Transovarial Transmission of Borrelia hermsii by Its Tick Vector and Reservoir Host Ornithodoros hermsi

Transovarial passage of relapsing fever spirochetes (Borrelia species) by infected female argasid ticks to their progeny is a widespread phenomenon. Yet this form of vertical inheritance has been considered rare for the North American tick Ornithodoros hermsi infected with Borrelia hermsii. A laboratory colony of O. hermsi was established from a single infected female and two infected males that produced a population of ticks with a high prevalence of transovarial transmission based on infection assays of single and pooled ticks feeding on mice and immunofluorescence microscopy of eggs and larvae. Thirty-eight of forty-five (84.4%) larval cohorts (groups of larvae originating from the same egg clutch) transmitted B. hermsii to mice over four and a half years, and one hundred and three single and one hundred and fifty-three pooled nymphal and adult ticks transmitted spirochetes during two hundred and fourteen of two hundred and fifty-six (83.6%) feedings on mice over seven and a half years. The perpetuation of B. hermsii for many years by infected ticks only (without acquisition of spirochetes from vertebrate hosts) demonstrates the reservoir competence of O. hermsi. B. hermsii produced the variable tick protein in eggs and unfed larvae infected by transovarial transmission, leading to speculation of the possible steps in the evolution of borreliae from a tick-borne symbiont to a tick-transmitted parasite of vertebrates.

Vector Specificity of the Relapsing Fever Spirochete Borrelia hermsii (Spirochaetales: Borreliaceae) for the Tick Ornithodoros hermsi (Acari: Argasidae) Involves Persistent Infection of the Salivary Glands

The relapsing fever spirochetes Borrelia hermsii and Borrelia turicatae are each maintained and transmitted in nature by their specific tick vectors, Ornithodoros hermsi Wheeler (Acari: Argasidae) and Ornithodoros turicata (Duges), respectively. The basis for this spirochete and vector specificity is not known, but persistent colonization of spirochetes in the tick’s salivary glands is presumed to be essential for transmission by these long-lived ticks that feed in only minutes on their warm-blooded hosts. To examine this hypothesis further, cohorts of O. hermsi and O. turicata were infected with B. hermsii and examined 7–260 d later for infection in their midgut, salivary glands, and synganglion. While the midgut from all ticks of both species at all time points examined were infected with spirochetes, the salivary glands of only O. hermsi remained persistently infected. The salivary glands of O. turicata were susceptible to an early transient infection. However, no spirochetes were observed in these tissues beyond the first 32 d after acquisition. Ticks of both species were fed on mice 112 d after they acquired spirochetes and only those mice fed upon by O. hermsi became infected. Thus, the vector competency for B. hermsii displayed by O. hermsi but not O. turicata lies, in part, in the persistent infection of the salivary glands of the former but not the latter species of tick. The genetic and biochemical mechanisms supporting this spirochete and vector specificity remain to be identified.

Borrelia hermsii Acquisition Order in Superinfected Ticks Determines Transmission Efficiency

ABSTRACTMultilocus sequence typing ofBorrelia hermsiiisolates reveals its divergence into two major genomic groups (GG), but no differences in transmission efficiency or host pathogenicity are associated with these genotypes. To compare GGI and GGII in the tick-host infection cycle, we first determined if spirochetes from the two groups could superinfect the tick vectorOrnithodoros hermsi. We infected mice with isolates from each group and fed ticks sequentially on these mice. We then fed the infected ticks on naive mice and measured GGI and GGII spirochete densities in vector and host, using quantitative PCR of genotype-specific chromosomal DNA sequences. Sequential feedings resulted in dual tick infections, showing that GGI or GGII primary acquisition did not block superinfection by a secondary agent. On transmission to naive mice at short intervals after acquisition, ticks with primary GGI and secondary GGII spirochete infections caused mixed GGI and GGII infections in mice. However, ticks with primary GGII and secondary GGI spirochete infections caused only GGII infections with all isolate pairs examined. At longer intervals after acquisition, the exclusion of GGI by GGII spirochetes declined and cotransmission predominated. We then examined GGI and GGII spirochetemia in mice following single inoculation and coinoculation by needle and found that GGI spirochete densities were reduced on multiple days when coinoculated with GGII. These findings indicate that dual GGI-GGII spirochete infections can persist in ticks and that transmission to a vertebrate host is dependent on the order of tick acquisition and the interval between acquisition and transmission events.

Cotransmission of Divergent Relapsing Fever Spirochetes by Artificially Infected Ornithodoros hermsi

ABSTRACTThe soft tickOrnithodoros hermsi, which ranges in specific arboreal zones of western North America, acts as a vector for the relapsing fever spirocheteBorrelia hermsii. Two genomic groups (genomic group I [GGI] and GGII) ofB. hermsiiare differentiated by multilocus sequence typing yet are codistributed in much of the vector's range. To test whether the tick vector can be infected via immersion, noninfected, colony-derivedO. hermsilarvae were exposed to reduced-humidity conditions before immersion in culture suspensions of several GGI and GGII isolates. We tested for spirochetes in ticks by immunofluorescence microscopy and in mouse blood by quantitative PCR of thevtplocus to differentiate spirochete genotypes. The immersed larval ticks were capable of spirochete transmission to mice at the first nymphal feeding. Tick infection with mixed cultures of isolates DAH (vtp-6) (GGI) and MTW-2 (vtp-5) (GGII) resulted in ticks that caused spirochetemias in mice consisting of MTW-2 or both DAH and MTW-2. These findings show that this soft tick species can acquireB. hermsiiby immersion in spirochete suspensions, that GGI and GGII isolates can coinfect the tick vector by this method, and that these spirochetes can be cotransmitted to a rodent host.

Identification of conserved antigens for early serodiagnosis of relapsing fever Borrelia

Borrelia hermsii is a blood-borne pathogen transmitted by the argasid tick Ornithodoros hermsi. Since spirochaete clearance in mice is associated with an IgM-mediated response, an immunoproteomic analysis was used to identify proteins reactive with IgM. We report that IgM from both mice and human patients infected with B. hermsii not only reacted with the previously identified variable membrane proteins but also identified candidate antigens including heat-shock proteins, an adhesin protein, ABC transporter proteins, flagellar proteins, housekeeping proteins, an immune evasion protein, and proteins with unknown function. Furthermore, IgM reactivity to recombinant glycerophosphodiester phosphodiesterase was detected during early spirochaete infection and prior to a detectable IgG response. Lastly, a conserved hypothetical protein was produced in Escherichia coli and tested with immune serum against B. hermsii and Borrelia recurrentis. These results identify a much larger set of immunoreactive proteins, and could help in the early serodiagnosis of this tick-borne infection.

Transmission Of Borreua Hermsii, The Agent Of Relapsing Fever, By The Tick Vector Ornithodoros Hermsi

Relapsing fever, a disease characterized by recurrent episodes of high fevers, is caused by geographically distinct spirochetes of the genus Borrelia,transmitted by ticks of the genus Ornithodoros. In the Northwestern United States, the soft tick Ornithodoros hermsi has been identified as the vector for the spirochete Borrelia hermsii. The life cycle of O.hermsi includes larval and multiple nymphal stages prior to full maturation into an adult male or female (Fig.1). Progression into each stage requires a blood-meal typically provided by squirrels and chipmunks, and incidentally humans. Feeding is rapid, lasting 10-60 minutes, and during this time an infected tick can transmit the agent of relapsing fever, B. hermsii. Following ingestion, spirochetes are initially found in the tick midgut. Within 1-3 weeks, they are found in other organs, including the central ganglion and salivary glands. Since saliva is the primary mode of transmission of these bacteria during tick feeding, we assessed by electron microscopy the structural and functional relationships between the spirochetes and the salivary glands.