Aggregation in the tick Ixodes ricinus (Acari: Ixodidae): use and reuse of questing vantage points

Ongoing work in oak woods in Killarney National Park in southwestern Ireland is focusing on the factors influencing the fine-scale aggregated distribution of Ixodes ricinus L. (Acari: Ixodidae) on the ground. The extent of reuse of stems of vegetation as questing points by adult ticks was determined by paint-marking stems on which ticks were found, counting and removing these ticks, and subsequently reexamining the same stems for ticks on two further occasions. Overall, an estimated 2,967 stems in 123 separate rush plants (Juncus effusus L.) were examined. Statistical analysis of the data demonstrated a highly significant reoccupancy by ticks of stems previously and recently used. Furthermore, it is shown that the extent of stem reuse by ticks is significantly and positively correlated with the numbers of ticks originally observed on those stems. Although other factors may be involved in generating clumping of ticks, the results are compatible with the proposition that aggregation of I. ricinus on the ground is pheromone-mediated. The findings are discussed in relation to what is known about the powers of lateral movement of I. ricinus on the ground and the possible implications for the performance of tick traps.     

Origin of alimentary tract in embryogenesis of Ixodes ricinus (Acari: Ixodidae)

We studied the embryos of Ixodes ricinus (L.) in the second and third trimester of embryonic development, by using light and transmission electron microscopy. At the beginning of the second trimester, the formation of the foregut and rectal sac, by a process of invagination, was observed. The invagination, which develops into the primordium of the hindgut, forms only in the third trimester. The rectum forms in the last phase of embryogenesis. The development of the midgut is incomplete during embryogenesis. The yolk is surrounded by a wall, formed of an amorphous basal lamina and flattened cells, that gradually accumulate deutoplasmic material. These cells do not acquire the typical features of the gut epithelium until after larval hatching. These features are, however, found in the cells forming the rectal sac.     

Ultrastructural localization of a sialic acid-specific hemolymph lectin in the hemocytes and other tissues of the hard tick Ixodes ricinus (Acari; Chelicerata)

 Lectins have been suggested to function as pattern-recognition molecules in invertebrate immune mechanisms. A lectin from the hemolymph of the tick Ixodes ricinus with main specificity for sialic acid was characterized and antibodies directed against this lectin were prepared. In this study, these antibodies were used to localize the lectin in the tissues of I. ricinus. Immunoreactivity with poly- and monoclonal antibodies was detected in the granules of both types of granular hemocytes, at the membrane of hemocytes, and at the basal laminae surrounding the hemocoel. Furthermore, cells attached to the midgut, invaginations of Géné’s organ, and granular inclusions of nephrocytes were labeled. The immunoreactivity detected in hemocytes and the hemocoel lining supports the idea that the hemolymph lectin may function as a recognition molecule in the immune system of I. ricinus. Another function could be protection of eggs that are coated with secretions by Géné’s organ. The lectin activity could also be involved in transmission of Borrelia burgdorferi, the causative agent of Lyme borreliosis, and the tick-borne encephalitis virus. Received: 7 June 1995 / Accepted: 29 August 1995     

Distribution of the common tick, Ixodes ricinus (Acari: Ixodidae), in different vegetation types in southern Sweden

The aim of this study was to investigate whether differences in Ixodes ricinus (L.) nymphal relative density exist among different vegetation types in southern Sweden. Nymphal I. ricinus were sampled in southeastern Scania in southern Sweden during June-August 1997. A total of 110-180 25-m2 samples were taken by blanket-dragging from each of nine different vegetation types. There was a highly significant difference in nymphal abundance between the open areas as one group and the forested areas as another group (P < 0.0001). Vegetation types that differed significantly in median nymphal abundance from all other vegetation types were pine forest (16 nymphs/100 m2) and beech forest (40 nymphs/100 m2). No significant differences in median nymphal density were revealed among mixed deciduous forest, alder forest, oak forest, and hazel forest (28-32 nymphs/100 m2), or among dry meadow, meadow, and heath (0 nymphs/100 m2). Forestation of open areas is likely to lead to increased I. ricinus abundance and disease risk.     

Subadult Ixodes ricinus (Acari: Ixodidae) on rodents in Berlin, West Germany

To identify hosts that may serve as reservoirs for the agent of Lyme disease in Central Europe, we determined whether Ixodes ricinus L. feed most frequently on certain rodents and whether the abundance of these hosts corresponds to the season of feeding activity of the tick in four sites in Berlin, Federal Republic of Germany. In addition, we correlated abundance of I. ricinus with that of particular rodent hosts. Two small rodents were more abundant than any others; a mouse, Apodemus flavicollis, predominated in a wooded site and a vole, Clethrionomys glareolus, in three brush- or grass-covered sites. The tick was most abundant in the mouse-infested site. Although A. flavicollis comprised only about a third of rodents collected, nearly 60% of all such rodent parasitizing I. ricinus fed on this mouse. These ticks were more abundant on mice than voles in each of the study sites and throughout the year, and more larvae fed on these rodents than did nymphs. Although larval as well as nymphal I. ricinus are most abundant during midsummer, they feed on rodents from April through October. Taken together, these observations suggest A. flavicollis as a potentially important reservoir host for I. ricinus-borne infections.     

Winter activity of Ixodes scapularis (Acari: Ixodidae) and the operation of deer-targeted tick control devices in Maryland

Host-seeking activity of adult blacklegged ticks, Ixodes scapularis Say was monitored by flagging during winter months in Beltsville, MD. Ticks were active many days during January and February, the coldest months, with some captures made when there was 70% snow cover and temperatures as low as -2 degrees C. Substantial numbers (70-90 ticks/h of flagging) of adult I. scapularis were captured on favorable days in January and February. The cost to treat white-tailed deer, Odocoileus virginianus (Zimmermann), using '4-poster' deer self-treatment devices, was estimated on a per female tick basis. We estimated deer abundance and tick attachment rates using data from the literature, tick activity levels using weather data and drag counts of ticks, and costs to operate the devices from experience. We found that self-treatment devices need not be operated continuously from late December until the third week of February. On average, savings of half the operating costs would be realized by not operating the devices when tick activity was low.     

Prevalence of spotted fever group rickettsiae in Ixodes ricinus (Acari: Ixodidae) in southern Germany

Host-seeking Ixodes ricinus (L.) (Acari: Ixodidae) ticks were collected systematically, from May to September 2006, at selected sites in southern Germany, including a large city park in Munich. Polymerase chain reactions for amplification of genes of the rickettsial citrate synthase (gltA), the outer membrane proteins A and B (ompA and ompB), and the 16S rDNA were used to investigate 2,861 specimens (adults and nymphs). GltA sequences of spotted fever group rickettsiae were detected in 151 of all samples (5.3%; 95% CI = 4.3-6.2%). Sequencing revealed Rickettsia helvetica in 91.4% of the samples and R. monacensis in 8.6%. Amplification of ompA was not possible for R. helvetica, but in all except one of the R. monacensis. The results were analyzed statistically to test the effects of season, location, developmental stage, and gender of the tick on prevalence of Rickettsia spp. Although rickettsial DNA was detected in all investigated sites, sites in natural forest areas had significantly higher prevalences than sites in landscaped city parks. Adult female and male ticks had a similar prevalence and were significantly more often infected than nymphs. Monthly differences were not statistically significant. These results clearly show that R. helvetica is widespread throughout the study region and could result in a threat to public health in areas of high prevalence.     

Distribution of 'Ehrlichia walkeri' in Ixodes ricinus (Acari: Ixodidae) from the northern part of Italy

The prevalence of 'Ehrlichia walkeri' in Ixodes ricinus in northern Italy was studied. A total of 456 ticks were collected from seven places around Belluno, and 292 living ticks were examined using polymerase chain reaction (PCR). Nineteen ticks were positive for gltA of 'E. walkeri', and the prevalence rate for each place ranged from 0 to 12.8%. The disparity of prevalence rates among areas examined suggests that 'E. walkeri' is not widespread in ticks and requires an animal host for its maintenance in nature.     

Infection of Ixodes ricinus (Acari: Ixodidae) by Borrelia burgdorferi sensu lato in North Africa

Free-living adult Ixodes ricinus L, were collected in Amdoun, situated in the Kroumiry mountains in northwestern Tunisia (North Africa). Using direct fluorescence antibody assay, the infection rate of field-collected I. ricinus by Borrelia burgdorferi sensu lato was 30.5% (n = 72). No difference in infection rate was observed between male and female ticks. Spirochetes that had been isolated from I. ricinus from Ain Drahim (Kroumiry Mountains) in 1988 were identified as Borrelia lusitaniae (formerly genospecies PotiB2). This is the first identification of a genospecies of Borrelia burgdorferi sensu lato from the continent of Africa.     

Comparative behavior of different life-cycle stages of Ixodes ricinus (Acari: Ixodidae) to human-produced stimuli

Nymphs of Ixodes ricinus (Linnaeus, 1758), I. scapularis Say, 1821, and I. pacificus Cooley & Kohls, 1943 are epidemiologically the most dangerous stage for transmission of Lyme disease to humans. Many factors play a role in the epidemiological significance of the nymphs. In this study, we address the question of whether nymphs show a greater tendency than adults to accept humans as their host. To evaluate this, we have compared the host acceptance behavior of nymphs and adults (males and females) with respect to a human in Rambouillet forest, a focus of Lyme disease. Individual ticks (nymph, male or female) located on a herbaceous stem were exposed to different stimuli (e.g., approach, stem movement, breathing), and the response of each individual to these stimuli was noted. Tick responses were categorised into classes (from 0 to 3) according to their intensity. Statistical analysis carried out on 22 ticks allowed us to compare the behavior of the nymph stage with respect to males and females. Despite the small sample sizes, it appears that nymphs are more responsive to a human than are the adults.     

Ixodes dammini (Acari: Ixodidae) in Indiana

Ixodes dammini Spielman, Clifford, Piesman and Corwin is reported from Indiana for the first time. Four specimens were taken from deer and one from a dog. All specimens are adult females and all were collected in 1987. The significance of this finding is discussed.     

Distribution, abundance, and habitat preferences of Ixodes ricinus (Acari: Ixodidae) in northern Spain

Ixodes ricinus (L.) was collected by standard dragging in 2,082 different sites in 18 broad vegetation categories in northern Spain to explore the influence of vegetation on its abundance. Of these, 785 sites were surveyed in 1995, 636 in 1996, and 661 in 1997. The impact of habitat features on differences in tick numbers is addressed. The tick was present in low numbers in areas of old, heterogeneous coniferous forests. Ticks appeared to prefer sites that had substantial secondary plant growth such as river canopies, heterogeneous Pinus uncinata forests, mixed forests, and deciduous heterogeneous woods. Highest tick abundance was recorded for sites that contained Quercus spp., as well as for mixed old forests that had many ecotones. I. ricinus was absent in open habitats, homogeneous young coniferous forests, and open hillsides. These differences were attributed to greater shrub cover and litter depth, which created more favorable microclimatic conditions for tick survival. The abundance of I. ricinus nymphs was not homogeneous in sites within the same habitat category and vegetation physiognomy at these sites did not appear to cause differences in tick abundance. Analysis of variance (ANOVA) indicated that variation in tick abundance could be explained by the exposure of the sampled site, at least for some zones within deciduous forest categories. However, this factor did not explain the variation observed in other habitats. Temperature and vegetation (normalized derived vegetation index) features were recorded daily by remotely sensed imagery throughout the study period and the data were used to obtain long-term mean and maximum values of the physical parameters considered. Multiple regression analysis performed between these long-term abiotic factors and nymphal abundance in positive sites showed high relationship (R2 coefficients) for every habitat category and explained >50% of the variation in tick abundance.     

Seasonal and spatial occurrence of glycerol-3-phosphate dehydrogenase variability in Ixodes ricinus (Acari: Ixodidae) populations

Previous results indicate that glycerol-3-phosphate dehydrogenase variability represents the adaptation of Ixodes ricinus L. (Acari: Ixodidae) to fluctuations of environmental conditions, particularly to temperature. Analysis of crucial polymorphisms in I. ricinus Gpdh gene was done by the restriction method, and three different haplotypes were obtained (GPDH441 1, GPDH441 2, and GPDH441 3), corresponding to GPDH alleles detected by allozyme electrophoresis. Differences in GPDH441 haplotype and genotype frequencies were found between samples from open and forest habitats. Significant seasonal variations of GPDH441 haplotype and genotype frequencies were detected in samples from the open habitats. No seasonal variations were observed at forest localities, probably because of the less pronounced amplitude of environmental factors. The possible role of host availability was discussed as an important factor that affects seasonal dynamics and genetic composition of tick populations.     

Rickettsiella-like bacteria in Ixodes woodi (Acari: Ixodidae)

We examined a parthenogenetic strain of the hard tick Ixodes woodi Bishopp for the presence of endosymbiotic bacteria. Electron microscopic examination revealed the ovarian tissues and Malpighian tubules were infected with pleomorphic bacteria. Two basic types were observed: a larger granular cell and a smaller condensed cell. Cloning and sequence analysis of polymerase chain reaction (PCR) amplified 16S rRNA gene yielded a single sequence from bacteria present in I. woodi tissues. Phylogenetic analysis of the nearly complete 16S rDNA indicated that the ticks were infected with an endosymbiont belonging to the gamma subdivision of the Proteobacteria. It clustered with the insect pathogenic species Rickettsiellagrylli (Vago and Martoja 1963) and the animal pathogen Coxiella burnetii (Derrick 1939) Philip 1948. Our results suggest that the I. woodi females harbored a single endosymbiotic bacterium related to selected Rickettsiella species and to C burnetii.     

Taxonomic status of Ixodes didelphidis (Acari: Ixodidae)

Ixodes didelphilis Fonseca & Arag?o was described in Brazil in 1952 as a new tick species that differed from Ixodes loricatus Neumann by the spiracular plate pattern. We have reared four tick colonies from different geographic areas in the laboratory that were started from single engorged females originally identified as I. didelphidis (BMG, colony ) and I. Iocicatus (CSP, PSP, and TRJ colonies). We analyzed the spiracular plate morphology of F1 adult ticks from each tick colony, compared their biological 11th, and performed a molecular analysis of the second internal transcribed rDNA spacer (ITS2) to test the validity of the species I. didelphidis. The spiracular plate analysis of laboratory F1 adult ticks yielded single females from the four colonies showed variations that invalidate morphological parameters for differentiation of loricatus and I. didelphidis. Biological data of the BMC, CSP, and TRJ colonies were similar. The biology ofthe PSP colony was not evaluated. The ITS2 sequence variations observed between the tick colonies ranged from 1.3 to 4.9%, and the similarity tree constructed by the neighbor-joining method with nucleotide distances showerl that the distances between the samples were similar to what is expected for intraspecific variations found in other ticks species. The morphological and biological results, in conjunction with the ITS2 analysis, supported the conspecificity I. loricatus and I. didelphidis.     

Host associations of the tick, Ixodes angustus (Acari: Ixodidae), on Alaskan mammals

Infestation parameters are presented for 227 ticks, all Ixodes angustus Neumann, collected from individual mammals (n = 531) in southeastern and south-central Alaska from 1996 to 1999. This tick was recovered from 12 of the 19 mammal species examined, with four species of shrews (Sorex spp.), two species of voles [Clethrionomys gapperi (Vigors) and Clethrionomys rutilus (Pallas)], one species of mouse [Peromyscus keeni (Rhoads)], and the red squirrel [Tamiasciurus hudsonicus (Erxleben)] being the most frequently parasitized hosts. Larvae (n = 146) of I. angustus were collected most often, followed by nymphs (n = 50), females (n = 30), and a single male. The leptinid mammal-nest beetle Leptinus occidentamericanus Peck (1 male, 5 females) was also recovered from five individual small mammals; three of these were C. rutilus.     

Characterization of a new spotted fever group rickettsia detected in Ixodes ricinus (Acari: Ixodidae) collected in Slovakia

Two previously undescribed rickettsiae were detected in Ixodes ricinus Ricketts by polymerase chain reaction. Ixodes ricinus Slovakia (IRS) 3 and IRS4 were identified in ticks collected in northeastern and southwestern Slovakia, respectively. Sequences of the 16S rRNA citrate synthase (gltA) and outer membrane protein rOmpA (ompA) encoding genes of both strains were nearly identical but were distinct from those of all other known rickettsiae. Phylogenetic relationships inferred from the comparison of these sequences with those of other members of the genus Rickettsia indicate that IRS3 and IRS4 constitute a new rickettsial genotype and form a separate cluster among the spotted fever group rickettsiae.     

Status of Ixodes dammini (Acari: Ixodidae) in Illinois

Ixodes dammini Spielman, Clifford, Piesman & Corwin was found for the first time in Illinois in November 1987, when two adult females were collected from two deer in Jo Daviess County in the northwestern corner of the state. In 1988, in a study of six state parks in northern Illinois, questing adults and nymphs were encountered in one park in Ogle County. During the firearm deer hunt in November 1988, adult female and male ticks were found in several counties, with a high rate of infestation (greater than 25%) in two counties (Ogle and Rock Island) along the Rock River, which flows from Wisconsin into the Mississippi River. Several cases in humans with no history of travel outside of the state have been reported, primarily from northern Illinois. We suspect that infiltration of infected ticks and wildlife from Wisconsin is resulting in the emergence of Lyme disease in Illinois. Because all the components necessary for the completion of the tick life cycle and for the transmission of Borrelia burgdorferi are available throughout much of the state, I. dammini and Lyme disease can spread and become established in large portions of Illinois.