Spirochetes in ticks and antibodies to Borrelia burgdorferi in white-tailed deer from Connecticut, New York State, and North Carolina

Ticks were screened for spirochetes and serum samples from white-tailed deer (Odocoileus virginianus) were assayed for antibodies to Borrelia burgdorferi during 1983-1984. Using fluorescein isothiocyanate-labeled rabbit antibodies produced to B. burgdorferi, the etiologic agent of Lyme disease, spirochetes were detected in Ixodes dammini (10.5% of 1,193) and Dermacentor albipictus (0.6% of 157) adults from Connecticut, I. dammini nymphs (49.1% of 108) and adults (64.7% of 99) from Armonk, New York, and in I. scapularis (0.4% of 531) and Amblyomma americanum (3.5% of 173) adults from North Carolina. Infected ticks were either seeking hosts or feeding on deer during the summer and fall. Direct fluorescent antibody staining also revealed spirochetes in two larvae of I. scapularis that emerged from eggs deposited by separate females in the laboratory. Using indirect immunofluorescence tests, antibodies to B. burgdorferi were identified in white-tailed deer living in tick-infested areas of all three states. Aside from minor cross-reactivity, there was no serologic evidence of Treponema or Leptospira infections. Ixodes dammini is a primary vector of B. burgdorferi in northeastern United States, but in North Carolina, other ixodid ticks may transmit this spirochete to humans and wildlife.     

Widespread dispersal of Borrelia burgdorferi-infected ticks collected from songbirds across Canada

Millions of Lyme disease vector ticks are dispersed annually by songbirds across Canada, but often overlooked as the source of infection. For clarity on vector distribution, we sampled 481 ticks (12 species and 3 undetermined ticks) from 211 songbirds (42 species/subspecies) nationwide. Using PCR, 52 (29.5%) of 176 Ixodes ticks tested were positive for the Lyme disease spirochete, Borrelia burgdorferi s.l. Immature blacklegged ticks, Ixodes scapularis , collected from infested songbirds had a B. burgdorferi infection prevalence of 36% (larvae, 48%; nymphs, 31%). Notably, Ixodes affinis is reported in Canada for the first time and, similarly, Ixodes auritulus for the initial time in the Yukon. Firsts for bird-parasitizing ticks include I. scapularis in Quebec and Saskatchewan. We provide the first records of 3 tick species cofeeding on passerines (song sparrow, Swainson's thrush). New host records reveal I. scapularis on the blackpoll warbler and Nashville warbler. We furnish the following first Canadian reports of B. burgdorferi-positive ticks: I. scapularis on chipping sparrow, house wren, indigo bunting; I. auritulus on Bewick's wren; and I. spinipalpis on a Bewick's wren and song sparrow. First records of B. burgdorferi-infected ticks on songbirds include the following: the rabbit-associated tick, Ixodes dentatus, in western Canada; I. scapularis in Quebec, Saskatchewan, northern New Brunswick, northern Ontario; and Ixodes spinipalpis (collected in British Columbia). The presence of B. burgdorferi in Ixodes larvae suggests reservoir competency in 9 passerines (Bewick's wren, common yellowthroat, dark-eyed junco, Oregon junco, red-winged blackbird, song sparrow, Swainson's thrush, swamp sparrow, and white-throated sparrow). We report transstadial transmission (larva to nymph) of B. burgdorferi in I. auritulus. Data suggest a possible 4-tick, i.e., I. angustus, I. auritulus, I. pacificus, and I. spinipalpis, enzootic cycle of B. burgdorferi on Vancouver Island, British Columbia. Our results suggest that songbirds infested with B. burgdorferi-infected ticks have the potential to start new tick populations endemic for Lyme disease. Because songbirds disperse B. burgdorferi-infected ticks outside their anticipated range, health-care providers are advised that people can contract Lyme disease locally without any history of travel.     

Isolation of Borrelia burgdorferi from Peromyscus leucopus in Oklahoma.

Borrelia burgdorferi was isolated from a field-caught Peromyscus leucopus from central Oklahoma (USA). The strain was identified as B. burgdorferi by reaction with monoclonal antibody H5332 specific for the outer surface protein OspA of B. burgdorferi. This represents the first isolation of B. burgdorferi from a wild mouse outside of the normal range of the known vectors Ixodes dammini and I. pacificus.     

Migratory songbirds disperse ticks across Canada, and first isolation of the Lyme disease spirochete, Borrelia burgdorferi, from the avian tick, Ixodes auritulus

During a 3-yr comprehensive study, 196 ixodid ticks (9 species) were collected from 89 passerine birds (32 species) from 25 localities across Canada to determine the distribution of avian-associated tick species and endogenous Lyme disease spirochetes, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt, and Brenner. We report the following first records of tick parasitism on avian hosts: the rabbit-associated tick, Ixodes dentatus Marx, from Manitoba and Ontario; the mouse tick, Ixodes muris Bishopp and Smith, from British Columbia; and the blacklegged tick, Ixodes scapularis Say, from New Brunswick. Moreover, we provide the first record of the Neotropical tick, Amblyomma humerale Koch (1 nymph), in Canada and its parasitism of any bird. This tick was compared morphologically with nymphs of other Neotropical Amblyomma spp., and genetically, using a 344-bp fragment of the 12S rDNA sequence of 41 New World Amblyomma species. The first collections of the western blacklegged tick, Ixodes pacificus Cooley and Kohls, from passerine species in Alberta and British Columbia, are also reported. Notably, we further report the first isolation of B. burgdorferi from the bird tick, Ixodes auritulus Neumann, collected from an American robin, Turdus migratorius L., on Vancouver Island. Furthermore, B. burgdorferi-positive I. auritulus larvae were collected from a reservoir-competent fox sparrow, Passerella iliaca (Merrem). Our findings indicate that ground-dwelling passerines, in particular, are parasitized by certain ixodid ticks and play an important role across Canada in the wide dispersal of B. burgdorferi-infected ticks and increased risk of Lyme disease exposure.     

Discovery of the Lyme disease spirochete and its relation to tick vectors.

The various hypotheses concerning the etiologic agent of erythema chronicum migrans of Europe and of Lyme disease in the United States are reviewed, and an account of events that led to the discovery of the causative spirochetal agent in Ixodes dammini is presented. Spirochetes morphologically and antigenically similar, if not identical to, the organism detected in I. dammini were also found for the first time in Ixodes pacificus and Ixodes ricinus, the vectors hitherto incriminated, respectively, in western United States and Europe. In most infected ticks, spirochetal development was found to be limited to the midgut. Ticks with generalized infections were shown to transmit spirochetes via eggs, but infections decreased in intensity and became restricted to the central ganglion as filial ticks developed to adults. Although the mechanisms of transmission to a host are still under investigation, the spirochetes may be transmitted by saliva of ticks with generalized infectious and possibly also by regurgitation of infected gut contents, or even by means of infected fecal material.     

Peritrophic matrix in the midgut of tick females of the genus Ixodes (Acari: Ixodidae)

The formation of the peritrophic matrix in the midgut of females of 5 ixodid tick species (Ixodes pacificus, I. pavlovskyi, I. persulcatus, I. ricinus and I. scapularis) was studied by means of light and electron microscopy in different periods of the feeding and after detachment. The formation of the peritrophic matrix started when the first food portions came into the gut lumen, 9-12 hours after the attachment. Renovation of the peritrophic matrix took place during the whole feeding period; every new generation of midgut cells synthesized their own matrix. It was deposited on the apical surface of every midgut cell in the beginning of differentiation, and was functioning during the life of the cell. The peritrophic matrix separates spaces of the cavitary and cytozoic digestions.     

Salivary gland antigens of Ixodes dammini are glycoproteins that have interspecies cross-reactivity.

Serum from rabbits and rats exposed to Ixodes dammini adults and larvae, respectively, contain antibodies to a large number of antigens in salivary gland homogenates from adult ticks that cross react with the antigens of a closely related species, Ixodes scapularis, and significantly with Dermacentor variabilis antigens. The salivary gland antigens of I. dammini are glycoproteins composed of both N- and O-linked carbohydrate chains. The antigenic determinants reside in both the polypeptide and carbohydrate chains.     

Species of Borrelia distinguished by restriction site polymorphisms in 16S rRNA genes

Abstract Three phyletic groups of Borrelia associated with Lyme disease, B. burgdorferi, B. garinii and group VS461 can be distinguished from each other and other species of Borrelia by Bfa I restriction site polymorphisms in PCR amplified 16S rRNA genes. One strain isolated from an Ixodes pacificus tick in California that was previously unclassifiable was distinguishable from B. burgdorferi by an Mnl I restriction site polymorphism.     

Avian and mammalian hosts for spirochete-infected ticks and insects in a Lyme disease focus in Connecticut.

Spirochetes and their vectors and reservoirs were studied in a Lyme disease focus in East Haddam, Connecticut, from mid-May through September 1983. Ixodes dammini subadults were comparable in number on white-footed mice (Peromyscus leucopus) (means = 2.9 +/- 3.6 SD) to those on 27 different species of birds (means = 2.3 +/- 4.2 SD) representing 11 families within the order Passeriformes. Less commonly found ticks on birds (means less than or equal to 0.1) were immature Ixodes dentatus and Haemaphysalis leporispalustris. Although spirochete-infected I. dammini larvae and nymphs were taken off eight and nine different species of birds, respectively, significantly fewer positive larvae were removed from birds than from white-footed mice. Spirochetes were detected in the midguts of I. dammini, Dermacentor variabilis, and H. leporispalustris and two species of insects (Cuterebra fontinella and Orchopeas leucopus). Possibly, arthropods other than I. dammini vector these spirochetes in northeastern United States. Spirochetes grew in a cell-free medium inoculated with bloods from four white-footed mice, one woodland jumping mouse (Napaeozapus insignis), one northern mockingbird (Mimus polyglottos), one gray catbird (Dumetella carolinensis), two prairie warblers (Dendroica discolor), one orchard oriole (Icterus spurius), one common yellowthroat (Geothlypis trichas), and one American robin (Turdus migratorius). We suggest that avian hosts, like mammals, develop spirochetemias of the causative agent of Lyme disease. Erythematous tissues from a white-footed mouse were infected with spirochetes.     

Host associations of ticks parasitizing rodents at Borrelia burgdorferi enzootic sites in South Carolina.

A total of 237 rodents was collected in 4 regions of South Carolina from July 1994 through December 1995. Eight species were collected, including cotton mouse, hispid cotton rat, eastern woodrat, marsh rice rat, white-footed mouse, eastern harvest mouse, golden mouse, and black rat. Of the 1,514 ticks recovered from these hosts, Ixodes minor Neumann, including larvae, nymphs, and adults, was the most abundant species, representing 54% of the total. Only immature stages of other tick species were found, including larvae and nymphs of Dermacentor variabilis (Say), Amblyomma maculatum Koch, Ixodes affinis Neumann, and Ixodes scapularis Say. All 5 tick species parasitized cotton mice, cotton rats, and woodrats, which were the most important small mammal hosts for ticks at the localities studied. Rice rats were hosts of A. maculatum, D. variabilis, and L. minor. Amblyomma maculatum was more strongly associated with cotton rats than other rodent species. Ixodes scapularis was most strongly associated with cotton mice, and I. minor was more strongly associated with both woodrats and cotton mice than other species of rodents. Ixodes minor parasitized hosts in the Coastal Zone only, where among spirochete-infected hosts, it was present in significantly greater numbers than other ticks. Furthermore, I. minor was the only tick species that showed a statistically significant positive association with spirochetal infection in rodents. More I. affinis parasitized spirochete-infected hosts than I. scapularis, but fewer than I. minor. The findings discussed herein provide evidence that implicates I. minor as the possible primary enzootic vector of the Lyme disease spirochete Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt and Brenner in the Coastal Zone of South Carolina. They also indicate that the high level of B. burgdorferi infection in rodents from this region may be a function of the combined involvement of I. minor, I. affinis, and I. scapularis in the enzootic transmission of the spirochete.     

Ticks feeding on northern pocket gophers (Thomomys talpoides) in central Saskatchewan and the unexpected detection of Ixodes scapularis larvae

Morphological examination of ticks feeding on northern pocket gophers, Thomomys talpoides, near Clavet (Saskatchewan, Canada) revealed the presence of two genera, Ixodes and Dermacentor. All adult ticks collected were identified as I. kingi. Single strand conformation polymorphism (SSCP) analyses and DNA sequencing of the mitochondrial 16S rRNA gene confirmed the species identity of most Ixodes immatures as I. kingi (two nymphs and 82 larvae), and the Dermacentor immatures as D. variabilis (one nymph and one larva) and D. andersoni (three larvae). Six Ixodes larvae feeding on three T. talpoides individuals were identified as four different 16S haplotypes of I. scapularis, which was unexpected because there are no known established populations of this species in Saskatchewan. However, flagging for questing ticks and further examination of the ticks feeding on T. talpoides in two subsequent years failed to detect the presence of I. scapularis near Clavet, suggesting that there is no established population of I. scapularis in this area. Nonetheless, since I. scapularis is a vector of pathogenic agents, passive and active surveillance needs to be conducted in Saskatchewan on an ongoing basis to determine if this tick species and its associated pathogens become established within the province.     

Molecular phylogenetic analysis of ixodid ticks based on the ribosomal DNA spacer, internal transcribed spacer 2, sequences

An internal transcribed spacer (ITS2) sequence between the 5.8S and 28S rRNA genes was used to estimate the phyletic relationships among Ixodes spp. tick vectors of Lyme disease-causing Borrelia spirochetes. Analysis indicates that Borrelia burgdorferi sensu lato species associated with Lyme disease are found mainly in ticks of the Ixodes ricinus species complex. Other closely related tick species are not known to transmit the Borrelia-that cause Lyme disease in humans, but they appear to have a specific association with other closely related Borrelia species. There is a high degree of concordance in the phylogenetics of Borrelia taxa and the phylogenetic relationships among Ixodes ticks.     

Tick infestations of the eastern cottontail rabbit (Sylvilagus floridanus) and small rodentia in northwest Alabama and implications for disease transmission.

Studies were conducted over a four-county area of northwest Alabama to determine the association of eastern cottontail rabbits with Dermacentor variabilis, the eastern United States vector of Rocky Mountain spotted fever. A secondary objective was to compare infestations of this tick on rabbits with infestations on commonly encountered rodent species as a means of determining the relative importance of each in the disease transmission cycle. These epidemiologic surveys were conducted in response to reported fatal cases of Rocky Mountain Spotted Fever in two counties of the study area. From 202 eastern cottontail rabbits, 3,956 ticks were collected. Of this total, 79.87% were Haemphysalis leporispalustris, 9.15% Amblyomma americanum, 8.22% Ixodes dentatus, and 2.76% D. variabilis. Only immature stages of D. variabilis were collected from cottontail rabbits. Ticks were collected on rabbits in all months except November, and only one specimen was taken in January. Based on the average number of ticks per host collected in each month, April was the peak month for D. variabilis and I. dentatus. High values for H. leporispalustris also occurred at this time, but even higher values occurred in October and December. The heaviest infestation of A. americanum occurred during the month ofAugust and coincides with the activity period for the larvae of this species. Two hundred sixty-nine of the smaller Rodentia, comprising 13 species, yielded 264 ticks, all D. variabilis, and all but two were immature stages. Five rodent species, Microtus ochragaster Orozomys palustris, Peromyscus gossypinus, Peromyscus leucopus, and Sigmodon hispidus accounted for 95.83% of the ticks collected, and appeared to be preferred hosts for D. variabilis; all five had higher infestation levels per host than did the eastern cottontail rabbit. Data on host relationships in association with seasonal activity are presented.     

Bird-feeding ticks transstadially transmit Borrelia burgdorferi that infect Syrian hamsters

Bird-feeding Ixodes dammini ticks were documented for the first time to successfully molt and transstadially pass Borrelia burgdorferi spirochetes that were indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from the type B31 strain. Forty-six of 73 blood-engorged larvae and 50 of 66 fully-fed nymphs, removed from wild-caught birds, successfully molted. Borreliae were isolated from 21 of 78 partially- and fully-fed larvae off birds, including six specimens that molted. Spirochete-positive cultures also were obtained from 35 of 60 partially- and fully-fed nymphs that had fed from birds, including 20 nymphs that molted into adult ticks. Transstadially passed borreliae by bird-feeding larval and nymphal I. dammini were infectious to hamsters, leading us to suggest that these ticks are capable of subsequently transmitting infectious spirochetes to mammals, including humans. An isolated of B. burgdorferi, recovered from a bird-feeding larval Ixodes dentatus, was indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from the B31 strain. This isolate, unlike another from I. dentatus off a cottontail rabbit (Sylvilagus floridanus), had a protein band with a molecular weight of approximately 31,000 that reacted with murine monoclonal antibodies H3TS and H5332 in western blot analysis. Thus, closely related borreliae are present in both I. dentatus and I. dammini.     

Development of the Ixodes ticks (Ixodidae) in the reclaimed parts of forest in the Novgorod region

Data of observations on the development of Ixodes persulcatus, I. ricinus and Dermocentor pictus in drained parts of small-leaved and coniferous forests with different types of soils are given. It has been established that microclimatic changes take place in sites of ticks development. The draining favours the improvement of ecological conditions of the development of ticks. In drained parts of the forest the survival of preimaginal phases is higher than that in undrained ones.     

The roles of birds, lizards, and rodents as hosts for the western black-legged tick Ixodes pacificus.

We compared the infestation by ixodid ticks of lizards, rodents, and birds collected simultaneously within areas representing common habitat types in Mendocino County, CA. Lizards were infested only by Ixodes pacificus Cooley and Kohls, birds by I. pacificus and Haemaphysalis leporispalustris (Packard), and rodents by I. pacificus, I. spinipalpis Hadwen and Nuttall, I. woodi Bishopp, Dermacentor occidentalis Marx, and D. variabilis (Say). Infestation by I. pacificus larvae and nymphs of lizards (Sceloporus occidentalis Baird and Girard; Elgaria spp.) and western gray squirrels (Sciurus griseus Ord) (means of 9-35 larvae and 5-6 nymphs per animal) was several times greater than for Neotoma fuscipes Baird woodrats, Peromyscus spp. mice, and birds (means of 0.9-3.5 larvae and 0-0.3 nymphs). Overall, Borrelia-refractory lizards accounted for 84% of I. pacificus larvae and 91% of nymphs collected from animals in dense woodlands. Bird species frequently utilizing tick-questing substrates such as leaf litter (guild I birds) were more heavily infested by I. pacificus subadults (5.2 larvae and 1.0 nymphs per bird) than guild IV birds with minimal perceived contact with tick-questing substrates (0.08 larvae and 0.06 nymphs per bird). Notably, guild I birds carried similar larval loads and at least 20-fold higher nymphal loads relative to woodrats and mice. Only guild IV birds carried as few I. pacificus nymphs as did these rodents. The ratios of larvae to nymphs suggest that, relative to birds, lizards, and squirrels (infested by 1.3-6.0 larvae per nymph), nocturnally active ground-dwelling rodents such as woodrats and mice are underutilized by the nymphal stage (69 to >100 larvae per nymph). The western gray squirrel and guild I-II birds (e.g., the dark-eyed junco, Junco hyemalis [L.]) were the only potential reservoirs of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt, and Brenner (the causative agent of Lyme disease in North America) that were frequently infested with both I. pacificus larvae and nymphs and commonly utilized dense woodland habitats.     

Borreliacidal activity of saliva of the tick Amblyomma americanum

Amblyomma americanum (Linneaus) (Acari: Ixodidae), an important tick vector of human and animal disease, is not a competent vector of the bacterial agent of Lyme disease, Borrelia burgdorferi, although its range overlaps the geographical distribution of Lyme disease within the United States. A possible mechanism that could prevent acquisition of B. burgdorferi spirochetes from infected hosts is the toxic effect of A. americanum saliva on B. burgdorferi. The data presented here indicate that after 24 and 48 h of exposure to A. americanum saliva, significantly fewer B. burgdorferi were alive compared to treatment controls as assessed by spirochete motility under dark-field microscopy and resistance to the dead stain, propidium iodide. After 48 h, fewer than 13% of saliva-exposed B. burgdorferi were alive. In contrast, significantly more B. burgdorferi exposed to Ixodes scapularis (Acari: Ixodidae) saliva survived after 24 or 48 h compared to A. americanum saliva or treatment controls.     

Ixodes (Pholeoixodes) hexagonus, an efficient vector of Borrelia burgdorferi in the laboratory

Borrelia burgdorferi Johnson et al. was first isolated from the midgut of Ixodes dammini Spielman et al. in the U.S.A. and from the midgut of I.ricinus (L.) in Europe. I.ricinus was considered to be the only tick vector of this borrelia, in Europe, until I.hexagonus Leach, the hedgehog tick, was found to harbour spirochaetes. This paper reports an evaluation of the vector competence of I.hexagonus for the spirochaete B.burgdorferi. Transovarial and trans-stadial survival were demonstrated and the spirochaete was transmitted to laboratory mice via the bites of trans-stadially infected I.hexagonus females.     

The New Zealand white rabbit: an experimental host for infecting ticks with Lyme disease spirochetes

Efficiency of the New Zealand white rabbit as a host for infecting larval Ixodes dammini, I. pacificus, and I. ricinus with Lyme disease spirochetes was evaluated. Rabbits inoculated with infected midgut suspensions of I. dammini from Shelter Island, New York, or fed upon by infected ticks from the same area, responded with spirochetemias of sufficient concentrations to infect as many as 30 percent of the ticks. When infected ticks were used as indicators, it appeared that spirochetemias persisting for up to ten days occurred as early as the tenth day after inoculation or feeding of ticks.