Borrelia burgdorferi sensu lato in the female cement plug of Ixodes persulcatus ticks (Acari,Ixodidae)

Borrelia burgdorferi sensu lato was detected in one out of five cement plugs of female Ixodes persulcatus ticks. The spirochetes were found by dark field microscopy as early as 18 h after attachment of the ticks to the skin of a white mouse. The relevance of this finding is discussed in relation to the epidemiology of Lyme Borreliosis.     

Questing behaviour of Ixodes ricinus ticks (Acari: Ixodidae)

The vertical distribution in the vegetation of questing Ixodes ricinus ticks was investigated in two different vegetation types (high and low vegetation) at two localities in south-central Sweden during 1992-1993 (Toro) and 1995 (Bogesund). Significant correlations were found between the vertical distribution of immature ticks and the height of the vegetation. The greatest mean availabilities of the larvae and nymphs in low vegetation were in the intervals 0-9 and 30-39 cm, respectively. The larval numbers were greatest close to the ground (0-29 cm) in both high and low vegetation. The larval : nymphal ratio, at ground level at localities free of ground vegetation, varied between 8 : 1 and 32 : 1. In high vegetation, the greatest mean numbers of nymphal and adult ticks were at height intervals of 50-59 and 60-79 cm, respectively. These ranges are within the estimated height interval (40-100 cm) of the main part of the body surface of their preferred host, the roe deer (Capreolus capreolus). The presence of most questing I. ricinus larvae at ground level would favour the transmission of Borrelia burgdorferi s.l., since this is where the highly reservoir-competent rodents and shrews usually occur.     

Differences in Borrelia infections in adult Ixodes persulcatus and Ixodes ricinus ticks (Acari: Ixodidae) in populations of north-western Russia

Dark-field microscopy was used to determine the number of Borrelia spirochetes in 630 standard preparations obtained from adult ixodid ticks (344 Ixodes persulcatus and 286 I. ricinus) collected in 1989-92 in the Leningrad region of Russia. The average numbers of Borrelia in I. persulcatus and I. ricinus preparations were 34.7 and 23.3 per 100 microscopic fields, respectively. The maximal individual values registered each year for ticks of both species were several hundred times greater than the minimal values. Ticks carrying relatively small numbers of Borrelia generally predominated. Proportions of more heavily infected ticks varied considerably from year to year. These parameters were significantly higher in foci with predominance of I. persulcatus ticks. As a consequence, risk to acquire Lyme borreliosis in such foci is considered greater than in foci where I. ricinus predominates.     

Physiological Age of Field-collected Female Taiga Ticks, Ixodes persulcatus (Acari: Ixodidae), and their Infection with Borrelia burgdorferi Sensu Lato

In some studies the prevalence of tick infection (infection rate) and the intensity of infection are negatively correlated with unfed tick age (in the broad sense of this term). However, no special research has been carried out to consider the phenomenon thoroughly. The infection indices of the female taiga ticks, Ixodes persulcatus, infected with Borrelia burgdorferi s.l. were related to tick physiological age, an index that more precisely reflects tick physiological state than the time of tick collection in the field or the duration of tick survival under laboratory conditions. A novel quantitative technique of physiological age determination based on the evaluation of the ratios between sizes of the stable (scutum) and the changing (alloscutum) structures of the tick body was used. The age was estimated in accordance with the classical age-grade scale introduced by Balashov and a more fractional scale determined by the new technique. In total, 131 female ticks were examined for their infection and physiological age, 46 of which were infected with B. burgdorferi s.l. (mean infection rate 35.1%). The minimal intensity of infection was 0.4 bacterial cells per 100 fields of view whereas the maximal infection was 172 cells. There was no difference between the prevalence of infection in ticks of different physiological age. The intensity of infection obviously differed between ticks of different age groups in the scale introduced by Balashov but did not significantly differ between ticks of different age groups according to the fractional age-grade scale. The data concerning the relationships between Borrelia burgdorferi and unfed Ixodes ticks are considered.     

Detection and typing of Borrelia burgdorferi sensu lato genospecies in Ixodes persulcatus ticks in West Siberia, Russia

The prevalence of Borrelia burgdorferi sensu lato (s.l.) genospecies in West Siberia as well as in many other regions of Russia remains insufficiently investigated. In the present study a total of 151 adult female ticks Ixodes persulcatus Schulze, collected at three localities in eastern regions of West Siberia, where Lyme disease is endemic, were examined for the presence of the spirochete B. burgdorferi s.l. by polymerase chain reaction targeting the 23S-5S rRNA intergenic spacer regions. Spirochetal DNA was detected in on average 15.2+/-3.0% of the ticks examined. The infection rate of adult ticks with B. burgdorferi s.l. at various localities ranged from 8.6+/-3.4% to 29.0+/-7.6%, being greatest in the northernmost site studied and decreasing southwards. The restriction patterns obtained after MseI digestion of the 23S-5S rRNA intergenic spacer amplicons assigned 23 DNA samples to the following genomic groups: 19 to B. garinii (12 to group NT29 and seven to group 20047(T)), three to B. afzelii, and one to mixed B. afzelii and B. garinii NT29. We have not detected other genospecies, which were found in ticks in Europe, the Russian Far East and Japan. Thus, the ticks examined were associated only with two genospecies of Borrelia burgdorferi s.l. pathogenic to humans (B. garinii and B. afzelii), and B. garinii was the major genospecies infecting adult I. persulcatus in eastern regions of West Siberia.     

Vector competence of Ixodes angustus (Acari: Ixodidae) for Borrelia burgdorferi sensu stricto

The vector competence of Ixodes angustus for Borrelia burgdorferi sensu stricto (s.s.) was investigated in the laboratory. The larval progeny of female ticks from Washington State were placed on Swiss-Webster mice that had been inoculated intravenously with 10⁸ spirochetes each of a Californian isolate of B. burgdorferi. Spirochetes were detected in 6 (12%) of 50 nymphs derived from larvae that had fed on these animals. Ten nymphs from the same cohort of experimentally infected ticks were placed on each of 4 naive deer mice (Peromyscus maniculatus). One of the mice seroconverted to B. burgdorferi and spirochetes were isolated from its ear tissues 4 weeks after exposure to ticks. Further vector competence trials were conducted with I. angustus ticks from California. Larvae were fed on deer mice that had been inoculated intradermally with B. burgdorferi along with larvae of I. spinipalpis as a comparison group. There was no significant difference in the prevalence of infection in nymphs of I. angustus (8.2%) versus those of I. spinipalpis (12.1%). We conclude that I. angustus is a competent experimental vector of B. burgdorferi s.s. and its efficiency for acquiring and transstadially passing such spirochetes is similar to that of I. spinipalpis.     

中华硬蜱、越原血蜱和青海血蜱经期传播莱姆病螺旋体的实验研究

以实验室培育的非感染中华硬蜱、越原血蜱和青海血蜱刺叮人工感染21天的阳性KM鼠,利用分离培养和PCR方法检测蜱对莱姆病螺旋体Borrelia garinii的保持能力以及体内螺旋体对敏感动物的感染能力。中华硬蜱、越原血蜱和青海血蜱非感染幼蜱均可以通过吸血获得莱姆病螺旋体, 然而,它们在饱血、消化、蜕皮和再次吸血过程中对莱姆病螺旋体的保持能力差异很大。只有中华硬蜱可以保持活的莱姆病螺旋体直至蜕化为下一发育阶段,蜕化后仍携带莱姆病螺旋体并对敏感KM鼠有感染能力。而越原血蜱和青海血蜱对莱姆病螺旋体的保持能力不能跨越消化、蜕皮阶段,蜕化后这两种血蜱不再携带莱姆病螺旋体,因此,这两种血蜱不具备经期传播能力,作为莱姆病媒介的可能性不大。中华硬蜱于幼蜱到若蜱以及若蜱到成蜱两个阶段均具备经期传播莱姆病螺旋体的能力,提示了中华硬蜱作为南方莱姆病的潜在媒介是可能的。     

Infestation of mammals by Ixodes ricinus ticks (Acari: Ixodidae) in south-central Sweden

Infestation by Ixodes ricinus ticks on rodents, hares and cervids was examined at Bogesund, 10 km north of Stockholm, in south-central Sweden during 1991-1994 and on varying hares (Lepus timidus) at Stora Karlso and Gotska Sandon in the Baltic Sea during 1992-1993. At Bogesund, there were great differences between two consecutive years in the number of I. ricinus larvae infesting bank voles (Clethrionomys glareolus). The seasonal pattern of infestation by I. ricinus larvae and nymphs on bank voles was unimodal in 1991, with peaks in June-July and bimodal in 1992, with peaks in June and August. Male bank voles, compared to females and older voles, compared to young voles, harboured greater numbers of I. ricinus ticks. Apodemus mice, compared to bank voles, harboured greater numbers of I. ricinus ticks. Ixodes ricinus larvae engorged on Apodemus mice were heavier than larvae engorged on bank voles and resulted in larger nymphs. However, there was no difference in the proportions of viable nymphs resulting from larvae engorged on mice or voles. The ranges in the numbers of I. ricinus ticks infesting individual hosts were 1-451 for rodents, 16-2374 for hares and 428-2072 for roe deer (Capreolus capreolus). These ranges of tick numbers are estimated to represent potential blood losses from individual hosts of approximately 0.2-65% for rodents, 0.2-13% for hares and 0.3-9.0% for roe deer. Within the populations of all host species examined, the distributions of all stages of I. ricinus were clumped, with most host individuals harbouring few ticks and only a few individuals harbouring many ticks. The data suggest that, even though a small proportion of tick hosts may be severely affected, the direct effects of feeding by I. ricinus are unlikely to play an important role on mammal population dynamics.     

Chemical composition of lipophylic compounds from the body surface of unfed adult Ixodes persulcatus ticks (Acari: Ixodidae)

Chemical compositions of ethereal extracts of the body surface of unfed male and female Ixodes persulcatus ticks (Acari: Ixodidae) were studied by gas chromatography using mass-spectrometric detection. More than 100 different organic compounds were detected. The predominant components were saturated fatty hydrocarbons, saturated and unsaturated fatty acids, aldehydes, squalene, cholesterol and cholesterol derivatives. A number of compounds found in I. persulcatus are known as components of pheromones or constituents of dermal gland secretions in tick species of the genus Amblyomma: nonanoic acid, saturated fatty acids having from 14 to 16 carbons, and squalene. Saturated fatty aldehydes have not been reported previously as body surface components of hard ticks. Substituted phenols were not found in the extracts, although they are known as common components of sex and attraction–aggregation–attachment pheromones in Amblyomma ticks. With a few exceptions (henicosanal, 2,4-holestadiene and two unidentified cholesterol derivatives), there was no marked difference in composition of surface components between male and female I. persulcatus. The possible role of the different chemical groups in communication between I. persulcatus ticks is discussed.     

Detection of Lyme disease spirochete,Borrelia burgdorferi sensu lato,including three novel genotypes in ticks (Acari: Ixodidae) collected from songbirds (Passeriformes) across Canada

Lyme disease is reported across Canada, but pinpointing the source of infection has been problematic. In this three‐year, bird‐tick‐pathogen study (2004–2006), 366 ticks representing 12 species were collected from 151 songbirds (31 passerine species/subspecies) at 16 locations Canada‐wide. Of the 167 ticks/pools tested, 19 (11.4%) were infected with Borrelia burgdorferi sensu lato (s.l.). Sequencing of the rrf‐rrl intergenic spacer gene revealed four Borrelia genotypes: B. burgdorferi sensu stricto (s.s.) and three novel genotypes (BC genotype 1, BC genotype 2, BC genotype 3). All four genotypes were detected in spirochete‐infected Ixodes auritulus (females, nymphs, larvae) suggesting this tick species is a vector for B. burgdorferi s.l. We provide first‐time records for: ticks in the Yukon (north of 60° latitude), northernmost collection of Amblyomma americanum in North America, and Amblyomma imitator in Canada. First reports of bird‐derived ticks infected with B. burgdorferi s.l. include: live culture of spirochetes from Ixodes pacificus (nymph) plus detection in I. auritulus nymphs, Ixodes scapularis in New Brunswick, and an I. scapularis larva in Canada. We provide the first account of B. burgdorferi s. l. in an Ixodes muris tick collected from a songbird anywhere. Congruent with previous data for the American Robin, we suggest that the Common Yellowthroat, Golden‐crowned Sparrow, Song Sparrow, and Swainson's Thrush are reservoir‐competent hosts. Song Sparrows, the predominant hosts, were parasitized by I. auritulus harboring all four Borrelia genotypes. Our results show that songbirds import B. burgdorferi s.l.‐infected ticks into Canada. Bird‐feeding I. scapularis subadults were infected with Lyme spirochetes during both spring and fall migration in eastern Canada. Because songbirds disperse millions of infected ticks across Canada, people and domestic animals contract Lyme disease outside of the known and expected range.     

Optimal sampling and spatial distribution of Ixodes pacificus, Dermacentor occidentalis and Dermacentor variabilis ticks (Acari: Ixodidae)

A common method for sampling tick populations is flagging, which is a method of dragging a white cloth over a plant substrate for a fixed distance along a transect. Flagging over rough physical surfaces or using long subtransect lengths could lead to the underestimation of tick densities. Using estimates of the drop-off rates of adult Ixodes pacificus in flag sampling, optimal sampling schemes (the length and number of subtransects) were examined using the relationships between the tick drop-off rates (c), the tick density and distribution patterns and the roughness of the sampled plant substrate. It was found that the optimal number of subtransects and Lmax, the longest subtransect length which did not significantly underestimate the tick density from c, were affected by the tick density, substrate roughness and tick distribution pattern. This study also showed that the density and distribution of I. pacificus and Dermacentor occidentalis varied greatly over time in the populations sampled, while the Dermacentor variabilis densities were low and showed no significant changes over time. Both I. pacificus and D. occidentalis had clumped distributions along trails and these clumps were aggregated. However, the clump sizes (or individual clump areas) changed significantly over time because of density fluctuation or the movement of ticks. Finally, a positive association between the number of I. pacificus and D. occidentalis adults was observed from 2 m subtransect flag collections in March 1995; no relationship was found in 1994. © Rapid Science Ltd. 1998     

First report of Anaplasma phagocytophilum and its co-infections with Borrelia burgdorferi sensu lato in Ixodes ricinus ticks (Acari: Ixodidae) from Republic of Moldova

We examined 198 questing Ixodes ricinus ticks collected in Chisinau City, Republic of Moldova by PCR assays for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and co-infection of both pathogens, which were detected in 9%, 25.2% and 2.5% of tested ticks, respectively. B. burgdorferi s.l. genotyping revealed the presence of five genospecies with dominance of B. garinii. Our preliminary study provides evidence about occurrence of both pathogens in this populated area, which represent a potential health risk for inhabitants.     

Recent discovery of widespread Ixodes affinis (Acari: Ixodidae) distribution in North Carolina with implications for Lyme disease studies

Ixodes affinis, which is similar morphologically to Ixodes scapularis, is widely distributed in North Carolina. Collections have documented this species in 32 of 41 coastal plain counties, but no piedmont or mountain counties. This coastal plain distribution is similar to its distribution in Georgia and South Carolina, where it is considered an enzootic vector of Borrelia burgdorferi sensu stricto. An updated list of hosts for I. affinis in the U.S.A. is included, increasing the number to 15 mammal and one bird species. The presence of questing adults of I. affinis from April to November reinforces the need for confirmed identifications of suspected tick vectors of Borrelia spirochetes collected during warm months.     

Horizontal and vertical movements of host‐seeking Ixodes pacificus (Acari: Ixodidae) nymphs in a hardwood forest

The nymph of the western black‐legged tick (Ixodes pacificus) is an important bridging vector of the Lyme disease spirochete (Borrelia burgdorferi) to humans in the far‐western United States. The previously unknown dispersal capabilities of this life stage were studied in relation to logs, tree trunks, and adjacent leaf‐litter areas in a mixed hardwood forest using mark‐release‐recapture methods. In two spatially and temporally well‐spaced trials involving logs, the estimated mean distances that nymphs dispersed ranged from ≈0.04 to 0.20 m/day on logs vs 0.11 to 0.72 m/day in litter. Prior to recapture in either trial and within the confines of the sampling grids, the greatest estimated dispersal distances by individual nymphs released on logs, and in litter 0.5 m or 1.5 m from logs, were 2.4, 3.0, and 3.0 m, respectively. Nymphs released on logs or litter tended to remain within the same biotopes in which they were freed while host‐seeking. In two simultaneous trials involving trunks spaced close‐at‐hand, nymphs released at the trunk/litter interface on all four aspects collectively dispersed a mean of 0.353 m/day on trunks vs 0.175 m/day in litter. In either trial, the greatest distances that recaptured nymphs climbed trunks, or dispersed in litter in an encircling 3‐m grid, were 1.55 m and 2.97 m, respectively. Nymphs ascending trunks did not exhibit a preference for any one aspect, and the B. burgdorferi‐infection prevalences in nymphs that climbed trunks (3.2–4.0%) did not differ significantly from those that moved horizontally into litter (10.5–17.6%). We conclude that I. pacificus nymphs use an ambush host‐seeking strategy; that they disperse slowly in all biotopes studied; that they usually continue to host‐seek in or on whatever substratum they access initially; and that B. burgdorferi‐infected nymphs are as likely to move horizontally as vertically when offered a choice.     

Whole-Genome Sequencing of Borrelia garinii BgVir,Isolated from Taiga Ticks (Ixodes persulcatus)

Most Lyme borreliosis cases in Russia result from Borrelia garinii NT29 group infection. Borrelias of this group circulate exclusively in Ixodes persulcatus ticks, which are seldom found beyond Russia and the far east. Here we report the whole-genome sequence of Borrelia garinii BgVir isolated from an I. persulcatus female.     

Antibiotic Treatment in Mice Infected with Japanese Borrelia garinii: Efficacy of Ceftriaxone for Eradicating the Infection Induced by Ixodes persulcatus Tick Bites

We compared the efficacy of three antibiotics (ceftriaxone, erythromycin and clarithromycin) against Borrelia garinii infection in mice. The nymphal ticks of Ixodes persulcatus infected with the strain JEM6 of Japanese B. garinii were allowed to feed on female C3H mice. The mice were treated with each of the antibiotics for 5 consecutive days 1, 3, or 7 weeks after tick detachment. The doses of antibiotics per day were as follows: 5 mg intraperitoneal injection of ceftriaxone, 2 mg intraperitoneal injection of erythromycin and 1 mg peroral administration of clarithromycin. The infection status in treated mice was monitored by culturing their earlobes, hearts and urinary bladders in BSK II medium. Ceftriaxone eliminated borreliae completely; however, a recurrence of infection was observed in mice treated with erythromycin and clarithromycin.     

A case of abnormal size in eggs and larvae of the Taiga tick,Ixodes persulcatus (Acari: Ixodidae)

Egg batches laid by females of Ixodes persulcatus (maintained under a photoperiod of L:D 20:4 before feeding) contained eggs of two sizes: numerous normal eggs of 0.578 × 0.421 mm and some giant eggs of 0.776×0.515 mm. Giant eggs were 32–34% greater in length and 22–24% greater in width than normal eggs. Females maintained under a short-day photoperiod (L:D 12:12) laid only normal eggs (0.588×0.417 mm). There were no differences between these eggs in viability and developmental time, but larvae hatched from giant eggs were larger (length of body 23–28% greater and of gnathosoma 12–15% greater) than larvae from normal eggs. This dimorphism of eggs and larvae was not sex determined, but may be a maternal photoperiodic effect.     

Survival rates of immature Ixodes pacificus (Acari: Ixodidae) ticks estimated using field‐placed enclosures

Granulocytic anaplasmosis (GA) and Lyme borreliosis are emerging tick‐borne diseases caused by infection with Anaplasma phagocytophilum and Borrelia burgdorferi, respectively, and maintained in rodent‐Ixodes spp. tick cycles, including I. pacificus in the western U.S. Ixodes pacificus has a multiple‐year life cycle and B. burgdorferi and A. phagocytophilum are transstadially, but not transovarially, transmitted within ticks, thus ticks function importantly in maintaining infection in nature. In this study, the survival of larval and nymphal I. pacificus was determined using ticks placed in tubes in leaf litter from June 2005 to September 2006 at two field sites in the California northern coast range mountains and a laboratory control. In all three sites, nymphal and larval survival ranged from 90–400 d, with differences in mean survival among sites. Fewer ticks died in the autumn in the moister field sites compared with the drier incubator control treatment. The first large die‐off event in late autumn occurred at all sites shortly before relative humidity increased from 80–100% and temperature declined from approximately 22–15° C. The concurrent die‐off in the incubator population, subject to relative humidity and temperature regimes that were invariant, suggests that survival time was dependent on other factors in addition to environmental conditions. These results suggested that many ticks exhausted resources or tolerance for relatively low humidity within six months of questing, and that higher humidity prolonged survival. Based on observed longevity, humans and other animals could acquire A. phagocytophilum infection from adult I. pacificus that were infected up to three years earlier.     

Ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum ticks to acquire and transstadially transmit Borrelia garinii

In China, the ability of Ixodes persulcatus, Haemaphysalis concinna and Dermacentor silvarum to transmit Borrelia spirochetes was determined under laboratory conditions. Results showed that all three tick species can acquire spirochetes by feeding on infected mice. However, the capability of the three species to maintain spirochetes was different. Only I. persulcatus is able to maintain spirochetes alive during molting, and subsequent tick stages transmitted the infection to naive mice. In H. concinna and D. silvarum ticks, spirochetes could not survive after the digestion period of blood and after the molting process was finished, spirochetes were no longer detected. Contrary to H. concinna and D. silvarum, I. persulcatus should be considered the principal vector of Lyme disease in north China.