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Numerous tick species are undergoing significant range expansion in Canada, including several Dermacentor spp Koch (Acari: Ixodidae). With the recent description of Dermacentor similis Lado in the western United States, additional research is required to determine the current range of this species. Five hundred ninety-eight Dermacentor spp. were collected from companion animals in the western Canadian provinces of British Columbia, Alberta, and Saskatchewan. Ticks were morphologically identified to species, followed by PCR and gel electrophoresis of the ITS-2 partial gene target (n = 595). Ninety-seven percent (n = 579/595) generated valid banding patterns. The banding pattern for the majority (74%, n = 206/278) of Dermacentor spp. from southern British Columbia was consistent with D. variabilis (Say), while 26% (n = 72/278) was consistent with D. andersoni Stiles. For samples from Alberta, 38% (n = 3/8) had banding patterns consistent with D. variabilis and 63% (n = 5/8) with D. andersoni. All (n = 293) ticks from Saskatchewan had banding patterns consistent with D. variabilis. After the description of D. similis was published, DNA sequencing of mitochondrial (16S rDNA gene, COI gene) and nuclear (ITS-2) markers was used to confirm the identity of 40 samples. Twenty-seven samples that had banding patterns consistent with D. variabilis from British Columbia were confirmed to be D. similis. One sample from Alberta and five from Saskatchewan were confirmed to be D. variabilis and seven samples from British Columbia were D. andersoni. The ITS-2 amplicons were not useful for differentiating between D. variabilis and D. similis. These results provide evidence of D. similis in western Canada and highlight that sequences of the mitochondrial genes are effective for distinguishing D. andersoni, D. variabilis, and D. similis.

Dermacentor marginatus is a medically important tick species due to its preference humans and domestic animals as hosts and its vectorial competence, yet it remains understudied in many regions. This study aimed to examine the population structure and demographic history of D. marginatus using the cox1 and ITS2 genes, focusing on populations from Central and Northeast Anatolia—two regions on either side of the Anatolian Diagonal, a natural biogeographical barrier. A total of 361 host-seeking adult D. marginatus ticks from 31 sampling sites were analyzed, revealing 131 haplotypes for cox1 and 104 genotypes for ITS2. Neutrality tests and mismatch distribution patterns rejected the null hypothesis of the neutral theory, indicating that the population of D. marginatus in Anatolia has undergone a recent demographic expansion. Significant genetic differentiation and population structuring were observed between the Central and Northeastern Anatolian populations of D. marginatus , correlating with geographic distance and suggesting that the Anatolian Diagonal acts as a potential barrier to gene flow. Intrapopulation gene flow was higher in Central Anatolian populations compared to Northeastern Anatolian populations. Bayesian phylogeny revealed a highly divergent D. marginatus haplotype within the Northeastern Anatolian population, clustering into a Central Asian clade. Additionally, phylogenetic trees of the subgenus Serdjukovia revealed taxonomic ambiguities, including the absence of a distinct clade for D. niveus and potential misidentifications of D. marginatus and D. raskemensis specimens. Furthermore, the monophyletic relationship between D. marginatus and D. raskemensis supports the likelihood of sympatric speciation. These findings enhance our understanding of the genetic structure, phylogeography, and evolutionary dynamics of D. marginatus while providing a framework for future research on tick populations.

Ticks are a group of blood-sucking ectoparasites that play an important role in human health and livestock production development as vectors of zoonotic diseases. The phylogenetic tree of single genes cannot accurately reflect the true kinship between species. Based on the complete mitochondrial genome analysis one can help to elucidate the phylogenetic relationships among species. In this study, the complete mitochondrial genome of Dermacentor steini (isolate Longyan) was sequenced and compared with the mitochondrial genes of 3 other Chinese isolates (Nanchang, Jinhua and Yingtan). In Dermacentor steini 4 isolates had identical or similar mitochondrial genome lengths and an overall variation of 0.76% between sequences. All nucleotide compositions showed a distinct AT preference. The most common initiation and stop codons were ATG and TAA, respectively. Fewer base mismatches were found in the tRNA gene of D. steini (isolate Longyan), and the vicinity of the control region and tRNA gene was a hot rearrangement region of the genus Dermacentor. Maximum likelihood trees and Bayesian trees indicate that D. steini is most closely related to Dermacentor auratus. The results enrich the mitochondrial genomic data of species in the genus Dermacentor and provide novel insights for further studies on the phylogeographic classification and molecular evolution of ticks.

In recent years, significant changes have been observed in the distribution and abundance of local Dermacentor reticulatus populations. However, changes in D. reticulatus dynamics have not been studied in southeastern Poland. Our objective was to enhance our understanding of the environmental factors influencing the occurrence and density of D. reticulatus in this area. Additionally, we sought to investigate the genetic diversity of the tick population and the prevalence of tick-borne pathogens (TBPs). To this end, we established 45 study sites in the Subcarpathian province. Ticks were collected during their peak activity in both spring and autumn. A subset of randomly selected specimens underwent molecular analysis for TBPs screening, using high-throughput microfluidic real-time PCR. Positive amplicons were then sequenced, and phylogenetic analyses were conducted. Our findings confirmed the presence of D. reticulatus ticks in 24 surveyed sites, primarily concentrated in the northern and eastern parts of the region. The mean density of D. reticulatus ticks in their compact range was 5.8 ± 6.4 specimens/100 m2. Notably, air temperature and altitude emerged as significant factors influencing the species’ activity. We also identified a high prevalence of Rickettsia raoultii infections in adult D. reticulatus, reaching up to 84.21%. Additionally, 9.52% of ticks were found to be infected with R. helvetica and 4.76% with Anaplasma phagocytophilum. Furthermore, our genetic analyses confirmed the identity of D. reticulatus in the Subcarpathian region, aligning with haplotypes found in other regions of Poland, Czechia, Croatia, and Portugal. In conclusion, our study suggests that the surveyed region represents the current boundary of the compact range of D. reticulatus in Poland in which this tick species exhibits low genetic diversity and a narrow spectrum of detected TBPs.

Hard ticks (Acari: Ixodidae) have been historically identified by morphological methods which require highly specialized expertise and more recently by DNA-based molecular assays that involve high costs. Although both approaches provide complementary data for tick identification, each method has limitations which restrict their use on large-scale settings such as regional or national tick surveillance programs. To overcome those obstacles, the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been introduced as a cost-efficient method for the identification of various organisms, as it balances performance, speed, and high data output. Here we describe the use of this technology to validate the distinction of two closely related Dermacentor tick species based on the development of the first nationwide MALDI-TOF MS reference database described to date. The dataset obtained from this protein-based approach confirms that tick specimens collected from United States regions west of the Rocky Mountains and identified previously as Dermacentor variabilis are the recently described species, Dermacentor similis . Therefore, we propose that this integrative taxonomic tool can facilitate vector and vector-borne pathogen surveillance programs in the United States and elsewhere.

Background Vietnam and its region are regarded as an ixodid tick biodiversity hotspot for at least two genera: Haemaphysalis and Dermacentor . To contribute to our knowledge on the tick fauna of this country, ticks from these two genera as well as an Ixodes species were analyzed morphologically and their molecular-phylogenetic relationships were examined in taxonomic and geographical contexts. Methods For this study, seven Haemaphysalis sp. ticks were removed from dogs and collected from the vegetation. These showed morphological differences from congeneric species known to occur in Vietnam. In addition, three Ixodes sp. ticks were collected from pygmy slow lorises ( Xanthonycticebus pygmaeus ), and a Dermacentor female had been previously collected from the vegetation. After DNA extraction, these were molecularly or phylogenetically analyzed based on the cytochrome c oxidase subunit I ( cox 1) and 16S rRNA genes. Results The three species were morphologically identified as (i) Ixodes granulatus , which had nearly or exactly 100% sequence identities to conspecific ticks reported from large (approximately 2000 km) geographical distances but was more different (having lower, only 94.2% cox 1 and 96.7% 16S rRNA sequence identity) from samples collected within 1000 km of Vietnam in Southern China and Malaysia, respectively; (ii) Haemaphysalis bispinosa , which showed 100% sequence identity to samples reported within both narrow and broad geographical ranges; and (iii) a new species, Dermacentor pseudotamokensis Hornok sp. nov., described here morphologically and shown to be phylogenetically a sister species to Dermacentor tamokensis . Conclusions Haemaphysalis bispinosa shows genetic homogeneity in the whole of South and Southeast Asia, probably owing to its frequent association with domestic ruminants and dogs (i.e. frequently transported hosts). However, I. granulatus , the Asian rodent tick, has a mixed geographical pattern of haplotypes, probably because it may associate with either synanthropic or wild-living rodents as primary hosts. This tick species is recorded here, for the first time to our knowledge, as parasitizing lorises in Vietnam and its region. Based on phylogenetic analyses, D. pseudotamokensis Hornok sp. nov., recognized and described here for the first time, was almost certainly misidentified previously as Dermacentor steini , drawing attention to the need to barcode all Dermacentor spp. in Southern Asia. Graphical Abstract

In the 1930s, R. A. Cooley noted that Dermacentor occidentalis (Acarina: Ixodidae) and Dermacentor andersoni were closely related and could hybridize. Decades later, James Oliver discovered that crosses of Dermacentor variabilis, D. andersoni, and D. occidentalis could, on occasion, produce hybrids. A recent molecular analysis (both mtDNA and nDNA) in our laboratory revealed that certain specimens of Dermacentor andersoni nested with Dermacentor parumapertus. Does this close relationship, along with the mito-nuclear discordance we have observed, mean D. andersoni and D. parumapertus are a single species? By contemporary taxonomic criteria, this seems improbable based on their distinctly different morphologies, host associations, and ecologies. This paper explores ideas related to mito-nuclear discordance, hybridization, and introgression (primarily) not only in these two species but also other members of the genus Dermacentor. Both D. andersoni and D. parumapertus can be found on the same hosts and have sympatric distributions, so introgression of genetic material by occasional cross-mating between these two species is possible. Further, the difficulty in applying specific species concepts in ticks has been recently pointed out and a unified agreement on an integrative species concepts could clearly be useful in this situation. With the discovery of D. parumapertus as a potential vector of Rickettsia parkeri and the historically recognized role of D. andersoni in transmission of Rickettsia rickettsii, understanding the specific status of each lineage of these species (and others in the genus) is extremely important from a public health perspective.

Background The tick Dermacentor silvarum Olenev (Acari: Ixodidae) is a vital vector tick species mainly distributed in the north of China and overwinters in the unfed adult stage. The knowledge of the mechanism that underlies its molecular adaptation against cold is limited. In the present study, genes of hsp70 and hsp90 cDNA, named Dshsp70 and Dshsp90 , and tubulin were cloned and characterized from D. silvarum , and their functions in cold stress were further evaluated. Methods The genome of the heat shock proteins and tubulin of D. silvarum were sequenced and analyzed using bioinformatics methods. Each group of 20 ticks were injected in triplicate with Dshsp90 -, Dshsp70 -, and tubulin-derived dsRNA, whereas the control group was injected with GFP dsRNA. Then, the total RNA was extracted and cDNA was synthesized and subjected to RT-qPCR. After the confirmation of knockdown, the ticks were incubated for 24 h and were exposed to − 20 °C lethal temperature (LT50), and then the mortality was calculated. Results Results indicated that Dshsp70 and Dshsp90 contained an open reading frame of 345 and 2190 nucleotides that encoded 114 and 729 amino acid residues, respectively. The transcript Dshsp70 showed 90% similarity with that identified from Dermacentor variabilis , whereas Dshsp90 showed 85% similarity with that identified from Ixodes scapularis . Multiple sequence alignment indicates that the deduced amino acid sequences of D. silvarum Hsp90, Hsp70, and tubulin show very high sequence identity to their corresponding sequences in other species. Hsp90 and Hsp70 display highly conserved and signature amino acid sequences with well-conserved MEEVD motif at the C-terminal in Hsp90 and a variable C-terminal region with a V/IEEVD-motif in Hsp70 that bind to numerous co-chaperones. RNA interference revealed that the mortality of D. silvarum was significantly increased after injection of dsRNA of Dshsp70 ( P  = 0.0298) and tubulin ( P  = 0.0448), whereas no significant increases were observed after the interference of Dshsp90 ( P  = 0.0709). Conclusions The above results suggested that Dshsp70 and tubulin play an essential role in the low-temperature adaptation of ticks. The results of this study can contribute to the understanding of the survival and acclimatization of overwintering ticks. Graphical abstract

Ticks are hematophagous vectors that transmit a variety of pathogens, posing significant threats to the health of both humans and animals. Tick midgut proteins play essential roles in blood digestion, feeding, toxic waste processing, and pathogen transmission. Dermacentor nuttalli is the primary vector of tick-borne pathogens, including rickettsioses in the Qinghai-Tibet Plateau. However, there is a lack of genomic, transcriptomic, and proteomic information regarding the biology of D. nuttalli . In this study, we assembled and compared the midgut transcriptomes of female D. nuttalli ticks at 0, 24, 48, 72, and 96 h during blood feeding, identifying the genes with differentially regulated expression following feeding. The obtained data were compiled and annotated in multiple databases including Nr, NT, PFAM, KOG, KEGG, and GO. The high-quality clean readings of midgut tissue at the different blood-feeding times were recorded as 22,524,912, 23,752,325, 20,377,718, 21,300,710, and 20,378,658, respectively. The transcripts were classified into eight large categories, including immunogenic proteases (8.37%), protease inhibitors (0.85%), transporters (3.96%), ligand binding proteins (1.98%), ribosomal function proteins (0.94%), heat shock proteins (0.30%), other proteases and miscellaneous proteins (57.61%), and unknown proteins (26.00%). Significant differences were observed in the genes obtained at 0, 24, 48, 72, and 96 h during blood feeding. The differentially expressed genes include catalytic proteins that play an important role in accelerating biochemical reactions, binding activity proteins which are involved in various molecular interactions, and proteins that actively participate in multiple metabolic pathways and cellular processes. Notably, the gene expression in the midgut of D. nuttalli shows dynamic changes every 24 h throughout the blood-feeding process. This change may represent an equivalent strategy of antigenic variation for ticks, designed to protect their essential feeding function against the host’s immune system. The tick antigens identified in this study may serve as promising candidates for the development of effective vaccines or as drug targets for acaricides.

Anaplasma spp. are obligate intracellular, tick-borne, bacterial pathogens that cause bovine and human anaplasmosis. We lack tools to prevent these diseases in part due to major knowledge gaps in our fundamental understanding of the tick–pathogen interface, including the requirement for and molecules involved in iron transport during tick colonization. We determine that iron is required for the pathogen Anaplasma marginale, which causes bovine anaplasmosis, to replicate in Dermacentor andersoni tick cells. Using bioinformatics and protein modeling, we identified three orthologs of the Gram-negative siderophore-independent iron uptake system, FbpABC. Am069, the A. marginale ortholog of FbpA, lacks predicted iron-binding residues according to the NCBI conserved domain database. However, according to protein modeling, the best structural orthologs of Am069 are iron transport proteins from Cyanobacteria and Campylobacterjejuni. We then determined that all three A. marginale genes are modestly differentially expressed in response to altered host cell iron levels, despite the lack of a Ferric uptake regulator or operon structure. This work is foundational for building a mechanistic understanding of iron uptake, which could lead to interventions to prevent bovine and human anaplasmosis.