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.     

Spatiotemporal patterns of host-seeking Ixodes scapularis nymphs (Acari: Ixodidae) in the United States

The risk of Lyme disease for humans in the eastern United States is dependent on the density of host-seeking Ixodes scapularis Say nymphal stage ticks infected with Borrelia burgdorferi. Although many local and regional studies have estimated Lyme disease risk using these parameters, this is the first large-scale study using a standardized methodology. Density of host-seeking I. scapularis nymphs was measured by drag sampling of closed canopy deciduous forest habitats in 95 locations spaced among 2 degrees quadrants covering the entire United States east of the 100th meridian. Sampling was done in five standardized transects at each site and repeated three to six times during the summer of 2004. The total number of adults and nymphs of the seven tick species collected was 17,972, with 1,405 nymphal I. scapularis collected in 31 of the 95 sites. Peak global spatial autocorrelation values were found at the smallest lag distance (300 km) and decreased significantly after 1,000 km. Local auto-correlation statistics identified two significant high-density clusters around endemic areas in the northeast and upper Midwest and a low-density cluster in sites south of the 39th parallel, where only 21 nymphs were collected. Peak nymphal host-seeking density occurred earlier in the southern than in the most northern sites. Spatiotemporal density patterns will be combined with Borrelia prevalence data as part of a 4-yr survey to generate a nationwide spatial risk model for I. scapularis-borne Borrelia, which will improve targeting of disease prevention efforts.     

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.     

Mitotic activity of the hemocytes in the tick Ixodes ricinus (Acari; Ixodidae)

 The blood cells, or hemocytes, of Ixodes ricinus have been shown to recognize, attack, and phagocytose microorganisms invading the body cavity, or hemocoel, of this tick. Regulated proliferation and differentiation of hemocytes, also referred to as immunocytes, is basic to an effective immune response to invading microorganisms. Therefore, this study dealt with hemopoiesis in I. ricinus, the vector tick of the Lyme disease spirochete Borrelia burgdorferi. Histological evidence for the presence of hemopoietic tissue, a preferential proliferation site of hemocytes, is presented. Mainly the mitotic activity of free-floating hemocytes was examined. By means of microscopical photometry and flow cytometry, all three types of hemocytes in engorging female I. ricinus were found in different stages of the cell cycle. In the engorging tick, up to 40% of the hemocytes counted were in the S phase or the G₂/M phase. From this study we conclude that the differentiated hemocyte types do not differentiate from stem cells in the adult tick. Moreover, microorganisms entering the hemocoel of engorging ticks are confronted with high numbers of hemocytes and, therefore, with an effective cellular immune response. Received: 20 October 1995 / Accepted: 30 January 1996     

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.     

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.     

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.     

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.     

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.     

Seasonal activity and host associations of Ixodes scapularis (Acari: Ixodidae) in southeastern Missouri

Based on tick collections recovered from wild vertebrates and by dragging, the seasonal occurrence of adult blacklegged ticks, Ixodes scapularis Say, extended from October through May in southeastern Missouri. Adult activity was bimodal with the higher peak occurring in November followed by a lower peak in February. The activity of immature I. scapularis had the general pattern of that found in the Northeast where Lyme disease is hyperendemic, with larval activity (July) peaking after that of nymphs (May and June). Vertebrates varied in their importance as hosts of I. scapularis. White-tailed deer, Odocoileus virginanus (Zimmerman), and coyotes, Canis latrans Say, were the primary hosts of adult I. scapularis. Broad-headed skinks, Eumeces laticeps (Schneider), and eastern fence lizards, Sceloporus undulatus (Latreille), were the primary hosts of nymphal I. scapularis. The broad-headed skink, 5-lined skink, Eumeces fasciatus (L.), and Carolina wren, Thryothorus ludovicianus (Latham), were the primary hosts of larval I. scapularis. Homeotherms were important hosts of immature I. scapularis, accounting for 30% of nymphs and 39% of larvae collected. The eastern cottontail rabbit, Sylvilagus floridanus (Allen), may play an important role in the epidemiology of Lyme disease in Missouri. Isolates of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner were made from ticks recovered from rabbits, making the cottontail rabbit a key species for further study of the epidemiology of Lyme borreliosis in Missouri.     

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.     

Detection of Anaplasma phagocytophilum and Babesia odocoilei DNA in Ixodes scapularis (Acari: Ixodidae) collected in Indiana

The blacklegged tick, Ixodes scapularis Say, first reported in Indiana in 1987, has now been detected in more than half of Indiana's counties. The first case of human granulocytic ehrlichiosis (human anaplasmosis) in Indiana was reported in 2002. We now report the detection of Anaplasma phagocytophilum and Babesia odocoilei (Emerson and Wright 1968) in I. scapularis ticks collected in northern Indiana. Using polymerase chain reaction analysis, 41 of 193 adult ticks (21.2%) collected from deer were positive for A. phagocytophylum, and 22 (11.4%) were positive for Babesia sp. Restriction fragment analysis of 12, and sequencing of another five of the amplified products identified these parasites as B. odocoilei. Five ticks (2.6%) were coinfected. Eight of 68 questing adult ticks (11.8%) were positive for A. phagocytophilum; seven (10.3%) were positive for Babesia sp. Six of the latter seven positive samples were determined to be B. odocoilei by restriction fragment analysis and sequencing of two samples. None of 39 pools of nymphs was positive for Babesia sp. Three of 15 ticks (20%) collected from a dog were positive for A. phagocytophilum and three ticks (20%) were positive for Babesia sp. One was confirmed as B. odocoilei. One tick was coinfected. This is the first report of the presence of these two agents in ticks in Indiana.     

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.     

Different activities and footwear influence exposure to host-seeking nymphs of Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae)

The relative potential for a person accidentally acquiring host-seeking nymphs of the blacklegged tick, Ixodes scapularis Say, and lone star tick, Amblyomma americanum (L.), while wearing either of two types of footwear, walking, crawling on hands and knees, and sitting on large fallen logs in deciduous woods, was evaluated. Although flag samples indicated substantial populations of I. scapularis nymphs and low to moderate numbers of A. americanum at the study sites, relatively few I. scapularis and fewer still A. americanum nymphs were acquired during 30-s and 5-min walks. Significantly fewer I. scapularis were picked up when boots were, worn with ankles taped (an anti-tick precaution) than when sneakers were worn with socks exposed during 5-min walks, but when thus attired, there was no significant difference between the number of nymphs acquired during 30-s walks. Nymphs of I. scapularis did not appear to accumulate incrementally on footwear or clothing during walks when boots were worn and ankles taped. Crawling for 30 s (approximately 3 m distance) yielded significantly more I. scapularis nymphs than walking for 30 s. During crawling, I. scapularis nymphs were picked up on 58% of the 30-s samples. Most ticks picked up during crawls were on pant legs. When a flannel flag cloth (0.5 by 0.5 m) was appressed to the upper surface of logs suitable to be sat upon by tired hikers, I. scapularis nymphs were found on 87% of the logs and in 36% of the samples. These data indicate that the potential for contact with host-seeking nymphs of I. scapularis occurring at these densities is greatly elevated by engaging in activities that involve contact with fallen logs and close contact of hands and knees with leaf litter.     

New host of Ixodes dentatus (Acari: Ixodidae), and first record in Tennessee.

Two specimens of Ixodes dentatus Marx were found in southwestern Tennessee in Shelby County during a 14-mo tick survey. This was the first record for this tick in Tennessee. An adult female was found on a dry ice trap and one larva was found on a live-trapped Peromyscus leucopus, the first time this tick has been reported from P. leucopus. This may be another combination of vector and host in the ecology and transmission of B. burgdorferi in the wild.     

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.     

Borrelia lusitaniae and spotted fever group rickettsiae in Ixodes ricinus (Acari: Ixodidae) in Tuscany, central Italy

Prevalence of infection by Borrelia burgdorferi s.l. and spotted fever group (SFG) rickettsiae was estimated in host-seeking ticks in an area in Tuscany, central Italy, where Lyme borreliosis was reported in a forestry worker. B. burgdorferi s.l. was identified by polymerase chain reaction in 16.7% (95% CI = 10.3, 24.8) of Ixodes ricinus (L.) nymphs and 39.6% (95% CI = 26.5, 54.0) of adults. Borrelia lusitaniae accounted for 82.9% of positive samples, followed by Borrelia garinii (9.8%), Borrelia afzelii (2.4%), and Borrelia burgdorferi s.s. (2.4%). One Rhipicephalus spp. adult was infected with B. garinii (prevalence = 8.3%; 95% CI = 0.21, 38.5). Prevalence of infection by SFG rickettsiae was 38.5% (95% CI = 26.7, 51.4) in I. ricinus nymphs, 34.6% (95% CI = 22.0, 49.1) in I. ricinus adults, and 50% (95% CI = 21.1, 78.9) in Rhipicephalus spp. adults. Phylogenetic analysis showed the similarity of B. lusitaniae strains that were identified in this study and of a strain that was previously isolated from a human patient in Portugal. Results of this study confirm the dominance of B. lusitaniae in areas in the Mediterranean basin and the infection by SFG rickettsiae in I. ricinus.