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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.

Animals↗

Prevalence of Borrelia burgdorferi (sensu lato) in Ixodes ricinus (L.) populations in France, according to a phytoecological zoning of the territory.

Ixodes ricinus is considered as the main vector of Lyme Borreliosis in France. The aim of our investigations was to obtain a comprehensive view of the spatial risk linked to the distribution of the species in our country. Previous studies [1] have provided strong evidence that the species populations are widely distributed, so the objective of the present work was to ascertain the bacteriological infection of the tick by the agent of the zoonosis (Borrelia burgdorferi, sensu lato), over the French territory, whatever the ecological conditions may be. For this purpose, we kept the same framework as that used in our acarological investigation, a phytoecological zoning of the territory into 54 geographically separate and ecologically distinct units distributed into three climatic zones. Batches of ticks, picked up in these different phytoecological units (only two thirds of which were sampled), were submitted for bacteriological investigation. A total of 4,673 ticks were examined, individually, for the presence of Borrelia burgdorferi, by immunofluorescence. Percentages of infection according to the various stages of Ixodes ricinus free stages, collected by flagging, were as follows: 4.95% in 3,247 nymphs, 11.2% in 699 males, 12.5% in 727 females. Larvae were ignored. Practically all the tested units harboured the bacterium. The percentage of tick samples (25 ticks or more) absolutely free of Borrelia, wherever they came from, is very low (not exceeding 10 percent of the sampled forests). Our study confirms the assessment of a widespread distribution of the zoonosis in France which was, until now, exclusively based on an approximate distribution of limited human cases observed in the country.

Animals↗

Rate of infection of Ixodes ricinus ticks with Borrelia burgdorferi sensu stricto, Borrelia garinii, Borrelia afzelii and group VS116 in an endemic focus of Lyme disease in Italy.

A study to evaluate the natural rate of infection of Ixodes ricinus with Borrelia burgdorferi sensu lato was carried out in an endemic focus of Lyme disease in the Trieste area in northern Italy. Two-hundred and twenty-seven ticks collected in ten different stations were tested individually for the presence of the spirochetes using polymerase chain reaction techniques able to identify both Borrelia burgdorferi sensu lato and the four genospecies (Borrelia burgdorferi sensu stricto, Borrelia garinii. Borrelia afzelii and group VS116). Multiple infection of individual ticks was found. The infection rate ranged from 0-70%. Infection of Ixodes ricinus with Borrelia burgdorferi group VS116 was found for the first time in Italy in both a high and a low endemic focus of Lyme disease.

Animals↗

Prevalence of Borrelia burgdorferi species and identification of Borrelia valaisiana in questing Ixodes ricinus in the Lyon region of France as determined by polymerase chain reaction-restriction fragment length polymorphism.

Many cases of Lyme borreliosis have been reported over the years in the region of Lyon, France. The identification and prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus were investigated by polymerase chain reaction (PCR) of the flagellin gene and restriction fragment length polymorphism (RFLP) analysis. Questing Ixodes ricinus larvae, nymphs and adults were collected by the flagging method from deciduous forests in four areas in the Lyon region of France between October 1994 and September 1995 and in June 1998. The overall prevalence of Borrelia burgdorferi sensu lato was 13.2% (91/688). No significant differences in prevalence were observed between the different stages and sex of the ixodids or between collection areas. The majority of infections were simple infections (82.4%; 75/91), most of which were due to Borrelia afzelii (41.4%), while coinfections (12.1%) were predominantly (54.5%) a combination of Borrelia valaisiana and Borrelia garinii. No tick was infected with more than two borrelial species, nor was Borrelia lusitaniae identified. The Borrelia valaisiana species was detected for the first time in France, confirming its widespread presence in Europe. This study confirms that the surroundings of Lyon are risk areas for contracting Lyme disease and that no particular clinical manifestations predominate due to the heterogeneous distribution of Borrelia genospecies. Moreover, the polymerase chain reaction restriction fragment length polymorphism analysis is a rapid and easy method for genotyping of Borrelia species.

Animals↗

Physiological age of field-collected female taiga ticks, Ixodes persulcatus (Acari: Ixodidae), and their infection with Borrelia burgdorferi sensu lato.

In some studies the prevalence of tick infection (infection rate) and the intensity of infection are negatively correlated with unfed tick age (in the broad sense of this term). However, no special research has been carried out to consider the phenomenon thoroughly. The infection indices of the female taiga ticks, Ixodes persulcatus, infected with Borrelia burgdorferi s.l. were related to tick physiological age, an index that more precisely reflects tick physiological state than the time of tick collection in the field or the duration of tick survival under laboratory conditions. A novel quantitative technique of physiological age determination based on the evaluation of the ratios between sizes of the stable (scutum) and the changing (alloscutum) structures of the tick body was used. The age was estimated in accordance with the classical age-grade scale introduced by Balashov and a more fractional scale determined by the new technique. In total, 131 female ticks were examined for their infection and physiological age, 46 of which were infected with B. burgdorferi s.l. (mean infection rate 35.1%). The minimal intensity of infection was 0.4 bacterial cells per 100 fields of view whereas the maximal infection was 172 cells. There was no difference between the prevalence of infection in ticks of different physiological age. The intensity of infection obviously differed between ticks of different age groups in the scale introduced by Balashov but did not significantly differ between ticks of different age groups according to the fractional age-grade scale. The data concerning the relationships between Borrelia burgdorferi and unfed Ixodes ticks are considered.

Age Factors↗

The transcriptome of the salivary glands of the female western black-legged tick Ixodes pacificus (Acari: Ixodidae).

Sequencing of an Ixodes pacificus salivary gland cDNA library yielded 1068 sequences with an average undetermined nucleotide of 1.9% and an average length of 487 base pairs. Assembly of the expressed sequence tags yielded 557 contigs, 138 of which appear to code for secreted peptides or proteins based on translation of a putative signal peptide. Based on the BLASTX similarity of these contigs to 66 matches of Ixodes scapularis peptide sequences, only 58% sequence identity was found, indicating a rapid divergence of salivary proteins as observed previously for mosquito and triatomine bug salivary proteins. Here we report 106 mostly full-length sequences that clustered in 16 different families: Basic-tail proteins rich in lysine in the carboxy-terminal, Kunitz-containing proteins (monolaris, ixolaris and penthalaris families), proline-rich peptides, 5-, 9.4- and 18.7-kDa proteins of unknown functions, in addition to metalloproteases (class PIII-like) similar to reprolysins. We also have found a family of disintegrins, named ixodegrins that display homology to variabilin, a GPIIb/IIIa antagonist from the tick Dermacentor variabilis. In addition, we describe peptides (here named ixostatins) that display remarkable similarities to the cysteine-rich domain of ADAMST-4 (aggrecanase). Many molecules were assigned in the lipocalin family (histamine-binding proteins); others appear to be involved in oxidant metabolism, and still others were similar to ixodid proteins such as the anticomplement ISAC. We also identified for the first time a neuropeptide-like protein (nlp-31) with GGY repeats that may have antimicrobial activity. In addition, 16 novel proteins without significant similarities to other tick proteins and 37 housekeeping proteins that may be useful for phylogenetic studies are described. Some of these proteins may be useful for studying vascular biology or the immune system, for vaccine development, or as immunoreagents to detect prior exposure to ticks. Electronic version of the manuscript can be found at.

Amino Acid Sequence↗

The paralogous salivary anti-complement proteins IRAC I and IRAC II encoded by Ixodes ricinus ticks have broad and complementary inhibitory activities against the complement of different host species.

Several observations suggest that inhibition of the host complement alternative pathway by Ixodes tick saliva is crucial to achieve blood feeding. We recently described two paralogous anti-complement proteins called Ixodes ricinus anti-complement (IRAC) proteins I and II co-expressed in I. ricinus salivary glands. Phylogenetic analyses suggested that these sequences were diversifying by a process of positive Darwinian selection, possibly leading to molecules with different biological properties. In the present study, we tested the hypothesis that each paralogue may have different inhibitory activities against the complement of different natural host species, thereby contributing to broaden the host range of I. ricinus ticks. IRAC I and IRAC II were tested against the complement of eight I. ricinus natural host species (six mammals and two birds). The results demonstrate that IRAC I and IRAC II have broad and complementary inhibition activities against the complement of different host species. This report is the first description of paralogous anti-complement molecules encoded by a pathogen with broad and complementary inhibitory activities against the complement of different host species.

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Relationship between Borrelia burgdorferi sensu lato species, red squirrels (Sciurus vulgaris) and Ixodes ricinus in enzootic areas in Switzerland.

The infection and reservoir status of red squirrels (Sciurus vulgaris) for Borrelia burgdorferi sensu lato were studied in Switzerland. B. burgdorferi sensu lato was isolated from 15 skin samples from 4/6 dead red squirrels, victims of road traffic. Isolates were identified using restriction fragment length polymorphism (RFLP): B. burgdorferi sensu stricto was present in 14 culture tubes containing skin samples and B. afzelii in two other tubes. A mixed infection was revealed in one case. A total of 227 ticks attached to squirrels were cultivated in BSKII medium and 90 isolates were obtained. Genotypic identification by RFLP showed that B. afzelii (59%) and B. burgdorferi sensu stricto (46%) dominated in ticks feeding on red squirrels. Data collected from one particular animal, highly infested with Ixodes ricinus and harbouring numerous Borrelia-infected Ixodes ricinus ticks, showed that transmission of B. burgdorferi sensu lato occurred from S. vulgaris to feeding ticks. More precisely, B. burgdorferi sensu stricto and B. afzelii were mainly transmitted from S. vulgaris to ticks. The present data emphasized the results obtained previously from small rodents and birds in Japan and in Switzerland, showing the occurrence of specific associations between host species and Borrelia genospecies.

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Epidemiological surveillance of tick populations: a model to predict the colonization success of Ixodes ricinus (Acari:Ixodidae).

A model that predicts the habitat availability for permanent populations of the tick Ixodes ricinus has been developed and applied to Spain. The model is based on the fuzzy logic rules and has a spatial resolution of about 4 km. Six long-term climate variables, which cover temperature and rainfall factors, drive the model by assigning the expected probability to find stable tick populations. Because the high accuracy of this inference tool, it could be used to map favourable sites for this tick in the epidemiological lookout of the Ixodes ricinus borne diseases.

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The distribution and prevalence of B. burgdorferi genomospecies in Ixodes ricinus ticks in Ireland.

Questing Ixodes ricinus ticks were collected from six locations throughout Ireland and 638 nymphs, 111 females and 118 males were investigated for infection with Borrelia burgdorferi sensulato (s.l.). The total prevalence of B. burgdorferi s.l. in the ticks was determined as 14.9% by polymerase chain reaction (PCR) amplification of the spacer region of 5S-23S rRNA genes. Infection prevalence was significantly higher in adult (20.1%) Ixodes ricinus compared to nymphs (13.1%). The prevalence of infection in adult male and female ticks was similar (19.5% and 20.7% respectively). The genomospecies B. burgdorferi sensu stricto, B. afzelii, B. garinii and group VS116 were identified by reverse line blot (RLB) using genomospecies specific oligonucleotide probes. The most prevalent B. burgdorferi genomospecies identified were VS116 (34.6%), B. garinii (24.3%) and B. burgdorferi sensu stricto (18.4%). B. afzelii was uncommon (6.6%). Multiple infections were observed in 13.2% of the infected ticks. The distribution of the genomospecies showed geographical variation and also seemed to be influenced by the nature of the habitat. A broad range of genomospecies seemed to be associated with the presence of a wide spectrum of potential reservoir hosts in the habitat and also with a high overall prevalence of B. burgdorferi s.l.

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High prevalence of Borrelia lusitaniae in Ixodes ricinus ticks in Tunisia.

To investigate whether ticks of the genus Ixodes are infected by Borrelia burgdorferi complex, 490 unfed Ixodes ricinus ticks were collected by flagging in three different areas of Tunisia in 1998. DNAs extracted from 81 adults, 60 nymphs and 38 larvae were analysed after genic amplification of the non-coding spacer between the two copies of the rrl-rrf genes of B. burgdorferi sl. The prevalence of B. burgdorferi sl. in adults, nymphs and larvae was found to be 34, 33.3 and 2.6%, respectively. All DNAs (n = 61) but one were identified as belonging to different genotypes of B. lusitaniae by analysis of the restriction fragment length polymorphism of amplification products. In addition, 290 adults, 14 nymphs and 7 larvae were used to inoculate BSK-H medium to isolate spirochetes. Fifteen strains were isolated from adult ticks in the humid areas of Tunisia, whereas only one was obtained from larvae. Isolates were identified as B. lusitaniae (15/16) and B. garinii (1/16). These results provide new evidence for the existence of Lyme borreliosis in North Africa.

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Infestation of mammals by Ixodes ricinus ticks (Acari: Ixodidae) in south-central Sweden.

Infestation by Ixodes ricinus ticks on rodents, hares and cervids was examined at Bogesund, 10 km north of Stockholm, in south-central Sweden during 1991-1994 and on varying hares (Lepus timidus) at Stora Karlsö and Gotska Sandön in the Baltic Sea during 1992-1993. At Bogesund, there were great differences between two consecutive years in the number of I. ricinus larvae infesting bank voles (Clethrionomys glareolus). The seasonal pattern of infestation by I. ricinus larvae and nymphs on bank voles was unimodal in 1991, with peaks in June-July and bimodal in 1992, with peaks in June and August. Male bank voles, compared to females and older voles, compared to young voles, harboured greater numbers of I. ricinus ticks. Apodemus mice, compared to bank voles, harboured greater numbers of I. ricinus ticks. Ixodes ricinus larvae engorged on Apodemus mice were heavier than larvae engorged on bank voles and resulted in larger nymphs. However, there was no difference in the proportions of viable nymphs resulting from larvae engorged on mice or voles. The ranges in the numbers of I. ricinus ticks infesting individual hosts were 1-451 for rodents, 16-2374 for hares and 428-2072 for roe deer (Capreolus capreolus). These ranges of tick numbers are estimated to represent potential blood losses from individual hosts of approximately 0.2-65% for rodents, 0.2-13% for hares and 0.3-9.0% for roe deer. Within the populations of all host species examined, the distributions of all stages of I. ricinus were clumped, with most host individuals harbouring few ticks and only a few individuals harbouring many ticks. The data suggest that, even though a small proportion of tick hosts may be severely affected, the direct effects of feeding by I. ricinus are unlikely to play an important role on mammal population dynamics.

Age Factors↗

Identification of four genes expressed by feeding female Ixodes scapularis, including three with sequence similarity to previously recognized genes.

We created a cDNA library from feeding, female Ixodes scapularis ticks and screened the library with a subtracted probe to eliminate most genes common to feeding female and mating male I. scapularis ticks. Four unique genes were identified in this screen. One gene, Is 9, (represented by 16 cDNAs) was more highly expressed in female ticks. This gene encodes a putative glycine-rich protein, which matched a number of glycine-rich proteins including attachment cement proteins from Rhipicephalus appendiculatus. A second gene, Is 10 (represented by one cDNA) was also more highly expressed in female ticks, but did not match any other sequences in the GenBank database. The third gene, Is 11 (represented by one cDNA) was very similar to Drosophila sp. hsp68 and hsp70 genes and was expressed about equally in male and female ticks. The fourth gene, Is 12 (represented by two cDNAs) was also about equally expressed in male and female ticks, and was similar to a salivary gland gene from Ixodes ricinis. This gene also showed limited similarity to some cuticle genes from insects.

Amino Acid Sequence↗

Redescription of larva, nymph and adults of Ixodes (I .) loricatus Neumann, 1899 (Acari: Ixodidae) based on light and scanning electron microscopy.

Ticks of the genus Ixodes are poorly known in the Neotropical zoogeographical region, from where only 45 species have been recorded. In Brazil, the genus is currently represented by eight species, four of which are known only from this country. This paper presents a redescription of all active stages in the life-cycle of I. loricatus , based on optical and scanning electron microscopy. In addition, the relationship of I. loricatus to other Neotropical Ixodes is presented and discussed.

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Ixodes rubicundus nymphs are short-day diapause-induced ticks with thermolabile sensitivity and desiccation resistance.

The Karoo Paralysis tick, Ixodes rubicundus Neumann (Acari: Ixodidae), is a semi-voltine ixodid that survives stressful environmental conditions using morphogenetic diapause (eggs and engorged nymphs) and desiccation resistance. Both photoperiod and temperature influence diapause induction in the engorged nymph. Ixodes rubicundus nymphs are typical long-day photoperiodic organisms. The critical photoperiod is approximately 13.5 h light, 10.5 h dark, and they display a thermolabile response. The period between detachment and apolysis in engorged nymphs is modified by photoperiod; however, apolysis to ecdysis is not affected by photoperiod. Thus, initiation of development, but not the actual process is controlled by photoperiod. Most engorged nymphs delayed metamorphosis when exposed to short-day regimen (LD 12 : 12 h) after feeding. Nymphs exposed to pre- and post-feeding long-day regimen (LD 14 : 10 h) developed. Times for 50% of nymphs to apolyse when exposed to photoperiods of LD 14 : 10 h, 13.5 : 10.5 h, 13 : 11 h and 12 : 12 h were 28, 36, 40 and 58 days, respectively. Times for 50% of engorged nymphs to ecdyse ranged from 38 to 40 days after apolysis. Nymphs were sensitive to photoperiodic exposures before, during and after feeding. Nymphs exposed to long day (LD 14 :10 h) before and during feeding, moulted at 20 degrees C; however, most exposed to 10 degrees C followed by 20 degrees C (post-feeding) went into diapause. Both short- (10 : 14 h) and long- (14 : 10 h) day exposed engorged nymphs survived 45 days at 0% r.h. (n = 73), but diapause-destined ticks kept at 13 degrees C lost the least mass (29.5+/-9.5%, SD), while nondiapause ticks at 23 degrees C lost the most (48.7+/-8.2%, SD). Termination of diapause and transition to development probably coincides with a definite increase of water vapour uptake by engorged nymphs. Comparatively, I. rubicundus engorged nymphs are more desiccation tolerant than a North-American counterpart, Amblyomma cajennense (Fabricius) (Acari: Ixodidae), which is also semi-arid- to xeric-adapted. Diapause conveys important survival attributes that enable engorged I. rubicundus nymphs to inhabit a semi-arid environment with great temperature extremes, and to synchronize their activity periods with seasons and host utilization patterns.

Animals↗

Sigmodontinae rodents as hosts for larvae and nymphs of Ixodes loricatus Neumann, 1899 (Acari: Ixodidae).

Larvae and nymphs of Ixodes loricatus Neumann, 1899 ticks (confirmed by morphological characters and by comparison of 16S mitochondrial rDNA sequences) were collected from Sigmodontinae Wagner, rodents in central and northern Argentina and Uruguay. A total of 100 larvae and 38 nymphs of I. loricatus were collected on the genera Akodon Meyen (n = 36 individuals), Calomys Waterhouse (n = 2), Oligoryzomys Bang in = 12), Oxymycterus Waterhouse (n = 9), and Scapteromys Waterhouse (n = 13). 72 larvae and 18 nymphs were collected on Akodon. Adults of I. loricatus were found in central Argentina and Uruguay on Didelphimorphia of the genera Didelphis Linnaeus and Lutreolina Thomas. Ixodes loricatus has been considered a species with strict total specificity to Didelphimorphia. Our results show that this statement may not be justified. Sigmodontinae rodents are sympatric and share habitats with the phylogenetically distant Didelphimorphia; infestation with I. loricatus seems to be its consequence. We tentatively consider I. loricatus moderately specific to Didelphimorphia.

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The tick Ixodes granulatus infests Rattus rattus populating a small island offshore of Singapore.

The ixodid tick Ixodes granulosus Supino 1897 was found infesting Rattus rattus in Semakau island, one of the small offshore islands fringing Singapore to the south. None of the examined R. rattus from the other islands fringing Singapore, or from Singapore island were found infested. Ixodes granulatus occurs, however, on Singapore island on Rattus annandalei, resident of undisturbed forested habitats. We speculate that invading black rats in Semakau replaced autochthonous sylvatic rodent population and contracted their associate tick population.

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Evidence of Bartonella sp. in questing adult and nymphal Ixodes ricinus ticks from France and co-infection with Borrelia burgdorferi sensu lato and Babesia sp.

Ticks are known vectors for a wide range of pathogenic microorganisms. Their role in the transmission of some others is so far only suspected. Ticks can transmit multiple pathogens, however, little is known about the co-existence of these pathogens within questing ticks. We looked for the presence of DNA from three micro-organisms, Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. which are known or suspected tick-borne pathogens, using a cohort of 92 questing Ixodes ricinus ticks collected from pastures in northern France. DNA was extracted from each individual tick and the presence of the three pathogens was investigated using Polymerase Chain Reaction (PCR) amplification. Nine among 92 samples (9.8%) demonstrated PCR products using Bartonella specific primers, 3 among 92 (3.3%) using Borrelia burgdorferi sensu lato specific primers and 19 among 92 (20.6%) using Babesia specific primers. Seven among 92 samples (7.6%) were PCR positive for at least two of the pathogens and one sample was positive for all three. Adult ticks (12/18; 67%) showed significantly higher infection rates compared to nymphs (11/74; 15%) for all three pathogens (P < 0.001). This study is the demonstration of the simultaneous presence of Bartonella sp., Borrelia burgdorferi sensu lato and Babesia sp. in questing Ixodes ricinus ticks.

Animals↗