Marginatus Ticks Research Articles

Tick survey and detection of Crimean-Congo hemorrhagic fever virus in tick species from a non-endemic area, South Marmara region, Turkey

Crimean-Congo hemorrhagic fever (CCHF) is an increasing health concern in Turkey since 2002. There were also some recent human cases from the South Marmara region of Turkey; thus, a tick survey was performed, and possible vector tick species for the CCHF virus were determined in the region. A total of 740 adult ticks were collected from infested livestock from five locations: Çanakkale-Biga, Bursa-Orhaneli, Bursa-Keles, Balıkesir and Bilecik. Total of 11 tick species from the genera Hyalomma, Rhipicephalus, Dermacentor, Ixodes and Haemaphysalis were identified. Rhipicephalus ticks were dominant in the region; the most frequently observed tick species was R. turanicus, (53.1 %), and only 15.4 % of the identified ticks were H. marginatum. The occurrence of H. rufipes infestation in the region fort he first time. A total of 73 pools of adult ticks were tested with both an antigen-detecting ELISA and RT real-time PCR (RT rt PCR). The presence of the CCHF virus was demonstrated in 9 (12.3 %) of the tested tick pools. Although seven of the tick pools were positive for the CCHF virus with both of the methods, one pool was positive only with RT rt PCR and the other pool was only positive with the ELISA. Positive results were obtained from ticks collected from cattle, sheep and goats from two locations, Bursa-Orhaneli and Bilecik. The CCHF virus was detected in R. turanicus (n = 3), R. bursa (n = 2), H. marginatum (n = 2) and D. marginatus (n = 2) ticks. The results of this study confirm the presence of the CCHF virus and present preliminary data on the vector tick species in the southern Marmara region of Turkey.

Guineafowl, ticks and Crimean–Congo hemorrhagic fever in Turkey: the perfect storm?

To control ticks carrying the emerging Crimean-Congo hemorrhagic fever (CCHF) in Turkey the government is introducing thousands of exotic helmeted guineafowl (Numida meleagris). However, not only do these birds eat negligible numbers of ticks, but they are also excellent nurseries for the young of Hyalomma marginatum ticks, the best vectors of CCHF.

Tick-borne rickettsioses in Europe

Rickettsioses are caused by obligate intracellular bacteria within the genus Rickettsia, mainly transmitted by arthropods. Until recently, Mediterranean spotted fever (MSF) caused by Rickettsia conorii was considered the only tick-borne rickettsiosis in Europe. However, ‘new’ TBR have been described in Europe during last years. For instance, other subspecies such as R. conorii caspia and R. conorii israelensis have been involved in MSF. Dermacentor-borne necrosis erythema and lymphadenopathy/tick-borne lymphadenopathy (DEBONEL/TIBOLA) cases caused by Rickettsia slovaca, Rickettsia raoultii, and Rickettsia rioja been described in several countries where Dermacentor marginatus ticks (the mainly implicated vector) are present. Rickettsia helvetica has also been involved as a human pathogen in cases of fever with and without rash and in patients with meningitis and carditis. Other TBR such as lymphangitis-associated rickettsioses (LAR), caused by Rickettsia sibirica mongolitimonae, have been diagnosed in different European countries (France, Spain, Portugal and Greece). Rickettsia massiliae is considered an etiological agent of MSF-like illness in the Mediterranean basin. Furthermore, Rickettsia monacensis that is distributed all along Europe has been isolated from patients with MSF-like illness in Spain. Although Rickettsia aeschlimannii has been associated with MSF-like in Africa and is distributed in the Mediterranean area, no autochthonous human cases have been reported for Europe. Other Rickettsia species detected in ticks and unrelated to human disease (Candidatus Rickettsia kotlanii, Candidatus Rickettsia barbariae, Candidatus Rickettsia vini) could be potentially involved in the next years. Climate changes, among other factors, may contribute to the emergence of other rickettsioses or change their distribution. Lastly, African tick-bite fever (ATBF), caused by Rickettsia africae, is frequently diagnosed in Europe in patients returning from endemic areas.

Spotted fever group rickettsiae identified in Dermacentor marginatus and Ixodes ricinus ticks in Algeria

Our study was carried out using Ixodes ricinus ticks collected from cattle from Tizi-Ouzou and Dermacentor marginatus ticks collected from the vegetation of the Blida region, a tourist site, both regions situated in northern Algeria. The results of real-time quantitative PCR (qPCR) specific for a partial sequence of the citrate synthase gene (gltA) indicate that Rickettsia spp. were present in 11/23 (48%) and 4/9 (44%) of the examined ticks from Tizi-Ouzou and Blida, respectively. The sequences of Rickettsia helvetica and Ri. monacensis were found in I. ricinus ticks using gltA primers. In addition, Ri. slovaca was detected based on the sequences of the gltA and the outer membrane protein (OmpA) genes in D. marginatus ticks. DNA sequencing to identify the species revealed for the first time the presence of Ri. helvetica in I. ricinus ticks and Ri. slovaca in D. marginatus ticks from Algeria and confirmed the presence of Ri. monacensis.

Rickettsia raoultii–like Bacteria inDermacentorspp. Ticks, Tibet, China

<i>Rickettsia raoultii</i>–like Bacteria in<i>Dermacentor</i>spp. Ticks, Tibet, China

CO2 flagging - an improved method for the collection of questing ticks

BackgroundMost epidemiological studies on tick-borne pathogens involve collection of ticks from the environment. An efficient collection method is essential for large sample pools. Our main aim was to evaluate the efficacy of a new method, where traditional flagging was enhanced by the use of CO2 dispersed into the white flannel. The CO2 was spread through a rubber hose network inserted into the flag blanket. The research was conducted in spring, in March-April 2011 in two locations from Cluj County, Romania.MethodsThe research was conducted in March-April 2011 in two locations from Cluj County, Romania. The flag to be tested contained a fine silicone rubber hose network which dispersed the CO2 in the shaft. On each collection site n=30 samplings were performed. Each sampling consisted in the simultaneous use of both flags (with and without CO2) by two persons. The CO2 concentration level on the flag canvas surface was measured. The efficacy of the method was determined by counting comparatively the total number of ticks and separate developmental stage count.ResultsUsing the CO2 improved flag, 2411 (59%) Ixodes ricinus and 100 (53.8%) Dermacentor marginatus ticks were captured, while the CO2-free flag accounted for the collection of 1670 I. ricinus (41%) and 86 (46.2%) D. marginatus ticks. The addition of CO2 prompted a concentration difference on the surface of the flag ranging between 756.5 and 1135.0 ppm with a mean value of 848.9 ppm.ConclusionThe study showed that the CO2 enhanced sweep flag increased the ability of I. ricinus (p < 0001) but not of D. marginatus to be attracted to the flag blanket.

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.

Coinfections of Rickettsia slovaca and Rickettsia helvetica with Borrelia lusitaniae in ticks collected in a Safari Park, Portugal

Borrelia and Rickettsia bacteria are the most important tick-borne agents causing disease in Portugal. Identification and characterization of these circulating agents, mainly in recreational areas, is crucial for the development of preventive measures in response to the gradually increasing exposure of humans to tick vectors. A total of 677 questing ticks including Dermacentor marginatus, Rhipicephalus sanguineus, Ixodes ricinus, Hyalomma lusitanicum, H. marginatum, and Haemaphysalis punctata were collected in a Safari Park in Alentejo, Portugal, to investigate the prevalences of infection and characterize Borrelia and Rickettsia species. From a total of 371 ticks tested by PCR for Borrelia burgdorferi sensu lato (s.l.), of which 247 were tested for Rickettsia, an infection prevalence of 18.3% was found for B. lusitaniae and 55.1% for Rickettsia spp. Sequence analysis of positive amplicons identified the presence of B. lusitaniae (18.3%), R. monacensis strain IRS3 (51.7%), and R. helvetica (48.3%) in I. ricinus. R. slovaca (41.5%), R. raoultii (58.5%), and also B. lusitaniae (21%) were identified in D. marginatus ticks. One (5.9%) H. lusitanicum was infected with B. lusitaniae, and R. massiliae was found in one Rhipicephalus sanguineus. Coinfection was found in 7 (20%) I. ricinus and 34 (23.3%) D. marginatus ticks. We report, for the first time, simultaneous infection with R. helvetica and B. lusitaniae and also R. slovaca, the agent of TIBOLA/DEBONEL, with B. lusitaniae. Additionally, 6 isolates of B. lusitaniae were established, and isolates of Rickettsia were also obtained for the detected species using tick macerates cultured in mammalian and mosquito cell lines. This report describes the detection and isolation of tick-borne agents from a Portuguese Safari Park, highlighting the increased likelihood of infection with multiple agents to potential visitors or staff.

Static and Dynamic Systems in Rickettsia slovaca Life Cycle Evaluated by Quantitative Real-Time Polymerase Chain Reaction

Quantitative real-time polymerase chain reaction was used to characterize the growth of Rickettsia slovaca, a tick-borne pathogen transmitted by Dermacentor reticulatus and D. marginatus ticks, in static (L929 and Vero cells) and dynamic (D. marginatus and Ixodes ricinus ticks) cultivation systems. The highest points of bacterial multiplication and the time-spans between the inoculum and the maximum of rickettsial copies were increased in consecutive order from eukaryotic cells, I. ricinus to D. marginatus systems. In dynamic system, multiplication maximum of R. slovaca was achieved 9 days earlier in I. ricinus; however, the number of rickettsial DNA copies was approximately 3.6 x 10(6) more in D. marginatus.

Detection of Rickettsia aeschlimannii in Hyalomma marginatum ticks in western Russia

Gamaleya Research Institute of Epidemiology andMicrobiology, Moscow, RussiaINTRODUCTIONSFG rickettsiae are obligate intracellular Gram-negative bacteria. These bacteria are associatedwith arthropods, mainly ticks, and can be trans-mitted to mammals and humans. Ixodid ticks ofthe Hyalomma marginatum species are widelyspread in semi-desert and steppe landscape areasof south-western Russia, in particular in theStavropol region where they have been reportedto bite humans [1]. The main hosts of the larvaeand nymphs of this tick species include severalspecies of wild and domestic birds, the Europeanbrown hare, and four-toed and eared hedgehogs.In contrast, adult ticks parasitise livestock anddomestic dogs, and may attack humans. InRussia, H. marginatum ticks are known to be themain vectors of the Crimean-Congo haemorrhagicfever (CCHF) virus in the southern part of thecountry. In addition, strains of West Nile fevervirus were isolated from H. marginatum tickscollected in the Volga delta in Astrakhan.However, to date, the presence of rickettsiae inH. marginatum has not been evaluated. In thepresent study, we investigated by PCR thepresence of rickettsiae in H. marginatum tickscollected in the Stavropol region of Russia.MATERIALS AND METHODS

Prospective evaluation of rickettsioses in the Trakya (European) Region of Turkey in 2005

Several rickettsial diseases have been reported in the Mediterranean basin, including Mediterranean spotted fever, caused by Rickettsia conorii and transmitted by Rhipicephalus sanguineus ticks, tick-borne lymphadenopathy, caused by Rickettsia slovaca and transmitted by Dermacentor marginatus ticks, Rickettsia mongolitimonae infection, transmitted by Hyalomma spp. ticks, Rickettsia helvetica infection, transmitted by Ixodes ricinus ticks, and flea-borne spotted fever, caused by Rickettsia felis and transmitted by cat fleas (Ctenocephalides felis) [1]. Since 2003, prospective surveillance and serological and molecular diagnosis of rickettsioses in the Trakya Region of Turkey has been conducted by the Trakya University in Edirne and the Universite de la Mediterranee in Marseille [1,2]. In 2003 and 2004, 22 patients were diagnosed as spotted fever group (SFG) rickettsioses, using molecular diagnostic methods [1,2]. R. conorii was previously isolated from the skin biopsy specimens of three of ten patients, and was also demonstrated by PCR in skin biopsy specimens of nine of ten patients [1]. From May to September 2005, 24 patients with SFG rickettsioses were admitted into the Trakya University Hospital, a tertiary-care hospital in Edirne. In this prospective study, our aim was to isolate and identify the rickettsial strains from skin biopsy specimens and blood samples of these 24 patients diagnosed with spotted fever in the Trakya Region of Turkey in 2005. M A T E R I A L S A N D M E T H O D S

&lt;I&gt;Rickettsia slovaca&lt;/I&gt; and &lt;I&gt;Rickettsia raoultii&lt;/I&gt; in &lt;I&gt;Dermacentor marginatus&lt;/I&gt; Ticks Collected on Wild Boars in Tuscany, Italy

During the hunting season 2007-2008, 494 Dermacentor marginatus (Sulzer) ticks were collected from 109 hunter-killed wild boars, Sus scrofa, in Lucca's province, Tuscany, Italy. Rickettsia slovaca, the causative agent of tick-borne lymphadenopathy (TIBOLA), was detected in 32.1% of ticks tested (n=112) by using polymerase chain reaction primers targeting gltA, ompA, and ompB rickettsial genes. Moreover, Rickettsia raoultii was found for the first time in Italy, with 1.8% infection prevalence. This study confirms the risk posed to humans by ticks and tick-borne pathogens in the study area, where cases of spotted fever rickettsiosis (TIBOLA) are reported.

Dermacentor marginatus and Ixodes ricinus ticks versus L929 and Vero cell lines in Rickettsia slovaca life cycle evaluated by quantitative real time PCR

Ticks transmit many different pathogens to animals, humans and their pets. Rickettsia slovaca, as a member of the spotted-fever-group rickettsiae is an agent of the human disease Tick-borne lymphadenopathy (TIBOLA), also called Dermacentor-borne necrosis erythema and lymphadenopathy (DEBONEL), which occurs from the Mediterranean to central Europe, transmitted by Dermacentor reticulatus and Dermacentor marginatus (Acari: Ixodidae). In this study, quantitative real time PCR was used to characterize the growth of R. slovaca, strain B in static (mammalian L929 and Vero cells without replacement of growth medium) and dynamic (D. marginatus and Ixodes ricinus ticks) cultivation systems. Curves of bacterial growth in static cultivations were modeled with exponential, stationary and death phases, whereas in dynamic systems the stationary phase was absent. The highest point of multiplication of R. slovaca was recorded on the 4th day post infection in both cell lines and the rickettsial DNA copy number in L929 and Vero cells at this point was 21 and 27 times greater than rickettsial DNA copy number of inoculum, respectively. In the dynamic system, the highest point of multiplication was on the 21th and 12th day after feeding of ticks and rickettsial DNA copy numbers were 7,482 and 865 times greater than the inoculum in D. marginatus and I. ricinus, respectively. Life cycle of R. slovaca in mammalian cell lines was shorter; supposedly, bacteria destroyed these cells and ticks, especially D. marginatus, were considered a more appropriate environment.

Rickettsia slovacaInfection in Humans in the Northeast of Spain: Seroprevalence Study

Catalonia is an endemic area of Mediterranean spotted fever. In 1997, A. Lakos described a new tick-borne infectious disease called tick-borne lymphadenopathy. The causative agent is Rickettsia slovaca, which is transmitted by Dermacentor marginatus ticks. We have diagnosed human cases in Catalonia. The objective of this study was to determinate seroprevalence of R. slovaca infection in humans in the northeast of Spain. The population included 217 subjects from Catalonia, northeast of Spain and was stratified by age and living place (rural, suburban, and urban). Age, gender, residence area, contact with animals, occupation, and history of rickettsioses was surveyed. Immunoglobulin G was measured by indirect immunofluorescence assay. Titers >or= 1/40 were considered. Seroprevalence of R. slovaca was 5.5% at titers of 1/40-1/320. Eight (3.7%) sera had antibodies against R. slovaca exclusively. Four sera reacted also against Rickettsia conorii and/or Bar29. Seroprevalence of R. slovaca would range from 3.7% to 5.5%. The only statistically significant association was that between R. slovaca seropositivity and age. We present serologic evidence of R. slovaca infection among population of Catalonia, northeast of Spain.

Arthropods (Pseudoscorpionidea, Acarina, Coleoptera, Siphonaptera) in nests of the bearded tit (Panurus biarmicus)

AbstractIn the period 1993–2006, during investigation of reproduction biology of the bearded tit, 106 deserted nests of the species were collected in Slovakia, Austria and Italy and their arthropod fauna was analyzed. Occasionally introduced individuals of the pseudoscorpion Lamprochernes nodosus, a frequent species in Central Europe, were recorded in the nests. Altogether 984 individuals and 33 species of mesostigmatic mites (Acari) were found in 46.2% of the nests examined. The ectoparasite Ornithonyssus sylviarum was most abundant and frequent; it represented almost 68.3% of all individuals. Due to it, the parasitic mites predominated (69.4% of individuals). Other ecological groups were less represented: edaphic detriticols − 11.6%, coprophils − 10.7%, species of vegetation stratum − 8.2%, and nidicols − 0.2%. Beetles (40 species, 246 individuals) were present in 57 nests. Most of the beetles were strongly hygrophilous species inhabiting soil surface in the reed stands or other types of wetlands and the shore vegetation. Predators represented 59% of all individuals. They might find food in the nests, but none of the species had a close relationship to bird nests and represented 35% of species. All beetle species penetrated the nests occasionally, when ascending on the vegetation or searching cover during periods of increased water level. Occasionally, larvae and nymphs of the Dermacentor marginatus tick were found. They were most probably introduced by insectivores of the genus Neomys. Only one species of fleas, Ceratophyllus garei — a parasite of birds nesting in humid environment, was recorded in the nests.

The Role of Wild Boar (Sus scrofa) in The Eco-epidemiology of R. slovaca in Northeastern Spain

Rickettsia slovaca is considered the etiological agent of tick-borne lymphadenopathy (TIBOLA), an emerging disease transmitted by tick bites. Dermacentor marginatus constitutes the most important vector and wild boar (Sus scrofa) is the main wild host in our area. The epidemiology of this tick-borne rickettsioses has not been completely clarified. During hunting season 2004, wild boar sera sample were collected from northeastern Spain. Ticks were collected both from wild boar and from flagging vegetation in the same areas where wild boar were hunted. Serologic study was carried out using the immunofluorescent antibody (IFA) technique. Ticks were identified as D. marginatus and Rhipicephalus uranicus. Based on sequence analysis on ompA gene, R. slovaca was identified in 30.5% D. marginatus ticks removed from wild boar and in 33.3% D. marginatus collected from flagging vegetation. Rickettsia sp. RpA4 was identified in 4 specimens of D. marginatus removed from wild boar. Twelve of 23 wild boar were seropositive to R. slovaca. Results suggested wild boar are exposed to R. slovaca infection and this pathogen is well established in the wild cycle of D. marginatus in our area.

Microevolution of tick-borne encephalitis virus in course of host alternation

Two tick-borne encephalitis (TBE) virus variants were studied: mouse brain-adapted strain EK-328 and its derivate adapted to Hyalomma marginatum ticks. The tick-adapted virus exhibited small-plaque phenotype and slower replication in PEK cells, higher yield in ticks, decreased neuroinvasiveness in mice, increased binding to heparin-sepharose. A total of 15 nucleotide substitutions distinguished genomes of these variants, six substitutions resulted in protein sequence alterations, and two were in 5′NTR. Two amino acid substitutions in E protein were responsible for the observed phenotypic differences. Data obtained during reverse passaging of the tick-adapted virus in vivo and in vitro suggest that TBE virus exists as a heterogeneous population that contains virus variants most adapted to reproduction in either ticks or mammals. Host switch results in a change in the ratio of these variants in the population. Plaque purification of the tick-adapted virus resulted in the prompt emergence of new mutants with different virulence for mammals.

Prevalence of Rickettsia slovaca in Dermacentor marginatus Ticks Removed from Wild Boar (Sus scrofa) in Northeastern Spain

Rickettsia slovaca, the causative agent of TIBOLA, is transmitted by Dermacentor ticks. Dermacentor marginatus is the most widely species distributed in northeastern Spain, and the wild boar constitutes the main host. D. marginatus ticks were collected from hunter-killed wild boar and were tested by PCR/RFLP. Rickettsial DNA-positive ticks were sequenced using the ompA PCR primers. The prevalence of R. slovaca in D. marginatus ticks was 17.7%. Other spotted fever group rickettsiae were detected in ticks, but these were not definitely identified.

Prevalence of Spotted Fever Group Rickettsia Species Detected in Ticks in La Rioja, Spain

Our objective was to learn the prevalence of spotted fever group (SFG) Rickettsia detected in ticks in La Rioja, in the north of Spain. From 2001 to 2005, 496 ticks representing 7 tick species were analysed at the Hospital de La Rioja. Ticks were removed from humans with or without rickettsial syndrome (n = 59) or collected from mammals (n = 371) or from vegetation by dragging (n = 66). The presence of SFG Rickettsia in these ticks was investigated by semi-nested PCR (ompA gene) and sequencing. A phylogenetic tree using Clustal method (neighbor-joining) was constructed with these data. Only 3 of 170 Hyalomma marginatum ticks carried SFG Rickettsia. Sequencing analysis demonstrated the presence of Rickettsia aeschlimannii (1.8%). Furthermore, Rickettsia massiliae and BAR29 were found in 3 of 120 Rhipicephalus sanguineus specimens (2.5%). In contrast, 81 of 83 tested Dermacentor marginatus ticks were PCR-positive (97%). Rickettsia slovaca (40.6%) and Rickettsia sp. strains RpA4, DnS14, DnS28 and JL-02 (59.3%) were found within this tick species. No SFG Rickettsia was detected using ompA primers when Ixodes ricinus, Rhipicephalus bursa, Rhipicephalus turanicus, Rhipicephalus eversti eversti, Hyalomma detritum scupense and Rhipicephalus sp. were analyzed. We detected 17.5% of ticks associated with different SFG Rickettsia: R. aeschlimannii, R. massiliae, BAR29, R. slovaca and Rickettsia sp. strains RpA4, DnS14, DnS28 and JL-02. Their presence has to be taken into account since most of them have been recognized as human pathogens.

Rickettsia slovacaInfection, France

<i>Rickettsia slovaca</i>Infection, France