Marginatus Ticks Research Articles

Insights into the virome of Hyalomma marginatum in the Danube Delta: a major vector of Crimean-Congo hemorrhagic fever virus in Eastern Europe

BackgroundTicks are significant vectors of pathogens, including viruses, bacteria, and protozoa. With approximately 900 tick species worldwide, many are expanding their geographical range due to changing socioeconomic and climate factors. The Danube Delta, one of Europe’s largest wetlands, is an ecosystem that, despite its ecological importance, remains understudied concerning the risk of introducing new tick-borne viruses. This region serves as a critical habitat for migratory birds, which can carry ticks over long distances, potentially introducing exotic tick species and their pathogens into the local ecosystem. Hyalomma marginatum ticks, the primary vector of Crimean-Congo hemorrhagic fever virus (CCHFV), are of particular concern due to their expanding presence in Europe and potential to spread other arboviruses. In addition to being the primary vector for CCHFV, Hyalomma sp. ticks are capable of transmitting other pathogens of medical and veterinary importance, including Dugbe virus, West Nile virus, African horse sickness virus, and Kyasanur forest disease virus. Therefore, it is essential to monitor the presence of Hyalomma sp. ticks while simultaneously surveilling arbovirus circulation in tick populations to mitigate the risk of arboviral outbreaks.MethodsIn this work, we used an RNA sequencing technique to analyze the virome of H. marginatum ticks collected from the Danube Delta Biosphere Reserve, Romania, one of the major bird migration hubs from Africa to Europe.ResultsAmong the viral taxa detected in H. marginatum ticks, sequences belonging to Volzhskoe tick virus (VTV), Balambala tick virus (BMTV) and Bole tick virus 4 (BTV4) were identified. In addition, we report the first identification of a novel Rhabdoviridae-related virus, Hyalomma marginatum rhabdovirus (HMRV). No CCHFV or any CCHFV-related nairovirus were detected in this study.ConclusionsTo summarize, detecting new viruses is essential for monitoring potential viral outbreaks. Our research expands the understanding of virus diversity in Eastern Europe, including the identification of novel viruses. This insight is crucial for monitoring viruses that may pose risks to both animal and human health, such as CCHFV.Graphical

Genomic characterization of Volzhskoe tick virus (Bunyaviricetes) from a Hyalomma marginatum tick, Hungary

Hyalomma marginatum, a vector for the high-consequence pathogen, the Crimean–Congo hemorrhagic fever virus (CCHFV), needs particular attention due to its impact on public health. Although it is a known vector for CCHFV, its general virome is largely unexplored. Here, we report findings from a citizen science monitoring program aimed to understand the prevalence and diversity of tick-borne pathogens, particularly focusing on Hyalomma ticks in Hungary. In 2021, we identified one adult specimen of Hyalomma marginatum and subjected it to Illumina-based viral metagenomic sequencing. Our analysis revealed sequences of the uncharacterized Volzhskoe tick virus, an unclassified member of the class Bunyaviricetes. The in silico analysis uncovered key genetic regions, including the glycoprotein and the RNA-dependent RNA polymerase (RdRp) coding regions. Phylogenetic analysis indicated a close relationship between our Volzhskoe tick virus sequences and other unclassified Bunyaviricetes species. These related species of unclassified Bunyaviricetes were detected in vastly different geolocations. These findings highlight the remarkable diversity of tick specific viruses and emphasize the need for further research to understand the transmissibility, seroreactivity or the potential pathogenicity of Volzhskoe tick virus and related species.

Detection of Crimean-Congo haemorrhagic fever virus in Hyalomma marginatum ticks, southern France, May 2022 and April 2023.

Crimean-Congo haemorrhagic fever (CCHF), a potentially severe zoonotic viral disease causing fever and haemorrhagic manifestations in humans. As the Crimean-Congo haemorrhagic fever virus (CCHFV) has been detected in ticks in Spain and antibodies against the virus in ruminant sera in Corsica, it was necessary to know more about the situation in France. In 2022-2023, CCHFV was detected in 155 ticks collected from horses and cattle in southern France.

Grazing system and Hyalomma marginatum tick infestation in cattle with high prevalence of SFG Rickettsia spp.

Tick-borne diseases (TBDs) represent a significant portion of infectious diseases of global public health interest. In Italy, knowledge about the occurrence of tick-borne pathogens (TBPs) in ticks parasitizing cattle is scarce. In this research, we focused on ticks infesting Maremmana cattle grazing in open pasture and silvopasture systems. After being morphologically identified, ticks were molecularly tested for the presence of pathogens of the genus Rickettsia. Of the 794 ticks detected, 117 were collected, being the majority Hyalomma marginatum (72.6%), followed by other Hyalomma species (23%), Rhipicephalus turanicus (1.7%), Rh. bursa (0.9%), Hy. lusitanicum (0.9%) and Dermatocentor marginatus (0.9%). All ticks were adults, 58.1% males and 41.8% females. The highest tick prevalence was noted in April for silvopasture system cattle (90%), and in May for open pasture ones (85%). TBPs were detected only in Hy. marginatum, and all belong to Rickettsia spp. of zoonotic interest. In particular, 21/40 (52.5%) ticks scored positive for Rickettsia spp. by gltA gene and of these 15/21 (71.4%) also to spotted fever group (SFG) rickettsiae by ompA gene. Of the total positive specimens, 19 were successfully sequenced and scored Rickettsia aeschilimannii (17/19, 89.5%), R. slovaca (1/19, 5%), and R. massiliae (1/19, 5%). This research highlights the potential impact of grazing systems on cattle parasitization by hard ticks. The molecular investigation of TBPs in ticks collected from Maremmana cattle shed light on the presence of pathogenic bacteria of SFG Rickettsia spp., pointing out the potential risk of TBPs transmission between livestock and humans.

Germany-wide citizen science study reveals spread of Babesia canis-infected Dermacentor reticulatus ticks by dogs travelling within the country

The ornate dog tick Dermacentor reticulatus, vector of Babesia canis, has shown a considerable range expansion in several European countries. Previously, only few areas in Germany were recognised as endemic for B. canis, but a marked increase in autochthonous canine babesiosis cases and spread to new areas has been noted recently. To better assess the current risk for dogs, the present study screened 5913 specimens of D. reticulatus from all over Germany, collected in the frame of a Citizen Science study during 2019–2023. Moreover, 343 Dermacentor marginatus ticks were also included. Babesia detection was achieved by quantitative real-time PCR (qPCR). Positive samples were confirmed by sequencing. Moreover, a MGB-probe-based triplex qPCR was established to detect and distinguish between the canine Babesia spp. relevant in Europe, i.e. B. canis, Babesia vogeli and Babesia gibsoni. Overall, B. canis DNA was detected in five D. reticulatus specimens (0.08%). Two of the B. canis-positive ticks originated from areas previously known as endemic for canine babesiosis, namely from the area of Freiburg im Breisgau, federal state of Baden-Wuerttemberg, and from the district St. Wendel, federal state of Saarland. Three further B. canis-positive ticks were detected in districts not yet recognised as endemic, one each in the district of Mansfeld-Suedharz, federal state of Saxony-Anhalt, the district of Ravensburg, federal state of Baden-Wuerttemberg and in the city of Fürth, federal state of Bavaria. However, the tick in Fürth was found on a dog who had returned from a trip to the Breisgau region on the previous day, indicating translocation of the specimen out of this well-known endemic focus. The geographical distribution of the positive samples shows that B. canis is currently spreading in Germany, particularly via dogs travelling within the country, increasing the infection risk throughout the country. Important measures to contain a further spread of the pathogen include comprehensive year-round tick prophylaxis with licensed acaricides, not only to protect the individual pet, but also the entire dog population. Moreover, screening of dogs entering Germany from B. canis-endemic countries is required and any treatment should aim at pathogen elimination by use of appropriate imidocarb dosages.

1765. Distribution of Rickettsia species among hospitalized cases and ticks in Pavlodar region, Kazakhstan, 2019

Abstract Background Spotted Fever Group Rickettsioses (SFGR) is a group of neglected, life-threatening zoonotic diseases caused by closely related bacteria spread primarily by ticks. Treatment is available if identified, but testing for Rickettsiae is limited. Little is known about SFGR prevalence and distribution of Rickettsiae species in Kazakhstan. This knowledge can help inform SFGR prevention and treatment efforts. Methods We conducted sentinel surveillance from April to October 2019 in six hospitals in the Pavlodar region, an area where several SFGR cases had previously been detected. We interviewed and collected samples from all consenting patients ≥ 6 years old with SFRG symptoms. Parents consented for patients < 18 years old. Samples included skin lesions and two serum samples at the acute stage (AS) and convalescent stage (CS). We classified participants as having acute SFGR if they had either: (1) positive PCR, or (2) a four-fold increase in IFA IgG titers from AS to CS samples, or (3) IgG titers (≥ 1:64) in an AS sample. We also collected ticks using dragging or flagging of vegetation and tested them using real-time PCR. Results Of 105 people enrolled, 69% had acute SFGR (fig. 1). Incidence rates per 100,000 population at sentinel sites ranged from 7 to 50. Common signs were fever (100%), headache (89%), rash (75%), and myalgia (58%). R. sibirica (40%), R. raoultii (39%), and R. slovaca (7%) were identified in participants (tab. 1). Participants with R. sibirica were more likely than participants with R. raoultii to have eschar (80% vs. 7%, p < 0.01, respectively) and rash (90% vs. 61%, p=0.01, respectively). Doxycycline was administered to 64% of participants, and nobody died. R. raoultii and R. spp were identified in 9% of Dermacentor reticulatus ticks (n=610) and 7% of Dermacentor marginatus ticks (n=349). R. sibirica was identified in 1% of D. marginatus ticks. S. slovaca was not detected in ticks (fig. 2).Figure 1.Patient enrollment, specimen collection and testing results, Pavlodar region, 2019. PCR: polymerase chain reaction; RSB: rash skin biopsy or /and eschar or/and swab from eschar samples; IFA: immunofluorescence assay; AS: acute serum collected at acute phase; CS: convalescent serum collected at convalescent phase; ND: not defined. *11 – PCR negative tested with IFA. Of which, 5 had both AS and CS (results: 2-positive, 2 -negative, 1-recent infection) and 6 had AS only (results: 2-probable, 4 not defined).Table 1.Rickettsia species identified from hospitalized cases (n=72)Figure 2.Rickettsia species in ticks by selected sites (Aksu and Pavlodar districts), Pavlodar region, 2019 Conclusion There was a two-fold increase in cases detected through active surveillance during our study than reported in previous years (105 in 2019 compared to 54 in 2018 and 40 in 2017). R. sibirica and R. raoultii are the dominant species associated with SFGR in Pavlodar and are also found in local ticks. Improved disease detection and regular testing of humans and ticks in areas with SFGR cases is needed. Disclosures All Authors: No reported disclosures

In vitro feeding of Hyalomma excavatum and Hyalomma marginatum tick species.

The rearing of ticks is an important technique for studies aiming to elucidate the course and pathogenesis of tick-borne diseases (TBDs). TBDs caused by protozoans (Theileria, Babesia) and bacteria (Anaplasma/Ehrlichia) impose a serious constraint upon livestock health and production in tropical and sub-tropical regions where the distributions of host, pathogen, and vector overlap. This study focuses on Hyalomma marginatum, one of the most important Hyalomma species in the Mediterranean region, being a vector of the virus that causes Crimean-Congo haemorrhagic fever in humans, together with H. excavatum, a vector of Theileria annulata, an important protozoan of cattle. The adaptation of ticks to feeding on artificial membranes allows the creation of model systems that can be put to use examining the underlying mechanisms of pathogen transmission by ticks. Silicone membranes, in particular, offer researchers the flexibility to adjust membrane thickness and content during artificial feeding. The aim of the present study was to develop an artificial feeding technique using silicone-based membranes for all developmental stages of H. excavatum and H. marginatum ticks. Attachment rates after feeding on silicone membranes for females H. marginatum and H. excavatum were 8.33% (8/96) and 7.95% (7/88), respectively. The use of cow hair as a stimulant increased the attachment rate of H. marginatum adults in comparison to other stimulants. The engorgement of H. marginatum and H. excavatum females took 20.5 and 23days with average weights of 307.85 and 260.64mg, respectively. Although both tick species could complete egg-laying, and this was followed by hatching of larvae; their larvae and nymphs could not be fed artificially. Taken together, the results of the present study clearly indicate that silicone membranes are suitable for feeding of H. excavatum and H. marginatum adult ticks, supporting engorgement, laying of eggs, and hatching of the larvae. They thus represent a versatile tool for studying transmission mechanisms of tick-borne pathogens. Further studies are warranted to examine attachment and feeding behaviours in order to increase the success of artificial feeding of larvae and nymphal stages.

ИССЛЕДОВАНИЯ КЛЕЩЕЙ РОДА DERMACENTOR (ACARI; IXODIDAE) НА ЕСТЕСТВЕННУЮ ВСТРЕЧАЕМОСТЬ ВОЗБУДИТЕЛЯ ЛИХОРАДКИ КУ В УСЛОВИЯХ ЦЕНТРАЛЬНОГО ПРЕДКАВКАЗЬЯ

The purpose of the research: to assess the natural occurrence of Coxiella burnetii in ticks of the genus Dermacentor; clarify of the relationship between the physiological age of ticks and their infection with the Q fever patogen. Materials and methods: In the period from 2017 to 2019, we examined 4908 specimens of Dermacentor marginatus (616 pools) 4208 specimens of Dermacentor reticulatus (566 pools) and 43 specimens of Dermacentor niveus (18 pools) were examined for infection with the Q fever patogen. To assess the dependence of the detection of Coxiella burnetii in ticks depending on the physiological age, we examined 3048 specimens D. reticulatus (381 pools) and 2319 specimens D. marginatus (366 pools). The study of ixodid ticks for the presence of Coxiella burnetii DNA of the causative agent of Q fever was carried out by the molecular genetic method, namely the polymerase chain reaction (PCR), in accordance with the regulatory guidelines. Results and discussion: The results of the research showed that the infection of ixodid ticks with Coxiella in the Central Ciscaucasian natural populations on D. reticulatus and D. marginatus fluctuates over the years and does not exceed one percent. When considering the dependence of the detection of Coxiella burnetii in ticks on their physiological age, statistically significant results were obtained only for D. reticulatus. There is a significant correlation between the more frequent detection of infected ticks of physiological age II compared with ticks of physiological age III and IV. For D. marginatus ticks, there was no statistically significant relationship between the detection of Q fever pathogen and physiological age. Key words: ixodid ticks, Dermacentor reticulatus, Dermacentor marginatus, Dermacentor niveus, Q fever, physiological age.

Epidemiological situation on Crimean-Congo Hemorrhagic Fever in the Russian Federation in 2021

The review presents an analysis of the epidemiological and epizootiological situation on Crimean-Congo hemorrhagic fever (CCHF) in the Russian Federation in 2021. 49 cases of CCHF were detected in 2021, which is 1.53 times higher than in 2020. The mortality rate was 6.1 %. Sporadic cases of CCHF were registered in the Stavropol Territory, Rostov, Volgograd Regions, the Republics of Dagestan and Kalmykia. The incidence rates of CCHF were below the long-term average annual values in the majority of the constituent entities. Epizootiological survey of stationary observation points has revealed that the number of Hyalomma marginatum imago corresponded to the average long-term indicators in 2021, the peak of H. marginatum activity was noted in the II–III decades of May. The proportion of Ixodidae tick pools positive for Crimean-Congo hemorrhagic fever (CCHF) virus markers exceeded the long-term average indexes in a number of regions. On the territory of the natural focus of CCHF, the circulation of the CCHF virus of the genetic lineages “Europe-1” and “Europe-3” was detected in 2021. Based on the analysis of the epidemiological data of the previous year and natural and climatic factors affecting the abundance and vital activity of H. marginatum ticks, risk-based quantitative forecast for the incidence of CCHF in the Stavropol Territory for 2022 has been compiled.

First detection of Rickettsia aeschlimannii in Hyalomma marginatum in Tibet, China

Objective: Hyalomma marginatum is an important arthropod vector in the transmission of various zoonoses. The aim of this study was to identify the tick-borne pathogens (TBPs) maintained in Hy. marginatum in Tibet and to estimate the risk of human tick-borne diseases. Methods: Adult Hy. marginatum ticks (n = 14) feeding on yaks were collected. The individual DNA samples of these ticks were sequenced with metagenomic next-generation sequencing to survey the presence of TBPs. TBPs in individual ticks were identified with nested polymerase chain reaction (PCR) combined with DNA sequencing. Results: The presence of Rickettsia, Anaplasma, and Ehrlichia in individual ticks was indicated by the taxonomic profiles at the genus level, but only Rickettsia aeschlimannii (100%, 13/13) was further detected in the ticks by nested PCR. Conclusion: This study provides information on the microbial communities of Hy. marginatum in Tibet, China, and provides the first report of R. aeschlimannii found in Hy. marginatum in Tibet. The results of this study indicated that yaks in Tibet are exposed to R. aeschlimannii.

Rearing of Hyalomma marginatum (Acarina: Ixodidae) under laboratory conditions in Morocco

Hyalomma marginatum Koch is one of the main tick vectors of human and animal tick-borne diseases. The objective of this study was to establish standard procedures for rearing H. marginatum under laboratory conditions. Such laboratory tick populations are required to study acaricide resistance of Hyalomma ticks. In our rearing program, larvae and nymphs were fed on New Zealand white rabbits, whereas adults were fed on sheep. Non-parasitic stages were held at 18 and 28 °C to study the effect of temperature on development and survival. In our experiments, H. marginatum ticks have maintained the characteristics of a two-host life cycle. The engorged larvae did not detach and moulted on the rabbit, after which the emerged nymphs continued to feed on the same animal. The life cycle duration of H. marginatum was influenced by temperature, with each non-parasitic stage—i.e., larva and nymph molting—developing faster at 28 than at 18 °C; preoviposition and oviposition periods were shorter at 28 than at 18 °C. At 18 °C, no eggs hatched. The whole cycle from the collection of an engorged field tick until the emergence of second-generation larvae took 189 days. One such tick on average results in 3500 eggs which over time, taking into account the losses at each developmental stage, develop into 1200 adult ticks. Rearing these ticks a second generation therefore could result in millions of larval ticks.

Efficacy of a single administration of afoxolaner (NexGard®) or fipronil plus permethrin (Frontline® Tri-Act) against Hyalomma marginatum ticks in dogs.

This study was conducted to assess the acaricidal efficacy of afoxolaner (NexGard®, Boehringer Ingelheim), and fipronil - permethrin (Frontline® Tri-Act, Boehringer Ingelheim) administered once to dogs experimentally infested with Hyalomma marginatum ticks. Twenty-four Beagle dogs were randomly allocated based on a pre-treatment H. marginatum infestation to an untreated control group, a NexGard® or a Frontline® Tri-Act treated groups. Treatments were administered once on Day 0 as per the products' labels. For the efficacy evaluation, dogs were experimentally infested with 30 adult H. marginatum ticks on Days -2, 7, 28 and 36. In-situ counts were performed at 48h post-treatment on Day 2 and post-infestations on Days 9, 30 and 38. Ticks were removed and counted at 72h post-treatment on Day 3 and after each tick infestation on Days 10, 31 and 39. The numbers of live ticks counted in the treated groups were significantly different than in the control group at all time-points (p≤0.0006). The efficacy was at least 97% after 48h, and at least 99% after 72h for both treatments. In this study both afoxolaner and fipronil/permethrin formulations demonstrated a high efficacy against adult H. marginatum ticks in treated dogs for at least five weeks.

High-Throughput Microfluidic Real-Time PCR for the Detection of Multiple Microorganisms in Ixodid Cattle Ticks in Northeast Algeria.

Ixodid ticks are hematophagous arthropods considered to be prominent ectoparasite vectors that have a negative impact on cattle, either through direct injury or via the transmission of several pathogens. In this study, we investigated the molecular infection rates of numerous tick-borne pathogens in ticks sampled on cattle from the Kabylia region, northeastern Algeria, using a high-throughput microfluidic real-time PCR system. A total of 235 ticks belonging to seven species of the genera Rhipicephalus, Hyalomma, and Ixodes were sampled on cattle and then screened for the presence of 36 different species of bacteria and protozoans. The most prevalent tick-borne microorganisms were Rickettsia spp. at 79.1%, followed by Francisella-like endosymbionts (62.9%), Theileria spp. (17.8%), Anaplasma spp. (14.4%), Bartonella spp. (6.8%), Borrelia spp. (6.8%), and Babesia spp. (2.5%). Among the 80.4% of ticks bearing microorganisms, 20%, 36.6%, 21.7%, and 2.1% were positive for one, two, three, and four different microorganisms, respectively. Rickettsia aeschlimannii was detected in Hyalomma marginatum, Hyalomma detritum, and Rhipicephalus bursa ticks. Rickettsia massiliae was found in Rhipicephalus sanguineus, and Rickettsia monacensis and Rickettsia helvetica were detected in Ixodes ricinus. Anaplasma marginale was found in all identified tick genera, but Anaplasma centrale was detected exclusively in Rhipicephalus spp. ticks. The DNA of Borrelia spp. and Bartonella spp. was identified in several tick species. Theileria orientalis was found in R. bursa, R. sanguineus, H. detritum, H. marginatum, and I. ricinus and Babesia bigemina was found in Rhipicephalus annulatus and R. sanguineus. Our study highlights the importance of tick-borne pathogens in cattle in Algeria.

Scalp eschar and neck lymphadenopathy by Rickettsia slovaca after Dermacentor marginatus tick bite case report: multidisciplinary approach to a tick-borne disease

BackgroundScalp Eschar and Neck LymphAdenopathy after Tick bite is a zoonotic non-pathogen-specific disease most commonly due to Rickettsia slovaca and Rickettsia raoultii. Diagnosis is mostly based only on epidemiological and clinical findings, without serological or molecular corroboration.We presented a clinical case in which diagnosis was supported by entomological identification and by R. slovaca DNA amplifications from the tick vector.Case presentationA 6-year-old child presented with asthenia, scalp eschar and supraclavicular and lateral-cervical lymphadenopathy. Scalp Eschar and Neck LymphAdenopathy After Tick bite syndrome following a Dermacentor marginatus bite was diagnosed. Serological test on serum revealed an IgG titer of 1:1024 against spotted fever group rickettsiae, polymerase chain reaction assays on tick identified Rickettsia slovaca. Patient was successfully treated with doxycycline for 10 days.ConclusionsA multidisciplinary approach including epidemiological information, clinical evaluations, entomological identification and molecular investigations on tick, enabled proper diagnosis and therapy.

First molecular survey of Anaplasma phagocytophilum in hard ticks (Ixodidae) from Southwestern Iran.

Anaplasma phagocytophilum is a gram-negative obligate intracellular tick-borne rickettsia with veterinary and public health importance worldwide. This organism is an etiologic agent of tick-borne fever (TBF) in domesticated animals and human granulocytic anaplasmosis (HGA) as well. Hard ticks (Ixodida: Ixodidae) are incriminated as the main biologic vectors for Anaplasma spp. Studies represent that Ixodes spp. are the main vectors for A. phago-cytophilum and few reports hinted that other tick species may play this role. So, the goal of the presented work was to investigate the A. phagocytophilum in 2000 hard ticks in Khuzestan province of Iran by specific nested-PCR performing two consecutive amplifications of 16SrRNA gene fragment with highly variable nucleotide region. Each reaction included 10 salivary glands of distinct tick species. Specific nested-PCR on accumulated salivary glands detected specific bands in 15.5% of reactions (31 of 200) in electrophoresis only in Rhipicephalus sanguineous and Hyalomma marginatum ticks. We concluded that the broad distribution of A. phagocytophilum infection is not only is due to the existence of Ixodes spp. but other hard ticks may also play a role in this issue.

Lymphadenopathy transmitted by tick. A pediatric case of rickettsiosis in primary care

The entity called tick-borne lymphadenopathy, also known as dermacentor-borne necrosis eritema lymphadenopathy, is included in the differential diagnosis of diseases transmitted by ticks, which have increased considerably in recent years due to greater clinical-epidemiological observation and diagnostic improvement. It is a zoonosis caused by Rickettsia slovaca and transmitted by the bite of the Dermacentor marginatus tick. Affected patients have a necrotic eschar surrounded by an erythematous halo in the scalp, as well as painful regional lymphadenopathy. We present a seven-year-old male who shows, after the extraction of a tick, a necrotic eschar on the scalp, accompanied by lymphadenopathy and low-grade fever. The study of polymerase chain reaction and culture of the lesion confirm the presence of Rickettsia slovaca. The patient received oral azithromycin with a good response. In Primary Care, it is important to monitor the tick bites, to detect possible diseases transmitted by them.

Uptake and fecal excretion of Coxiella burnetii by Ixodes ricinus and Dermacentor marginatus ticks

BackgroundThe bacterium Coxiella burnetii is the etiological agent of Q fever and is mainly transmitted via inhalation of infectious aerosols. DNA of C. burnetii is frequently detected in ticks, but the role of ticks as vectors in the epidemiology of this agent is still controversial. In this study, Ixodes ricinus and Dermacentor marginatus adults as well as I. ricinus nymphs were fed on blood spiked with C. burnetii in order to study the fate of the bacterium within putative tick vectors.MethodsBlood-feeding experiments were performed in vitro in silicone-membrane based feeding units. The uptake, fecal excretion and transstadial transmission of C. burnetii was examined by quantitative real-time PCR as well as cultivation of feces and crushed tick filtrates in L-929 mouse fibroblast cells and cell-free culture medium.ResultsTicks successfully fed in the feeding system with engorgement rates ranging from 29% (D. marginatus) to 64% (I. ricinus adults). Coxiella burnetii DNA was detected in the feces of both tick species during and after feeding on blood containing 105 or 106 genomic equivalents per ml blood (GE/ml), but not when fed on blood containing only 104 GE/ml. Isolation and cultivation demonstrated the infectivity of C. burnetii in shed feces. In 25% of the I. ricinus nymphs feeding on inoculated blood, a transstadial transmission to the adult stage was detected. Females that molted from nymphs fed on inoculated blood excreted C. burnetii of up to 106 genomic equivalents per mg of feces.ConclusionsThese findings show that transstadial transmission of C. burnetii occurs in I. ricinus and confirm that I. ricinus is a potential vector for Q fever. Transmission from both tick species might occur by inhalation of feces containing high amounts of viable C. burnetii rather than via tick bites.

Results of investigation of ticks in Volga river delta (Astrakhan region, 2017) for Crimean-Congo hemorrhagic fever virus (Nairoviridae, Orthonairovirus, CCHFV) and other tick-borne arboviruses.

There are natural foci of Crimean-Congo hemorrhagic fever (CCHF) that vectored by Hyalomma marginatum ticks in Volga river delta (Astrakhan region, South of Russia). The circulation of Dhori virus (DHOV) (Thogotovirus: Orthomyxoviridae) has been also shown here. We hypothesized that other tick-borne arboviruses are also likely to circulate in the region. In particular, Bhanja virus (Phlebovirus: Phenuiviridae), Wad Medani virus (Orbivirus: Reoviridae), and Tamdy virus (Orthonairovirus: Nairoviridae), which were found to circulate in neighboring regions and are vectored by Haemaphysalis spp., Dermacenter spp., and Hyalomma spp. ticks. The aim of the study was to examine ixodid ticks in Volga river delta for the presence of CCHFV, DHOV, Bhanja virus, Wad Medani virus, and Tamdy virus. Ticks were collected in Volga river delta in 2017. We used molecular genetic methods for the detection and analysis of nucleic acids (PCR, sequencing, phylogenetic analysis). We detect CCHFV and DHOV RNA in H. marginatum ticks. The rate of infected H. marginatum ticks was 1.98% for CCHFV and 0.4% for DHOV. The results of genetic analysis showed that found DHOV strains are almost identical (99-100% in the M gene) and forms a separate genetic lineage alongside of Batken virus from Central Asia. At the same time, Bhanja virus, Wad Medani virus, and Tamdy virus were not found in ticks, collected in this region. DHOV is circulating in the natural foci of CCHF in the Volga river delta. The ratio of infection of H. marginatum with CCHFV and DHOV was determined for the first time.

Abnormal Development of Hyalomma Marginatum Ticks (Acari: Ixodidae) Induced by Plant Cytotoxic Substances.

The increasing application of toxic plant substances to deter and fight ticks proves the need for investigations focused on the elucidation of their impact on the developmental stages and populations of these arthropods. We examined the course of embryogenesis and egg hatch in Hyalomma marginatum ticks under the effect of cytotoxic plant substances. The investigations demonstrated that the length of embryonic development of egg batches treated with 20 μL of a 0.1875% colchicine solution did not differ significantly from that in the control group. Colchicine caused the high mortality of eggs (16.3%) and embryos (9.7%), disturbances in larval hatch (8.1%), and lower numbers of normal larval hatches (65.6%). In 0.2% of the larvae, colchicine induced anomalies in the idiosoma (67.6%) and gnathosoma (22.5%) as well as composite anomalies (8.5%). The study demonstrates that cytotoxic compounds with an effect similar to that of colchicine can reduce tick populations and cause teratological changes, which were observed in the specimens found during field studies. Since there are no data on the toxic effects of active plant substances on other organisms and the risk of development of tick resistance, a strategy for the use of such compounds in tick control and the management of plant products should be developed.

Molecular characterization and phylogenetic analysis of Babesia and Theileria spp. in ticks from domestic and wild hosts in Sardinia

Piroplasmoses are tick-borne protozoan diseases caused by hemoprotozoan parasites with considerable economic, veterinary and medical impact worldwide. Here, the presence and diversity of piroplasmids was investigated in ticks collected from domestic and wild hosts in a typical subtropical environment of Sardinia island by standard PCR, sequencing, and phylogenetic analyses. We demonstrate the presence of strains closely related to the Theileria buffeli/sergentii/orientalis complex in Rhipicephalus sanguineus s.l., Rh. bursa, Rh. annulatus, Hyalomma marginatum, Dermacentor marginatus and Haemaphysalis punctata ticks. A strain detected in two Rh. sanguineus s.l. ticks collected from dogs grouped with T. equi, the agent of equine piroplasmosis. T. ovis, the main etiological agent of ovine theileriosis, was detected in one Rh. bursa tick from a mouflon. Babesia bigemina, the causative agent of bovine babesiosis, was detected in two Rh. sanguineus s.l. ticks from dogs. Our findings expand the knowledge on the repertoire of tick-borne pathogens present in Mediterranean ticks. Further analyses are needed to determine the role of ticks in the biological or mechanical transmission of piroplasmoses in this area.