Klíšťata sají obrovské množství krve, která je jejich jediným zdrojem živin a energie. Přesný enzymatický mechanismus zpracování krve ve střevě klíšťat však kupodivu nebyl donedávna vůbec znám. Náš komplexní molekulární model trávení hostitelského hemoglobinu u klíštěte obecného (Ixodes ricinus) poprvé odhalil analogii enzymatického aparátu s krevsajícími ploštěnci a hlísticemi, a zároveň tato znalost představuje zásadní poznatek pro účinný boj s klíšťaty a jimi přenášenými patogeny., Ticks (in this case Ixodidae and Argasidae) feed on enormous amounts of host blood, which provides their ultimate source of energy and nutrients. There has been only limited evidence on the exact molecular mechanisms of blood digestion in ticks. For the first time, our complex enzymatic model of proteolytic digestion in the Common Tick (Ixodes ricinus) reveals the analogy of tick intestinal proteolysis with bloodfeeding platyhelminthes and nematodes and presents a future application potential in tick or tick-borne pathogen interventions., and Daniel Sojka.
The contribution of woodiniee (Apodemus sylvaticus), yellow-necked mice (Apodemus flavicollis) and bank voles (Clethrionomys glareolus) was compared in a focus of Lyme borreliosis in Switzerland during a 7 months’ study. All three species of mice and one species of shrews (Sorex araneus) were shown parasitized by infected Ixodes ricinus immatures. About 14% of larvae and 50% of nymphs collected on small mammals were infected with B. burgdorferi. Spirochetes were isolated from blood of 3 woodmice and one yellow-necked mouse. The infectious status of rodents was estimated by tick xenodiagnosis. Prevalence of infected rodents ranged from 20% to 44%. Mice presented a higher potential infectivity than voles. The prevalence of infected rodents showed a seasonal variation.
Salivary gland extracts (SGE) from unfed and 5 days led adult female Ixodes ricinus (Linnaeus, 1758); Haemaphysalis inermis (Birula, 1895) and Dermacentor reticulatus (Fabricius, 1794) ticks were prepared. The protein content after feeding increased by 10.6, 8.7 and 6.8 limes, respectively. Extracts were equilibrated to the same protein content and submitted to SDS-polyacrylamide gel electrophoresis followed by computer analysis of the scanned gels. Relative differences in protein profiles of extracts obtained from unfed and partially fed ticks were found in all species and some of them were similar in all three species used in the study. Results demonstrate that the increase of the protein content in salivary glands during the feeding does not occur proportionally. Some proteins are synthesised preferentially (67.1 kDa, 13.5 kDa) but other bands (in range of 15-16 kDa) present in the SGE derived from unfed ticks arc less discernible in that of fed ticks.
Borrelia burgdorferi sensu lato (s.l.) is the etiological agent of Lyme disease, transmitted by ticks of the genus Ixodes Latreille. Diagnosis of Lyme disease in humans is often difficult and a detailed knowledge of the circulation of B. burgdorferi s.l. in tick hosts is therefore fundamental to support clinical procedures. Here we developed a molecular approach for the detection of B. burgdorferi s.l. in North Italian Ixodes ricinus (Linnaeus). The method is based on the amplification of a fragment of the groEL gene, which encodes a heat-shock protein highly conserved among B. burgdorferi s.l. species. The tool was applied in both qualitative and Real-time PCR approaches testing ticks collected in a North Italian area. The obtained results suggest that this new molecular tool could represent a sensitive and specific method for epidemiological studies aimed at defining the distribution of B. burgdorferi s.l. in I. ricinus and, consequently, the exposure risk for humans.
Saliva-activated transmission (SAT) of Borrelia burgdorferi sensu stricto was demonstrated using real-time PCR and salivary gland extract (SGE) from partially fed Ixodes ricinus ticks. C3H/HeN mice were injected intradermally with 1.5 × 103 spirochetes mixed with 40 µg of SGE per mouse. The control group was inoculated with the same dose of spirochetes without SGE. The accelerating effect of SGE on spirochete proliferation was demonstrated on day 1 post infection, when a 4.2-fold increase in spirochetes was found in the skin and a 10-fold increase in the blood, compared with control mice. The data represent the first direct evidence of a SAT effect of I. ricinus SGE on infection with the Lyme disease agent B. burgdorferi.
Previous studies have demonstrated that both tick saliva and Borrelia burgdorferi sensu lato antigens modulate the cytokine response of the host. In this paper, the effect of salivary gland extract (SGE) from Ixodes ricinus (L., 1758) ticks on cytokine production by primary cultures of mouse epidermal cells stimulated with Borrelia afzelii Canica, Nato, du Merle, Mazie, Baranton et Postic, 1993 spirochetes was analysed. Epidermal cells were derived from C3H/HeN mice, susceptible to Lyme disease, and BALB/c mice, which are resistant. In cultures from C3H/HeN mice, SGE down regulated production of tumour necrosis factor alpha (TNF-α) and up regulated Th2 cytokine, interleukin 4 (IL-4). Cultures from BALB/c mice produced higher basal levels of monitored cytokines, but their production was affected by SGE a different way. While Th2 cytokines IL-6 and IL-10 were down regulated, the effect on TNF-α and IL-4 was ambiguous. These results indicate that the effect of tick saliva on the epidermal cells of Lyme disease-susceptible C3H/HeN mice mirrors its effect on other cells of the immune system.
Ixodid ticks remain attached to their hosts for several days to weeks. During this extended feeding process new proteins involved in the modulation of host immune responses are expressed in tick salivary glands. In our study a stimulatory or inhibitory effect of salivary gland extracts (SGE) of unfed and partially fed female Ixodes ricinus (Linnaeus, 1758), female and male Amblyomma variegatum (Fabricius, 1794) and Rhipicephalus appendiculatus Neumann, 1901 ticks on human lymphocyte proliferation induced by Concanavalin A (ConA) and phytohaemagglutinin (PHA), respectively, was investigated. SGE of all female ticks examined suppressed proliferation of ConA-induced lymphocytes; highly significant suppression was observed in the presence of unfed I. ricinus and 9-day fed A. variegatum SGE. SGE of partially fed A. variegatum and I. ricinus females suppressed PHA responses of lymphocytes. Lymphocytes showed reduced PHA and ConA responses in the presence of SGE of unfed and 2-day fed R. appendiculatus females, while SGE of 6-day fed females enhanced PHA responses, but reduced their ConA responses; generally SGE of 2-day fed females displayed the strongest inhibition. Amblyomma variegatum male SGE slightly enhanced PHA, but significantly reduced ConA responses of lymphocytes and their inhibitory effect increased during feeding. SGE of unfed and 2-day fed R. appendiculatus males enhanced PHA and ConA responses and those of 6-day fed males suppressed lymphocyte proliferation. The results suggest that (i) species- and sex-specific differences exist in the effects of tick salivary gland antigens on human lymphocyte proliferation and (ii) effect of SGE on human lymphocyte responses to mitogens varies depending on the tick feeding status.
Haemagglutination activity (HA) was found and characterized in a midgut homogenate of Ixodes ricinus (L.). HA was induced by tick feeding; it was not detected in starved ticks. In a haemagglutination inhibition test, HA showed an affinity for some carbohydrates (N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, rhamnose, and dulcit) and glycoconjugates (especially lipopolysaccharides). Midgut protein components of 37, 60, 65, and 73 kDa were identified by immunoblotting as potential structural subunits of the new agglutinin.
Unfed nymphs of Ixodes ricinus (L.) can be divided into two morphological groups according to the length of idioso-ma, scutum, hypostome and palpal segment III, and the number of dorsal alloscutal setae. Specimens of greater body dimensions and more numerous dorsal alloscutal setae moulted predominantly into females. The frequency of different nymphal length categories in field-collected ticks followed a normal distribution. The length of unfed nymphs correlates well with the length (r = 0.7248 ± 0.0711, P < 0.001) and weight (r = 0.6519 ± 0.0782, P < 0.001) of engorged nymphs, however, it varies in ticks of different origin. In field-collected ticks, freshly engorged female nymphs were 2.30-2.94 mm long, male nymphs 2.14-2.46 mm long. Feeding period (P < 0.05) and premoulting period (P < 0,001 ) were significantly longer in female nymphs both in field-collected and laboratory-derived I. ricinus. The engorgement weight was found to be the best criterion for differentiation of male and female nymphs of ixodid ticks. In field-collected nymphs engorged on BALB/c mice, 98.6 % of females moulted from nymphs weighting more than 3.60 mg, while in laboratory-derived ticks, 98.4 % of females emerged from nymphs of 3.42 mg body mass or more.
In Britain, grey squirrels (Sciurus curolinensis Gmelin) and pheasants (Phasianus colchicus Linnaeus) are important hosts of larvae and nymphs of Ixodes ricinus L., the principal F.uropean vector of the Lyme disease spirochaete, Borrelia burgdorferi sensu lalo, T о test whether squirrels are competent hosts of B. burgdorferi s. I., three females were trapped in the wild and then held in captivity. Following treatment, each animal was exposed to uninfected xenodiagnostic I. ricinus ticks. Squirrel A (an adult) which was inoculated experimentally with B. burgdorferi s.l., transmitted the infection to xenodiagnostic ticks. In contrast, squirrel В (a juvenile that was not inoculated)-showed no evidence of infection. Xenodiagnostic ticks that fed on control squirrel С (an adult) became infected and subsequently transmitted the infection experimentally to an uninfected hamster. The results indicated that squirrel С had a disseminated infection acquired in the wild and which persisted for at least 11 weeks. These data clearly demonstrate that grey squirrels are amplifying and reservoir hosts of B. burgdorferi s.l. The strain associated with squirrels was related to the B. afzelii genotype. Two observations implicated pheasants in a similar role; (i) a high prevalence of infection in engorged larvae collected from trapped pheasants, and (ii) the detection of B. burgdorferi s.l. (В. garinii genotype) in the wattle of 1/10 pheasants using PCR. Xenodiagnostic experiments similar to those undertaken with the squirrels are needed to confirm the role of pheasants in the transmission cycle of Lyme disease spirochaetes.