Survey of ticks collected in Mississippi for Rickettsia, Ehrlichia, and Borrelia species

Jerome Goddard, John W Sumner, William L Nicholson, Christopher D Paddock, John Shen, Joseph Piesman
Journal of Vector Ecology 2003, 28 (2): 184-9
From November 1999 through October 2000, we tested ticks collected from vegetation as well as from deer, dogs, and humans for spotted fever group (SFG) rickettsiae, Ehrlichia chaffeensis, and Borrelia spp. spirochetes. A total of 149 adult ticks representing four species was collected from 11 collection sites from southwestern to northern Mississippi. Amblyomma americanum was most commonly collected (n=68), followed by Ixodes scapularis (n=53). The bird tick, Ixodes brunneus (usually rare), was the third most commonly collected tick (n=17). Eleven Dermacentor variabilis were also collected. Ticks were cut longitudinally to make smears on three microscope slides. The remaining body parts were frozen at -65 degrees C for additional testing. Tick smears were stained by direct immunofluorescence assays (DFA) for Rickettsia spp. and Borrelia spp., while indirect immunofluorescence assays (IFA) were used for Ehrlichia spp. The corresponding tick for each positive smear was evaluated using PCR analysis. None of the 149 ticks tested was DFA positive for Borrelia spp. However, smears of 30 (20%) and 32 (22%) ticks reacted with anti-E. chaffeensis sera and anti-R. rickettsii conjugate (known to react with several members of the spotted fever group), respectively. None of the ticks staining with the IFA for Ehrlichia was positive for E. chaffeensis using PCR. However, 23 (72%) of 32 FA-positive ticks for SFG rickettsiae yielded amplicons of the appropriate size when tested using a PCR assay for SFG rickettsiae, corresponding to an overall infection rate with SFG rickettsiae among the collected ticks of 15%. Smears of 12 (71%) of 17 I. brunneus revealed abundant bacilliform bacteria. PCR amplification of DNA from a single I. brunneus containing these bacteria was performed using universal primers for the 16S rRNA gene as well as Borrelia-specific primers. The predominant sequence obtained using the universal primers did not match any sequence in GenBank, but it showed 91% identity with an endosymbiont of Acanthoamoeba. Other sequences represented in the top 50 Basic Local Alignment Search (BLAST) scores were primarily from soil bacteria, although some similarity to several Anaplasma species and Ehrlichia risticii was indicated. The significance of this finding remains undetermined.

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