From 1997 to 2009, the Tick-Borne Disease Laboratory of the U.S. Army Public Health Command (USAPHC) (formerly the U.S. Army Center for Health Promotion and Preventive Medicine) screened 5286 Dermacentor variabilis ticks removed from Department of Defense (DOD) personnel, their dependents, and DOD civilian personnel for spotted fever group rickettsiae using polymerase chain reaction and restriction fragment length polymorphism analysis. Rickettsia montanensis (171/5286 = 3.2%) and Rickettsia amblyommii (7/5286 = 0.1%) were detected in a small number of samples, but no ticks were found positive for Rickettsia rickettsii, the agent of Rocky Mountain spotted fever (RMSF) until May 2009, when it was detected in one D. variabilis male removed from a child in Maryland. This result was confirmed by nucleotide sequence analysis of the rickettsial isolate and of the positive control used in the polymerase chain reaction, which was different from the isolate. Lethal effects of rickettsiostatic proteins of D. variabilis on R. rickettsii and lethal effects of R. rickettsii infection on tick hosts may account for this extremely low prevalence. Recent reports of R. rickettsii in species Rhipicephalus sanguineus and Amblyomma americanum ticks suggest their involvement in transmission of RMSF, and other pathogenic rickettsiae have been detected in Amblyomma maculatum. The areas of the U.S. endemic for RMSF are also those where D. variabilis exist in sympatry with populations of A. americanum and A. maculatum. Interactions among the sympatric species of ticks may be involved in the development of a focus of RMSF transmission. On the other hand, the overlap of foci of RMSF cases and areas of A. americanum and A. maculatum populations might indicate the misdiagnosis as RMSF of diseases actually caused by other rickettsiae vectored by these ticks. Further studies on tick vectors are needed to elucidate the etiology of RMSF.

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Polymerase chain reaction analysis of Amblyomma americanum adults, nymphs, and larvae from Aberdeen Proving Ground, MD (APG), revealed a very high prevalence of a spotted fever group (SFG) rickettsia. Restriction fragment length polymorphism (RFLP) and sequence analysis identified "Rickettsia amblyommii." This organism is not yet described or well studied, and its pathogenicity is unknown; however, investigations of the organism are warranted because of its high prevalence in A. americanum. This tick is extremely abundant at military training facilities in the south, central, and Mid-Atlantic United States, and many soldiers experience multiple concurrent tick bites. Bites by R. amblyommii-infected A. americanum may account for rates of SFG rickettsia seropositivity that are higher than reported rates of Rocky Mountain spotted fever (RMSF) cases from the same location. Seroconversion to SFG rickettsia following bites of A. americanum may suggest that R. amblyommii is infectious in humans. Subclinical infection in the numerous A. americanum tick bite victims could contaminate donated blood and compromise immunodeficient recipients. Detection of R. amblyommii in questing A. americanum larvae suggests transovarial transmission. The absence of R. rickettsii, the agent of RMSF, in A. americanum may be due to transovarial interference by R. amblyommii. The likelihood of pathogen transmission by larvae is magnified by their habit of mass attack. The very small size of the larvae is also a risk factor for pathogen transmission. High R. amblyommii prevalence in populations of A. americanum presage co-infection with other A. americanum-borne pathogens. A. americanum nymphs and adults from APG were found to be co-infected with R. amblyommii and Borrelia lonestari, Ehrlichia chaffeensis and Ehrlichia ewingii, respectively, and larval pools were infected with both R. amblyommii and B. lonestari. Co-infections can compound effects and complicate diagnosis of tick-borne disease.

No abstract

The predominant human-biting tick throughout the southeastern United States is Amblyomma americanum. Its ability to transmit pathogens causing Lyme disease-like illnesses is a subject of ongoing controversy. Results of previous testing by the Department of Defense Human Tick Test Kit Program and other laboratories indicated that it is highly unlikely that A. americanum transmits any pathogen that causes Lyme disease. In contrast, a recent publication by Clark and colleagues (K. L. Clark, B. Leydet, and S. Hartman, Int. J. Med. Sci. 10:915-931, 2013) reported detection of Lyme group Borrelia in A. americanum using a nested-flagellin-gene PCR. We evaluated this assay by using it and other assays to test 1,097 A. americanum ticks collected from humans. Using the Clark assay, in most samples we observed nonspecific amplification and nonrepeatability of results on subsequent testing of samples. Lack of reaction specificity and repeatability is consistent with mispriming, likely due to high primer concentrations and low annealing temperatures in this protocol. In six suspect-positive samples, Borrelia lonestari was identified by sequencing of an independent gene region; this is not a Lyme group spirochete and is not considered zoonotic. B. burgdorferi was weakly amplified from one pool using some assays, but not others, and attempts to sequence the amplicon of this pool failed, as did attempts to amplify and sequence B. burgdorferi from the five individual samples comprising this pool. Therefore, B. burgdorferi was not confirmed in any sample. Our results do not support the hypothesis that A. americanum ticks are a vector for Lyme group Borrelia infections. T he vectors and etiologic agents of Lyme-like diseases in the southeastern United States are a subject of ongoing controversy (1, 2). In the United States, most Lyme disease is caused by infection with Borrelia burgdorferi sensu stricto, a bacterium that is phylogenetically within the B. burgdorferi sensu lato "Lyme group" of spirochetes vectored by hard ticks. The Lyme group also includes genospecies implicated as the etiologic agents of Lyme disease in other geographic regions, including B. garinii, B. afzelii, B. spielmanii, and B. valaisiana in Europe. Additional Lyme group genospecies continue to be described. In the United States, these include B. americana, B. andersonii, and B. carolinensis, all of unknown pathogenicity, and B. bissettii, which has been implicated in cases of human illness (1). In contrast to the Lyme group, the relapsing fever (RF) group spirochetes, many of which are vectored by soft ticks, are a separate phylogenetic cluster that includes agents associated with RF disease in humans. In the United States, genospecies within the RF group include B. hermsii, B. turicatae, and B. parkeri, vectored by soft ticks, and B. miyamotoi, B. davisii, and B. lonestari, vectored by hard ticks. B. hermsii is the main cause of tick-borne relapsing fever in the United States; B. turicatae and B. parkeri have also been associated with human disease (3). B...