Dermacentor Reticulatus Ticks Research Articles

Efficacy of an imidacloprid/flumethrin collar against fleas, ticks, mites and lice on dogs

BackgroundThe studies reported here were conducted to ascertain the efficacy of imidacloprid/flumethrin incorporated in a slow-release matrix collar, against infestations of dogs by fleas, ticks, mites and lice. Efficacy was evaluated against the flea Ctenocephalides felis felis, the ticks Rhipicephalus sanguineus, Ixodes ricinus, Ixodes scapularis, Dermacentor reticulatus and Dermacentor variabilis, the mite Sarcoptes scabiei and the biting louse Trichodectes canis.MethodsGroups of collar-treated dogs (n = 7–10) were infested with fleas and/or ticks at monthly intervals at least, over a period of up to 8 months. Efficacy against fleas was evaluated 24 to 48 h after treatment and 24 h after each re-infestation. Efficacy against ticks was evaluated at 48 h (acaricidal), 6 h (repellent) and 48 h (sustained) after infestation. The effect of regular shampooing or immersion in water on the efficacy of the collars was also tested. Efficacy against flea larvae was assessed by incubating blanket samples after dog contact with viable flea eggs. Effectiveness against lice and mites was evaluated after treatment of naturally infested animals. With the exception of the mites, efficacy was calculated by comparison with untreated negative control groups.ResultsEfficacy against fleas (24 h) generally exceeded 95%, and against flea larvae it exceeded 99% for 8 months. Sustained acaricidal (48 h) efficacy, covering a period of 8 months was 100% against I. ricinus, starting 2 days after treatment (in vivo), and 100% against I. scapularis (in vitro), above 97% against R. sanguineus, generally above 97% against D. reticulatus and above 90% for D. variabilis.Repellent (6 h) efficacy 2 days after treatment and continuing for 8 months was consistently 100% against I. ricinus, and above 90% against R. sanguineus.Regular shampooing affected efficacy against fleas and ticks to a lesser extent than regular immersion in water.The collars eliminated Trichodectes canis within 2 days and Sarcoptes scabiei within 3 months.ConclusionThe rapid insecticidal and acaricidal properties of the medicated collars against newly-acquired infestations of fleas and ticks and their sustained high levels of preventive efficacy have been clearly shown. Consequently they have the potential to prevent the transmission of vector-borne diseases and other conditions directly associated with infestation throughout an entire season of parasite abundance.

Molecular detection and characterization of Anaplasma phagocytophilum strains

The zoonotic rickettsial pathogen Anaplasma phagocytophilum has a broad geographic distribution and a high degree of biological and clinical diversity. To investigate the genetic diversity of A. phagocytophilum strains in the Baltic region and Norway, three species of Ixodidae ticks were examined for A. phagocytophilum infection, and two genes of the pathogen genome were analyzed. Analysis of partial 16S rRNA and partial major surface protein (msp4) gene sequences was accomplished through nested PCR and sequence analysis. Strains identified in this study were compared with those originating from other European countries and the United States. Seven 16S rRNA gene variants and fifteen msp4 gene variants of A. phagocytophilum were detected. Nine sequences had unique nucleotide polymorphisms and therefore differed from other A. phagocytophilum sequences previously submitted to GenBank. The present study represents the first molecular characterization of A. phagocytophum strains circulating in Lithuania and describes the strains detected in Ixodes persulcatus ticks in Latvia and Estonia. This is also the first report describing A. phagocytophilum strains isolated from Dermacentor reticulatus ticks.

Intraurban Near-Water Areas and Their Role in Distribution of Ticks (Acari, Ixodidae) in Megapolis the Example of Kyiv

Intraurban Near-Water Areas and Their Role in Distribution of Ticks (Acari, Ixodidae) in Megapolis the Example of Kyiv In 2002-2009, we studied 12 islands within the city of Kyiv, and found four indigenous species of ticks (Dermacentor reticulatus Fabricius, 1794; Ixodes ricinus Linnaeus, 1758; I. apronophorus Schulze, 1924; Rhipicephalus rossicus Jakimov et Kohl-Jakimova, 1911) and one invasive species (Hyalomma marginatum Koch, 1844). For A-E-gradient, established by man's impact degree, we revealed atypical distribution of species: their majority is noted in two pole areas - where the man impact is maximum and minimum. The investigated near-water areas are a kind of survival areas and sources of ixodides diversity in megalopolis.

The distribution and spreading pattern of Dermacentor reticulatus over its threshold area in the Czech Republic—How much is range of this vector expanding?

Host-seeking Dermacentor reticulatus ticks were detected by flagging method at 46 localities at south-east part of the Czech Republic, in the basins of rivers Morava and Dyje. Exact north-west distribution limits of D. reticulatus were defined in this area for the first time. Detailed prediction map of probabilities of D. reticulatus occurrence was obtained using GIS analysis. Spatial model delimited a south–north gradient in probability across the studied area, with highest probabilities above 0.8 in its southernmost part. Abundance of D. reticulatus varied markedly between localities in interval 0.33–222 of ticks per flag per hour. The highest abundances were in flooded areas at lower streams, towards upper streams abundance and density of these ticks decreased. Females prevailed in samples with population sex ratio of 0.413, significantly deviating from parity. Larvae and nymphs of this species were not detected by flagging. Although D. reticulatus range expansion probably did not reach such a degree as reported in other countries, these ticks became very abundant in some parts of studied area. Since spreading of vector-borne diseases became a problem in Europe, the knowledge of their exact recent geographic ranges is important for future modelling of their shift predictability.

A hidden beneficial: biology of the tick-wasp Ixodiphagus hookeri in Germany

The parasitic wasp Ixodiphagus hookeri parasitizes hard ticks and is therefore considered as a potential candidate for biological control of ticks. However, there are still considerable gaps of knowledge about the biology of I. hookeri, especially for European populations. Thus, the present study was performed to assess important life-history parameters of the parasitoid in Germany. Field studies accomplished in three successive years revealed that unfed Ixodes ricinus nymphs are infested by the parasitoid at a low but constant rate of 1.9–3.8% and that adult wasps are present only during a short period in late summer. The mean developmental time of wasps in I. ricinus nymphs ranged from 28 to 70 days under constant laboratory conditions and was prolonged in the second half of the year. Bioassays on parasitization and host preference behaviour showed that unfed nymphs of the host species I. ricinus are significantly preferred in experiments, in which unfed and engorged larvae as well as fully engorged nymphs were offered as alternatives. The marsh tick Dermacentor reticulatus was not accepted as an alternative host. Our data show that the investigated I. hookeri populations differ markedly from populations in other regions of the world in many aspects. The adaptation of different strains to local conditions explains the limited success of imported strains in earlier biological control attempts and highlights the importance of doing research to enhance the control potential of native strains.

Prevalence of tick-borne encephalitis virus in Ixodes ricinus and Dermacentor reticulatus ticks collected from the Lublin region (eastern Poland)

A total of 875 nymphal and adult Ixodes ricinus ticks and 148 adult Dermacentor reticulatus ticks were collected by flagging lower vegetation in the Lublin region (eastern Poland) and examined for the presence of RNA of tick-borne encephalitis virus (TBEV) by nested RT-PCR. The minimum infection rate of I. ricinus ticks with TBEV amounted to 1.6% while the infection rate of D. reticulatus ticks was 10.8%. The results suggest that D. reticulatus may be a potential vector of TBEV in Central Europe.

Potential role of ticks as vectors of bluetongue virus

When the first outbreak of bluetongue virus serotype 8 (BTV8) was recorded in North-West Europe in August 2006 and renewed outbreaks occurred in the summer of 2007 and again in 2008, the question was raised how the virus survived the winter. Since most adult Culicoides vector midges are assumed not to survive the northern European winter, and transovarial transmission in Culicoides is not recorded, we examined the potential vector role of ixodid and argasid ticks for bluetongue virus. Four species of ixodid ticks (Ixodes ricinus, Ixodes hexagonus, Dermacentor reticulatus and Rhipicephalus bursa) and one soft tick species, Ornithodoros savignyi, ingested BTV8-containing blood either through capillary feeding or by feeding on artificial membranes. The virus was taken up by the ticks and was found to pass through the gut barrier and spread via the haemolymph into the salivary glands, ovaries and testes, as demonstrated by real-time reverse transcriptase PCR (PCR-test). BTV8 was detected in various tissues of ixodid ticks for up to 21 days post feeding and in Ornithodoros ticks for up to 26 days. It was found after moulting in adult Ixodes hexagonus and was also able to pass through the ovaries into the eggs of an Ornithodoros savignyi tick. This study demonstrates that ticks can become infected with bluetongue virus serotype 8. The transstadial passage in hard ticks and transovarial passage in soft ticks suggest that ticks have potential vectorial capacity for bluetongue virus. Further studies are required to investigate transmission from infected ticks to domestic livestock. This route of transmission could provide an additional clue in the unresolved mystery of the epidemiology of Bluetongue in Europe by considering ticks as a potential overwintering mechanism for bluetongue virus.

16S rRNA gene-based identification of cultured bacterial flora from host-seeking Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks, vectors of vertebrate pathogens

A total of 151 bacterial isolates were recovered from different developmental stages (larvae, nymphs and adults) of field-collected ticks (67 strains from Ixodes ricinus, 38 from Dermacentor reticulatus, 46 from Haemaphysalis concinna). Microorganisms were identified by means of 16S rRNA gene sequencing. Almost 87 % of the strains belonged to G(+) bacteria with predominantly occurring genera Bacillus and Paenibacillus. Other G(+) strains included Arthrobacter, Corynebacterium, Frigoribacterium, Kocuria, Microbacterium, Micrococcus, Plantibacter, Rhodococcus, Rothia, and Staphylococcus. G(-) strains occurred less frequently, comprising genera Advenella, Pseudomonas, Rahnella, Stenotrophomonas, and Xanthomonas. Several strains of medical importance were found, namely Advenella incenata, Corynebacterium aurimucosum, Microbacterium oxydans, M. schleiferi, Staphylococcus spp., and Stenotrophomonas maltophilia. Data on cultivable microbial diversity in Eurasian tick species D. reticulatus and H. concinna are given, along with the extension of present knowledge concerning bacterial flora of I. ricinus.

Spatial distribution of Dermacentor reticulatus tick in Slovakia in the beginning of the 21st century

A new field survey monitoring the spatial distribution of Dermacentor ( D.) reticulatus (Fabricius, 1794) tick in Slovakia was carried out in 2005–2008 in order to record changes in its distribution when compared to former studies. Last surveys on the geographical distribution were conducted in 1950s and 1970s and the presence of D. reticulatus was determined along the rivers in the south-east (Latorica) as well as in the south-west (Morava, Dunaj) Slovakia. In the present survey new areas with D. reticulatus occurrence were detected, providing evidence that this tick species has extended its range in the surroundings of its former habitats but also by at least 200 km further North and by 300 m of elevation into higher altitudes. D. reticulatus is known to transmit Babesia spp. causing babesiosis in cattle and dogs. Expansion of D. reticulatus range is therefore likely to bring a spread of babesiosis, which can be severe or fatal especially for dogs.

Utilisation of rodent species by larvae and nymphs of hard ticks (Ixodidae) in two habitats in NE Poland

The impact of host identity and habitat type on the density of hard ticks (Ixodes ricinus and Dermacentor reticulatus) infections on rodents in forest and abandoned field habitats in NE Poland was investigated. Ixodes ricinus was most abundant in the forest system, but D. reticulatus, although rarer, was most abundant in the field system. Environmental humidity and the much lower density of rodents probably limited the abundance of I. ricinus larvae in the field system, although this tick was still common on Microtus oeconomus from around small ponds. Nymphs of I. ricinus were comparatively rare in the forest, probably because of infection of non-rodent hosts. Dermacentor reticulatus nymphs on the other hand were very much more common in the ears of Microtus species than would have been predicted based on larval densities. The impact of habitat change (in this case successional change following field abandonment) on tick densities is emphasised, and the role of Apodemus as an epidemiological bridge, linking woodland and field habitats, is highlighted.

Emerging arthropod-borne diseases of companion animals in Europe

Vector-borne diseases are caused by parasites, bacteria or viruses transmitted by the bite of hematophagous arthropods (mainly ticks and mosquitoes). The past few years have seen the emergence of new diseases, or re-emergence of existing ones, usually with changes in their epidemiology (i.e. geographical distribution, prevalence, and pathogenicity). The frequency of some vector-borne diseases of pets is increasing in Europe, i.e. canine babesiosis, granulocytic anaplasmosis, canine monocytic ehrlichiosis, thrombocytic anaplasmosis, and leishmaniosis. Except for the last, these diseases are transmitted by ticks. Both the distribution and abundance of the three main tick species, Rhipicephalus sanguineus, Dermacentor reticulatus and Ixodes ricinus are changing. The conditions for such changes involve primarily human factors, such as travel with pets, changes in human habitats, social and leisure activities, but climate changes also have a direct impact on arthropod vectors (abundance, geographical distribution, and vectorial capacity). Besides the most known diseases, attention should be kept on tick-borne encephalitis, which seems to be increasing in western Europe, as well as flea-borne diseases like the flea-transmitted rickettsiosis. Here, after consideration of the main reasons for changes in tick vector ecology, an overview of each “emerging” vector-borne diseases of pets is presented.

Efficacy of a single application of a 12.5% pyriprole topical solution against laboratory infestations with ticks ( Ixodes ricinus, Dermacentor reticulatus, and Rhipicephalus sanguineus) on dogs

In three separate studies, the efficacy of a single treatment with a 12.5% pyriprole spot-on solution was investigated against induced infestation with Ixodes ricinus, Dermacentor reticulatus and Rhipicephalus sanguineus on dogs (both sexes; Beagles in Studies 1 and 2, mixed-breed in Study 3). For each tick species, one group of 8 dogs left untreated (Studies 1 and 2) or treated with a placebo solution (Study 3) was compared with another group treated once with the spot-on solution at a dose rate of at least 12.5 mg/kg. The dogs were infested with 50 unfed adult ticks of the respective species at various time-points before and after treatment and the surviving attached and unattached ticks were recorded 48 h after re-infestation. For each tick species, efficacy was assessed for each time-point and cumulatively for the whole evaluation period. The dogs were submitted to general health observations and clinical assessments during the study. Efficacy against I. ricinus and R. sanguineus was 100% during the whole evaluation period of 30 days. For D. reticulatus cumulative efficacy for the 30 days after treatment was 98.9%. The product was well tolerated by all the animals.

Effects of host diversity and the community composition of hard ticks (Ixodidae) on Babesia microti infection

Babesia microti (Apicomplexa, Piroplasmida) is one of the aetiological agents of human babesiosis, maintained in nature by complex zoonotic transmission cycles, involving small wild mammals as hosts and hard ticks (Ixodidae) as vectors. We previously reported that the B. microti strain commonly encountered among Microtus rodents in Poland is not zoonotic and that it is therefore most unlikely that it represents a risk to public health. Here, we analysed the contribution of four rodent species, Myodes glareolus, Apodemus flavicollis, Microtus arvalis, and Mi. oeconomus as hosts for two species of locally abundant ticks ( Ixodes ricinus and Dermacentor reticulatus) and the rodent infective B. microti. Studies were carried out in the Mazury Lakes District of north-eastern Poland in 2005 and 2006. Infestation prevalence and abundance of both species of ticks (altogether 5674) collected from 781 rodent hosts, including 61 A. flavicollis, 495 My. glareolus, 99 Mi. arvalis, and 126 Mi. oeconomus from two different habitats, were determined. A. flavicollis and My. glareolus from the woodland habitat were more frequently infested with larvae and nymphs of I. ricinus ticks (96.7% and 87.9%, respectively) than the other species of hosts. In contrast, a higher percentage (>30%) of the other two species of rodents ( Mi. arvalis, Mi. oeconomus) from fallow lands were infested with larvae and nymphs of D. reticulatus ticks (31.3% and 68.3%, respectively). Pronounced co-infestation rates (36.3%) of I. ricinus and D. reticulatus ticks (larvae and nymphs) were recorded on Mi. oeconomus. B. microti infections in blood samples of hosts and in ticks collected from hosts were examined by PCR, followed by sequence analysis. The prevalence of B. microti in hosts from the fallow lands ( Mi. arvalis and Mi. oeconomus) was much higher than that in hosts from woodland ( A. flavicollis and My. glareolus). However, the overall prevalence of B. microti in ticks collected from rodents was quite low, and varied from 3.4% for larvae to 4.8% for nymphs of I. ricinus and 11.8% for larvae to 4.0% for nymphs of D. reticulatus. The present study revealed for the first time the presence of B. microti DNA in partly engorged immature I. ricinus, but also in partly engorged larvae and nymphs of D. reticulatus. Furthermore, our results suggest widely differing roles of various rodent host species from two different habitats in maintaining zoonotic transmission of B. microti.

Ticks and Associated Pathogens Collected from Domestic Animals in the Netherlands

Following an outbreak of autochthonous canine babesiosis in the Netherlands, a request made to veterinarians and the public to collect ticks from companion animals resulted in 4298 ticks submitted between July 2005 and October 2006 to our center. Ticks were identified as Ixodes ricinus adults (2907/4298, 67.6%), Ixodes sp. nymphs (529/4298, 12.3%) and Ixodes sp. larvae (385/4298, 9.0%), I. hexagonus adults (328/4298, 7.6%), Dermacentor reticulatus (72/4298, 1.7%), and several other exotic tick species such as Amblyomma flavomaculatum (formerly Aponomma flavomaculatum), Hyalomma marginatum rufipes, Rhipicephalus sanguineus, and R. turanicus (55/4298, 1.3%). Eight localities were surveyed for the presence of local D. reticulatus, a tick not indigenous to the Netherlands, based on multiple submissions of D. reticulatus ticks from these areas. D. reticulatus was collected from the vegetation in six of these localities, confirming the presence of populations of this tick in the Netherlands. Adult I. ricinus (n=251), I. hexagonus (n=237), and D. reticulatus (n=344) ticks were selected at random and subsequently screened by polymerase chain reaction (PCR) and reverse line blot (RLB) hybridization for the presence of Borrelia, Babesia, Theileria, Anaplasma, Ehrlichia, and Rickettsia species. I. ricinus ticks were infected with Rickettsia helvetica (24.7%), spirochetes belonging to the Borrelia burgdorferi sensu lato group (7.2%), the Ehrlichia-like "Schotii" variant (2.4%), Anaplasma phagocytophilum (1.6%), Babesia sp. (EU1) (1.2%), Babesia divergens (0.4%), and Babesia microti (0.4%). A. phagocytophilum (5.9%) and R. helvetica (0.8%) were also detected in adult I. hexagonus ticks. Spotted fever group Rickettsiae, previously reported as Rickettsia sp. DnS14/RpA4 (14.0%), and Borrelia burgdorferi sensu lato (0.3%) were detected in the D. reticulatus ticks, which appeared to be free from B. canis infection. We concluded that a much broader spectrum of ticks and tick-borne pathogens is present in the Netherlands than previously thought, including several potential zoonotic pathogens.

Temporal distribution of Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna in Hungary

A survey was carried out over a 4-year period to describe the temporal distribution of three ‘anthropophilic’ tick species, Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna in Hungary. Altogether 4658 adult ticks belonging to the three species were collected from 1931 red foxes ( Vulpes vulpes) killed in an area of about 70,000 km 2 representing all major climatic areas of the country. The seasonal activity of the three species was different. I. ricinus ticks were most active between April and June with an activity peak in May. A less marked increase of activity was also observed in September and October. The highest activity of D. reticulatus ticks was seen between September and November with an activity peak in October, nevertheless, a marked increase of activity could also be observed in April. Small number of I. ricinus and D. reticulatus were collected in all other months. H. concinna ticks were active from May to July with an activity peak in June and completely disappeared between October and March. The temporal distribution of the three tick species might be used for predictions on the seasonality of tick-borne diseases in Hungary.

Evidence for an increased geographical distribution of Dermacentor reticulatus in Germany and detection of Rickettsia sp. RpA4

Two studies were performed to elucidate the current distribution of the tick Dermacentor reticulatus in Germany. In the first one in 2003, a total of 365 dogs from 171 sites in the states of Berlin and Brandenburg was screened for ticks, and the corresponding outdoor sites that the dogs usually visited were searched for host-seeking ticks by the flagging method. A total of 1155 ticks was removed from the dogs. The majority were Ixodes ricinus (88.5%), followed by D. reticulatus (9.1%) and I. hexagonus (2.4%). Altogether, 222 dogs carried I. ricinus (60.8%), 41 D. reticulatus (11.2%) and 15 I. hexagonus (4.1%) ticks. Based on scutal index determination, the removed I. ricinus and D. reticulatus had been feeding on the dogs for a mean of 4.0 and 4.5 days, respectively. The dogs infested with D. reticulatus lived at 26 different sites, all previously unknown as Dermacentor sites. Seven of the sites could be confirmed subsequently by flagging the vegetation for ticks. In the second study, a total of 721 deer was shot at 201 different sites from a total of 160 districts all over Germany during the autumn hunting season 2004. A total of 23 deer (3.2%) originating from 14 sites was infested with D. reticulatus. Hereby, significantly more red deer ( Cervus elaphus elaphus) than roe deer ( Capreolus capreolus) or fallow deer ( Dama dama) harboured D. reticulatus ticks. Only two of the sites found had already been known as D. reticulatus areas, whereas all other sites in Brandenburg, Saxony Anhalt, Hesse and Bavaria had been unknown. The results of both studies show that D. reticulatus presently occurs at far more sites than previously known in Germany and thus most likely has expanded its range. Additionally, a total of 135 D. reticulatus removed from deer was screened for Babesia canis and Rickettsia sp. by PCR. A total of 31 D. reticulatus (23%) were positive for Rickettsia. Sequencing revealed in all cases 100% identity with the strain RpA4 that was first isolated from Rhipicephalus ticks in Russia.

Manipulation of host cytokine network by ticks: a potential gateway for pathogen transmission

Ticks are obligatory blood-feeding arthropods that secrete various immunomodulatory molecules to antagonize host inflammatory and immune responses. Cytokines play an important role in regulating these responses. We investigated the extent to which ticks interact with the sophisticated cytokine network by comparing the effect of salivary gland extracts (SGE) of 3 ixodid tick species, Dermacentor reticulatus, Amblyomma variegatum and Ixodes ricinus, all of which are important vectors of tick-borne pathogens. Using specific ELISAs, anti-cytokine activity was demonstrated with 7 cytokines: IL-8, MCP-1, MIP-1alpha, RANTES, eotaxin, IL-2 and IL-4. The results varied between species, and between adult males and females of the same species. Relatively high activity levels were detected in saliva of female D. reticulatus, confirming that the observed anti-cytokine activities are an integral part of tick saliva secreted into the host. Results with fractionated SGE indicated that from 2 to 6 putative cytokine binding molecules are produced, depending on species and sex. Binding ability of SGE molecules was verified by cross-linking with radio-isotope labelled MIP-1alpha. By targeting different cytokines, ixodid ticks can manipulate the cytokine network, which will greatly facilitate blood-feeding and provide a gateway for tick-borne pathogens that helps explain why ticks are such efficient and effective disease vectors.

Evidence for the reproductive isolation of Dermacentor marginatus and Dermacentor reticulatus (Acari: Ixodidae) ticks based on cross-breeding, morphology and molecular studies

The species status of Dermacentor marginatus and Dermacentor reticulatus was evaluated by scanning electron microscope (SEM) examination of adult ticks, cross-breeding experiments and molecular biological analysis of eggs derived from transspecific pairings. The SEM investigations including the morphometric quantification of phenotypic features resulted in an unequivocal differentiation of adult D. marginatus and D. reticulatus ticks. The cross-breeding experiments demonstrated that irrespective of whether female ticks of both species were applied with con- or transspecific male ticks or without males to sheep, they engorged and laid eggs. The larvae, however, developed only in eggs which originated from conspecific matings. A nested polymerase chain reaction (PCR) of the second internal transcribed spacer (ITS2) using the DNA of eggs from transspecific pairings and sequencing of the PCR products revealed two different genotypes. The genotypes of eggs originating from D. marginatus and D. reticulatus females of these pairings differed. However, the eggs deposited by D. marginatus always possessed the same two genotypes as did the eggs produced by D. reticulatus. These results argue for a strict reproductive isolation of D. marginatus and D. reticulatus and, therefore, for a separate species status.

Use of partially engorged female ticks as laboratory animals in microbiological research

Partially engorged female ticks were used as laboratory animals in microbiological research. The ticks, which were inoculated intracoelomally, became a convenient substrate for the detection of viruses, rickettsiae and protozoal parasites. This research concerned the isolation of newly recovered micro-organisms, the study of development, structure and distribution of microbial agents in ticks, and the study of their interaction with other pathogens or symbionts during mixed infection in a tick body. The isolation and maintenance of Rickettsiella phytoseiuli, the organism not of tick-borne origin, was achieved. For use in Central Europe the tick Dermacentor reticulatus is recommended for the above investigations.

Enhancement of tick-borne encephalitis virus transmission by tick salivary gland extracts.

To investigate the role of ticks in TBE virus transmission, salivary gland extract (SGE) was derived from partially fed female Ixodes ricinus, Dermacentor reticulatus and Rhipicephalus appendiculatus ticks. Guinea-pigs were infested with uninfected R. appendiculatus nymphs and inoculated with a mixture of TBE virus and SGE or with virus alone. The number of ticks which on average acquired virus from feeding on animals inoculated with TBE virus and SGE from partially fed ticks was 4-fold greater than the number that became infected by feeding on animals inoculated with virus alone or virus plus SGE from unfed I. ricinus. Viraemia was detected in 67% of guinea-pigs inoculated with virus plus SGE compared to 30% of guinea-pigs inoculated with virus alone. Virus titres in the blood were similar for both groups of animals [range 2.0-2.8 log10 plaque-forming units (PFU)/ml of blood]; however, the number of ticks that became infected was significantly higher on animals inoculated with virus plus SGE from partially fed ticks. No significant difference was observed with respect to the tick species used to derive SGE. The results indicate that TBE virus transmission is enhanced by factor(s) associated with the salivary glands of feeding ticks, and that these factor(s) may facilitate efficient transmission of TBE virus between infected and uninfected ticks even when they feed on hosts that have no detectable viraemia.