New Records of Ixodes affinis (Acari: Ixodidae) Parasitizing Avian Hosts in Southeastern Virginia.

Ixodes affinis Neumann (Acari: Ixodidae) is a hard-bodied tick species distributed throughout much of the southeastern United States. Although I. affinis does not parasitize humans, it is a competent vector of Borrelia burgdorferi sensu stricto, the causative-agent of Lyme disease, and thus contributes to the enzootic maintenance of this pathogen. This study presents evidence of I. affinis parasitizing five new host passerine species. During 2012-2014, 1,888 birds were captured and examined for ticks, and 18 immature I. affinis were collected from 12 birds-six Carolina Wrens (Thyrothorus ludovicianus); two Brown Thrashers (Toxostoma rufum); and one American Robin (Turdus migratorius), Eastern Towhee (Pipilo erythrophthalmus), Northern Cardinal (Cardinalis cardinalis), and White-throated Sparrow (Zonotrichia albicollis). Of 15 larvae and 3 nymphs collected, one nymph tested positive for B. burgdorferi DNA. I. affinis was found co-feeding on birds with immature Amblyomma americanum (L.), Ixodes brunneus Koch, Ixodes dentatus Marx, Ixodes scapularis Say, and Haemaphysalis leporispalustris Packard. The results of this research provide a better understanding of I. affinis hosts and identify avian taxa that may play a role in the maintenance and dispersal of this tick species.

Rickettsia parkeri Transmission to Amblyomma americanum by Cofeeding with Amblyomma maculatum (Acari: Ixodidae) and Potential for Spillover.

Amblyomma americanum (L.) is a human-biting ixodid tick distributed throughout much of the southeastern United States. Rickettsia parkeri is a member of the spotted fever group rickettsiae and causes a febrile illness in humans commonly referred to as "Tidewater spotted fever" or "R. parkeri rickettsiosis." Although the Gulf Coast tick, Amblyomma maculatum Koch, is the primary vector of R. parkeri, a small proportion of A. americanum have also been shown to harbor R. parkeri. The purpose of this investigation was to determine whether R. parkeri is spilling over into A. americanum in eastern Virginia and also to determine through laboratory experiments, whether A. americanum can acquire R. parkeri by cofeeding alongside infected ticks. Of 317 wild-caught, flat adult A. americanum tested from 29 counties and independent cities in coastal Virginia, a single female A. americanum was positive for R. parkeri, suggesting that R. parkeri is spilling over into this species, but at very low rates (<1.0%). Laboratory studies using guinea pigs indicated that nymphal A. americanum were able to acquire R. parkeri while feeding alongside infected A. maculatum and then transstadially maintain the infection. Nymphal A. americanum infected with Rickettsia amblyommii, however, were less likely to acquire R. parkeri, suggesting that infection with R. amblyommii may prevent R. parkeri from establishing infection in A. americanum.

Experimental vertical transmission of Rickettsia parkeri in the Gulf Coast tick, Amblyomma maculatum.

Rickettsia parkeri, an obligate intracellular bacterium, is a member of the spotted fever group of rickettsiae (SFGR), and is transmitted to humans and other animals by invertebrate vectors. In the United States, the primary vector of R. parkeri is the Gulf Coast tick, Amblyomma maculatum Koch. This study investigates the vertical transmission dynamics of R. parkeri within a field-derived, naturally infected colony of A. maculatum. Transovarial and transstadial transmission of the pathogen was observed over three generations, with transovarial transmission efficiency averaging 83.7% and transstadial transmission rates approaching 100%. Fitness costs were determined by comparing reproduction values of the R. parkeri-infected A. maculatum colony to values from a R. parkeri-free colony. No significant reproductive fitness costs to the host ticks were detected in the R. parkeri-infected A. maculatum colony. Significantly fewer engorged F1 nymphs and F2 larvae of the R. parkeri-free colony succeeded in molting, suggesting that there may be some advantage to survival conferred by R. parkeri. The results of this study indicate that R. parkeri is maintained in A. maculatum populations efficiently by transovarial and transstadial transmission without any noticeable effects on tick reproduction or survival.

Prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in Amblyomma americanum and Dermacentor variabilis collected from southeastern Virginia, 2010-2011.

Amblyomma americanum is the most commonly encountered tick species in southeastern Virginia, representing approximately 95% of the human-biting tick population in this area. Here we investigated the prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in questing Amblyomma americanum and Dermacentor variabilis ticks collected from multiple sites in southeastern Virginia from 2010 to 2011. Although both Ehrlichia species were detected in Amblyomma americanum, no evidence of either pathogen was found in Dermacentor variabilis. Prevalence of E. chaffeensis varied by location, ranging from 0 to 5.08% among Amblyomma americanum populations. Ehrlichia ewingii prevalence was slightly higher, ranging from 0 to 8.20% among A. americanum populations. We conclude that both pathogens are established in southeastern Virginia A. americanum populations, and that although there are no apparent temporal trends in Ehrlichia prevalence, there is variation among locations, suggesting the potential for disease hotspots.

Ticks and spotted fever group rickettsiae of southeastern Virginia.

The incidence of tick-borne rickettsial disease in the southeastern United States has been rising steadily through the past decade, and the range expansions of tick species and tick-borne infectious agents, new and old, has resulted in an unprecedented mix of vectors and pathogens. The results of an ongoing 4-year surveillance project describe the relative abundance of questing tick populations in southeastern Virginia. Since 2009, more than 66,000 questing ticks of 7 species have been collected from vegetation in a variety of habitats, with Amblyomma americanum constituting over 95% of ticks collected. Other species represented included Ixodes scapularis, Dermacentor variabilis, Amblyomma maculatum, Ixodes affinis, Haemaphysalis leporispalustris, and Ixodes brunneus. We found that 26.9-54.9% of A. americanum ticks tested were positive for Rickettsia amblyommii, a non-pathogenic symbiont of this tick species. We also found no evidence of R. rickettsii in D. variabilis ticks, although they did show low infection rates of R. montanensis (1.5-2.0%). Rickettsia parkeri and Candidatus R. andeanae were found in 41.8-55.7% and 0-1.5% A. maculatum ticks, respectively. The rate of R. parkeri in A. maculatum ticks is among the highest in the literature and has increased in the 2 years since R. parkeri and A. maculatum were first reported in southeastern Virginia. We conclude that tick populations in southeastern Virginia have recently undergone dramatic changes in species and abundance and that these populations support a variety of rickettsial agents with the potential for increased risk to human health.

Single-tube real-time PCR assay for differentiation of Ixodes affinis and Ixodes scapularis.

Ixodes affinis Neumann (1899) and Ixodes scapularis Say (1821) are tick vectors of the etiologic agent of Lyme disease, Borrelia burgdorferi sensu stricto. Ixodes affinis and I. scapularis are morphologically very similar, and as they are sympatric in the mid- and south-Atlantic U.S. coastal states, their accurate identification is crucial to studies of disease and vector ecology in this area. This work describes a rapid, single-tube SYBR(®) Green-based real-time PCR assay for differentiation of I. affinis and I. scapularis at all life stages. The assay employs 2 pairs of species-specific primers directed against the internal transcribed spacer 2 (ITS2) region of the nuclear rRNA operon. Amplification products for these primer pairs differ in size and may be differentiated with a melt curve analysis. This tool is intended as a supplement to morphological methods for accurate identification of these ticks.

Rickettsia parkeri in gulf coast ticks, southeastern Virginia, USA.

We report evidence that Amblyomma maculatum tick populations are well established in southeastern Virginia. We found that 43.1% of the adult Gulf Coast ticks collected in the summer of 2010 carried Rickettsia parkeri, suggesting that persons living in or visiting southeastern Virginia are at risk for infection with this pathogen.