Ticks in the metropolitan area of Berlin,Germany

A high-resolution city map showing the geographic distribution of 12 tick species (Acari: Argasidae, Ixodidae) that have been recorded from the metropolitan area of Berlin, Germany is presented. A total of 237 tick locations was mapped. These include ten ixodid tick species: Dermacentor reticulatus, Haemaphysalis concinna, Hyalomma rufipes, Ixodes ricinus, Ixodes canisuga, Ixodes hexagonus, Ixodes arboricola, Ixodes frontalis, Ixodes trianguliceps and Rhipicephalus sanguineus sensu lato. The two tick species Hy. rufipes and R. sanguineus s.l. are not endemic to Berlin. Hyalomma rufipes ticks are introduced in Europe with migratory birds from Africa every spring. Rhipicephalus sanguineus s.l. are introduced to Central Europe with dogs that had travelled to or were imported from countries where this tick is endemic. In Germany, they are able to develop and reproduce inside heated buildings. Occurrences of two soft tick species, the pigeon tick Argas reflexus and the short-legged bat tick Carios vespertilionis were also mapped. Other tick species that are likely to be endemic to Berlin and its environs, but for which documented findings or geographical coordinates are lacking, are mentioned. These include the long-legged bat tick I. vespertilionis and the marten tick I. rugicollis documented in Brandenburg, the federal state surrounding Berlin. It can be assumed that if appropriate field studies are carried out, these tick species will also be found in the metropolitan area of Berlin. The high-resolution mapping of all tick species found in a city (like Berlin) forms the basis for further investigations into the impact of climate change and changing land use on ticks and tick-borne diseases, precisely in those habitats where most people will live in the future.     

DNA barcoding of hard ticks (Ixodidae), notes on distribution of vector species and new faunal record for Croatia

Molecular methods are increasingly being utilized for accurate identification of ticks (Acari: Ixodidae), especially in cases of morphologically highly similar species. In this study, we performed molecular research of the tick fauna in Croatia using DNA barcoding method. Ticks were sampled in three biogeographical regions and thirteen species were recorded: Dermacentor marginatus, Dermacentor reticulatus, Haemaphysalis concinna, Haemaphysalis inermis, Haemaphysalis punctata, Hyalomma marginatum, Ixodes frontalis, Ixodes hexagonus, Ixodes kaiseri, Ixodes ricinus, Rhipicephalus bursa, Rhipicephalus sanguineus s.l. and Rhipicephalus turanicus. Ixodes kaiseri is for the first time recorded in the fauna of Croatia. Of the thirteen hard tick species analyzed in this study, pathogens from different groups (bacteria, protozoa and viruses) have been detected in eight species in Croatia so far. For the important vector species R. sanguineus s.s., new distributional data for Croatia are given. The standard COI barcoding region was amplified, and the sequences were analyzed by species delimitation methods together with the sequences of conspecific and congeneric species from the public BOLD database. Our specimens of H. punctata represent a new, genetically distinct MOTU. A brief overview of the available public DNA barcoding data for Ixodidae is presented, highlighting the need for an integrative approach for the clarification of the taxonomic status of problematic Ixodid taxa. The results provide a basis for the establishment of a molecular data platform for the Ixodidae of the Croatian fauna.     

Tick (Acari: Ixodoidea) fauna and zoogeographic division of Jiangxi Province,China

Tick fauna and zoogeographic distribution of Jiangxi Province remain largely unknown due to the lack of data on distribution, occurrence, and host associations of ticks. Considering this, we collected 1,817 individual samples from natural hosts, humans, and vegetation in 18 counties/districts throughout Jiangxi Province, China, from 2015 to 2021. These 1,817 individuals were found to 13 tick species, 4 genera, and 1 family. In addition, the tick sample data from 8 sampling localities (counties and districts) reported in previous studies were also included in our data. A total of 4,021 individuals, including our sample collection and the previously reported data, were assigned to at least 18 species, 6 genera, and 2 families. One newly recorded species Dermacentor sp. (near D. steini Schulze) was found; three misidentified species (Ixodes acuminatus, Haemaphysalis spinigera, and Haemaphysalis verticalis) reported previously were deleted; and one misidentified species Dermacentor auratus Supino was revised as Dermacentor steini Schulze. In addition, we divided the tick fauna in Jiangxi Province into 5 zoogeographic areas and assigned the 18 tick species collected from 26 localities to these 5 zoogeographic areas. To summarize, our findings provide valuable information on the distribution, tick-host associations, and zoogeographic division of ticks in Jiangxi Province, China. Their molecular characterizations, phylogenetic relationships, and tick-borne pathogens that they may transmit should be further explored.     

Identification and functional analysis of ferritin 2 from the Taiga tick Ixodes persulcatus Schulze

Ferritin 2 (FER2) is an iron storage protein, which has been shown to be critical for iron homeostasis during blood feeding and reproduction in ticks and is therefore suitable as a component for anti-tick vaccines. In this study, we identified the FER2 of Ixodes persulcatus, a major vector for zoonotic diseases such as Lyme borreliosis and tick-borne relapsing fever in Japan, and investigated its functions. Ixodes persulcatus-derived ferritin 2 (Ip-FER2) showed concentration-dependent iron-binding ability and high amino acid conservation, consistent with FER2s of other tick species. Vaccines containing the recombinant Ip-FER2 elicited a significant reduction of the engorgement weight of adult I. persulcatus. Interestingly, the reduction of engorgement weight was also observed in Ixodes ovatus, a sympatric species of I. persulcatus. In silico analyses of FER2 sequences of I. persulcatus and other ticks showed a greater similarity with I. scapularis and I. ricinus and lesser similarity with Hyalomma anatolicum, Haemaphysalis longicornis, Rhipicephalus microplus, and R. appendiculatus. Moreover, it was observed that the tick FER2 sequences possess conserved regions within the primary structures, and in silico epitope mapping analysis revealed that antigenic regions were also conserved, particularly among Ixodes spp ticks. In conclusion, the data support further protective tick vaccination applications using the Ip-FER2 antigens identified herein.     

Tick species infesting humans in the United States

The data for human tick encounters in the United States (US) presented in this paper were compiled with the goals of: (i) presenting quantitative data across the full range of native or recently established human biting ixodid (hard) and argasid (soft) tick species with regards to their frequency of infesting humans, based on published records of ticks collected while biting humans or crawling on clothing or skin; and (ii) providing a guide to publications on human tick encounters. Summary data are presented in table format, and the detailed data these summaries were based on are included in a set of Supplementary Tables. To date, totals of 36 ixodid species (234,722 specimens) and 13 argasid species (230 specimens) have been recorded in the published literature to infest humans in the US. Nationally, the top five ixodid species recorded from humans were the blacklegged tick, Ixodes scapularis (n=158,008 specimens); the lone star tick, Amblyomma americanum (n=36,004); the American dog tick, Dermacentor variabilis (n=26,624); the western blacklegged tick, Ixodes pacificus (n=4,158); and the Rocky Mountain wood tick, Dermacentor andersoni (n=3,518). Additional species with more than 250 ticks recorded from humans included Ixodes cookei (n=2,494); the Pacific Coast tick, Dermacentor occidentalis (n=809); the brown dog tick, Rhipicephalus sanguineus sensu lato (n=714); the winter tick, Dermacentor albipictus (n=465); and the Gulf Coast tick, Amblyomma maculatum (n=335). The spinose ear tick, Otobius megnini (n=69), and the pajaroello tick, Ornithodoros coriaceus (n=55) were the argasid species most commonly recorded from humans. Additional information presented for each of the 49 tick species include a breakdown of life stages recorded from humans, broad geographical distribution in the US, host preference, and associated human pathogens or medical conditions. The paper also provides a history of publications on human tick encounters in the US, with tables outlining publications containing quantitative data on human tick encounters as well as other notable publications on human-tick interactions. Data limitations are discussed. Researchers and public health professionals in possession of unpublished human tick encounter data are strongly encouraged to publish this information in peer-reviewed scientific journals. In future papers, it would be beneficial if data consistently were broken down by tick species and life stage as well as host species and ticks found biting versus crawling on clothing or skin.     

Comprehensive screening of tick-borne microorganisms indicates that a great variety of pathogens are circulating between hard ticks (Ixodoidea: Ixodidae) and domestic ruminants in natural foci of Anatolia

Grazing domestic ruminants serve as important reservoirs and/or amplificatory hosts in the ecology of tick-borne pathogens (TBPs) and tick vectors in the natural foci; however, many enzootic life cycles including ruminants and ticks are still unknown. This study investigated a wide range of TBPs circulating among ticks and grazing ruminants in the natural foci of Anatolia, Turkey. Tick specimens (n = 1815) were collected from cattle, sheep, and goats in three ecologically distinct areas (wooded, transitional, and semi-arid zones) of Anatolia and identified by species: Dermacentor marginatus, Dermacentor reticulatus, Hyalomma anatolicum, Hyalomma excavatum, Hyalomma marginatum, Hyalomma scupense, Haemaphysalis inermis, Haemaphysalis parva, Haemaphysalis punctata, Haemaphysalis sulcata, Ixodes ricinus, Rhipicephalus bursa, and Rhipicephalus turanicus. PCR-sequencing analyses revealed TBPs of great diversity, with 32 different agents identified in the ticks: six Babesia spp. (Babesia occultans, Babesia crassa, Babesia microti, Babesia rossi, Babesia sp. tavsan1, and Babesia sp. Ucbas); four Theileria spp., including one putative novel species (Theileria annulata, Theileria orientalis, Theileria ovis, and Theileria sp.); one Hepatozoon sp.; four Anaplasma spp., including one novel genotype (Anaplasma phagocytophilum, Anaplasma marginale, Anaplasma ovis, and Anaplasma sp.); six unnamed Ehrlichia spp. genotypes; Neoehrlichia mikurensis; nine spotted fever group rickettsiae, including one putative novel species (Rickettsia aeschlimannii, Rickettsia slovaca, Rickettsia hoogstraalii, Rickettsia monacensis with strain IRS3, Rickettsia mongolitimonae, Rickettsia raoultii, Candidatus Rickettsia goldwasserii, Candidatus Rickettsia barbariae, and Rickettsia sp.); and Borrelia valaisiana. Detailed phylogenetic analyses showed that some of the detected pathogens represent more than one haplotype, potentially relating to the tick species or the host. Additionally, the presence of Neoehrlichia mikurensis, an emerging pathogen for humans, was reported for the first time in Turkey, expanding its geographical distribution. Consequently, this study describes some previously unknown tick-borne protozoan and bacterial species/genotypes and provides informative epidemiological data on TBPs, which are related to animal and human health, serving the one health concept.     

Extensive examination of different tick species for infestation with Coxiella burnetii in Slovakia

Between 1987–1989 almost 7000 adult Ixodes ricinus, Dermacentor reticulatus, Dermacentor marginatus, Haemaphysalis concinna, Haemaphysalis punctata and Haemaphysalis inermis ticks collected in all 38 districts of Slovakia were screened for the presence of Coxiella burnetii. The proportion of ticks containing C. burnetii as indicated by the haemocyte test was less than 3%. Attempts to recover C. burnetii by inoculation of yolk sacs of embryonated hen eggs from pools of 1–6 specimens of haemocyte test positive ticks resulted in the isolation of 10 rickettsial strains. Six strains were recovered from I. ricinus, the remaining ones from single pools of D. reticulatus, D. marginatus. H. concinna and H. inermis ticks.In addition to the previous recovery of C. burnetii from H. punctata ticks, the agent was thus isolated from all important ticks living in Slovakia. The agent was found in tick habitats regardless of the latitude and altitude in the entire country. These results are not consistent with the negligible number of Q fever cases occurring in past years in Slovakia.Presented at the 4th International Symposium on Rickettsiae and Rickettsial Diseases, Pietany, C.S.F.R., 1–6 October, 1990.     

Genetic diversity of Theileria orientalis in tick vectors detected in Hokkaido and Okinawa,Japan

In the present study, we investigated the possible tick vectors that can transmit Theileria orientalis in eastern Hokkaido, Japan. Questing ticks collected from three different districts, Taiki, Otofuke, and Shin-Hidaka, of Hokkaido included Ixodes persulcatus, Haemaphysalis megaspinosa, Haemaphysalis douglasi, and Ixodes ovatus, while all the ticks collected from Yonaguni island of Okinawa were identified as Haemaphysalis longicornis. When the ticks were screened by polymerase chain reaction (PCR) for T. orientalis, the parasite was commonly detected among all tick species. Genotype-specific PCR assays revealed that all tick species in Hokkaido were predominantly detected with type 2, while ticks collected from Okinawa (H. longicornis) were predominantly detected with type 1. Consistent with the genetic diversity of T. orientalis in ticks, genotyping PCR assays from cattle grazed in the same Hokkaido sampling locations identified type 2 as the most prevalent genotype. This study provides the first identification of I. persulcatus, H. megaspinosa, H. douglasi, and I. ovatus as possible tick vectors of T. orientalis, and finds that the variety of vectors apparently capable of transmitting T. orientalis is wider in Japan than expected. The authors suggest that tick control strategies should be modified in Hokkaido based on the seasonal activities of ticks identified in the present study.     

Winter activity of questing ticks (Ixodes ricinus and Dermacentor reticulatus) in Germany − Evidence from quasi-natural tick plots,field studies and a tick submission study

Changing climatic conditions and other anthropogenic influences have altered tick distribution, abundance and seasonal activity over the last decades. In Germany, the two most important tick species are Ixodes ricinus and Dermacentor reticulatus, the latter of which has expanded its range across the country during the past three decades. While I. ricinus was rarely found during the colder months in the past, D. reticulatus is known to be active at lower temperatures. To quantify tick appearance during winter, specimens were monitored in quasi-natural tick plots three times a week. Additionally, the questing activities of these two tick species were observed throughout the year at nine field collection sites that were regularly sampled by the flagging method from April 2020 to April 2022. Furthermore, tick winter activity in terms of host infestation was analysed as part of a nationwide submission study from March 2020 to October 2021, in which veterinarians sent in ticks mainly collected from dogs and cats. All three study approaches showed a year-round activity of I. ricinus and D. reticulatus in Germany. During the winter months (December to February), on average 1.1% of the inserted I. ricinus specimens were observed at the tops of rods in the tick plots. The average questing activity of I. ricinus amounted to 2 ticks/100 m² (range: 1-17) in the flagging study, and 32.4% (211/651) of ticks found infesting dogs and cats during winter 2020/21 were I. ricinus. On average 14.7-20.0% of the inserted D. reticulatus specimens were observed at the tops of rods in the tick plots, while the average winter questing activity in the field study amounted to 23 specimens/100 m² (range: 0-62), and 49.8% (324/651) of all ticks collected from dogs and cats during winter 2020/21 were D. reticulatus. Additionally, the hedgehog tick Ixodes hexagonus was found to infest dogs and cats quite frequently during the winter months, accounting for 13.2% (86/651) of the collected ticks. A generalized linear mixed model identified significant correlations of D. reticulatus winter activity in quasi-natural plots with climatic variables. The combined study approaches confirmed a complementary main activity pattern of I. ricinus and D. reticulatus with climate change-driven winter activity of both species. Milder winters and a decrease of snowfall, and consequently high winter activity of D. reticulatus, among other factors, may have contributed to the rapid spread of this tick species throughout the country. Therefore, an effective year-round tick control is strongly recommended to not only efficiently protect dogs and cats with outdoor access from ticks and tick-borne pathogens (TBPs), but also to limit the further geographical spread of ticks and TBPs to so far non-endemic regions. Further measures, including information of the public, are necessary to protect both, humans and animals, in a One Health approach.     

绥芬河口岸蜱类调查报告

目的 调查绥芬河口岸蜱种组成及生态习性.方法 采用人工小时布旗法采集蜱类.结果 2012年共捕获蜱类779只,1科3属4种.优势种群是全沟硬蜱,其次是嗜群血蜱、日本血蜱、森林革蜱.4-8月份均有蜱类活动,5、6月份为蜱类活动高峰,其蜱密度分别为63.3只/人工小时.旗、41.5只/人工小时.旗.结论 本次调查掌握了绥芬河口岸蜱类种群及季节消长情况,为绥芬河口岸蜱类的防控工作提供了依据.     

Ticks (Acari: Ixodidae,Argasidae) associated with wild birds in Argentina

The aim of this study was to report tick infestations on wild birds from four Phytogeographic Provinces of Argentina. A total of 1085 birds was captured (124 species, 97 genera, 29 families and 13 orders), and ticks were collected from 265 birds (48 species, 40 genera and five orders). A total of 1469 ticks (1102 larvae, 363 nymphs and 4 females) belonging to 15 tick species (Amblyomma calcaratum, Amblyomma dubitatum, Amblyomma nodosum, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma tigrinum, Amblyomma triste, Haemaphysalis juxtakochi, Haemaphysalis leporispalustris, Ixodes auritulus sensu lato, Ixodes pararicinus, Ixodes silvanus, Ixodes sp. cf. I. affinis and Ornithodoros sp. cf. O. mimon). Eighty-one new associations between bird species and stages of tick species are detected. The families Thamnophilidae, Turdidae, Thraupidae, Passerellidae, Furnariidae and Troglodytidae were the most prevalent. According to the Phytogeographic Provinces involved in this study, the prevalence of infection for each of them in birds was: (1) Chaco: 28.2% (11 tick species); (2) Yungas: 22.0% (8 tick species); (3) Espinal: 11.1% (2 tick species); and (4) Pampa: 3.9% (1 tick species). This study provided information on the diversity of tick species that parasitize wild birds, the variability of the specific tick-bird associations between the different Phytogeographic Provinces and the relevance of some families of birds as hosts of different tick species.     

Cercopithifilaria spp. in ticks of companion animals from Asia: new putative hosts and vectors

Cercopithifilaria bainae, Cercopithifilaria grassi, and Cercopithifilaria sp. II sensu Otranto et al., 2013 tick borne filarioids are typically found in dogs. Among them, Cercopithifilaria bainae has a worldwide distribution according to the occurrence of its tick vector, Rhipicephalus sanguineus sensu lato (s.l.). Nevertheless, in Asian countries, despite the wide presence of this tick species, data on Cercopithifilaria spp. are scant. Therefore, this study aimed to assess the occurrence of these dermal filarioids in ixodid ticks collected on dogs and cats from Asian countries, providing a better epidemiological picture on their distribution in this continent. Ticks (n = 687) of the species Rhipicephalus sanguineus s. l. (n = 667), Rhipicephalus haemaphysaloides (n = 8), Haemaphysalis longicornis (n = 7), Haemaphysalis campanulata (n = 1), Haemaphysalis wellingtoni (n = 2), Haemaphysalis hystricis (n = 1), and Ixodes sp. (n = 1) were collected on dogs and cats under the frame of previous studies in China, India, Indonesia, Malaysia, Singapore, Taiwan, Thailand, the Philippines and Vietnam. Tick samples were molecularly screened for Cercopithifilaria spp. by conventional PCR and real-time PCR using two pair of primers targeting partial sequences of cytochrome c oxidase 1 gene. Overall, Cercophitifilaria spp. DNA was detected in 9.5% (n = 65/687) of the tick specimens tested, with C. bainae being the most prevalent species (8.9%), followed by C. grassii (0.6%). Most Cercophitifilaria spp. positive ticks were collected on dogs (92.3%; 60/65); whereas ticks collected on cats represented 7.7% of the positive specimens. In addition, Cercopithifilaria spp. were mostly detected in R. sanguineus s.l. ticks (96.9%; 63/65), followed by Rhipicephalus haemaphysaloides (3.1%; 2/65). Data herein presented demonstrate the occurrence of dermal tick borne filarioids of the genus Cercopithifilaria in several Asian countries, with C. bainae being the most prevalent species. We also report for the first time the molecular detection of C. bainae in R. sanguineus s.l. ticks collected on cats, as well as in R. haemaphysaloides ticks, suggesting that the biological cycle of this filarioid species may involve other intermediate and definitive hosts than R. sanguineus s.l. and dogs. However, confirmatory studies on the role of other tick species and domestic cats on the biology of C. bainae are advocated.     

Wild Birds and Urban Ecology of Ticks and Tick-borne Pathogens, Chicago, Illinois, USA, 2005�C2010

Bird-facilitated introduction of ticks and associated pathogens is postulated to promote invasion of tick-borne zoonotic diseases into urban areas. Results of a longitudinal study conducted in suburban Chicago, Illinois, USA, during 2005–2010 show that 1.6% of 6,180 wild birds captured in mist nets harbored ticks. Tick species in order of abundance were Haemaphysalis leporispalustris, Ixodes dentatus, and I. scapularis, but 2 neotropical tick species of the genus Amblyomma were sampled during the spring migration. I. scapularis ticks were absent at the beginning of the study but constituted the majority of ticks by study end and were found predominantly on birds captured in areas designated as urban green spaces. Of 120 ticks, 5 were infected with Borrelia burgdorferi, spanning 3 ribotypes, but none were infected with Anaplasma phagocytophilum. Results allow inferences about propagule pressure for introduction of tick-borne diseases and emphasize the large sample sizes required to estimate this pressure.     

福建莱姆病蜱类媒介及其宿主的调查研究

１９９０～１９９５年间，作者对福建已发现莱姆病地区的蜱类媒介及其宿主进行调查，查出蜱类２０种，宿主包括家野鼠类、野猪、野兔、獾、山羊等野生动物和狗、牛等家畜。从蜱类、鼠类分离出多株莱姆病病原体，从台湾角血蜱查见病原体，粒形硬蜱可能是闽北莱姆病的主要蜱媒。     

Ticks (Ixodidae) on birds migrating from Europe and Asia to Africa 1959-61

The need for imaginative thinking and research in the epidemiology of diseases transmitted by arthropods is made manifest by new views of the longevity and host ranges of arthropod-borne viruses, as well as by other biological and medical phenomena. Among these is the intercontinental transport of ticks by migrating birds. During the fall migration periods of 1959, 1960 and 1961, 32 086 birds (comprising 72 forms) were examined for ticks in Egypt while en route from Asia and eastern Europe to tropical Africa. Of these, 40 forms, represented by 31 434 birds, were tick-infested. The bird hosts, numbering 1040 (3.31% of the tick-infested bird forms examined), bore 1761 ticks, or 1.69 ticks per host. Common ticks taken were Hyalomma m. marginatum, Haemaphysalis punctata, and Ixodes ricinus. Ixodes frontalis and Hyalomma aegyptium were less common and Haemaphysalis sulcata, H. otophila, and H. pavlovskyi were rare. The common tick species are known to be reservoirs and vectors of pathogens causing a number of human and animal diseases in Europe and Asia. Several of the bird hosts have also been incriminated as reservoirs in their summer ranges. Over 20 strains of pathogenic viruses were isolated from these birds and their ticks in Egypt in the 1961 fall migration period.     

黑龙江省东部边境口岸莱姆病生物媒介调查及病原分型研究

目的调查黑龙江省东部中俄边境口岸蜱的种类、分布、携带莱姆病螺旋体情况及病原基因型。方法捕蜱采用布旗法;蜱带莱姆病螺旋体率采用直接荧光法检查;病原体分离培养采用BSKⅡ培养基,对分离到的螺旋体用单克隆抗体(H_(5332)、H_(6831)、H_(9724))、SDS-PAGE蛋白图谱分析、PCR-RFLP分析以及基因测序等方法进行表型和基因分型研究。结果黑龙江省东部7个中俄边境口岸蜱的种类为1科3属4种,即硬蜱科硬蜱属全沟硬蜱,革蜱属森林革蜱,血蜱属嗜群血蜱和日本血蜱。不同生态环境中蜱的分布不同,各口岸优势蜱种不同。全沟硬蜱带莱姆病螺旋体率较高,平均为31.4%,森林革蜱和嗜群血蜱的带菌率较低,平均为2.2%和3.8%,日本血蜱中未检出阳性者。首次在中俄边境口岸——东宁、同江采集的全沟硬蜱中分离到伯氏疏螺旋体并鉴定为B.garinii基因型。结论黑龙江省东部边境口岸为莱姆病的自然疫源地。     

Constituting a glutathione S-transferase-cocktail vaccine against tick infestation

Cocktail vaccines are proposed as an attractive way to increase protection efficacy against specific tick species. Furthermore, such vaccines made with different tick antigens have the potential of cross-protecting against a broad range of tick species. However, there are still limitations to the selection of immunogen candidates. Acknowledging that glutathione S-transferases (GSTs) have been exploited as vaccines against ticks and other parasites, this study aimed to analyze a GST-cocktail vaccine as a potential broad-spectrum tick vaccine. To constitute the GST-cocktail vaccine, five tick species of economic importance for livestock industry were studied (Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Amblyomma variegatum, and Haemaphysalis longicornis). Tick GST ORF sequences were cloned, and the recombinant GSTs were produced in Escherichia coli. rGSTs were purified and inoculated into rabbits, and the immunological response was characterized. The humoral response against rGST-Rd and rGST-Av showed a stronger cross-reactivity against heterologous rGSTs compared to rGST-Hl, rGST-Ra, and rGST-Rm. Therefore, rGST-Rd and rGST-Av were selected for constituting an experimental rGST-cocktail vaccine. Vaccination experiment in rabbits showed that rGST-cocktail caused 35% reduction in female numbers in a Rhipicephalus sanguineus infestation. This study brings forward an approach to selecting immunogens for cocktail vaccines, and the results highlight rGST–Rd and rGST-Av as potentially useful tools for the development of a broad-spectrum tick vaccine.     

Detection of Coxiella burnetii in ticks collected in Slovakia and Hungary

A total of 235 adult ticks collected from vegetation in Slovakia and Hungary in 1998–2000 were tested for Coxiella burnetii by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). C. burnetii was identified in six ticks of Ixodes ricinus, Dermacentor marginatus, and Haemaphysalis concinna species.     

Characterization of ferritin 2 for the control of tick infestations

Ixodes ricinus is one the most abundant tick species in Europe and these ticks transmit pathogens causing human and animal diseases. The cattle ticks, Rhipicephalus (Boophilus) spp., affect cattle production in tropical and subtropical regions of the world. Development of vaccines directed against tick proteins may reduce tick infestations and the transmission of tick-borne pathogens. However, a limiting step in tick vaccine development has been the identification of tick protective antigens. Herein, the tick iron metabolism pathway was targeted in an effort to identify new tick protective antigens. Recombinant I. ricinus (IrFER2) and Rhipicephalus microplus (RmFER2) ferritin 2 proteins were expressed in Escherichia coli and used to immunize rabbits and cattle, respectively. Vaccination with IrFER2 reduced I. ricinus tick numbers, weight and fertility in rabbits with an overall vaccine efficacy (E) of 98%. Control of cattle tick, R. microplus and Rhipicephalus annulatus infestations was obtained in vaccinated cattle with overall E of 64% and 72%, respectively. Notably, the efficacy of the RmFER2 vaccine was similar to that obtained with Bm86 against R. microplus. These collective results demonstrated the feasibility of using ferritin 2 to develop vaccines for the control of tick infestations.     

A practical protocol to prepare paraffin-embedded whole tick histology sections

Ticks are important ectoparasites that are capable of transmitting multiple classes of pathogens and are currently linked with many emerging tick-borne diseases worldwide. With increasing occurrences of tick-borne diseases in both humans and veterinary species, there is a continuous need to further our understanding of ticks and the pathogens they transmit. Whole tick histology provides a full scope of the tick internal anatomy, allowing researchers to examine multiple organs of interest in a single section. This is in contrast to other techniques that are more commonly utilized in tick-borne disease research, such as electron microscopy and light microscopy of individual organs. There is a lack of literature describing a practical technique to process whole tick histologic sections. Therefore, the current study aims to provide researchers with a workable protocol to prepare high quality paraffin-embedded whole tick histology sections. Amblyomma americanum adults were used as an example species for this study. After a series of pilot experiments using a combination of various fixatives, softening agents and processing techniques, we elected to compare two common fixatives, 10% neutral-buffered formalin (NBF) and Bouin's solution for whole ticks. Equal numbers of A. americanum unfed adults (n = 10/fixative) were processed identically and their whole tick histology coronal sections were individually scored. Higher scores were assigned to whole tick sections that contained more internal organs that are crucial for tick-borne disease research (e.g. salivary glands and midgut), high integrity of tissues and exoskeleton on the section, and good fixation and staining quality of the tissues. The mean total scores for Bouin's-fixed ticks were significantly higher compared to NBF-fixed ticks (p = 0.001). To further assess our preferred technique, we also demonstrated the feasibility of producing high quality whole tick sections for three other common tick species of medical importance (Rhipicephalus sanguineus, Ixodes scapularis, and Dermacentor variabilis) using Bouin's solution. While this technique may require further optimization for other tick species, we described a feasible protocol that uses commonly available tools, reagents and standard histologic equipment. This should allow any investigator to easily make adjustments to this protocol as needed based on their experimental goals.