruckler†, Robert Benson†, Timothy Rowbotham‡, Mahmoud Halablab*, and Barry Fields†
Author affiliations: *Kings’ College, London, UK; †Centers for Disease Control and Prevention, Atlanta, Georgia, USA; ‡Public Health Laboratory, Leeds, UK

Main Article

Table 2

16S rRNA similarities (%) between members of the genus Legionella and different LLAP strains.

L1
L2
L3
L4
SL
L6
L7
L8
L9
L10
L11
L12
Lpn
Lfl
Lgr
Lts
Lpr
Lst
Ldm
Lwd
Cox
L1 94.1 93.6 93.9 93.7 94.0 93.7 94.1 93.7 94.4 94.1 94.1 93.8 96.6 93.7 94.0 93.9 94.3 94.1 94.0 85.4
L2 98.9 96.9 99.2 99.8 99.4 95.4 99.3 96.6 97.4 97.4 95.8 94.5 96.2 96.3 96.3 96.7 96.8 96.3 85.6
L3 96.9 99.4 99.0 99.5 94.9 99.5 96.3 97.0 97.0 95.8 93.9 95.7 96.2 96.1 96.5 96.4 96.0 85.6
L4 97.0 97.0 97.2 95.1 97.1 96.0 98.5 98.5 95.5 93.4 95.5 95.9 95.9 96.0 95.9 96.0 85.2
SL 99.1 99.9 94.9 99.8 96.3 97.2 97.2 95.8 94.1 95.5 96.1 95.9 96.4 96.3 96.0 85.6
L6 99.3 95.1 99.2 96.8 97.3 97.3 95.6 94.4 96.1 96.4 96.3 96.5 96.5 96.1 85.5
L7 95.0 99.9 96.5 97.4 97.4 95.9 94.1 95.7 96.3 96.0 96.6 96.5 96.1 85.8
L8 94.9 94.9 95.7 95.7 96.1 95.1 95.9 95.9 95.9 95.8 96.1 95.4 85.5
L9 96.4 97.3 97.3 95.8 94.0 95.6 96.2 95.9 96.5 96.4 96.0 85.8
L10 96.3 96.3 96.7 94.4 96.3 95.9 97.0 96.3 96.4 96.6 85.9
L11 100.0 96.3 94.0 96.3 97.0 96.7 96.9 96.5 96.5 85.0
L12 96.3 94.0 96.3 97.0 96.7 96.9 96.5 96.5 85.0
Lpn 94.3 96.8 96.9 97.3 97.5 97.0 96.8 85.5
Lfl 94.4 94.9 95.0 95.2 95.4 95.1 86.2
Lgr 97.1 97.2 97.4 97.6 96.4 85.5
Lts 97.8 98.1 97.3 96.8 85.8
Lpr 98.1 98.4 98.0 86.3
Lst 98.4 97.6 85.7
Ldm 98.0 86.1
Lwd 85.8

L. LLAP, Lpn. L. pneumophila, Lfl. L. feeleii, Lgr. L. gratiana, Lts. L. tusconensis, Lpr. L. parisiensis, Lst. L. steigerwaltii, Ldm. L. dumoffii, Lwd. L. wadsworthii, Cox. Coxiella burnetii.

Main Article

1Genomic DNA was isolated by standard methods and extracted with phenol/chloroform. A 1400-bp 16S rDNA segment was amplified by using primers specific for Eubacteria and based on a consensus of Legionella 16S rRNA sequences. Each strain was sequenced at least 3 times to ascertain accuracy of obtained results. 16S rRNA sequences for all Legionella species, those of some previously sequenced LLAPs and Coxiella burnetii were obtained from GenBank/EMBL and aligned with sequences generated in this study, using the GCG sequence analysis package (version 8.0). Comparisons were performed at NCBI using the BLAST network service (32). Ambiguous and hypervariable regions were removed before phylogenetic analysis, which was carried out on 53 strains for 1303 nucleotides by the neighbor-joining method from the Phylogeny Inference Package (Phylip) (33), version 3.5. Multiple datasets (X100) were analyzed, and different distance models were compared to ensure reliability. Some of the 16S rRNA sequences included in the data analysis are of unpublished and undescribed stains. Accession numbers for sequences included in this study are as follows: LLAP-1 U64034, LLAP-2 U44909, LLAP-3 X60080, LLAP-4 X97357, LLAP-6 X97357, LLAP-7 U44910, LLAP-8 U64035, LLAP- 9 U44911, LLAP-10 X97363, LLAP-11 X97362, LLAP-12 X97366. L. adelaidensis Z49716, L.anisa X733394, L. birminghamensis Z49717, L. brunensis X73403, L. cherrii X73404, L. cincinnatiensis X73407, Coxiella burnetii M21291, L. donaldsonii Z49724, L.dumoffii X73405, L.erythra Z32638, L. fairfieldensis Z49722, L. feelii X73395, L. geestiana Z49723, L. gormanii Z32639, L. gratiana Z49725, L. hackeliae M36028, L. israelensis X73408, L. jamestowniensis X73409, L. jordanis X73396, L. lansingensis Z49727, L. londiniensis Z49728, L. longbeachae M36029, L. maceachernii X60081, L. micdadei M36032, L. moravica Z49729, L. nautarum Z49730, L. oakridgensis X73397, L. parisiensis Z49731, L. pneumophila M59157, L. quateirensis Z49732, L. worsliensis Z49739, L. quinlivanii Z49733, L. rubrilucens X73398, L. santicrucis Z49735, Sarcobium lyticum X66835, L. shakespearei Z49736, L. spiritensis M36030, L. steigerwaltii X73400, L. sainthelensi Z49734, L. tusconensis Z32644, L. wadsworthii X73401.

2The Remel Poly-ID kit used in this study is designed to identify 22 species and 31 serogroups of Legionella (34,35).

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