Temporal Distribution of the Annual Nymphal Stock of Ixodes Ricinus Ticks

The human risk of contracting Lyme disease or other tick borne diseases transmitted by the tick species Ixodes ricinus is broadly linked to the tick nymph density. The study was performed in Rambouillet forest (Yvelines, France), a known focus of Lyme borreliosis, from January 1997 to December 1999. We used a nymph sampling methodology which permitted us to obtain a monthly nymph density index (from 0 to 5). Studying the seasonal nymph and larval activity patterns and estimating the larval developmental duration, we demonstrate the existence of an annual nymphal stock. Secondly, we elucidate how this stock is distributed throughout the year, month by month. Its distribution is principally dependent on two factors: the monthly mean ambient temperature and the proportion of active nymphs which find a host each month. Expected monthly nymph densities derived from a theoretical model describing the temperature-dependent stock distribution gave a good fit to the observed densities, accounting for between 76–86% of the monthly variation in observed nymph densities. Predicting the temporal distribution of nymph activity within a stable Lyme borreliosis focus enables more precise identification of risk periods.     

Interstadial and Infestation Level-dependent Variation in the Transmission Efficiency of Borrelia Burgdorferi from Mice to Ixodes Ricinus Ticks

The efficiency with which the spirochaete Borrelia burgdorferi sensu stricto was transmitted from laboratory mice to larval and nymphal Ixodes ricinus ticks was assessed, using the polymerase chain reaction. The transmission efficiency to nymphs was significantly greater than to larvae when both fed together on the same host. Increased tick infestation levels of mice were correlated with significantly greater engorgement weights and higher B. burgdorferi transmission coefficients from mice to nymphs. These observations indicate that both the feeding success of ticks and the transmission coefficients from host to tick may be influenced by the tick infestation level of an infected host. The infestation level and the relative numbers of each life stage of the tick are factors which should be considered in the design of transmission experiments.     

Understanding the Relationships between Landscape Connectivity and Abundance of Ixodes ricinus Ticks

The density of Ixodes ricinus ticks in a heterogeneous landscape of northern Spain was determined and associated with some aspects of habitat topology. The habitat mosaic was used to quantify connectivity between patches of different tick density. The analysis revealed that patches with high tick abundance are ‘stepping-stone’ territories that, when removed from the landscape, cause large changes in connectivity. Sites with medium tick abundance do not cause such a critical transition in connectivity. Patches with low tick abundance, but optimal abiotic conditions for survival are located within the minimum cost corridors network joining the patches, while those sites where the tick has been intermittently collected are located at variable distances from this network. These results suggest that tick distribution in a zone is highly affected not only by abiotic variables (vegetation and weather) but also by host movements. Whether these high-density ‘stepping-stone’ patches occur in other tick species needs to be evaluated because of the potential implications of these foci for human health. This revised version was published online in July 2006 with corrections to the Cover Date.     

Entomopathogenic fungi associated with Ixodes ricinus ticks

The objective of this study was to demonstrate the occurrence of entomopathogenic fungi on Ixodes ricinus ticks in relation to the tick stage, engorgement and season. Ticks were collected from the vegetation, from small rodents and from deer. All entomopathogenic fungi found belonged to the Hyphomycetes. Paecilomyces farinosus and Verticillium lecanii were the predominant species. Other species, found only on engorged females were: Beauveria bassiana, B. brongniartii, P. fumosoroseus and V. aranearum. Eight out of 1833 ticks collected from the vegetation and three out of 269 engorged nymphs were infected with fungi. Thirty-three out of 149 engorged females were infected, whereas males and engorged larvae were not infected. Throughout the season, a significantly higher proportion of ticks collected in autumn were infected. Entomopathogenic fungi may have a significant impact on the size of the I. ricinus population, since females were the most frequently infected stage.     

Seasonal incidence of Ixodes ricinus ticks (Acari: ixodidae) on rodents in western France

Data collected from a longitudinal survey carried out over 2 years on four farms in western France were used to assess the incidence and infestation of Ixodes ricinus on rodents. Once a month, on each farm, 25 Sherman live traps were set in hedges bordering selected pastures. A total of 799 micromammals were examined, including Apodemus sylvaticus, Clethrionomys glareolus, Microtus agrestis, Microtus arvalis, and Crocidura spp. Larvae and nymphs of I. ricinus were found. Small numbers of Ixodes (Exopalpiger) trianguliceps were also recovered from each farm. The mean infestation rate of the I. ricinus larvae (1.6–5.9) among all animals examined varied between farms Most animals were infested by only a single tick, but one M. agrestis harboured 43 I. ricinus larvae. Larvae or nymphs were found throughout the year, with peaks from March to October.     

Development of the salivary glands in embryos of Ixodes ricinus (Acari: Ixodidae)

We examined Ixodes ricinus embryos between 18 and 28 days of development with light, scanning and transmission electron microscopy. The differences in inner structure attested to establish three successive developmental stages: days 18–20, day 23, and days 26–28. Between 18 and 20 days the embryos are at early stages of organogenesis. Salivary glands cannot be identified at that stage. In 23-day-old embryos salivary glands are already outlined but the structure of alveoles is still different from that in larvae in which the embryonic development has been completed. Gland cells start to form alveoles and become active between 26 and 28 days of the development. This revised version was published online in July 2006 with corrections to the Cover Date.     

Seasonal incidence and ecology of the tick Ixodes ricinus (Acari: Ixodidae) on grazing pastures in Western France

A longitudinal survey was carried out during a 2 year period in Western France to assess the infestation level of grazing pastures byIxodes ricinus ticks. Four farms were visited once a month and each of the grazing pastures was sampled in the centre and at the border using the blanket dragging method. A total of 3562I. ricinus (34 adults, 900 nymphs and 2628 larvae) were collected and the infestation was significantly higher during the first year (p<0.0001). The infestation level byI. ricinus varied between grazing pastures and farms. Grazing pastures in the vicinity of forest were more infested than the others, all through the study. The seasonal distribution of ticks showed peaks, with low fluctuations between farms, years and stages. Tick abundance could not be related to vegetation, but only to the vicinity of woods.     

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.     

Acquisition of Borrelia burgdorferi by Ixodes ricinus ticks fed on the European hedgehog,Erinaceus europaeus L

A hedgehog, Erinaceus europaeus, was found to be heavily infested with larval and nymphal Ixodes ricinus in a forest park in Co. Galway, Ireland. A large proportion of the ticks that engorged and detached were infected with the spirochacte, Borrelia burgdorferi, the causative agent of human Lyme borreliosis. The identity of these spirochaetes was confirmed by immunofluorescent assay with B. burgdorferi-specific monoclonal antibody and by polymerase chain reaction test and they were transmitted from the hedgehog to laboratory-reared ticks and from the ticks obtained from the hedgehog to gerbils (Meriones unguiculatus). The high infection rate of the larvae that fed on the hedgehog in comparison with unfed larvae from the same habitat was interpreted as strong evidence that this host species is reservoir competent. Since hedgehogs can evidently feed adult ticks as well as many immature stages, they may well have an important role in the ecology of Lyme borreliosis in some habitats.     

Densities of Ixodes ricinus ticks (Acari: Ixodidae) on moorland vegetation communities in the UK

Unfed (questing) Ixodes ricinus ticks were collected by blanket dragging on a monthly basis from heather-dominated, Vaccinium-dominated and bracken-dominated vegetation communities from two different biogeographical regions of the UK (the Quantock Hills in Somerset, south west England and the North York Moors, north east England) throughout the spring and summer months of 1991 and 1992. Eighteen sites were monitored across the two regions and a total of 1920 blanket drags were carried out. Vaccinium sites showed high tick densities at all life stages, as did bracken sites. Significantly lower numbers of larval and nymphal ticks per drag were collected on heather sites than were collected on either Vaccinium (bilberry/whortleberry) or bracken sites, while similar numbers of adult ticks per drag were collected from each of the three vegetation communities. There was no significant difference between the mean numbers of any tick life stage collected on the Quantock Hills and those collected on the North York Moors on these vegetation communities or between the mean numbers of any tick life stage collected in 1991 and those collected in 1992 on these vegetation communities.     

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.     

Cuticular hydrocarbon composition and phenotypic variability in sympatric populations of Ixodes ricinus ticks from Poland

Gas chromatography of cuticular hydrocarbons was used to assess genetic similarity and heterozygosity between 18 sympatric populations of adult Ixodes ricinus ticks collected from Poland. Sixteen n-alkanes, 45 monomethylalkanes, and 36 dimethylalkanes were identified. Forty-one compounds were detected in 90% of the specimens and nine hydrocarbons were present in all the specimens studied. Visual inspection of chromatograms revealed major differences in the relative abundance of hydrocarbons between populations of I. ricinus. The average proportion of heterozygotic hydrocarbons was 50.68%, while the average genetic similarity among all populations was 0.967. Two other methods were used to measure the intrapopulation variability: the number of isomeric forms for each hydrocarbon and the range of hydrocarbon abundance. It was found that some hydrocarbons manifested an unexpectedly high number of isomeric patterns, such as nonacosane (21 patterns), triacontane (23), pentacosane (26), and hexacosane (28). Our data suggest that I. ricinus ticks from Poland consist of several interbreeding, non-isolated populations, with a high degree of genetic flow between them.     

Variability in cuticular hydrocarbons and phenotypic discrimination of Ixodes ricinus populations (Acarina: Ixodidae) from Europe

The cuticular hydrocarbon composition and a stepwise discriminant analysis are used to elucidate the phenotypic relationships of 66 populations of Ixodes ricinus in Europe. The method correctly allocates Ixodes persulcatus (outgroup) populations away from the main cluster of I. ricinus samples and separates the samples into ten relatively defined clusters of specimens. Populations from Poland are inseparable from samples collected in Germany, Switzerland and the Italian Alps, while individuals from Slovakia and the Czech Republic come into separate groups of phenotypic similarity. Irish and British specimens are separated but highly related and Spanish populations show an unexpectedly high distance from the remaining clusters.     

The spatial distribution of Borrelia burgdorferi-infected Ixodes ricinus in the Connemara region of County Galway,Ireland

Studies were carried out in the Connemara area of County Galway in the west of Ireland in order to determine the abundance and distribution of the tick, Ixodes ricinus and the prevalence of its infection with Borrelia burgdorferi. The tick was very abundant locally, in particular when associated with cattle, sheep and enclosed red deer. Large numbers of ticks not only occurred on the pastures, but also on adjacent roadside verges. No infections with B. burgdorferi could be demonstrated when nymphal ticks were sampled from central areas of the pastures, suggesting that livestock and red deer are probably not significant reservoirs of the spirochaete. Small numbers of infected nymphal and adult ticks were associated with hedges, dry stone walls, the margins of woodland adjoining infested pastures and in woodland from which livestock were excluded. Woodmice (Apodemus sylvaticus) were most numerous in such habitats and the majority were infected with B. burgdorferi.     

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.     

Dynamics of Borrelia burgdorferi infection in nymphal Ixodes ricinus ticks during feeding

We report the sequential developmental events of Borrelia burgdorferi in histological sections of Ixodes ricinus nymphs before, during and after feeding. During the blood meal a decrease of approximately 50% in the number of infected ticks was recorded (eight out of 76, 11%) in comparison with the infection rate of unfed ticks (12 out of 56, 21%). Spirochetes were detected in tick salivary glands only after 2 days of attachment. From day 3 until drop-off, the number of infected ticks increased to 31% (15 out of 49). A quadratic logistic regression analysis showed that the variation in the number of infected ticks was significant, but only during the blood meal. The drop in the percentage of infected ticks during the first hours following attachment to the host is explained by our observation of spirochetes in the faeces of the ticks. The increase in the infection rate of replete ticks may be due to an uptake of spirochetes from the host skin at the feeding site.     

The effect of feeding status on sexual attractiveness of Ixodes ricinus (Acari: Ixodidae) females

Sexual attractiveness of unengorged, semiengorged and fully engorged females of both field and laboratory populations of Ixodes ricinus was studied under laboratory conditions by means of a computerised video tracking system. A male and a female were allowed to walk freely in a glass arena during 1 h and their behavioural interactions were observed. Obtained results revealed that the feeding status of I. ricinus females affects their sexual attractiveness. The highest attractiveness was observed in engorged females, the lowest in unengorged females of the field population. Copulation occurred in all experimental groups with the highest frequency observed in the unengorged virgin laboratory females. Despite engorged females being highly attractive for males on distance, the frequency of copulation was low. Differences between field and laboratory populations were found in both behavioural parameters and frequency of copulation. This revised version was published online in July 2006 with corrections to the Cover Date.     

The impact of gene knock-down and vaccination against salivary metalloproteases on blood feeding and egg laying by Ixodes ricinus

Two cDNAs coding homologous putative metalloproteases (Metis 1 and Metis 2, expected molecular weights of 55.6 and 56.0kDa, respectively) were identified from the hard tick Ixodes ricinus. The expression of Metis genes was induced in salivary glands during tick blood meal. RNA interference was used to assess the role of both Metis 1 and Metis 2 in tick feeding. It was found that salivary gland extracts lacking Metis 1-2 had a restricted ability to interfere with fibrinolysis. RNAi against Metis 1-2 also induced a high mortality rate. An immune reaction was raised in repeatedly bitten animals against Metis 1 and 2. Vaccination of hosts with the recombinant Metis 1 protein produced in a eukaryotic system partially interfered with completion of the blood meal. Although vaccination did not alter the survival rate or feeding time of ticks, their weight gain and oviposition rate were reduced. This will affect their reproductive fitness in the field. We believe this is the first report of an anti-tick vaccine trial using a metalloprotease derived from I. ricinus.     

The Dynamics of an Avoidance Behavior by Ixodid Ticks to Liquid Water

Life stages of different tick species avoid walking on a wet surface surrounding a dry patch by systematically returning to the dry each time they contact the wet surface beyond the border with the tip of a first leg tarsus. Sequential analysis of the border behaviors shows that repetitive contact with the water increases the probability of walks astride the border. Ticks accept this unilateral contact with the water for longer intervals and eventually walk on to the wet surface after a combination of a short patch walk followed by a border walk which is longer than the foregoing ones. Staying time on a small circular patch is shorter than on a large one, arising probably from faster adaptation of peripheral receptors following a higher frequency of border contacts. However, an equal number of border reactions on patches of different sizes and shapes suggests that a counter in the CNS may also influence dry patch departure.     

Lyme borreliosis spirochetes in Ixodes ricinus and Haemaphysalis punctata ticks (Acari: Ixodidae) on three islands in the Baltic Sea

The prevalence of infection with Lyme borreliosis spirochetes in nymphal Ixodes ricinus and Haemaphysalis punctata was examined on three Swedish islands in the Baltic Sea. The proportion of H. punctata nymphs infected with Borrelia spirochetes (2%) was lower than that of I. ricinus nymphs (11–16%), even on the island of Stora Karlsö where both tick species feed on the single mammalian host present, the varying hare, Lepus timidus. Ten of the 12 infected questing H. punctata nymphs harboured very few spirochetes, whereas the remaining two harboured as many as 30–40 spirochetes. However, it remains to be seen whether H. punctata nymphs are capable of transmitting spirochetes during their blood meal and, thus, serving as vectors for Lyme borreliosis spirochetes. On Gotland and Fårö, I. ricinus was more abundant than H. punctata in deciduous and coniferous forest, whereas H. punctata was equally or more abundant than I. ricinus in juniper brush areas on open land. Host preference and biotope usage of H. punctata seems, in general, to prevent this tick from feeding on highly infective Borrelia reservoirs, such as Apodemus mice.