Tick Transmission Research Articles

Importance of Localized Skin Infection in Tick-Borne Encephalitis Virus Transmission

Arboviruses are transmitted to vertebrates by the ”bite“ of infected arthropods. Events at the site of virus deposition are largely unknown despite increasing evidence that blood-sucking arthropods immunomodulate their skin site of feeding. This question is particularly relevant for ixodid ticks that feed for several days. To examine events under conditions mimicking tick-borne encephalitis (TBE) virus transmission in nature (i.e., infected and uninfectedIxodes ricinusticks feeding on the same animal), infected adult and uninfected nymphal ticks were placed in one retaining chamber (skin site A) and uninfected nymphs were placed within a second chamber posteriorly (skin site B) on two natural host species, yellow-necked field mice (Apodemus flavicollis) and bank voles (Clethrionomys glareolus). Virus transmission from infected to uninfected cofeeding ticks was correlated with infection in the skin site of tick feeding. Furthermore, virus was recruited preferentially to the site in which ticks were feeding compared with uninfested skin sites. Viremia did not correspond with a generalized infection of the skin; virus was not detected in an uninfested skin site (C) of 12/13 natural hosts that had viremia levels ≥2.0 log10ic mouse LD50/0.02 ml blood. To characterize infected cells, laboratory mouse strains were infested with infected ticks and then explants were removed from selected skin sites and floated on culture medium. Numerous leukocytes were found to migrate from the skin explants of tick feeding sites. Two-color immunocytochemistry revealed viral antigen in both migratory Langerhans cells and neutrophils; in addition, the migratory monocyte/macrophages were shown to produce infectious virus. The results indicate that the local skin site of tick feeding is an important focus of viral replication early after TBE virus transmission by ticks. Cellular infiltration of tick feeding sites, and the migration of cells from such sites, may provide a vehicle for transmission between infected and uninfected cofeeding ticks that is independent of a patent viremia. The data support the hypothesis that viremia is a product, rather than a prerequisite, of tick-borne virus transmission.

Demonstration of vertical transmission of Cowdria ruminantium, the causative agent of heartwater, from cows to their calves

One of the most important questions about the epidemiology of heartwater in the field is how Cowdria ruminantium is transmitted within vertebrate host populations. In this study vertical transmission of C. ruminantium from cows to their calves was demonstrated. Twelve mixed-breed calves, born to dams living in a heartwater-endemic area of Zimbabwe, were tested post-natally for the presence of C. ruminantium. Vertical transmission was demonstrated to occur under natural field conditions using tests in which uninfected laboratory-reared Amblyomma ticks were fed on neonatal calves and subsequently either fed on, or inoculated into, susceptible small ruminants or tested by a polymerase chain reaction (PCR) assay. Prior to natural Amblyomma tick infestation, C. ruminantium infection in 5 of the 12 calves was confirmed by tick transmission to small ruminants and 11 of the 12 calves tested positive based on PCR analysis of ticks fed on them. The role of colostral cells, as one mode of infection, was demonstrated by the transmission of C. ruminantium to three out of five goats inoculated intravenously with viable colostral cells collected from dams living in a heartwater-endemic area. The significance of vertical transmission is presented in relation to the epidemiology and control of heartwater.

Quantifying parameters in the transmission of Babesia microti by the tick Ixodes trianguliceps amongst voles (Clethrionomys glareolus).

The estimation of two parameters in the transmission of Babesia microti by the tick Ixodes trianguliceps amongst small mammals, (1) the duration of infectivity in natural hosts and (2) the probability of transmission from an infected to a susceptible vole, is described. When B. microti was maintained by direct tick transmission, the probability of a complete cycle of transmission via the larval-nymphal and nymphal-adult transstadial routes was 1.0 and 0.71 respectively, but only if the larvae or nymphs had engorged, as distinct from feeding slowly, while the source parasitaemia exceeded 2 or 0.2% respectively, but had not yet passed the peak level. The duration of this condition for infectivity in voles infected by nymphal bites was only 1-4 days, whilst infections delivered by adult ticks barely reached the threshold level necessary for successful transmission. When syringe passage was introduced into the parasite maintenance schedule (a) the probability of transmission declined markedly and (b) the time-course of the parasitaemia was altered. If these parameter values are put into a simple model, together with field data on tick and host survival rates, it becomes apparent that additional factors, such as the highly aggregated distribution of ticks on their hosts, must account for the maintenance of B. microti at the levels seen in wild small mammal populations.

Theileria parva carrier state in naturally infected and artificially immunised cattle

Rhipicephalus appendiculatus adult ticks, collected in the field from zebu (Bos indicus) and exotic (Bos tarus) cattle with high antibody titres to Theileria parva schizont antigen, transmitted Theileria parva infection typical of East Coast fever to susceptible cattle. Uninfected R. appendiculatus nymphs applied to naturally recovered zebu and exotic cattle kept under tick-free conditions in the laboratory for 16 and 7 months respectively, transmitted fatal theileriosis to susceptible cattle. Cattle immunised by the infection and treatment method were shown to be carriers of Theileria parva by examination of the salivary glands of ticks applied to them and by tick transmission. Three and 7 months after immunisation, Theileria parva infected lymphocytes were established in vitro from peripheral blood lymphocytes. This was the first demonstration, in vitro, of the existence of schizonts in Theileria parva (East Coast fever) infection carrier status. These studies show that cattle from endemic and epidemic areas of East Coast fever (ECF) become carriers thereby maintaining the T. parva population. The relevance of the findings in this study to the control of ECF by dipping, immunisation and treatment is discussed.

The Lyme disease vaccine candidate outer surface protein A (OspA) in a formulation compatible with human use protects mice against natural tick transmission of B. burgdorferi

Development of a vaccine for the Lyme disease spirochete, Borrelia burgdorferi, has focused on the bacterial lipoprotein, major outer surface protein A (OspA). With few exceptions, testing of OspA vaccines in animal models has involved challenge with needle inoculation of cultured spirochetes. Recombinant OspA proteins from two OspA divergent strains of B. burgdorferi were tested for their vaccine potential in three different strains of mice challenged with laboratory reared ticks with a high rate of B. burgdorferi infection. All formulations of the B. burgdorferi sensu stricto derived OspA vaccine protected all strains of mice when challenged by ticks infected with an OspA homologous strain of the spirochete, whereas heterologous OspA from B. afzelii did not protect. Furthermore, ticks feeding on protected mice had reduced OspA levels compared to unvaccinated controls.

Tick transmission of Borrelia burgdorferi to inbred strains of mice induces an antibody response to P39 but not to outer surface protein A

Natural tick transmission of infection by Borrelia burgdorferi induces a very different serum antibody response than needle inoculation of spirochetes. We present data, obtained by using the mouse model, that show that the OspA response was barely detectable, whereas all animals developed significant anti-P39 titers after exposure to B. burgdorferi-infected ticks.

Impact of persistent Anaplasma marginale rickettsemia on tick infection and transmission

Anaplasma marginale, an intraerythrocytic rickettsia of cattle, is transmitted biologically by ticks. Because of the brevity of acute A. marginale infection, transmission may rely on the tick's ability to acquire the organism from persistently infected cattle with low rickettsemia levels. By using a nucleic acid probe to quantitate low-level infection, we found that rickettsemia levels in persistently infected cattle fluctuated at approximately 5-week intervals during a 24-week period, from < 10(4) infected erythrocytes per ml of blood to high levels of approximately 10(7) infected erythrocytes per ml of blood. Cattle maintained very low rickettsemia levels (< 10(4.3) infected erythrocytes per ml of blood) for approximately 4 to 8 days of every 5-week cycle. The effect of fluctuations in rickettsemia in persistently infected cattle on acquisition by Dermacentor andersoni nymphal and adult male ticks was examined. A positive correlation was observed between rickettsemia levels in cattle and the resulting infection rates of ticks. At high rickettsemia levels, up to 80% of ticks acquired infection, but even at extremely low rickettsemia levels, 27% of adult male ticks became infected. Moreover, once ticks acquired infection, biological replication of the organism within the ticks appeared to make up for initial differences in the infecting dose. The high infection rates in adult males, combined with their intermittent feeding behavior and the observation that only a few infected ticks were required for transmission to a susceptible host, suggest that adult male D. andersoni ticks are epidemiologically important in A. marginale transmission. Because cattle with all levels of rickettsemia were capable of efficient transmission to ticks, population control efforts must include decreasing transmission from persistently infected individuals.

Isolation and transmission of the Lyme disease spirochete from the southeastern United States.

The isolation of the Lyme disease spirochete (Borrelia burgdorferi) from the southeastern United States is reported. Three isolates, two from cotton mice (Peromyscus gossypinus) and one from the black-legged tick (Ixodes scapularis), were recovered from Sapelo Island, Georgia, in July and September 1991. The spirochetes were characterized by indirect fluorescent antibody assay using a battery of five monoclonal antibodies, by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE) of whole cell lysates, and by the polymerase chain reaction (PCR) assay using primers for three DNA target sequences found in B. burgdorferi reference strain B-31. Transmission experiments indicate that the three Georgia isolates can infect experimentally inoculated hamsters and mice. Tick transmission of one of the isolates has been attempted so far; I. scapularis transmitted isolate SI-1 from hamsters to mice, but the lone-star tick, Amblyomma americanum, did not.

The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi.

The causative agent of Lyme disease, Borrelia burgdorferi, is transmitted by ticks of the Ixodes ricinus complex. In this study, we report the antibody response of recombinant inbred strains of mice of the H-2, b, d, and k haplotypes, infected with B. burgdorferi as a result of exposure to infected I. dammini. The patterns of antibody response assayed by Western blot analysis indicate significant major histocompatibility complex (MHC) restriction to bacterial antigens within the first 2 mo of infection in mice. Other bacterial antigens induce a significant response across the MHC haplotypes tested when assayed on the same bacterial strain used to transmit the infection, but do not crossreact with the same proteins derived from heterologous strains of B. burgdorferi. No response to outer surface protein A was detected at any time during the 60-d period we analyzed this infection. A third group of bacterial antigens appear to generate a MHC-nonrestricted response, and this lack of restriction is maintained when assaying the crossreactivity of the response with other strains of B. burgdorferi. These proteins may provide more accurate diagnostic probes than those currently in use. Finally, there appears to be a significant difference in the expression of most bacterial antigens when the spirochete is cultured for many passages since the same strain of bacterium isolated from low-passage and high-passage preparations exhibit different banding patterns in Western blots when assayed with the same sera.

Experimental infection of the white-footed mouse with Borrelia burgdorferi

SUMMARY Laboratory raised white-footed mice (Peromyscus leucopus) were inoculated experimentally with live spirochetes (Borrelia burgdorferi), the etiologic agent of Lyme disease (borreliosis). Prior to inoculation, mouse sera were tested with an indirect fluorescent antibody test, and all mice were seronegative. All inoculated mice seroconverted. In tick transmission studies, immature stages of Ixodes dammini and Dermacentor variabilis attached and fed to repletion on mice, but only I dammini transferred spirochetes to uninfected mice. Mice were susceptible to oral infection and transmitted infection to each other through direct contact. Infection did not affect reproduction or development of young born from infected dams, nor did spirochetes appear in the tissues of neonates. Borrelia burgdorferi spirochetes were identified in the kidneys, liver, and spleen of infected mice by the use of the modified Steiner silver stain and a tissue indirect fluorescent antibody test. Spirochetes also were isolated in culture of the heart blood of 1 mouse. Regardless of the source of infection, no mice developed clinical signs or had any pathologic change resulting from infection. Spirochetes were always observed extracellularly within interstitial spaces. Antibody titers persisted for over 4 months in some mice and spirochetes were found in the tissues of 1 mouse that had been infected 1 year earlier.

Computer Simulation of Management Strategies for American Dog Ticks (Acari: Ixodidae) and Rocky Mountain Spotted Fever

Computer models were developed to simulate the effects of management technologies on populations of the American dog tick, Dermacentor variabilis (Say), principal vector of Rocky Mountain spotted fever (RMSF) in eastern North America. The technologies modeled were area-wide acaricide application, acaricide-food-baited tubes for self-treatment by small mammals, dipping of dogs in acaricides, acaricide-impregnated plastic dog collars, reduction of small mammal host populations (host management), and removal of vegetation that protects free-living tick stages (vegetative management). Submodels for each of these technologies were incorporated into a model (ADTSIM) for the population dynamics of the tick and RMSF transmission. Comparisons of simulated and observed data were used to verify reasonable accuracy of the submodels. Repetitive simulations were made to identify levels and timing of each control method (alone or combined) required to reduce tick populations below a RMSF transmission threshold of 252 unfed adults/ha. Eight to 30 acaricide applications, depending on acaricide and percentage of population treated, were needed during a 10-yr period to reduce densities of ticks below the threshold. The baited-tube method, host management, and vegetative management (depending on level and frequency of treatment) also were capable of reducing tick density below the threshold. However, acaricide-impregnated plastic dog collars did not reduce tick density below the threshold unless at least 50% of the hosts of adult ticks were domestic dogs. Integrated strategies were developed for management of ticks and RMSF in six selected states. These strategies reduced numbers of human cases of RMSF 90% or more by year 20 by maintaining tick densities between 100 and 252 unfed adults/ha.

Babesia bovis: Evidence for selection of subpopulations during attenuation

DNA probes were used to detect variation in subpopulations of virulent and serially passaged Babesia bovis. Two distinct patterns were evident after hybridization to genomic DNA; the first was a basic profile typical of virulent B. bovis and the second, a more variable array, was characteristic of B. bovis after various stages of attenuation. Tick transmission of avirulent B. bovis causes reversion to the virulent genomic pattern, suggesting that selective enrichment of a small residual subpopulation caused reversion to a virulent profile of subpopulations. Certain genomic fragments, predominant in either virulent or avirulent parasite forms, are putative “markers” or actual elements responsible for these biological characteristics.

Effect of different methods of maintenance on the development and morphology of Babesia bigemina in the gut of Boophilus microplus

The development and morphology of Babesia bigemina in the gut of Boophilus microplus ticks were studied during laboratory maintenance of the babesia by four methods: tick transmission in unsplenectomised (intact) calves; tick transmission in splenectomised calves; syringe passage at intervals of five days or less in splenectomised calves; and syringe passage at intervals of six weeks or more in intact calves. The first method had no apparent effect on the development of B bigemina in ticks compared to that of the original isolate. The other three methods had obvious effects, the most pronounced being increased numbers of babesial forms with processes, particularly during early stages of development. The findings emphasise the importance of maintaining laboratory strains of parasites by natural means in life cycle studies.

The use of tick transmission by Boophilus microplus to isolate pure strains of Babesia bovis, Babesia bigemina and Anaplasma marginale from cattle with mixed infections

Pure strains of Babesia bovi, Babesia bigemina and Anaplasma marginale were isolated from cattle infected with all 3 species as well as a Theileria sp. and Eperythrozoon teganodes, using only transmission by the tick, Boophilus microplus. Unengorged adult ticks transferred to susceptible cattle transmitted A. marginale, but not Babesia. Engorged adults gave rise to progeny that transmitted Babesia, B. bovis by larvae and B. bigemina by male ticks. The Theileria and E. teganodes were not transmitted by the ticks and thus did not appear in calves used for isolating the pure strains of Babesia and A. marginale.

Babesia bovis: Proteins of virulent and avirulent parasites passaged through ticks and splenectomized or intact calves

Passage of the avirulent vaccine (K) strain of Babesia bovis (K A) through either Boophilus microplus ticks, intact calves, or intact calves and then ticks, resulted in two distinct protein and protein antigen profiles as analyzed by two-dimensional gel electrophoresis of biosynthetically labeled proteins and immunoprecipitates. Different degrees of expression of two major acidic antigens of K A designated Ka 1 ( M r 47,500) and Ka 2 ( M r 43,000) were observed. Ka 1 was apparently lost following passage of K A B. bovis through intact calves but was strongly represented in the parasite population following a single tick passage. In contrast, passage through ticks of the virulent K V B. bovis (from which K A was derived by passage in splenectomized calves) did not lead to strong representation of the Ka 1 protein although there was increased representation of another major acidic protein antigen, designated Kv ( M r 35,000). These data suggest that the previously recognized reversion to a strain-dependent basal antigenic type in the tick vector depends also on intrastrain characteristics such as virulence and strain heterogeneity. The data suggest that K A is a more heterogeneous population than K V although cloned isolates are required to establish this point. Comparable syringe passage of another strain of B. bovis, designated C strain, through splenectomized calves resulted in less marked differences between the putative C A and C V B. bovis. This may explain the less stable avirulence of C A compared to K A B. bovis. Various selection pressures must act, in either the tick or the vertebrate host, on subpopulations in heterogeneous isolates to produce the changes described in protein antigen profiles of B. bovis. The possible relevance of changes in representation of proteins to biological characteristics of B. bovis (such as virulence and tick transmissibility) is discussed.

Tick transmission of Babesia microti to rhesus monkeys (Macaca mulatta).

To determine whether Ixodes dammini is capable of transmitting Babesia microti to primates, infected nymphal ticks were allowed to feed on five Macaca mulatta. The monkeys were then followed for at least 60 days with daily thick blood smears for evidence of infection. Patent B. microti parasitemia developed in four of the five animals. Prepatent periods were 13, 18, 20, and 28 days. Maximum parasitemia ranged from 83 to 7,068 organisms/mm3 blood. Splenectomy 15-17 months after exposure to ticks results in recurrences of parasitemia in three of the four infected monkeys.

Experimental Tick Transmission of Theileria Species between Eland

Experimental Tick Transmission of Theileria Species between Eland

Investigations on a Theileria species from an impala.

An immature impala (Aepyceros melampus), captured in the Machakos district of Kenya, had Theileria-like parasites in its erythrocytes. Blood from this animal was inoculated into a Theileria-free impala born in captivity and a Theileria-free, splenectomized steer. Theilerial piroplasms were detected in the erythrocytes of the recipient impala 10 days after inoculation, and a peak parasitaemia of 2% was reached after 48 days, but no infection was detected in the steer. Anaemia developed in the recipient impala in association with the theilerial infection. The morphology of the theilerial piroplasm is described. Possibly dividing parasites were identified. Attempted tick transmission of the impala Theileria to a steer using the ixodid tick, Rhipicephalus appendiculatus, was unsuccessful. With an antigen prepared from the impala blood and the indirect fluorescent antibody test, this parasite was serologically distinct from other theilerial parasites isolated from Bovidae in East Africa.

Transmission of Theileria lawrencei (Serengeti) by the ixodid tick, Rhipicephalus appendiculatus.

The transmission by the tickRhipicephalus appendiculatus ofTheileria lawrencei (Serengeti) from buffalo to cattle, and thence between cattle, is described. Buffalo were shown to act as carriers ofT. lawrencei (Serengeti) piroplasms infective for ticks for up to 26 months. Tick transmission ofT. lawrencei (Serengeti) from buffalo to cattle was efficient, but its transmission between cattle was less so.

Attempts to transmit Babesia (Nuttallia) microti in laboratory mice by the tick Rhipicephalus appendiculatus

Irvin A. D. and Brocklesby D. W. 1972. Attempts to transmit Babesia (Nuttallia) microti in laboratory mice by the tick Rhipicephalus appendiculatus. International Journal for Parasitology, 2, 99–103. The rodent piroplasm Babesia microti is easily maintained in laboratory mice by syringe passage, but experimental tick transmission has not been successfully achieved. Attempts were made to effect experimental transmission using a tick species ( Rhipicephalus appendiculatus) readily maintained in the laboratory. Larval ticks fed on parasitaemic mice failed to transmit infection when ground up and inoculated into clean mice. Nymphal ticks from the same batches also failed to transmit infection when applied to mice. Larval ticks applied to a rabbit into which parasitaemic blood was inoculated failed to become infected, even though the rabbit's blood remained infective for 3 days on mechanical passage into clean mice. Piroplasms were detected in the gut of engorged larvae but not in ova laid by female ticks which had engorged on parasitaemic mice.