Reservoir competence of white-footed mice for Lyme disease spirochetes

Using the vector tick, Ixodes dammini, we described the reservoir competence of the white-footed mouse, Peromyscus leucopus, for the Lyme disease spirochete, Borrelia burgdorferi. Nymphal I. dammini were used to infect mammals, and larval ticks were used to diagnose infection (a form of xenodiagnosis). One tick was nearly as efficient as more than 1 in transmitting the spirochete to mice. The duration of the prepatent period was about 1 week. Prevalence of infection approached 100% in ticks that fed as larvae on mice infected 2 or 3 weeks previously. Thereafter, infectivity gradually decreased, but duration exceeded 200 days. Hamsters, too, became infectious for larval I. dammini. This report formally demonstrates the life cycle of B. burgdorferi as it seems to occur in nature. We conclude that the white-footed mouse is a competent reservoir for the Lyme disease spirochete.     

Lyme disease and babesiosis: acaricide focused on potentially infected ticks

Permethrin-treated cotton, intended as rodent nesting material, was distributed in wooded sites in which the agents of Lyme disease and babesiosis were enzootic, in order to kill immature Ixodes dammini, the ticks that transmit these human pathogens. Such ticks feed most abundantly on white-footed mice (Peromyscus leucopus), apparently the main reservoir hosts of these agents, and tend to concentrate in mouse burrows. Mice captured after permethrin-treated cotton was distributed, were infested by a tenth as many ticks as were those captured in adjacent nontreated sites, a difference that continued throughout the 4-month period of observation. On average, 72% of all mice captured in treated sites were free of ticks, while virtually all mice captured in nontreated sites were infested. Voles (Microtus pennsylvanicus), however, were tick-infested, regardless of site of capture. Laboratory-reared I. dammini failed to attach to mice captured in treated sites, and most such exposed ticks died. Distribution of permethrin-treated cotton appears to be a means for preventing transmission of the pathogens that cause human babesiosis and Lyme disease.     

Travel and disease vector ticks

There are approximately twenty species of hard (ixodid) ticks worldwide that frequently affect human populations, many of which are associated with serious, sometimes fatal disease(s). When a tick travel souvenir is presented in the clinic, the risk must be immediately assessed by identifying the tick in question, ascertaining its disease vector status and determining if there has been the opportunity for the transfer of potential pathogens. This short review on identification of disease vector ticks and aspects of blood feeding and disease transmission includes the results of an examination of 59 specimens removed from UK domestic travellers and international travellers between 2002 and 2010. Sixteen tick species belonging to six genera were recorded and almost all showed evidence of blood feeding, which appears to contradict the view that because of their size, adult ticks are found early and therefore present an insignificant risk.     

Growth kinetics of the Lyme disease spirochete (Borrelia burgdorferi) in vector ticks (Ixodes dammini)

Lyme disease spirochetes (Borrelia burgdorferi) multiplied rapidly in larval Ixodes dammini, reaching a mean density of 2,735 spirochetes/tick on day 15 post-repletion. A 5-fold drop in spirochete levels occurred during the subsequent premolting period. Recently molted nymphs contained a mean of less than 300 spirochetes/tick. Following nymphal repletion, spirochete multiplication renewed, reaching a mean abundance of 61,275 spirochetes/nymph on day 75 post-repletion. A 10-fold drop in spirochete abundance occurred again when ticks molted to the adult stage. Tick-derived spirochetes proved to be infectious when greater than 10(4) spirochetes were inoculated ip into hamsters (4 of 4 animals infected). Inocula of 10(3-4) spirochetes were not always infectious (8 of 23 animals infected), and inocula of less than 10(3) spirochetes were insufficient for establishing infection (0 of 8 animals infected).     

Absence of Lyme disease spirochetes in larval Ixodes ricinus ticks

To determine which kind of spirochete infects larval Ixodes ricinus, we examined questing larvae and larvae derived from engorged females for the presence of particular spirochetal DNA that permitted species differentiation. Borrelia miyamotoi was the sole spirochete detected in larval ticks sampled while questing on vegetation. Questing nymphal and adult ticks were infected mainly by Borrelia afzelii, whereas larval ticks resulting from engorged females of the same population were solely infected by B. miyamotoi. Since larvae acquire Lyme disease spirochetes within a few hours of attachment to an infected rodent, questing larvae in nature may have acquired Lyme disease spirochetes from an interrupted host contact. Even if transovarial transmission of Lyme disease spirochetes may occasionally occur, it seems to be an exceedingly rare event. No undisputable proof exists for vertical transmission of Lyme disease spirochetes, whereas B. miyamotoi appears to be readily passed between generations of vector ticks.     

Chemotactic migration of the Lyme disease spirochete (Borrelia burgdorferi) to salivary gland extracts of vector ticks

The migration of Lyme disease spirochetes (Borrelia burgdorferi) toward salivary gland extracts (SGEs) of vector ticks was determined by a modified U-tube chemotaxis assay. Responses of cultured spirochetes to the SGEs were measured by dark-field microscopy at intervals after the initial inoculation. The average numbers of spirochetes that migrated were compared between U-tubes containing either SGEs or phosphate-buffered saline (PBS). Spirochetes showed increased migration in tubes containing SGE, and migration was approximately 10-20-fold higher than that for spirochetes observed in tubes containing PBS. In addition, the span of migration showed no significant difference relative to the origin of spirochete isolation. These results demonstrate that spirochete migration can be enhanced by SGEs from fed vector ticks. This mechanism may contribute to the novel transmission of Lyme disease spirochetes between cofeeding infected and uninfected ticks in nature.     

The western black-legged tick, Ixodes pacificus: a vector of Borrelia burgdorferi

To determine the significance of the western black-legged tick, Ixodes pacificus, as a vector of the Lyme disease spirochete, Borrelia burgdorferi, a tick/spirochete survey was conducted in northern California and southwestern Oregon from 1982 to 1984. Of 1,687 adult ticks collected off vegetation, 25 (1.48%) contained spirochetes. Of 715 ticks from Oregon, 14 (1.96%) were infected whereas 11 (1.13%) of 972 ticks from California harbored spirochetes. An isolate of 1 of the spirochetes reacted specifically when treated with monoclonal antibodies to B. burgdorferi. Polyacrylamide gel electrophoresis of a lysate of the isolate showed it to be nearly identical with 2 isolates of B. burgdorferi. Of the 25 infected I. pacificus, 17 had spirochetes in their midgut only; the remaining 8 ticks showed a generalized infection of all the tissues, with midgut, central ganglion and ovary or testes showing heavy spirochetal infections. Decreased immunofluorescent staining reactivity of spirochetes in tissues other than midgut in 6 of 8 I. pacificus with generalized infection may reflect adverse physiologic conditions for the development of spirochetes in the hemocele.     

Outer-surface protein C of the Lyme disease spirochete: a protein induced in ticks for infection of mammals

Environmentally responsive synthesis of surface proteins represents a hallmark of the infectious cycle of the Lyme disease agent, Borrelia burgdorferi. Here we created and analyzed a B. burgdorferi mutant lacking outer-surface protein C (OspC), an abundant Osp that spirochetes normally synthesize in the tick vector during the blood meal and down-regulate after transmission to the mammal. We demonstrate that B. burgdorferi strictly requires OspC to infect mice but not to localize or migrate appropriately in the tick. The induction of a spirochetal virulence factor preceding the time and host in which it is required demonstrates a developmental sequence for transmission of this arthropod-borne pathogen.     

The ecology of infectious disease: effects of host diversity and community composition on Lyme disease risk

The extent to which the biodiversity and community composition of ecosystems affect their functions is an issue that grows ever more compelling as human impacts on ecosystems increase. We present evidence that supports a novel function of vertebrate biodiversity, the buffering of human risk of exposure to Lyme-disease-bearing ticks. We tested the Dilution Effect model, which predicts that high species diversity in the community of tick hosts reduces vector infection prevalence by diluting the effects of the most competent disease reservoir, the ubiquitous white-footed mouse (Peromyscus leucopus). As habitats are degraded by fragmentation or other anthropogenic forces, some members of the host community disappear. Thus, species-poor communities tend to have mice, but few other hosts, whereas species-rich communities have mice, plus many other potential hosts. We demonstrate that the most common nonmouse hosts are relatively poor reservoirs for the Lyme spirochete and should reduce the prevalence of the disease by feeding, but rarely infecting, ticks. By accounting for nearly every host species' contribution to the number of larval ticks fed and infected, we show that as new host species are added to a depauperate community, the nymphal infection prevalence, a key risk factor, declines. We identify important "dilution hosts" (e.g., squirrels), characterized by high tick burdens, low reservoir competence, and high population density, as well as "rescue hosts" (e.g., shrews), which are capable of maintaining high disease risk when mouse density is low. Our study suggests that the preservation of vertebrate biodiversity and community composition can reduce the incidence of Lyme disease.     

Elimination of Borrelia burgdorferi from vector ticks feeding on OspA-immunized mice.

Although recombinant outer surface protein A (OspA) of Borrelia burgdorferi protects mice against injected Lyme disease spirochetes, the mode of protection has not yet been explored. Indeed, the efficacy of vaccine-induced immunity against a realistic vector-mediated challenge remains unexplored. Accordingly, we determined whether this immunogen protects mice against spirochetes delivered by nymphal Ixodes dammini ticks. Following challenge by tick bite, no spirochetes could be cultured from immunized mice, and no characteristic histopathology was found. The spirochete was not detected in ticks that fed on immunized animals and was present in virtually all ticks that fed on nonimmunized mice. We conclude that OspA-immunized mice are protected from spirochetal infection, at least in part, because the spirochete is destroyed in the infecting tick.     

Ixodes sinensis: competence as a vector to transmit the Lyme disease spirochete Borrelia garinii

In order to determine the principal vectors of Lyme disease in South China, the capability of Ixodes sinensis to transmit Lyme disease spirochetes transstadially was estimated in the laboratory. Results suggest that I. sinensis can acquire active Lyme disease spirochetes by feeding on infected mice of Kunming strains (KM), and the positive rates of larvae and nymph I. sinensis infected were 94.0% and 92.0%, respectively. I. sinensis maintained active spirochetes during blood digestion and molting periods; the subsequent questing tick stages were also found to be infected and infectious to naive KM mice, which resulted in more than 66.7% spirochete-positive KM mice. I. sinensis can transmit the Lyme disease spirochete Borrelia garinii from infected KM mice to naive KM mice during larvae-nymph and nymph-adult periods. Therefore, I. sinensis might be considered as a competent vector of Lyme disease in South China.     

Chronic Lyme disease: a review

Studies have shown that most patients diagnosed with chronic Lyme disease either have no objective evidence of previous or current infection with Borrelia burgdorferi or are patients who should be classified as having post-Lyme disease syndrome, which is defined as continuing or relapsing nonspecific symptoms (such as fatigue, musculoskeletal pain, and cognitive complaints) in a patient previously treated for Lyme disease. Despite extensive study, there is currently no clear evidence that post-Lyme disease syndrome is caused by persistent infection with B burgdorferi. Four randomized placebo-controlled studies have shown that antibiotic therapy offers no sustained benefit to patients who have post-Lyme disease syndrome. These studies also showed a substantial placebo effect and a significant risk of treatment-related adverse events. Further research to elucidate the mechanisms underlying persistent symptoms after Lyme disease and controlled trials of new approaches to the treatment and management of these patients are needed.     

Learned host preference in a Chagas disease vector, Rhodnius prolixus

It has been largely assumed that the individual experience of disease vector insects may affect host choice and, as a consequence, have an important influence on parasite transmission. In particular, it is speculated that vector insects should be able to learn and remember the most and/or less defensive hosts, shifting their preference accordingly. Nevertheless, despite the invested efforts in testing the capacity to learn and remember information of blood-sucking insects, only little conclusive information has been obtained hitherto. Recently, the ability of Rhodnius prolixus to associate a behaviourally neutral odour to the perspective of either obtaining a blood-meal or being punished has been demonstrated, the same odour becoming attractant or repellent for the bugs, respectively, according to the individual previous experience. The present work represents a step forward in the study of the cognitive abilities of Chagas disease vectors and their influence on host choice. We tested whether or not bugs bias their choice for a host based on the association of its odour with a negative experience. Our results show that whereas naïve bugs presented no preference when confronted to the odour of two different hosts, bugs previously exposed to the contingency of the odour of one host and a mechanical perturbation mimicking defensive behaviour, biased their preference towards the other host. This constitutes the first evidence of olfactory conditioning to host odours in triatomine bugs, vectors of Chagas disease and one of the few available up to date on haematophagous insects. The epidemiological significance of this finding is discussed.     

Interference between the agents of Lyme disease and human granulocytic ehrlichiosis in a natural reservoir host

Agents of Lyme disease (Borrelia burgdorferi) and human granulocytic ehrlichiosis (Ehrlichia phagocytophila) are perpetuated in a natural cycle involving the black-legged tick (Ixodes scapularis) and its vertebrate hosts. Using I. scapularis nymphs as the mode of infectious challenge, we studied how infection with one pathogen in white-footed mice (Peromyscus leucopus) affects their ability to acquire the other agent and subsequently to infect larvae, which these agents would do in nature. Two groups of mice were infected with either B. burgdorferi or E. phagocytophila. One week later, B. burgdorferi-infected mice were challenged with E. phagocytophila, and E. phagocytophila-infected mice were challenged with B. burgdorferi. Simultaneously, two control groups of uninfected mice were infected with each agent from the same tick cohorts used on the first groups of mice. Uninfected I. scapularis larvae were fed on all mice for xenodiagnosis at weekly intervals lasting 2 months. For the B. burgdorferi challenge, all control and E. phagocytophila-infected mice acquired B. burgdorferi. However, fewer xenodiagnostic larvae acquired B. burgdorferi from mice with mixed infections compared with mice infected with B. burgdorferi only. For the E. phagocytophila challenge, all five control mice acquired E. phagocytophila, but only two of five mice infected with B. burgdorferi subsequently acquired E. phagocytophila. Consequently, mice with both infections produced fewer xenodiagnostic ticks infected with E. phagocytophila than mice infected with E. phagocytophila only. Thus, a primary infection with either B. burgdorferi or E. phagocytophila in mice inhibited transmission of a second agent, suggesting interference between these two agents.