Tick infection rates with Borrelia Ixodes ricinus versus Haemaphysalis concinna and Dermacentor reticulatus in two locations in eastern Germany

Unfed nymphal Ixodes ricinus, Haemaphysalis concinna, and adult Dermacentor reticulatus were collected in two locations of Saxony in July and September 1991 by flagging. In July, the abundance of nymphal I. ricinus was about 2-3 times higher than that of nymphal H. concinna, a time of the year when nymphs of both species are reported to have a seasonal peak of activity. No D. reticulatus were flagged concurrently. In September, host-seeking activity of nymphal I. ricinus was again quite high as was that of adult D. reticulatus but only low numbers of nymphal H. concinna were collected. The flagged ticks were individually examined for Borrelia by an indirect immunofluorescence assay (I. ricinus: n = 414; H. concinna: n = 96; D. reticulatus: n = 116). The prevalence of Borrelia (probably B. burgdorferi) in I. ricinus varied from 12.1% to 21.0%. No borreliae were found in H. concinna. Of the examined D. reticulatus from one site (n = 97) 11.3% contained either B. burgdorferi or a related Borrelia. This may be the first finding of Borrelia in an Eurasian Dermacentor species.     

Ixodes ricinus is the dominant questing tick in forest habitats in Romania: the results from a countrywide dragging campaign

In 2010 and 2011, questing ticks were collected from 188 forested locations in all the 41 counties of Romania using the dragging method. The total of 13,771 ticks collected belonged to eleven species: Ixodes ricinus (86.9?%), Dermacentor marginatus (9.5?%), Haemaphysalis punctata (2.6?%), H. concinna (0.6?%), H. sulcata (0.3?%), H. parva (0.1?%), Hyalomma marginatum (0.02?%), D. reticulatus (0.02?%), I. crenulatus (0.007?%), I. hexagonus (0.007?%) and I. laguri (0.007?%). Ixodes ricinus was present in 97.7?% (n?=?180) of locations, occurring exclusively in 41.7?% of the locations, whereas it was the dominant species in 38.8 % of the other locations, accounting for over 70?% of the total tick community. The following most common questing ticks were D. marginatus, H. punctata and H. concinna. Ixodes ricinus co-occurred with one, two or three sympatric species. The occurrence of D. reticulatus in forested habitats from Romania was found to be accidental.     

Distribution and molecular detection of Theileria and Babesia in questing ticks from northern Spain

Abstract.  A total of 562 questing adult ixodid ticks, collected during 2003−05 in 10 recreational mountain areas in northern Spain, were analysed for piroplasm infection. Reverse line blot (RLB) analysis using a panel of probes for 23 piroplasm species identified 16 different piroplasms, with an overall prevalence of 9.3%. Most were Theileria spp.-positive (7.7%), 3.0% were positive for Babesia spp. and 1.4% of ticks harboured both genera. Ixodes ricinus (Linnaeus, 1758), the most abundant tick in the vegetation, ranked third with regard to piroplasm infection prevalence (11.4%) after Rhipicephalus bursa (Canestrini & Fanzago, 1878) (16.0%) and Haemaphysalis punctata (Canestrini & Fanzago, 1878) (13.5%). Infection was detected in 6.2% of Dermacentor reticulatus (Fabricius, 1794) and in 1.1% of Haemaphysalis inermis (Birula, 1895), but was absent from Haemaphysalis concinna (Koch, 1844). Ixodes ricinus carried more piroplasm species (13), followed by H. punctata (10), D. reticulatus (8), R. bursa (3) and H. inermis (1). Although most of the positive ticks harboured a single infection (76.9%), mixed infections with two or three different piroplasm species were also detected (23.1%). The various tick−pathogen associations found are discussed and prevalences of infection in ticks are compared with previous results on piroplasms infecting animals in the same region.     

Vector roles of Fennoscandian mosquitoes attracted to mammals, birds and frogs

1. Mosquitoes were sampled with five suction traps, from May to October 1983, at a forest site in south-central Sweden. 2. Twenty-three species of mosquitoes were identified among the total of 3108 females collected: 4% of them in an unbaited trap, 3% in a trap baited with two frogs, 24% with a guinea-pig, 28% with a hen and 40% in a trap baited with a rabbit. 3. The dominant species of Culicidae trapped were 39% Aedes communis (De Geer), 21% Ae. cinereus Meigen, 14% Coquillettidia richiardii (Ficalbi), 8% Ae. punctor (Kirby) and 4% Culiseta morsitans (Theobald). 4. Aedes annulipes (Meigen), Ae. cantans (Meigen), Ae. cinereus, Ae. communis and Ae. punctor were mostly attracted to the rabbit, whereas Culiseta morsitans, Culex pipiens L. and/or Cx torrentium Martini were strongly ornithophilic. 5. Based on these and previously published data the ecological and behavioural potential of the mosquitoes to transmit Sindbis, Inkoo, Tahyna and Batai viruses, tularaemia (caused by Francisella tularensis) and Ixodes-borne borreliosis (caused by Borrelia burgdorferi) in Fennoscandia is discussed. 6. The data support the hypothesis that Sindbis virus, which is enzootic in bird populations in Fennoscandia, is vectored between birds by Cx pipiens/torrentium and Cs. morsitans, and that abundant Aedes spp., particularly Ae. cinereus, which feed on both birds and mammals, are primary link vectors from infective birds to man and other mammals.     

Foci report on indigenous Dermacentor reticulatus populations in Belgium and a preliminary study of associated babesiosis pathogens

The occurrence of autochthonous clinical cases of canine and equine babesiosis in Belgium during the last two decades suggests that the vector of the pathogens responsible for these diseases, Dermacentor reticulatus (Ixodida: Ixodidae), may be present in this country. Consequently, evidence for the presence of this tick species in different locations within Belgium was investigated. Four different locations were monitored by flagging in 2010; these included the locations at which D. reticulatus was previously found on a dog in 2009 and on two red deer in 2007. Two different species of tick were identified, Ixodes ricinus (Ixodida: Ixodidae) and D. reticulatus. A total of 282 D. reticulatus adult ticks (98 males, 184 females) were collected from the four sites. Ticks were found mainly from early March until the end of May and a peak in activity was apparent in March. A Babesia spp. (Piroplasmida: Babesiidae) genus-specific polymerase chain reaction test based on the amplification of a fragment of the 18S rRNA gene was used to investigate the potential presence of Babesia spp. All DNA extracts isolated from the total tick samples yielded negative results. Additional studies to accurately determine the distribution and vectorial capacity of this important tick species in Belgium are warranted.     

Immunity of dogs against Babesia canis, its vector tick Dermacentor reticulatus, and Ixodes ricinus in endemic area

Previous epidemiological studies allowed us to accurately define endemic areas of canine babesiosis and tick distribution in southeastern France (Martinod, 1983). Using a micro-ELISA test 100 dogs sera were tested with 3 antigens: Babesia canis, Dermacentor reticulatus and Ixodes ricinus. Antibodies against B. canis and its vector D. reticulatus were detected in an endemic area, sometimes with high levels (optical density 1.38 and 0.80 respectively). A correlation factor and regression lines were found between ELISA activity of B. canis and vector tick antigens, even for dogs which never showed any babesiosis symptoms. These results were compared with those of an area without any babesiosis. Furthermore I. ricinus antigens detected ELISA activity in sera of dogs; some cross reactions were observed between I. ricinus and D. reticulatus antigen.     

Evaluation of the Possible Role of Ixodid Ticks in Natural Tularemia Foci in the Forest-Steppe Zone of European Russia

Entomological Review - Adult ixodid ticks Dermacentor reticulatus (Fabricius, 1894), D. marginatus (Sulzer, 1776) and Ixodes ricinus (Linnaeus, 1758), collected from vegetation in April–May...     

Acarological studies in two protected areas of Central Italy

In the present note are reported the results of preliminary studies on tick distribution in two wildlife areas of Abruzzo (National Park of Abruzzo, a mountainous area, approximately 40,000 ha, in Central Apennines, interesting Abruzzo, Latium and Molise regions) and Latium (Capocotta-Castel Porziano Presidential reserve, on the Tyrrhenian coast, 30 km from Rome). Sampling of ticks from domestic and wild mammals as well as from vegetation, was performed in three different areas of the National Park in 1995. Tick species identified include Rhipicephalus bursa, R. sanguineus, Hyalomma marginatum, Haemaphysalis punctata, Dermacentor marginatus, Ixodes ricinus, I. hexagonus. The presence of I. ricinus was discontinuous and sporadic. In Capocotta estate, samplings were performed bimonthly from March to November 1997 in an restricted area (1 ha) with typical Mediterranean flora and fauna. The following species were collected: I. ricinus, Haemaphysalis concinna, D. marginatus, R. bursa, Hy. marginatum. There was a high dominance of I. ricinus and H. concinna.     

Ixodid ticks of Pavlodar Province and their participation in the circulation of tularemia infection

Investigations carried out in Pavlodar Province have shown that 7 species of ixodid ticks, Ixodes crenulatus, I. lividus, I. persulcatus, I. laguri laguri, Dermacentor marginatus, D. reticulatus, Haemaphysalis concinna, and one brought species, Hyalomma asiaticum, parasitize domestic animals and wild mammals. Maximum activity of pasture ticks of the genus Dermacentor falls on the end of April--beginning of May, that of H. concinna on the beginning of June. The growth in the abundance of pasture ticks, recorded during the last years, is favoured by the increase in the cattle live stock, its seasonal movements and pasture area reduction. Natural nidi of tularemia become more active. Since 1977 221 cultures of tularemia agent have been isolatid from pasture ticks. H. concinna takes the most active part in the infection circulation.     

Results of the 20-year study of tick-borne encephalitis in Crimea

The history of tick-borne encephalitis (TBE) study in the Crimea was retraced and the possibility of introduction of TBE virus to the Crimea in the process of the acclimatization of vertebrates from areas enzootic for TBE is discussed. The foci of TBE were found throughout the whole of the mountain forest zone of the Crimea and coincided with the habitat area of lxodes ricinus, the main vector of TBE. Such ticks as Dermacentor reticulatus, D. marginatus and Hyalomma marginatum were also found to take part in the circulation of the virus. Among the residents of the mountain forest zone, 13.9% were found to have immunity to TBE, testifying to the wide contact of the population with the pathogen. TBE morbidity had pronounced seasonal character and the morbidity rate was low with the prevalence of mild clinical forms. The characteristic feature of the virus population was polymorphism: strains identical to the Far-Eastern strains Sofyin (about 60-70%) and strains differing in their antigenic structure circulated here.     

Rickettsia slovaca and Rickettsia raoultii in Dermacentor marginatus and Dermacentor reticulatus ticks from Slovak Republic

Rickettsiae, obligate intracellular Gram-negative bacteria, responsible for mild to severe diseases in humans are associated with arthropod vectors. Dermacentor marginatus and Dermacentor reticulatus are known vectors of Rickettsia slovaca and Rickettsia raoultii distributed across Europe. A total of 794 D. marginatus, D. reticulatus and Ixodes ricinus adult ticks were collected from the vegetation, removed from horses, sheep, goats and dogs in Slovakia. The DNA of Rickettsia sp. was found in 229 ticks by PCR amplifying parts of gltA, ompA and sca4 genes. Next analyses of Rickettsia-positive samples by PCR-RFLP and/or sequencing showed D. reticulatus ticks were more infected with R. raoultii and D. marginatus were more infected with R. slovaca. The prevalence of R. raoultii was 8.1-8.6% and 22.3-27% in D. marginatus and D. reticulatus, respectively. The prevalence of R. slovaca was 20.6-24.3% in D. marginatus and 1.7-3.4% in D. reticulatus. Intracellular growth of R. raoultii isolate from D. marginatus tick was evaluated by rOmpA-based quantitative SybrGreen PCR assay. The highest point of multiplication was recorded on the 7th and 8th day postinfection in Vero and L929 cells, respectively. R. raoultii was transmitted during feeding of R. raoultii-positive ticks to guinea pigs and subsequently rickettsial infection was recorded in all organs, the highest infection was in spleen, liver and heart. Our study describes the detection and isolation of tick-borne pathogens R. raoultii and R. slovaca, show that they are spread in Slovakia and highlight their risk for humans.     

Francisella tularensis prevalence and load in Dermacentor reticulatus ticks in an endemic area in Central Europe

A total of 7778 host‐seeking adult Dermacentor reticulatus (Ixodida: Ixodidae) ticks were examined for the prevalence of Francisella tularensis holarctica (Thiotrichales: Francisellaceae) in a natural focus of tularaemia in the floodplain forest–meadow ecosystem along the lower reaches of the Dyje (Thaya) river in South Moravia (Czech Republic) between 1995 and 2013. Ticks were pooled (10 specimens per pool) and their homogenates inoculated subcutaneously in 4‐week‐old specific pathogen‐free mice. Dead mice were sectioned, their spleens cultivated on thioglycollate–glucose–blood agar and impression smears from the spleen, liver and heart blood were Giemsa‐stained. Sixty‐four pools were positive for F. tularensis: the overall minimum infection rate (MIR) was 0.82%. Overall MIRs for the 4714 female and 3064 male D. reticulatus examined were 0.89 and 0.72%, respectively; MIRs fluctuated across years between 0.0 and 2.43%. The estimated bacterial load in infected ticks varied from 0.84 to 5.34 log₁₀ infectious F. tularensis cells per tick (i.e. from about seven to 220 000 cells). Ticks with low loads were more prevalent; more than 1000 infectious cells were detected in 24 ticks (0.3% of all ticks and 37.5% of infected ticks). Monitoring of D. reticulatus for the presence and cell numbers of F. tularensis may be a valuable tool in the surveillance of tularaemia.     

Francisella tularensis Subspecies holarctica Occurs in Swedish Mosquitoes,Persists Through the Developmental Stages of Laboratory-Infected Mosquitoes and Is Transmissible During Blood Feeding

In Sweden, mosquitoes are considered the major vectors of the bacterium Francisella tularensis subsp. holarctica, which causes tularaemia. The aim of this study was to investigate whether mosquitoes acquire the bacterium as aquatic larvae and transmit the disease as adults. Mosquitoes sampled in a Swedish area where tularaemia is endemic (Örebro) were positive for the presence of F. tularensis deoxyribonucleic acid throughout the summer. Presence of the clinically relevant F. tularensis subsp. holarctica was confirmed in 11 out of the 14 mosquito species sampled. Experiments performed using laboratory-reared Aedes aegypti confirmed that F. tularensis subsp. holarctica was transstadially maintained from orally infected larvae to adult mosquitoes and that 25 % of the adults exposed as larvae were positive for the presence of F. tularensis-specific sequences for at least 2 weeks. In addition, we found that F. tularensis subsp. holarctica was transmitted to 58 % of the adult mosquitoes feeding on diseased mice. In a small-scale in vivo transmission experiment with F. tularensis subsp. holarctica-positive adult mosquitoes and susceptible mice, none of the animals developed tularaemia. However, we confirmed that there was transmission of the bacterium to blood vials by mosquitoes that had been exposed to the bacterium in the larval stage. Taken together, these results provide evidence that mosquitoes play a role in disease transmission in part of Sweden where tularaemia recurs.     

Diel activity patterns of blood-seeking anthropophilic mosquitoes in central Sweden

1. Blood-seeking diel activity cycles of mosquitoes are described from 24 h landing catches on human bait in a field and a forest habitat during June-August in central Sweden. 2. Daytime activity in the field was low or absent, whilst in the forest activity was relatively even throughout the diel. 3. In both habitats, activity peaks generally occurred about sunset and sunrise and were more distinct in the field than in the forest. 4. Data from this and other studies show that, in general, Aedes cinereus Meigen feeds at night in field-type habitats and Ae.communis (DeGeer) during the diurnal-crepuscular period in forest and woodland habitats. 5. Although abundant in the study area, Anopheles maculipennis Meigen sensu lato, Culex pipiens L., Cx torrentium Martini and Culiseta morsitans (Theobald) were not caught on the human bait. This is because, in central Sweden, An.maculipennis s.l. is mainly feeding in animal sheds whereas the others are ornithophagous. 6. Ae.geniculatus (Olivier) was recorded from central Sweden for the first time. 7. Blood feeding success (proportion of mosquitoes that managed to obtain blood from the human bait) was significantly greater in Ae.cinereus (14.9%) than in Ae.punctor (Kirby), Ae.communis and Ae.annulipes (Meigen) (2.8-0.7%). 8. The data are interpreted in relation to the transmission of Ockelbo disease (caused by Sindbis virus) and tularaemia in Sweden.     

Distribution of Borrelia burgdorferi s.l. spirochaete DNA in British ticks (Argasidae and Ixodidae) since the 19th Century, assessed by PCR

The distribution of Borrelia burgdorferi sensu lato , the Lyme borreliosis agent, was surveyed in British ticks in the collection of the Natural History Museum, London. Alcohol-preserved specimens of eight species of ticks known to attack humans were studied: Ixodes ricinus , I. hexagonus , I. uriae , I. trianguliceps , Dermacentor reticulatus , Haemaphysalis punctata , Rhipicephalus sanguineus and Argas vespertilionis. The sample comprised all life stages and originated from a wide range of host species, collection dates (1896–1994) and geographical localities in England, Scotland and Wales.
Borrelia burgdorferi s. l. DNA, detected by a polymerase chain reaction that targeted the outer surface protein A gene, was found in all eight species. The overall proportion of PCR-positive specimens ranged from 7.8% for I. hexagonus (mostly from mustelids and hedgehogs) to 98.3% for I. uriae (mostly from seabirds). Borrelia burgdorferi s. l. DNA was found for the first time in the bat parasite A. vespertilionis (85.3%). The spirochaete is newly recorded in British populations of I. trianguliceps (97.4%, mostly from voles, mice and shrews), D. reticulatus (12.5% from dog and man) and R. sanguineus (30% from dogs and human dwellings). Of the four tick species with larvae available for testing, examples of I. ricinus, I. uriae and A. vespertilionis were PCR positive, as were significantly more nymphs than adults of I. ricinus, I. hexagonus and A. vespertilionis. Analyses showed that B. burgdorferi s. l. has been consistently present in British tick populations since at least 1897. Ticks positive for B. burgdorferi s. l. DNA were collected in all months of the year, throughout Britain, and were found on a wide range of mammal and bird species. PCR positivity does not prove vector or reservoir competence, but the use of archived material has demonstrated an extensive range of host–tick relationships involving B. burgdorferi s.l. in Britain for >100 years.     

Ticks and mosquitoes as vectors of Borrelia burgdorferi s. l. in the forested areas of Szczecin

The aim of the study was to determine the infection level of adult forms and larvae of ticks and mosquitoes with Borrelia burgdorferi in the forested areas of Szczecin. A total of 1699 ticks Ixodes ricinus, including 1422 nymphs, 277 adult forms and 2862 mosquito females representing the genera Aedes (89.6%) and Culex (10.4%) were collected between the years 2004 and 2005. A further 3746 larvae and 1596 pupae of Culex pipiens pipiens were colleted from water bodies. Borrelia burgdorferi s. l. was detected in the arthropods by the method of indirect immunofluorescence assay (IFA). A positive immunological reaction was detected in 16.6% of the adult forms and in 16.5% of the nymphs of Ixodes ricinus. Spirochetes were also detected in 1.7% of mosquito females, 3.2% of larvae and in 1.6% of pupae of Culex pipiens pipiens. The results of the present study confirm that contact with ticks constitutes the main risk of contracting Lyme disease, although mosquitoes play a role as vectors as well.     

The ultrastructure of the interaction of Rickettsia sibirica and R. slovaca with the cells of ixodid ticks

The ultrastructural aspects of the interaction of R. sibirica and R. slovaca with cells of mites of the species Dermacentor reticulatus, D. marginatus and Ixodes ricinus after their parenteral infection, as well as in the organs of D. marginatus infected naturally in the environment, have been studied. Both rickettsial species have similar morphology in different organs of the vector. These rickettsiae not only multiply, their populations are also partly destroyed in phagolysosomes. The natural mixed infection of R. sibirica and orbivirus in cells of D. reticulatus is described. As shown in this study, both associates pass through the complete ontogenetic cycle of development on the level of the host body and also on the level of an individual cell.     

Immunomodulatory arsenal of nymphal ticks

Ticks have developed their own immunomodulatory mechanisms to inhibit the host inflammatory response. One of them involves the ability to subvert the cytokine network at the site of tick feeding by secreting cytokine binding molecules. Most studies have focused on the immunomodulatory prowess of adult female ticks. Here we describe anti-cytokine activity in salivary gland extracts (SGEs) prepared from 2-day-fed nymphs of Dermacentor reticulatus Fabricius, Ixodes ricinus L., Rhipicephalus appendiculatus Neumann and Amblyomma variegatum Fabricius. Anti-CXCL8 activity was detected in nymphs of all species. Relatively high activity against CCL2, CCL3 and CCL11 was observed in SGEs of R. appendiculatus and A. variegatum nymphs, whereas SGEs of I. ricinus nymphs showed comparatively high anti-interleukin-2 (-IL-2) and anti-IL-4 activities. These data show that nymphs, which epidemiologically are usually more important than adults as disease vectors, possess a range of anti-cytokine activities that may facilitate pathogen transmission.     

Drosophila melanogaster as a model for elucidating the pathogenicity of Francisella tularensis

Drosophila melanogaster is a widely used model organism for research on innate immunity and serves as an experimental model for infectious diseases. The aetiological agent of the zoonotic disease tularaemia, Francisella tularensis, can be transmitted by ticks and mosquitoes and Drosophila might be a useful, genetically amenable model host to elucidate the interactions between the bacterium and its arthropod vectors. We found that the live vaccine strain of F. tularensis was phagocytosed by Drosophila and multiplied in fly haemocytes in vitro and in vivo. Bacteria injected into flies resided both inside haemocytes and extracellularly in the open circulatory system. A continuous activation of the humoral immune response, i.e. production of antimicrobial peptides under control of the imd/Relish signalling pathway, was observed and it may have contributed to the relative resistance to F. tularensis as flies defective in the imd/Relish pathway died rapidly. Importantly, bacterial strains deficient for genes of the F. tularensis intracellular growth locus or the macrophage growth locus were attenuated in D. melanogaster. Our results demonstrate that D. melanogaster is a suitable model for the analysis of interactions between F. tularensis and its arthropod hosts and that it can also be used to identify F. tularensis virulence factors relevant for mammalian hosts.     

The ixodid tick species attaching to domestic dogs and cats in Great Britain and Ireland

The species of ixodid ticks, attached to dogs and cats presented to veterinary practices in Great Britain and Ireland were identified. Most host animals carried only one tick species with Ixodes ricinus Linné (Acari: Ixodidae) being the most common, identified on 52% of animals, Ixodes hexagonus Leach (Acari: Ixodidae) the second most common (on 39%) and Ixodes canisuga Johnston (Acari: Ixodidae) the third most common (on 11%). A significantly higher proportion of dogs than cats carried I. ricinus, while I. hexagonus was more frequently carried by cats. One animal carried a single specimen of Haemaphysalis punctata Canestrini & Fanzago (Acari: Ixodidae), one carried a Dermacentor reticulatus Fabricius (Acari: Ixodidae) but none carried Rhipicephalus sanguineus Latreille (Acari: Ixodidae). This indicates that the latter two species, vectors of 'exotic' tick-borne diseases, remain at low densities in Great Britain and Ireland. Retrospective information on exposure of the animals to different habitats and geographic regions was collected by questionnaire and subject to contingency table and logistic regression analysis. Woodlands and moorlands were habitats significantly associated with I. ricinus attachment. Exposure to urban parks was significantly associated with I. hexagonus attachment and exposure to boarding kennels and catteries was significantly associated with I. canisuga attachment. Ixodes hexagonus, rather than I. ricinus, was the ixodid tick species most likely to be encountered by urban populations of dogs and cats and, by inference, possibly also humans. The implications of these findings, for the transmission of tick-borne pathogens to dogs, cats and humans are discussed.