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. 2005 May;71(5):2616-25.
doi: 10.1128/AEM.71.5.2616-2625.2005.

Molecular Identification and Analysis of Borrelia Burgdorferi Sensu Lato in Lizards in the Southeastern United States

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Molecular Identification and Analysis of Borrelia Burgdorferi Sensu Lato in Lizards in the Southeastern United States

Kerry Clark et al. Appl Environ Microbiol. .
Free PMC article

Abstract

Lyme borreliosis (LB) group spirochetes, collectively known as Borrelia burgdorferi sensu lato, are distributed worldwide. Wild rodents are acknowledged as the most important reservoir hosts. Ixodes scapularis is the primary vector of B. burgdorferi sensu lato in the eastern United States, and in the southeastern United States, the larvae and nymphs mostly parasitize certain species of lizards. The primary aim of the present study was to determine whether wild lizards in the southeastern United States are naturally infected with Lyme borreliae. Blood samples obtained from lizards in Florida and South Carolina were tested for the presence of LB spirochetes primarily by using B. burgdorferi sensu lato-specific PCR assays that amplify portions of the flagellin (flaB), outer surface protein A (ospA), and 66-kDa protein (p66) genes. Attempts to isolate spirochetes from a small number of PCR-positive lizards failed. However, PCR amplification and sequence analysis of partial flaB, ospA, and p66 gene fragments confirmed numerous strains of B. burgdorferi sensu lato, including Borrelia andersonii, Borrelia bissettii, and B. burgdorferi sensu stricto, in blood from lizards from both states. B. burgdorferi sensu lato DNA was identified in 86 of 160 (54%) lizards representing nine species and six genera. The high infection prevalence and broad distribution of infection among different lizard species at different sites and at different times of the year suggest that LB spirochetes are established in lizards in the southeastern United States.

Figures

FIG. 1.
FIG. 1.
Unrooted neighbor-joining phylogenetic tree based on 362 bp of the flaB gene obtained from lizards in Florida and South Carolina and B. burgdorferi sensu lato reference strains. The relapsing fever group species Borrelia lonestari was included as an outgroup. Numbers at the branch nodes represent bootstrap values as percentages of 1,000 replications. All sequences obtained in this study are identified by a square symbol preceding the sequence name, and names end with the state, either Florida or South Carolina. Reference strain sequences end with the country name. The strain name for reference sequences follows the species name (e.g., B.bissettii.DN127). Letter symbols representing the specific host and/or source follow the sequence and species or strain name. Abbreviations: Aa, Amblyomma americanum; Ac, Anolis carolinensis; As, Anolis sagrei; B.b.s.s., B. burgdorferi sensu stricto; B.sp., Borrelia species; Ei, Eumeces inexpectatus; El, Eumeces laticeps; Hs, human subject; Id, Ixodes dentatus; Im, Ixodes minor; Io, Ixodes ovatus; Ip, Ixodes pacificus or Ixodes persulcatus; Ir, Ixodes ricinus; I.s.N., Ixodes scapularis nymphs; Ov, Ophisaurus ventralis; Pg, Peromyscus gossypinus; Sf, Sylvilagus floridanus; Sh, Sigmodon hispidus; Sl, Scincella lateralis; Su, Sceloporus undulatus; and Sw, Sceloporus woodi. Examining the tree from top to bottom, the first major clade, beginning with strain FL204 and ending with B.b.s.s.B31, delineates B. burgdorferi sensu stricto strains. Below this group, the next major clade groups B. andersonii strains. Below this is a clade grouping B. bissettii strains.
FIG. 2.
FIG. 2.
Unrooted maximum parsimony bootstrap consensus phylogenetic tree based on 334 bp of the ospA gene obtained from lizards in Florida and South Carolina and B. burgdorferi sensu lato reference strains. Numbers at the branch nodes represent bootstrap values as percentages of 100 replications. Sequences obtained in this study are identified by a square symbol preceding the sequence name. Ia, Ixodes affinis. See the legend to Fig. 1 for other abbreviations used in descriptive information for the sequences.
FIG. 3.
FIG. 3.
Unrooted neighbor-joining bootstrap consensus phylogenetic tree based on 233 bp of the p66 gene obtained from lizards in Florida and South Carolina and B. burgdorferi sensu lato reference strains. The relapsing fever group species Borrelia hermsii was included as an outgroup. Numbers at the branch nodes represent bootstrap values as percentages of 1,000 replications. Sequences obtained in this study are identified by a square symbol preceding the sequence name. Abbreviations: Cs, Cnemidophorus sexlineatus; Is, Ixodes scapularis or Ixodes spinipalpis; Oh, Ornithodoros hermsii; and On, Ochrotomys nuttalli. See the legend to Fig. 1 for other abbreviations used in descriptive information for the sequences.

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