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PubMed1999

Influence of the genetic background and parasite load of mice on the immune response developed against nymphs of Ixodes ricinus.

The immune response of BALB/c (H-2d), DBA (H-2d), C57BL/6 (H-2b), C3H (H-2k), CBA (H-2k), SJL (H-2s), and FVB (H-2q) mice infested once with 15 nymphs of Ixodes ricinus is polarized toward Th2 as suggested by cytokines produced by lymph node cells stimulated with concanavalin A. The parasite load does not influence the polarization of the immune response as observed in BALB/c mice, which developed a Th2 response when infested with 5 or 45 nymphs. As assessed by attachment and weights of engorged nymphs, no resistance was acquired by BALB/c, C57BL/6, or C3H mice undergoing three successive infestations. However, these mice produced a gradual increase in IgE.

Animals↗
PubMed2000

Cytokines (IL-4 and IFN-gamma) and antibodies (IgE and IgG2a) produced in mice infected with Borrelia burgdorferi sensu stricto via nymphs of Ixodes ricinus ticks or syringe inoculations.

Mice were tolerant to tick bites during three infestations with nymphs of Ixodes ricinus infected with Borrelia burgdorferi sensu stricto. To determine whether tick bites influence the immune response against B. burgdorferi, we examined the production of cytokines IL-4 and IFN-gamma by lymph node cells of BALB/c mice and IL-4 deficient BALB/c mice after tick inoculation versus syringe inoculation of B. burgdorferi. We also measured IgG2a anti-borrelial antibodies and total IgE in these mice. Results showed that BALB/c mice developed a Th2 immune response against B. burgdorferi after tick inoculation and a mixed Th1/Th2 response after syringe inoculation of B. burgdorferi. IL-4 deficient mice produced a Th1 immune response in both cases. IL-4 produced following tick bites greatly decreased the production of anti-borrelial IgG2a antibodies by comparison with the production of anti-borrelial IgG2a antibodies produced following syringe injection of B. burgdorferi.

Animals↗
PubMed2006

Prevalence of Borrelia burgdorferi sensu lato in the tick Ixodes ricinus in the Styrian mountains of Austria.

A total of 691 Ixodes ricinus (22 male, 39 female, 501 nymphs and 129 larvae), the tick vector of Lyme borreliosis, were collected by flagging from vegetation in 11 areas at altitudes between 789 m and 1350 m above sea level in mixed woodland with pasture land (cattle) in the province of Styria in Austria. The ticks were individually examined for presence of Borrelia burgdorferi sensu lato by dark-field microscopy and 107 of them by real-time PCR. Attempts to cultivate borreliae were made in BSK-H medium. The overall positivity rate of all collected ticks (excepting larvae) was 10.9%: 9.1% in males, 17.9% in females and 10.4% in nymphs. The 129 larvae examined showed no presence of B. burgdorferi s.l. The mean infection rate of I. ricinus collected at the highest altitude in this study, Gaberl at 1350 m a.s.l.--and at the same time the highest one reported in Europe--was 6.4%: 1/9 males, 2/18 females and 6/114 (5.3%) nymphs were positive. Culture attempts were positive in 12 cases and species identification showed eight isolates were B. afzelii and four B. garinii. Three additional positive results found by PCR (negative by culture) were identified twice as B. afzelii and once as B. garinii. This study shows that the risk of acquiring Lyme borreliosis in habitats at higher altitudes is limited, because of the lower density of I. ricinus and its lesser infection rate than at lower altitudes in central Europe, but nevertheless the risk does exist.

Animals↗
PubMed1997

Definitive identification of louping ill virus by RT-PCR and sequencing in field populations of Ixodes ricinus on the Lochindorb estate.

Rapid and precise virus detection procedures are an important component of any epizootiological study. An automated one tube reverse transcriptase and nested primer polymerase chain reaction (RT-PCR) followed by nucleotide sequencing of the cDNA product, was used for the rapid detection and identification of louping ill (LI) virus in field caught Ixodes ricinus and compared with a classical isolation method i.e. infectivity in cell culture. The results establish the genetic identity of LI virus on the Lochindorb Estate. There was a high correlation between the results obtained by RT-PCR and infectivity assays. RT-PCR and sequencing proved to be a rapid and accurate system for identifying LI virus in field specimens. Development of this system should improve the capacity to undertake detailed epizootiological studies of LI virus.

Animals↗
PubMed2003

Prevalence of four species of Borrelia burgdorferi sensu lato and coinfection with Anaplasma phagocytophila in Ixodes ricinus ticks in central Germany.

A total of 305 Ixodes ricinus ticks collected from three areas of Thuringia in central Germany were investigated for infection with Borrelia burgdorferi sensu lato species and Anaplasma phagocytophila. Overall, 11.1% were infected with Borrelia burgdorferi and 2.3% with Anaplasma phagocytophila. Adult ticks showed a significantly higher rate of infection with both borreliae and Anaplasma phagocytophila. Borrelia garinii (55.9%) was detected most frequently, followed by Borrelia burgdorferi sensu stricto (32.4%), Borrelia afzelii (17.6%), and Borrelia valaisiana (5.9%). Four ticks had dual infection with Borrelia garinii and Borrelia burgdorferi sensu stricto. Two of the Borrelia-positive ticks were coinfected with Anaplasma phagocytophila.

Adult↗
PubMed1999

Incidence of Borrelia garinii and Borrelia afzelii in Ixodes ricinus ticks in an urban environment, Prague, Czech Republic, between 1995 and 1998.

The presence of Borrelia burgdorferi sensu lato and its individual genospecies was determined using a single-step polymerase chain reaction in a total of 779 Ixodes ricinus ticks collected from an urban park in Prague, Czech Republic. The incidence of Borrelia burgdorferi sensu lato was 9.2% in 1995, 3.4% in 1996, 4.5% in 1997 and 2.8% in 1998. The Borrelia garinii to Borrelia afzelii ratio was 1.4:1 and it did not differ significantly throughout the study period. Borrelia burgdorferi sensu stricto was not detected. Borrelia garinii/Borrelia afzelii coinfection was found in 5.7% of positive ticks. The results indicate it is possible for the urban population to come in contact with the causative agent of Lyme disease in an urban park.

Animals↗
PubMed2005

Ontogeny of tick hemocytes: a comparative analysis of Ixodes ricinus and Ornithodoros moubata.

Hemocytes of two tick species, Ixodes ricinus and Ornithodoros moubata, were investigated with the aim to determine their ultrastructural characteristics and developmental relationships. Only a limited number of ultrastructural features was shown to be unequivocally homological across all hemocyte types. The two species, representing distant groups of ticks, differ in the composition of their circular cell populations. In I. ricinus, three groups of distinct morphological types of hemocytes could be determined according to well-defined ultrastructural features: a typical non-phagocytic granular cell with electron-dense granula and homogeneous cytoplasm (Gr II), and two different types of phagocytic hemocytes, namely plasmatocytes with a low number of granula and phagocytic granolocytes, designated as Gr I. In contrast, an additional cell type resembling insect spherulocytes was determined in O. moubata. This cell type does not seem to be homologous to any I. ricinus hemocyte and may represent a cell type typical of soft ticks only. Possible ontogenetic lineages of the hemocytes of both tick-species were inferred.

Animals↗
PubMed2005

Pathogenicity of Metarhizium anisopliae (Deuteromycetes) and permethrin to Ixodes scapularis (Acari: Ixodidae) nymphs.

Effectiveness of the entomopathogenic fungus Metarhizium anisopliae, for controlling nymphal Ixodes scapularis, was tested in laboratory and field trials. In the laboratory, M. anisopliae (Metschnikoff) Sorokin strain ESC1 was moderately pathogenic, with an LC50 of 10(7) spores/ml and induced 70% mortality at 10(9) spores/ml. In a field study, however, 10(9) spores/ml M. anisopliae did not effectively control questing I. scapularis nymphs, and significant differences were not detected in pre- and post-treatment densities. For nymphs collected and returned to the laboratory for observation, mortality was low in treatment groups, ranging from 20 to 36%. To assess whether a chemical acaricide would synergistically enhance pathogenicity of the fungus, we challenged unfed nymphal I. scapularis with combinations of M. anisopliae and permethrin, a relatively safe pyrethroid acaricide, in two separate bioassays. Significant interactions between M. anisopliae and permethrin were not observed, supporting neither synergism nor antagonism.

Animals↗
PubMed2005

The parasitism of immature stages of Ixodes loricatus (Acari: Ixodidae) on wild rodents in Argentina.

Ixodes loricatus has long been considered with strict-total specificity to New World Marsupials. However, frequent findings of its immature stages on rodents suggest that these vertebrates play an important role in the tick's life cycle. Aspects dealing with the ecology of Sigmodontinae rodents infestation by I. loricatus are unknown. To contribute to the knowledge of the ecology of this tick species, environmental factors, as well as host species, sex and age, were evaluated to find associations of immature I. loricatus infestation of the most abundant wild rodent species from riparian locations of Buenos Aires province (Argentina). A total of 290 hosts belonging to the subfamily Sigmodontinae were captured and examined for ticks at six locations. The data analysis showed that there was a preference of I. loricatus towards Akodon azarae, but only for those hosts captured in lands not prone to flooding. The results suggest that the host preference of I. loricatus is mainly related to factors such as habitat type and host density.

Age Factors↗
PubMed2005

Hosts and pathogen detection for immature stages of Ixodes ricinus (Acari: Ixodidae) in North-Central Spain.

To determine hosts of the immature stages of a southern population of Ixodes ricinus, we trapped rodents and birds in an area of north-central Spain in May-June and August-September of 1998 and 1999. The most frequently trapped rodents were Apodemus sylvaticus (230 specimens) and Clethrionomys glareolus (99), with a larval infestation prevalence of 49% and 81%, respectively (in spring) and 21% and 41% (in summer). C. glareolus was always more heavily parasitized by larvae (mean numbers 19.8 in spring, 3.4 in summer) than A. sylvaticus (6.1 and 1.2, respectively). No nymphs were collected from the rodents. The larval distribution pattern differed considerably between rodent host species but not according to the season of the year. The most commonly trapped birds were members of the Turdidae, which had infestation incidences of about 70% (larvae) and 10% (nymphs) in spring and 20-15% in summer. We also used molecular methods for the identification of reservoir hosts of the larval ticks and of the pathogens they contained, from nymphs collected in the same site in June of 2002 and 2003. The study showed that most of the ticks had fed on birds and few on rodents and ruminants. Pathogens detected in these ticks included Babesia microti, Borrelia garinii, B. valaisiana and B. afzelii. Borrelia afzelii was detected in a tick that had apparently fed on a wild boar.

Animals↗
PubMed2006

Allozyme polymorphism of Mdh and alpha-Gpdh in Ixodes ricinus populations: comparison of borreliae-infected and uninfected ticks.

Ixodes ricinus Linnaeus (Acari: Ixodidae) ticks are vectors of numerous infectious diseases in humans and animals. The allozyme variability of MDH and alpha-Gpdh was detected by native polyacrylamide gel electrophoresis in I. ricinus natural populations in three localities in Serbia. Four alleles of Mdh locus (MDH 1, MDH 2, MDH 3 and MDH X) and four alleles of alpha-Gpdh locus (VS, S, F and VF) were detected. Interpopulation differences in Mdh and alpha-Gpdh allele frequencies were statistically insignificant. Significant difference in alpha-Gpdh allele frequencies between males and females was recorded in the largest sample only. Differences in allele frequencies, detected between borreliae-infected and uninfected I. ricinus ticks, were close to the level of statistical significance, especially for alpha-Gpdh locus. Clear significant difference appeared in females when sexes were tested separatelly (P = 0.037). It is interesting that genotypes containing rarer alleles (MDH 1 and S) were infected in higher proportion in comparison to other genotypes. Our results point towards a possible role of Mdh and alpha-Gpdh loci in I. ricinus ticks in the determination of energy requirements for host seeking. Sex differences in alpha-Gpdh allele frequencies suggest that selective pressure, concerning efficiency of reserve materials utilisation, points to alpha-Gpdh rather than to Mdh locus.

Animals↗
PubMed2006

Repellency of MyggA Natural spray (para-menthane-3,8-diol) and RB86 (neem oil) against the tick Ixodes ricinus (Acari: Ixodidae) in the field in east-central Sweden.

In the field in south-central Sweden, we tested by randomised, standardised methodology the potential anti-tick repellent activity of two concentrations of MyggA Natural spray (containing PMD) (4.2 and 3.2 g/m2) and one of RB86 (with 70% neem oil containing azadirachtin) (3 g/m2) to host seeking nymphs of Ixodes ricinus. Each substance was applied separately to 1 m2 cotton flannel cloths. Nymphal ticks on the cloths, pulled over the vegetation, were recorded at 10-m stops. Nymphal numbers recorded differed significantly between treated cloths [4.2 or 3.2 g MyggA Natural spray/m2 and 3 g RB86/m2] and the untreated control (df = 3, chi2 = 112.74, P < 0.0001). Nymphal numbers also differed significantly among collectors (df = 3, chi2 = 15.80, P < 0.001). Repellency of treated cloths, i.e., 4.2 or 3.2 g MyggA Natural spray/m2 and 3 g RB 86/m2 declined from day 0 (i.e. the day of impregnation) to day 3 after impregnation from 77 to 24%, 58 to 16% and 47 to 0.5%, respectively. This study suggests that all three treatments have significant repellent activities against I. ricinus nymphs.

Animals↗
PubMed2006

Genetic variation of ticks (Ixodes ricinus L.) in the Lithuanian and Norwegian populations.

RAPD markers were used to measure the genetic diversity of 119 individuals of Ixodes ricinus collected from Lithuania and Norway. The samples were analysed within and also between the populations. We analysed 74 loci in each of 6 populations. Our results show high levels of diversity within the populations. The percentage of polymorphic loci of the six analysed populations: Birzai, Vilnius, Kretinga, Tjore, Kjosvik and Odderoya were 68.9%, 58.1%, 78.38%, 62.2%, 44.6% and 68.9%, respectively. The percentage of polymorphic loci in the Lithuanian populations was 93.2%, and in the Norwegian populations 81.08%. The genetic distance ranged from 0.019 to 0.079 within Norwegian populations and from 0.005 to 0.0967 within Lithuanian populations and between the countries from 0.022 to 0.146. The genetic variation of I. ricinus among Norwegian populations was lower than among Lithuanian populations. The highest part of genetic variation in I. ricinus ticks depends on variation within Kretinga (Lithuania) and Odderoya (Norway) populations situated in coastal areas where many migratory and sea birds are aggregated.

Animals↗
PubMed1996

Interbreed differences in the efficacy of 1% deltamethrin pour-on to protect small livestock against infestation with Ixodes rubicundus (Acari:Ixodidae).

Synthetic pyrethroid pour-on compounds have been developed for use on cattle but many are now also registered for the control of ectoparasites on small ruminants. The breed of small livestock has been known to influence the efficacy of various pour-ons and we hypothesized that differences in the duration of protection afforded by a synthetic pyrethroid pour-on against ticks on small livestock, would be greatest between hairy and woolly breeds. Angora goats with mohair, Dorper sheep with hairy wool, and Merino sheep with wool were treated with 1% m/v deltamethrin pour-on with an oil base at a dosage rate of 1 ml 5 kg-1 body mass. The duration of effective protection against Ixodes rubicundus, the Karoo paralysis tick, was determined under field conditions. Angora goats proved to be the highest risk group, but were afforded the least protection. Efficacy never reached 100% in this group and within 3 weeks the goats were already at risk of paralysis. Dorper sheep were afforded above 90% protection for 3 weeks and 85% after 4 weeks. In the case of the Merino sheep efficacy of protection remained above 90% up to 7 weeks post-treatment and was still 87% during Week 8. Differences in the amount of sebum and suint in the epidermis of the different breeds may be responsible for differences in the rate of spread and the rate of dilution of the active compound in the epidermis.

Administration, Topical↗
PubMed1996

Nested polymerase chain reaction for detection of Ehrlichia equi genomic DNA in horses and ticks (Ixodes pacificus).

A nested polymerase chain reaction for detecting Ehrlichia equi in horses and ticks (Ixodes pacificus) was developed. A major second-round PCR product of 928 bp could be readily visualized in ethidium bromide-stained agarose minigels. An internal probe was used to verify the identity of the amplified product by non-radioactive (digoxigenin-based) Southern blotting; additional confirmation was provided by DNA sequence analysis. A dilution study testing the sensitivity of the PCR indicated that DNA derived from < = 7.6 but > 3 infected neutrophils was sufficient to generate a PCR signal. The specificity of the PCR was examined using a panel of rickettsiae, of which only E. equi and the closely-related human granulocytotropic ehrlichia produced PCR bands. In an in vivo infection study, E. equi DNA was detected in blood buffy-coat cells of an experimentally-infected horse on days three through 14 post-inoculation. In a separate study, three of six adult I. pacificus that as nymphs had been fed on an experimentally infected horse were found to be PCR-positive for E. equi.

Animals↗
PubMed1996

Acquired resistance and antibody response of raccoons (Procyon lotor) to sequential feedings of Ixodes scapularis (Acari: Ixodidae).

Captive-bred raccoons (Procyon lotor) developed immune resistance to infestation by the larval stage of the ixodid tick, Ixodes scapularis, the vector of Borrelia burgdorferi, following repeated applications of both nymphs and larvae. Resistance was expressed as a significant decrease in the proportion of engorged larvae recovered from each cohort. Resistance to nymphs was not noted, but, only two such cohorts were applied. Utilizing an enzyme-linked immunosor-bent assay (ELISA) developed to detect raccoon serum antibodies to tick salivary gland antigens, raccoons evidenced a two to ten-fold increase in anti-tick salivary extract antibody titer following the application of two cohorts of nymphs and eights cohorts of larvae. The tick saliva antigens recognized by both pre- and post-exposure raccoon sera were evaluated by Western blotting. The production of antibodies correlated with the development of resistance to infestation, suggesting that the resistance was immune-mediated and could be measured by anti-tick salivary extract antibody titers. Resistance in exposed raccoons prevents nearly 90% of larvae from prolonged feeding. Prolonged feeding is required for engorgement and the transmission of various infectious agents, such as B. burgdorferi.

Animals↗
PubMed2004

TROSPA, an Ixodes scapularis receptor for Borrelia burgdorferi.

The Lyme disease agent Borrelia burgdorferi naturally persists in a cycle that primarily involves ticks and mammals. We have now identified a tick receptor (TROSPA) that is required for spirochetal colonization of Ixodes scapularis. B. burgdorferi outer surface protein A, which is abundantly expressed on spirochetes within the arthropod and essential for pathogen adherence to the vector, specifically bound to TROSPA. TROSPA mRNA levels in ticks increased following spirochete infestation and decreased in response to engorgement, events that are temporally linked to B. burgdorferi entry into and egress from the vector. The blockade of TROSPA by TROSPA antisera or by the repression of TROSPA expression via RNA interference reduced B. burgdorferi adherence to the I. scapularis gut in vivo, thereby preventing efficient colonization of the vector and subsequently reducing pathogen transmission to the mammalian host. Identification of an I. scapularis receptor for B. burgdorferi is the first step toward elucidating arthropod ligands that are required for survival of spirochetes in nature.

Amino Acid Sequence↗
PubMed2006

Ixodes ricinus: the potential of two-dimensional gel electrophoresis as a tool for studying host-vector-pathogen interactions.

Ixodes ricinus is a three-host tick, with three active instars. For moulting to occur the tick has to find a host where it can take a blood meal. Throughout feeding I. ricinus can be infected or infect the host with different pathogens, e.g., Tick-Borne Encephalitis virus or Borrelia burgdorferi. The host-vector-pathogen interaction is very complex, making a detailed study difficult. Here we analyse the potential of two-dimensional gel electrophoresis (2DE) to study the host-vector-pathogen interaction. We examined 20 nymphs, which as larvae parasitised either mouse or hen. After moulting, they were kept alive for up to 30 weeks, to analyse whether tick ageing influenced host determination, and for comparison of the 2D-gels. Even though the number of proteins in the gel decreased during ageing, some proteins of the host determination persisted for all 30 weeks. We also discovered persisting proteins in relation to nymphs. These findings showed that 2DE is suitable as a tool for studying host-vector-pathogen interactions.

Actins↗