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Objective: We sought to explore the lived experience of people with Debilitating Symptom Complexes Attributed to Ticks (DSCATT) to inform the development of a potential treatment intervention., Methods: We conducted one-to-one in-depth, semi-structured interviews with 13 people living in Australia affected by DSCATT. Interviews were transcribed and analysed using thematic analysis., Results: Although participants attributed the origin of their illness to tick bites, not all were adamant they had Lyme disease. Negative experiences in conventional healthcare were marked and were reported to exacerbate the impact of the illness and affect mental health. Further, these negative experiences propelled participants to seek unapproved treatments (by Australian standards). The desire for the illness to be acknowledged and causative agents identified was pronounced among the participant group., Conclusions: Individuals with DSCATT experience significant challenges amid a contentious healthcare landscape surrounding chronic symptoms attributed to ticks in Australia. Our findings suggest the need for empathetic, supportive and patient-centred treatments for this cohort., Implications for Public Health: DSCATT results in a considerable burden across multiple domains for those affected. Negative experiences with healthcare exacerbate the suffering of people with DSCATT in Australia. New approaches that acknowledge the illness experience of people with DSCATT, alongside evidence-based treatments that encompass biopsychosocial models of care, are needed to tackle this debilitating condition., Competing Interests: Conflicts of interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: GO, VY & AD receive salary payments from the NHMRC grant funding this project. TC reports grants from UK NIHR, UKRI and Guy’s and St Thomas’ Charity on persistent physical symptoms. She has delivered workshops on persistent physical symptoms in the context of long-term conditions for which she has received payment during the course of the study. She is the author of self-help books on fatigue. KBG receives salary support from a Medical Research Futures Fund (MRFF) investigator grant (MRF1193727). VLO, SW & RK report no conflicts of interest., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Ticks are arachnid blood-feeding parasites, which infest livestock, wildlife, and humans, transmitting medically and veterinary significant pathogens. Their biodiversity and distribution in wild animals remains complex. This study analysed archived tick samples (n = 48) from the South African Biodiversity Institute (SANBI) Wildlife Biobank utilizing morphology and genetic analyses of the 16S rRNA and COI (DNA barcoding) mitochondrial genes to identify ticks collected among 13 vertebratesavian, reptilian, and mammalian host species. The specimens came from nine localities including nature reserves and captive facilities (zoological garden) in South Africa, Namibia, and Botswana. These ticks were also assessed for associated pathogens with the reverse line blot (RLB) hybridization assay. Seven tick genera, Amblyomma, Hyalomma, Haemaphysalis, Ixodes, Rhipicephalus, Rhipicentor, and Otobius were identified, with Amblyomma being the most prevalent (22.9 %) in our sample set. Obtained sequences were 95-100 % similar to published records of tick species collected from wild and domestic animals, as well as those collected from vegetation, from different southern African areas. However, tick specimens (n = 3) identified morphologically as Hyalomma truncatum, Rhipicephalus e. evertsi, and R. simus, were, on a molecularly level, more closely related to their sister taxa (H. glabrum, R. e. mimeticus, and R. gertrudae, respectively) suggesting a need for taxonomic verification. With the RLB hybridization assay, six samples reacted with the Ehrlichia/Anaplasma genus-specific probe, while two reacted with the Theileria/Babesia genus-specific probe. Sequencing of the RLB amplicons targeting the 18S rRNA gene (n = 2) indicated 100 % similarity to Hepatozoon fitzsimonsi, while one was closely related to He. ingwe with 99.39 % similarity. The results show that wildlife harbour different tick species, and pathogen detection identified novel genotypes, indicating wildlife as potential pathogens reservoirs. This study enhances our understanding of tick biodiversity, distribution and highlights wildlife's role in harbouring diverse tick species and novel pathogens., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Tick-borne rickettsial pathogens pose significant threats to public and animal health. In Upper Egypt, limited information exists regarding the prevalence and diversity of such tick-borne pathogens. Therefore, this study aimed to conduct a comprehensive investigation to elucidate the presence and variety of tick-borne rickettsial pathogens in Upper Egyptian camels. Our results revealed a prevalence of 2.96 % for Anaplasma marginale and 0.34 % for Candidatus Anaplasma camelii among Hyalomma ticks. However, Ehrlichia spp. weren't detected in our study. The identification of Ca. A. camelii in H. dromedari ticks was documented for the first time, suggesting a potential mode of transmission in camels. Notably, this study marks the first documentation of Rickettsia aeschlimannii with a prevalence of 6.06 % in the study area. Furthermore, we detected Coxiella burnetii in a prevalence of 8.08 % in Hyalomma ticks, indicating a potential risk of Q fever transmission. Molecular techniques results were confirmed by sequencing and phylogenetic analysis and provided valuable insights into the epidemiology of these pathogens, revealing their diversity. This study is vital in understanding tick-borne rickettsial pathogens' prevalence, distribution, and transmission dynamics in Upper Egypt. In conclusion, our findings emphasize the importance of continued research to enhance our understanding of the epidemiology and impact of these pathogens on both animal and human populations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Tick-borne Apicomplexan parasites pose a significant threat to both public health and animal husbandry. Identifying potential pathogenic parasites and gathering their epidemiological data are essential for prospectively preventing and controlling infections. In the present study, genomic DNA of ticks collected from two goat flocks (Goatflock1 and Goatflock2) and one dog group (Doggroup) were extracted and the 18S rRNA gene of Babesia/Theileria/Colpodella spp. was amplified by PCR and sequenced. Phylogenetic analysis was conducted based on the obtained sequences. The differences in pathogen positive rates between ticks of different groups were statistically analyzed using the Chi-square or continuity-adjusted Chi-square test. As a result, two pathogenic Theileria (T.) luwenshuni genotypes, one novel pathogenic Colpodella sp. HLJ genotype, and two potential novel Colpodella spp. (referred to as Colpodella sp. struthionis and Colpodella sp. yiyuansis in this study) were identified in the Haemaphysalis (H.) longicornis ticks. Ticks of Goatflock2 had a significantly higher positive rate of Colpodella spp. than those from Goatflock1 (χ2=92.10; P = 8.2 × 10 -22 ) and Doggroup (χ2=42.34; P = 7.7 × 10 -11 ), and a significantly higher positive rate of T. luwenshuni than Doggroup (χ2=5.38; P = 0.02). However, the positive rates of T. luwenshuni between Goatflock1 and Goatflock2 were not significantly different (χ2=2.02; P = 0.16), and so as the positive rates of both pathogens between Goatflock1 and Doggroup groups (P > 0.05). For either Colpodella spp. or T. luwenshuni, no significant difference was found in prevalence between male and female ticks. These findings underscore the potential importance of Colpodella spp. in domestic animal-attached ticks, as our study revealed two novel Colpodella spp. and identified Colpodella spp. in H. longicornis for the first time. The study also sheds light on goats' potential roles in the transmission of Colpodella spp. to ticks and provides crucial epidemiological data of pathogenic Theileria and Colpodella. These data may help physicians, veterinarians, and public health officers prepare suitable detection and treatment methods and develop prevention and control strategies., (Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Amblyomma sculptum (formerly Amblyomma cajennense) ticks have been implicated in the transmission of pathogens that cause diseases in animals and humans. Their wide geographic distribution and high impact on animal health and zoonotic disease transmission highlight the importance of studying and implementing effective control measures to mitigate the risks associated with this tick species. The aim of this study was to quantify and characterize the morphology and the ultrastructure of different types of hemocytes in the hemolymph in engorged A. sculptum females fed on rabbits. The hemolymph samples were collected by perforation of the cuticle in the dorsal region. Hemocyte types, sizes, and differential counts were determined using light microscopy, while ultrastructural analysis of hemocytes was performed using transmission electron microscopy. The average number of total hemocytes in the hemolymph was 1024 ± 597.6 cells µL -1 . Five morphologically distinct cell types were identified in A. sculptum females: prohemocytes (6 % ± 8.8), plasmatocytes (10 % ± 7.7), granulocytes (78 % ± 12.2), spherulocytes (5 % ± 4.48), and oenocytoids (1 % ± 1.6). In general, prohemocytes were the smallest hemocytes. The ultrastructural morphology of A. sculptum hemocytes described in the present study agrees with the findings for other hard ticks. This is the first study to investigate ultrastructural characteristics of hemocytes of female A. sculptum ticks., (Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

In several urban and peri‑urban areas of Brazil, populations of Amblyomma sculptum and Amblyomma dubitatum ticks are maintained by capybaras (Hydrochoerus hydrochaeris). In some of these areas, this host and these tick species are associated with Brazilian spotted fever (BSF), a lethal human disease caused by the bacterium Rickettsia rickettsii. In this work, we evaluated the risk of human exposure to these tick species using four collection techniques to discern host-seeking behavior. The study was carried out in 10 urban sites inhabited by capybaras in Uberlândia, a BSF-free municipality in southeastern Brazil. Ticks were collected in areas of 400 m 2 at each site and at three seasons. Within the same municipality, the distance and speed of A. sculptum nymphs moving towards the CO 2 traps were evaluated. In a sample of ticks Rickettsia DNA was investigated. During the study period, 52,953 ticks were collected. Among these, 83.4 % were A. sculptum (1,523 adults, 10,545 nymphs and 32,104 larvae) and 16.6 % were A. dubitatum (464 adults, 2,153 nymphs and 6,164 larvae). An average annual questing tick density of 4.4/m² was observed, with the highest density recorded at one site in autumn (31.8/m²) and the lowest in summer at another site (0.03/m²). The visual search yielded the highest proportion of A. sculptum larvae, constituting 47 % of the total and 63.6 % of all A. sculptum larvae. In contrast, CO 2 traps collected a greater proportion of nymphs and adults of A. sculptum ticks. In the case of A. dubitatum, the CO 2 trap was the most efficient technique with 57.7 % of captures of this species, especially of nymphs (94.5 % of captures) and adults (97.8 % of captures). Ticks' ambush height on vegetation (9 to 77 cm), observed by visual search 30 times, yielded a total of 20,771 ticks. Of these, 28 (93 %) were A. sculptum ticks, with only two (7 %) identified as A. dubitatum ticks. Among A. sculptum ticks, the nymph was the most attracted stage to humans and larva in the case of A. dubitatum. Amblyomma sculptum adults and nymphs were significantly more attracted to humans than those of A. dubitatum, but A. dubitatum larvae were significantly more attracted than the same stage of A. sculptum. The maximum distance and speed of horizontal displacement for A. sculptum nymphs were five meters and 2.0 m/h, respectively. The only species of Rickettsia detected in ticks, exclusively in A. dubitatum, was R. bellii. Importantly, it was observed that the higher the proportion of A. sculptum in the community of ticks, the lower the rate of infection of A. dubitatum by R. bellii. In conclusion, host-seeking behavior differed between the two tick species, as well as between stages of the same species. A greater restriction of A. dubitatum ticks to the soil was observed, while larvae and nymphs of A. sculptum dispersed higher in the vegetation. The behavior presented by A. sculptum provides greater opportunities for contact with the hosts, while A. dubitatum depends more on an active search for a host, the hunter behavior. Taken together, these observations show that a human being crossing an area infested with A. sculptum and A. dubitatum ticks will have almost exclusive contact with A. sculptum larvae and/or nymphs. Humans in a stationary position (sitting, lying or immobile) are exposed to both tick species, but they are more attractive to adults and mainly nymphs of A. sculptum compared to the corresponding stages of the tick A. dubitatum. The negative effect of A. sculptum on A. dubitatum infection by R. bellii deserves further studies., Competing Interests: Declaration of competing interest The authors declared that no competing interests exist., (Copyright © 2024. Published by Elsevier GmbH.)

Tick-borne microorganisms in many tick species and many areas of China are still not thoroughly investigated. In this study, 224 ticks including two species (Haemaphysalis longicornis and Haemaphysalis qinghaiensis) were collected from four cities in Hebei, Shandong, and Qinghai provinces, China. Ticks were screened for the presence of tick-borne bacterial microorganisms including Rickettsia, Anaplasmataceae (Anaplasma, Ehrlichia, Neoehrlichia, etc.), Coxiella, Borrelia, and Bartonella. Two Anaplasma species (Anaplasma ovis and Anaplasma capra) were detected in H. longicornis from Xingtai City of Hebei Province, with a positive rate of 3 % and 8 %, respectively. A Coxiella species was detected in H. longicornis ticks from all three locations in Hebei and Shandong provinces, with the positive rate ranging from 30 to 75 %. All the 16S and rpoB sequences were very similar (99.77-100 % identity) to Coxiella endosymbiont of Haemaphysalis ticks. An Ehrlichia species was detected in H. qinghaiensis (6/66, 9 %) from Xining City, Qinghai Province. The 16S and groEL sequences had 100 % and 97.40-97.85 % nucleotide identities to "Candidatus Ehrlichia pampeana" strains, respectively, suggesting that it may be a variant of "Candidatus Ehrlichia pampeana". All the ticks were negative for Rickettsia, Borrelia, and Bartonella. Because all the ticks were removed from goats or humans and were partially or fully engorged, it is possible that the microorganisms were from the blood meal but not vectored by the ticks. Our results may provide some information on the diversity and distribution of tick-borne pathogens in China., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Beef cattle production in Portugal is an important sector of national agricultural production, with half of the herd being in the Alentejo region. Despite this, animal health is essential for its productivity, which may be compromised by ticks and tick-borne diseases. So far, no study has been conducted in Portugal to assess knowledge, attitudes, and practices (KAP) on ticks and tick-borne diseases in cattle, which the authors are aware of. This type of questionnaire is a very useful tool in the development and application of effective and sustainable prevention and control measures. Therefore, a KAP questionnaire was applied to 44 cattle breeders of autochthonous Portuguese breeds, namely 14 breeders of the Alentejana breed and 30 of the Mertolenga breed, between January 1 and May 9, 2023. Based on the analysis criteria of these surveys, 64% of the Alentejana breeders and 63% of the Mertolenga breeders have an average level of knowledge about ticks and tick-borne diseases, and 21% of the Alentejana breeders and 33% of the Mertolenga breeders have a high level of knowledge. Although only 21.4% of the Alentejana and 36.7% of the Mertolenga breeders consider tick infestation as a major animal health problem, 71.4% of the Alentejana and 63.3% of breeders of the Mertolenga state that one of the main reasons for veterinary consultations on their farm is deworming of animals, and 92.9% of breeders of the Alentejana and 96.7% of breeders of the Mertolenga refer the use of dewormers as a strategy to control tick infestation. The results of this study contribute to highlighting the importance of correcting some identified knowledge gaps and improving knowledge, especially on the life cycle of this parasite, its local distribution and seasonality, resistance to acaricides, and alternative control strategies., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests. Jacinto Gomes reports financial support was provided by Foundation for Science and Technology. Jacinto Gomes reports financial support was provided by LEAP-Agri., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The Amblyomma genus is represented on the African continent by 24 species, out of which 17 are known to occur in different ecological niches of southern Africa. Amblyomma, known for their aggressive hunting behaviour and aptitude as pathogen vectors, are of main concern to travellers, mainly in rural and conservation areas of Africa. In this study, we highlight the overlapping distribution of Amblyomma eburneum and Amblyomma variegatum found on African buffaloes (Syncerus caffer) at Coutada 11, Central Mozambique. In total, 1,039 Amblyomma ticks were collected and morphologically identified using taxonomic keys, and genomic DNA was extracted. They were subjected to reverse line blotting for pathogen identification followed by molecular analysis (COI sequencing) of both tick species. Pathogens such as Ehrlichia ruminantium, Anaplasma centrale, Theileria sp., Babesia sp. and Rickettsia africae were detected, of which R. africae is zoonotic. Ehrlichia ruminantium, R. africae, Theileria mutans and Theileria velifera are well-established pathogens transmitted by Amblyomma ticks; however, Anaplasma spp. and Babesia spp. are not, suggesting residual parasite DNA in the bloodmeal. Little is mentioned in the literature about A. eburneum, including its role as a vector and reservoir for pathogens. In Mozambique A. eburneum is currently restricted to wildlife but the spread of the tick may be observed given the climate change that is occurring. The infection rates for the pathogens in both Amblyomma tick species were lower than expected, but this may be due to the low host density in the forest niche and the innate immunity of these hosts. With the propensity of ticks of the Amblyomma genus to form parapatric distributions, the mechanisms that allows for the overlapping distribution of these two Amblyomma species while maintaining tick species identity is of great interest., Competing Interests: Declaration of Competing Interest None, (Copyright © 2023. Published by Elsevier GmbH.)

New technologies have led to the discovery of novel tick-borne and tick-associated viruses. Dabieshan tick virus (DaTV) and Okutama tick virus (OkTV), which belong to the family Phenuiviridae, were discovered in ticks in China and Japan, respectively, in the 2010s. Although it is unknown whether these viruses cause disease in animals or humans, all tick-associated viruses have the potential to become etiological agents of infectious diseases through gene reassortment. Therefore, it is important to elucidate the ecology of these viruses, regardless of their pathogenicity. In this study, ticks were collected year-round in Cape Toi, Miyazaki Prefecture, Japan, and an epidemiological survey of tick-associated phenuiviruses was performed. A total of 516 ticks collected from the vegetation by dragging flannel sheets were used for analysis. Pan-phenuivirus reverse transcription PCR was performed on the tick samples, and DaTV and OkTV were detected. We found that 37.0% (85/230) and 23% (16/71) of nymphal and adult Haemaphysalis longicornis were infected with DaTV, respectively, and 10% (6/62) and 13% (1/8) of nymphal and adult Haemaphysalis flava were infected with OkTV, respectively. Phylogenetic analysis indicated that the DaTV identified in this study formed a unique clade that was distinct from the strains identified in China. The survey revealed that DaTV is distributed not only in China, but also in Japan. We believe that this study contributes to our understanding of the prevalence of tick-associated viruses., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest associated with this manuscript., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Ticks are blood-feeding ectoparasites that devastate cattle farming and are an omnipresent nuisance to pets and humans, posing a threat of pathogen transmission. Laboratory experimental models can be instrumental in the search for molecular targets of novel acaricides or vaccines. Mainly, though, the experimental models represent invaluable tools for broadening our basic understanding of key processes of tick blood-feeding physiology and vector competence. In order to understand the function of a single component within the full complexity of a feeding tick, genetic or biochemical interventions are used for systemic phenotypisation. In this work, we summarise current experimental modalities that represent powerful approaches for determining biological functions of tick molecular components., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

In spring, migratory birds reach Europe, mainly from sub-Saharan Africa or from northern African countries. Avian species may be implicated in the spread of pathogens, either as reservoirs, hosts or carriers of infected ectoparasites. In 2021, on Ventotene Island (Latium region, Italy) within a project focused on the potential incoming pathogens via migratory birds from Africa, we found two larvae of Argas sp., on the redstart Phoenicurus phoenicurus, that shared morphological features with the African Argas (Argas) africolumbae. Comparison of the tested larval DNA sequences to the adult reference sequences showed the highest identity (> 92%) with homologous sequences of A. africolumbae collected in South Africa and in Spain. This study reports the first detection of Argas africolumbae-like specimens in Italy., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Ticks pose a major threat to the health of humans and animals. The use of synthetic acaricides and repellents has raised the concerns of potential health and environmental risks and increasing resistance in ticks. This article highlights the importance of the research on tick chemosensation in developing novel control agents. It provides a review on our current understanding of tick chemosensory system and proposes using chemosensory receptor (CR) genes as molecular targets to discover novel tick control agents. The releases of high-quality tick genomes provide unprecedented opportunities to explore CR gene repertoires. Further functional characterization is necessary to identify the receptors for key chemical cues and signals and unravel whether tick chemosensation involves ionotropic and/or metabotropic mechanisms., Competing Interests: Declaration of Competing Interest The author declares no conflicts of interest., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Spotted fever caused by the bacterium Rickettsia parkeri, and canine rangeliosis caused by the protozoan Rangelia vitalii, are emerging or re-emerging tick-borne diseases in Brazil, where the main tick vectors are Amblyomma ovale and Amblyomma aureolatum, respectively. In the present study, we evaluated exposure to R. parkeri and R. vitalii infection among domestic (Canis lupus familiaris) and wild (Cerdocyon thous) canids sharing living areas within an Atlantic Forest area of southern Brazil. Four different domestic dogs and three different wild canids were captured. Most of the ticks collected from either domestic or wild canids were identified as A. ovale and A. aureolatum. DNA of R. parkeri was amplified from 14% of the A. ovale specimens. Two domestic dogs and two wild canids were seroreactive to R. parkeri antigens with high endpoint titers (>1024). Rangelia vitalii DNA was detected in two wild canids; one of them was resampled 93 days later, again infected by R. vitalii. We report exposure/infection of domestic dogs and wild canids to R. vitalii and/or R. parkeri-infected ticks in an Atlantic Forest area shared by both canid species, indicating that they also shared the same populations of the tick vectors, A. aureolatum and A. ovale. While A. ovale, A. aureolatum, R. parkeri, R. vitalii and C. thous, are all native to the Atlantic Forest, further studies are needed to evaluate the impact of an exotic canine species - C. lupus familiaris - on the enzootic cycles of R. vitalii and R. parkeri, as well as the possible role of domestic dogs in emergence and re-emergence of R. parkeri-spotted fever in humans and canine rangeliosis, respectively., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Ticks are vectors for several pathogens, including bacteria belonging to the Rickettsia genus, such as Rickettsia rickettsii and Rickettsia parkeri, the causative agents of spotted fever. The aim of the present study was to investigate the tick species richness and rickettsial agents associated with wild birds captured in the Humaita Forest Reserve, Acre, in the Western Amazon region. Wild birds were captured with ornithological nets for visual inspection with the purpose of collecting ticks, which were identified through morphological analyses and molecular tests for several genes (12S rDNA, 16S rDNA, gltA, ompA, and sca4). A total of 607 wild birds were captured, 12% of which were parasitized by 268 ticks of the Amblyomma genus, with new host-parasite associations reported for Amblyomma calcaratum, Amblyomma geayi, Amblyomma longirostre, Amblyomma naponense, Amblyomma nodosum, and Amblyomma varium. Of the total ticks collected, 113 were tested for the presence of rickettsial DNA fragments, with 19 testing positive for R. parkeri in A. geayi, Rickettsia tamurae-like in Amblyomma sp., and Rickettsia amblyommatis in A. geayi, A. longirostre, and Amblyomma sp. We detected R. tamurae-like in Amblyomma larvae for the first time in the Western Brazilian Amazon biome, and registered spotted fever group rickettsiae, although the relevance of the detected species in a public health context should be further explored in South America, as well as new host-parasite interactions in this underexplored region., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Tick eggs contain all essential proteins for embryogenesis, and egg proteins are a potential reservoir of tick-protective antigens. However, the protein profile and dynamics during embryonic development remain unknown. This study aimed to depict the protein profile and dynamics in tick embryogenesis, further providing protein candidates for targeted interventions. Eggs from Haemaphysalis flava ticks were incubated at 28 °C and 85% relative humidity. On days 0 (newly laid eggs without incubation), 7, 14 and 21, eggs were collected, dewaxed and subject to protein extraction. Extracted proteins were digested by filter-aided sample preparation and analyzed by liquid chromatography-tandem mass spectrometry (LC/MS-MS). MS data were searched against an in-house H. flava protein database for tick-derived protein identification. Abundances of 40 selected high-confidence proteins were further quantified by LC-parallel reaction monitoring (PRM)/MS analysis throughout egg incubation. A total of 93 high-confidence proteins were identified in eggs on 0-day incubation. Identified proteins belonged to seven functional categories: transporters, enzymes, proteinase inhibitors, immunity-related proteins, cytoskeletal proteins, heat shock proteins and uncharacterized proteins. The enzyme category contained the most types of proteins. Neutrophil elastase inhibitors represented the most abundant proteins in terms of intensity-based absolute-protein-quantification. LC-PRM/MS revealed that the abundances of 20 proteins increased including enolase, calreticulin, actin, GAPDH et cetera, and the abundances of 11 proteins decreased including vitellogenins, neutrophil elastase inhibitor, carboxypeptidase Q, et cetera from 0- to 21-day incubation. This study provides the most comprehensive egg protein profile and dynamics during tick embryogenesis. Further investigations are needed to test the tick-control efficacy by targeting the egg proteins., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Amblyomma is an important tick genus for animal and human health, with some species being the vectors of zoonotic pathogens, such as Rickettsia rickettsii, in the Neotropical region. Knowing their hosts may help to understand the distribution of these agents and decrease the occurrence of clinical cases. Primates are intelligent and adaptable animals that can get close to humans in the search for food. So, they may be an important epidemiological link for the spread of these ticks. Beyond that, primates also suffer from these infections, serving as sentinels for different diseases. Thus, the present study aims to report the parasitism by Amblyomma spp. on six species of Neotropical primates from different locations in Brazil. The 337 collected ticks were morphologically identified using stereomicroscopes and taxonomic keys, and six distinct species of ticks were identified. We report here the first record of nymphs of the tick species Amblyomma cajennense sensu stricto on Alouatta belzebul, a male of Amblyomma fuscum on Alouatta guariba clamitans, nymphs of Amblyomma sculptum on Leontopithecus chrysopygus and Callithrix aurita, as well as nymphs of Amblyomma geayi on Saimiri collinsi. Of the 337 tick specimens collected, 256 (75,96%) were nymphs. The importance of primates in the life cycle of these species remains to be elucidated., Competing Interests: Declaration of Competing Interest The aforementioned authors declare that there is no conflict of interest in the production of this manuscript., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Maps of the distribution of medically-important ticks throughout the US remain lacking in spatial and temporal resolution in many areas, leading to holes in our understanding of where and when people are at risk of tick encounters, an important baseline for informing public health response. In this work, we demonstrate the use of Bayesian Experimental Design (BED) in planning spatiotemporal surveillance of disease vectors. We frame survey planning as an optimization problem with the objective of identifying a calendar of sampling locations that maximizes the expected information regarding some goal. Here we consider the goals of understanding associations between environmental factors and tick presence and minimizing uncertainty in high risk areas. We illustrate our proposed BED workflow using an ongoing tick surveillance study in South Carolina parks. Following a model comparison study based on two years of initial data, several techniques for finding optimal surveys were compared to random sampling. Two optimization algorithms found surveys better than all replications of random sampling, while a space-filling heuristic performed favorably as well. Further, optimal surveys of just 20 visits were more effective than repeating the schedule of 111 visits used in 2021. We conclude that BED shows promise as a flexible and rigorous means of survey design for vector control, and could help alleviate pressure on local agencies by limiting the resources necessary for accurate information on arthropod distributions. We have made the code for our BED workflow publicly available on Zenodo to help promote the application of these methods to future surveillance efforts., (Copyright © 2024. Published by Elsevier GmbH.)

The Amblyomma marmoreum complex includes afrotropical species, such as Amblyomma sparsum, a three-host tick that parasitizes reptiles, birds, and mammals, and is a recognized vector of Ehrlichia ruminantium. However, the lack of morphological, genetic and ecological data on A. sparsum has caused considerable confusion in its identification. In this study, we used microscopy and metagenomic approaches to analyze A. sparsum ticks collected from a puff adder snake (Bitis arietans) in southwest Senegal (an endemic rickettsioses area) in order to supplement previous morphological descriptions, provide novel genomic data for the A. marmoreum complex, and describe the genome of a novel spotted fever group Rickettsia strain. Based on stereoscope and scanning electron microscopy (SEM) morphological evaluations, we provide high-quality images and new insights about punctation and enameling in the adult male of A. sparsum to facilitate identification for future studies. The metagenomic approach allowed us assembly the complete mitochondrial genome of A. sparsum, as well as the nearly entire chromosome and complete plasmid sequences of a novel Rickettsia africae strain. Phylogenomic analyses demonstrated a close relationship between A. sparsum and Amblyomma nuttalli for the first time and confirmed the position of A. sparsum within the A. marmoreum complex. Our results provide new insights into the systematics of A. sparsum and A. marmoreum complex, as well as the genetic diversity of R. africae in the Afrotropical region. Future studies should consider the possibility that A. sparsum may be a vector for R. africae., (Copyright © 2024. Published by Elsevier GmbH.)

The Internet is an important gateway for accessing health-related information, and data generated through web queries have been increasingly used as a complementary source for monitoring and forecasting of infectious diseases and they may partially address the issue of underreporting. In this study, we assessed whether tick-borne encephalitis (TBE)-related Internet search volume may be useful as a complementary tool for TBE surveillance in Italy. Monthly Google Trends (GT) data for TBE-related information were extracted for the period between January 2017 and September 2022, corresponding to the available time series of TBE notifications in Italy. Time series modeling was performed by applying seasonal autoregressive integrated moving average (SARIMA) models with or without GT data. The search terms relative to tick bites reflected best the observed temporal distribution of TBE cases, showing a correlation coefficient of 0.81 (95 % CI: 0.71-0.88). Particularly, both the reported number of TBE cases and GT searches occurred mainly during the summer. The peak of disease notifications coincided with that of Google searches in 4 of 6 years. Once calibrated, SARIMA models with or without GT data were applied to a validation set. Retrospective forecast made by the model with GT data was associated with a lower prediction error and accurately predicted the peak timing. By contrast, the traditional SARIMA model underestimated the actual number of TBE notifications by 65 %. Timeliness, easy availability, low cost and transparency make monitoring of the TBE-related Internet search queries a promising addition to the traditional methods of TBE surveillance in Italy., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

The Third International Conference on Crimean-Congo Hemorrhagic Fever (CCHF) was held in Thessaloniki, Greece, September 19-21, 2023, bringing together a diverse group of international partners, including public health professionals, clinicians, ecologists, epidemiologists, immunologists, and virologists. The conference was attended by 118 participants representing 24 countries and the World Health Organization (WHO). Meeting sessions covered the epidemiology of CCHF in humans; Crimean-Congo hemorrhagic fever virus (CCHFV) in ticks; wild and domestic animal hosts; molecular virology; pathogenesis and animal models; immune response related to therapeutics; and CCHF prevention in humans. The concluding session focused on recent WHO recommendations regarding disease prevention, control strategies, and innovations against CCHFV outbreaks. This meeting report summarizes lectures by the invited speakers and highlights advances in the field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier B.V.)

The bites of hard ticks are the major route of transmission of tick-borne infections to humans, causing thousands of cases of diseases worldwide. However, the characteristics of the human population that is exposed to tick bites are still understudied. This work is aimed at characterizing both the structure of the population directly contacting ticks and the human behavioral features associated with tick bites. We studied 25,970 individuals who sought medical help after a tick bite at the Centre for Diagnostics and Prevention of Tick-borne Infections (CDPTBI) in Irkutsk City (Russian Federation). The demographic and behavioral characteristics of the human population were analyzed using z-tests for proportions, the Mann-Whitney U test, and the Spearman rank correlation coefficient. The majority of bitten people were urban residents (70 %), and most of them were either of active ages between 30 and 74 years old (62 %), or children between 0 and 9 years old (approximately 20%). Tick bites occurred mostly in the range of 150 km around the location of the diagnostic facility (83 %). In comparison to the general population, significant differences were revealed in the representation of different age groups among bitten people. The population affected by tick bites included fewer men and women in the ages of 10-29 and over 75 years old than would be predicted based on the demographics of the general population. Vice versa, the proportions of people in the ages of 5-9 and 60-74 increased among bitten people. Among men, such activities (in order of occurrence) as "leisure and recreation", "visiting allotments", "foraging for forest food", and "fulfilling work duties" tend to be more associated with tick bites. Among women, tick bites occurred mainly during "visiting allotments", "leisure and recreation", "visiting cemeteries" and "contact with pets and plants at home". The overall vaccination rate was 12 %; however, significantly more men than women were vaccinated against tick-borne encephalitis (up to 20 % vs. approximately 7 % respectively). The structure of the tick bite - affected population suggests that it is age-specific human behavior that mainly determines the frequency of contact between people and ticks. However, in several age groups, especially among children from 5 to 9 and people aged 30-39 years old, gender-related factors could significantly change the exposure of people to tick bites., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Tick infestation poses a serious threat to animal health, leading to significant losses in terms of vector-borne disease transmission, reduced live weight, lower quality hides, decreased milk production, and impaired reproduction in tropical and subtropical regions worldwide. This study aimed to determine the prevalence, seasonal variation, distribution pattern, and associated risk factors of Ixodid family tick species in the cattle and sheep population of three different districts in Balochistan, Pakistan. This study employed a convenient sampling method, collecting 4080 adult ticks from 816 cattle and sheep of various breeds, ages, and sexes. Specific morphological keys were used to identify the ticks up to the genus and species level. Among cattle, the highest prevalence was recorded for R. (B) annulatus (27.01%), followed by R. (B) microplus (24.02%), and H. anatolicum (20.54%). H. dromedarii (5.29%) was the least prevalent species observed in cattle. In the sheep population, H. anatolicum (30.34%) showed the highest prevalence, followed by H. marginatium (22.99%), and R. (B) annulatus (20.88%). H. dromedarii (6.96%) was the least prevalent species observed in sheep. The prevalence of R. (B) decoloratus, H. anatolicum and H. dromedarii was found to be significantly associated (P < 0.05) with the breed, age, and sex of both cattle and sheep. However, the presence of R. (B) annulatus, R. (B) microplus and H. marginatium tick species showed no significant association (P > 0.05) with these factors. In addition, the prevalence of ticks was higher in younger, female, and crossbred Friesian cattle compared to adults, males, and other breeds. Conversely, the prevalence of ticks was higher in adult, female and Hernai breed of sheep in the studied area. In conclusion, R. (B) annulatus and H. anatolicum are the dominant tick species infesting the cattle and sheep population in Balochistan. Consequently, this study provides valuable insights for developing practical and effective control measures against ticks and tick-borne diseases in the sheep and cattle population of Balochistan, Pakistan., Competing Interests: Declaration of competing interest All other authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier B.V.)

As obligate blood-feeders, ticks serve as vectors for a variety of pathogens that pose threats on both human and livestock health. The microbiota that ticks harbor play important roles in influencing tick nutrition, development, reproduction, and vector. These microbes also affect the capacity of ticks to transmit pathogens (vector competence). Therefore, comprehending the functions of tick microbiota will help in developing novel and effective tick control strategies. Here, we summarize the effects of main tick symbiotic bacteria on tick physiology and vector competency., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Ticks are obligatory hematophagous arachnids, serving as vectors for a wide array of pathogens that can be transmitted to humans or animals. The ability of tick-borne pathogens to maintain within natural reservoirs is intricately influenced by the attractiveness of ticks to their animal hosts, including humans. However, the complex dynamics of tick behavior and host-seeking strategies remain understudied. This review aims to summarize the impact of volatiles or odors on tick behavior and vector competence. Our literature review has identified a selection of compounds, such as 1-octen-3-ol, hexanal, heptanal, nonanal, 6-methyl-5-hepten-2-one, acetone, and octanal, as having the potential to impact both ticks' and mosquitos' behaviors. In addition, carbon dioxide (CO 2 ) is a universal attractant for hematophagous arthropods. Moreover, we have gathered some clues indicating that volatiles emitted by infected animal hosts might play a role in the transmission of tick-borne pathogens. Nonetheless, our understanding of this phenomenon remains largely inadequate, particularly with regarding to whether the tick microbiome or the skin microbiota of the feeding mammals, including humans, can actively modulate tick-host-seeking behavior. Further investigations in this emerging field hold immense promise for the development of innovative strategies aimed at controlling vectors and curtailing the spread of tick-borne diseases., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interest., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Ticks pose a serious threat to public health as carriers and often vectors of zoonotic pathogens. There are few systematic studies on the prevalence and genetic diversity of tick-borne bacterial pathogens in Western China. In this study, 465 ticks were collected from free-ranging sheep in Gansu Province in China. Ticks were divided into 113 pools and tick DNA was extracted from these ticks. PCR assays were performed using specific primers to screen for tick-borne pathogens as well as sequence analysis based on the 16S rRNA (rrs), ompB, gltA, ompA genes for Rickettsia, rrs, groEL genes for Anaplasma, and ssrA and rpoB genes for Bartonella. The PCR results showed that the minimum infection rates with Rickettsia, Anaplasma, and Bartonella were 16.8% (78/465), 18.9% (88/465), and 0.9% (4/465), respectively. Sequence analysis based on the concatenated sequences of rrs-ompB-gltA-ompA indicated that the Rickettsia species identified in the ticks belonged to Rickettsia raoultii, Rickettsia slovaca, and Rickettsia sibirica, respectively; phylogenetic analysis based on the groEL gene showed that all Anaplasma strains identified were Anaplasma ovis; and phylogenetic analysis based on the ssrA and rpoB genes indicated that all Bartonella strains in the ticks belonged to Bartonella melophagi. The results of this study showed that ticks in Gansu Province harbored multiple pathogens that may cause rickettsial diseases and bartonellosis. These diseases were neglected in the area and physicians and public health workers need to pay attention to their diagnoses to prevent human infection., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Tick saliva is a reservoir of bioactive proteins. Saliva protein compositions change dynamically during blood-feeding. Decipherment of protein profiles in different blood-feeding stages may bring deeper insight into tick feeding physiology and provide targets for immunologic control alternatives. However, having the infancy of tick genome sequencing, assembly, annotation, and limited knowledge of tick salivary proteins restrain the data interpretation. Here, we aimed to depict the saliva protein profile in partially- (PE) and fully-engorged (FE) Haemaphysalis flava ticks, with a special focus on the analysis of those uncharacterized proteins. Saliva was collected from PE and FE adult female H. flava ticks. Saliva proteins were analyzed by high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS-MS). MS data were searched against an in-house salivary gland transcriptome library for identification of tick-derived proteins. Abundances of proteins were compared between PE and FE ticks. The uncharacterized proteins detected in saliva were further bioinformatically analyzed. In total, 614 proteins were identified including 94 host proteins and 520 tick-derived proteins. The 226 tick-derived high-confidence proteins were classified into 10 categories: transporters, enzymes, protease inhibitors, immunity-related proteins, lipocalins, glycine-rich proteins, muscle proteins, secreted proteins, uncharacterized proteins and others. A total of 98 proteins were shared in both PE and FE with 74 only in PE and 54 only in FE. Abundances of 24 shared proteins were significantly higher in PE. The profile of top 15 most abundant proteins was also different between PE and FE ticks. The 65 uncharacterized proteins detected in tick saliva were branched into subclusters 1 A, 1B, 2, 3 A, 3B and 3 C based on particular motifs like RGD, LRR, indicating their diverse predicted functions like anti-coagulation, regulation of innate immune, or other functions. This study provides and compares saliva proteomes of H. flava ticks in two feeding stages with special cluster analysis on the uncharacterized proteins. Further investigations are needed to confirm the roles of these uncharacterized proteins in ticks., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Immature hard ticks from the genus Amblyomma feed on blood from a wide range of Neotropical avian hosts. They serve as vectors for pathogens of medical and veterinary importance, such as Rickettsia agents of the spotted fever group (SFG). Hence, determining ecological factors that increase encounter rates between immature ticks and their avian hosts may contribute to the understanding of tick-borne diseases transmission. Here, we used 720 individual birds from 96 species surveyed in the Brazilian Pantanal to test whether host breeding season influenced tick infestation probabilities. Additionally, collected ticks were screened for Rickettsia agents to describe new avian-tick-bacteria associations. Our models revealed that the probability of an individual bird being infested with immature ticks was similar during the breeding and pre-breeding season, but higher loads of immature tick stages were found during the breeding season. Host sex did not predict infestation probability, but Rickettsia agents recovered from ticks were more prevalent during the pre-breeding season. The new records of host usage by larvae and nymphs of Amblyomma in Pantanal and the growing body of tick surveys in Neotropical avian communities, suggest that immature ticks may benefit from avian blood sources during their annual cycle. The low number of infected ticks with Rickettsia agents on Pantanal birds suggest that this vertebrate group are likely not acting as reservoirs for these microorganisms. However, long-term surveys at the same site are imperative to determine which tick species are acting as reservoirs for Rickettsia agents in Pantanal and determine whether birds are playing a role in dispersing ticks and tick-borne pathogens., (Copyright © 2023. Published by Elsevier GmbH.)

Haemaphysalis species are medically important hard ticks (Ixodidae) that parasitize nearly all domestic as well as wild hosts, mostly in the Oriental region. Genetic diversity of Haemaphysalis ticks in South Asia has gained little attention, despite its economic, medical, and veterinary importance. The purpose of this study was to genetically characterize Haemaphysalis indica and Haemaphysalis montgomeryi ticks to obtain their preliminary phylogenetic relationship. A total of 1171 ticks were collected from Indian mongooses (Herpestes javanicus and Herpestes edwardsii), sheep and goats in Pakistan. The collected ticks were morphologically identified as H. indica (n = 121, 10.3%) from mongooses and H. montgomeryi (n = 1050, 89.7%) from sheep and goats. Phylogenetic trees inferred from 12S rDNA, 16S rDNA, cox1, and ITS2 sequences revealed an evolutionary relationship of H. indica with the tick species belonging to the subgenus Rhipistoma, and H. montgomeryi with the tick species belonging to the subgenus Segalia, validating the described taxonomy based on the morphological descriptions of these ticks. Further genetic studies are essential to overcome morphology-based systematic issues associated with Haemaphysalinae ticks., (Copyright © 2022. Published by Elsevier GmbH.)

We report on the presence of the ixodid tick Amblyomma helvolum on three species of reptiles from Timor-Leste. Among a total of 21 host specimens (lizards: 18 four-fingered skinks, Carlia sp. 'Meleotegi' and two forest skinks, Sphenomorphus sp. 'Meleotegi'; snake: one Coelognathus subradiatus) four were parasitized by ticks. Whereas nymphs were associated with the lizards, an adult male was retrieved from the snake. This report is the first of A. helvolum for Timor-Leste, the first for the skink genera Carlia and Sphenomorphus, and the first for C. subradiatus. We present a comprehensive table with updated taxonomy of known associations between A. helvolum and other vertebrates., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Severe fever with thrombocytopenia syndrome virus (SFTSV), a tick-borne Bunyavirus, causes an emerging hemorrhagic fever in humans with a high fatality in Asia. The tick vectors and hosts of SFTSV are not well studied. We evaluated SFTSV transmission in laboratory reared Haemaphysalis flava ticks. RT-PCR demonstrated that after acquisition feeding in SFTSV-infected rabbits, 10 % (4/40) engorged larvae, 25% (5/20) engorged nymphs, and 50% (5/10) engorged females of H. flava became SFTSV RNA positive; after engorged larvae and nymphs molted into nymphs and adults, respectively, 12.5% (3/24) newly molted nymphs and 20% (2/10) newly molted adults were SFTSV RNA positive. Among 30 engorged females that oviposited, 10% (3/30) clutches of eggs and 3.3% (1/30) colonies of larvae were RNA positive for SFTSV. RT-PCR also showed that 6 days after being infested with SFTSV-infected ticks, 100% (3/3) rabbits infested with larvae, 100% (2/2) rabbits infested with nymphs, and 100% (2/2) rabbits infested with adult ticks became SFTSV RNA positive. In conclusion, H. flava can acquire SFTSV from infected rabbits by feeding; there is transstadial and transovarial transmission of the virus and all three stages of H. flava can transmit SFTSV to rabbits by feeding. Thus, H. flava tick is an effective vector of SFTSV and may play a role in the transmission of SFTSV in wild animals and humans., (Copyright © 2022. Published by Elsevier GmbH.)

In the past two decades, new species of Rickettsia have been detected and described worldwide, some of them considered pathogenic for humans. Although Costa Rica is considered a biodiversity hotspot, the knowledge about rickettsiae in sylvatic ecosystems and wild animals is scarce. The aim of this preliminary study was to detect and identify species of Rickettsia in ticks collected from wild animals in Costa Rica. A total 119 ticks were collected from 16 animal host species belonging to diverse vertebrate families (Didelphidae, Procyonidae, Felidae, Choloepodidae, Bradypodidae, Myrmecophagidae, Tayassuidae, Tapiridae, Phyllostomidae, Bufonidae, Geoemydidae, Boidae, Colubridae), and they were grouped into 43 pools to detect the presence of Rickettsia spp. DNA by PCR targeting the gltA gene. In positive pools, amplicons of the ompA, sca5 (ompB), and/or htrA genes were also amplified to identify the species present. The identification of some ticks was also confirmed by molecular methods. Four species of Rickettsia were detected in eight (19%) tick pools: Rickettsia amblyommatis in four pools of Amblyomma geayi (host: Caluromys derbianus) and one pool of Amblyomma cf. parvum (host: Nasua narica), Rickettsia rhipicephali in one pool of Dermacentor latus (host: Tayassu pecari), 'Candidatus Rickettsia colombianensi' in one pool of Amblyomma sp. nymphs (host: Boa constrictor), and Rickettsia sp. genotype IbR/CRC in one pool of Ixodes cf. boliviensis (host: Puma concolor). This is the first molecular detection of R. rhipicephali in Central America, and of 'Candidatus R. colombianensi' in Costa Rica. Results show that diverse wild animals and their ticks are associated with several species of rickettsiae in Costa Rica, which may come in contact with humans and other domestic animals in sylvatic environments., Competing Interests: Declaration of Competing Interest Authors declare no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Heartland and Bourbon viruses are pathogenic tick-borne viruses putatively transmitted by Amblyomma americanum, an abundant tick species in Missouri. To assess the prevalence of these viruses in ticks, we collected 2778 ticks from eight sampling sites at Tyson Research Center, an environmental field station within St. Louis County and close to the City of St. Louis, from May - July in 2019 and 2021. Ticks were pooled according to life stage and sex, grouped by year and sampling site to create 355 pools and screened by RT-qPCR for Bourbon and Heartland viruses. Overall, 14 (3.9%) and 27 (7.6%) of the pools were positive for Bourbon virus and Heartland virus respectively. In 2019, 11 and 23 pools were positive for Bourbon and Heartland viruses respectively. These positives pools were of males, females and nymphs. In 2021, there were 4 virus positive pools out of which 3 were positive for both viruses and were comprised of females and nymphs. Five out of the 8 sampling sites were positive for at least one virus. This included a site that was positive for both viruses in both years. Detection of these viruses in an area close to a relatively large metropolis presents a greater public health threat than previously thought., (Copyright © 2022 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Phleboviruses are emerging pathogens of public health importance. However, their association with ticks is poorly described, particularly in Africa. Here, adult ticks infesting cattle, goats and sheep were collected in two dryland pastoralist ecosystems of Kenya (Baringo and Kajiado counties) and were screened for infection with phleboviruses. Ticks mainly belonged to the species Rhipicephalus appendiculatus, Hyalomma impeltatum, and Hyalomma rufipes. A fragment of the RNA-dependent RNA polymerase (RdRp) gene was identified in thirty of 671 tick pools, of which twenty-nine were from livestock sampled in Baringo county. Phylogenetic analyses revealed that twenty-five sequences were falling in three clades within the group of tick-associated phleboviruses. The sequences of the three clades showed nucleotide distances 8%, 19% and 22%, respectively, to previously known viruses suggesting that these sequence fragments may belong to three distinct viruses. Viruses of the group of tick-associated phleboviruses have been found in several countries and continents but so far have not been associated with disease in humans or animals. In addition, five sequences were found to group with the sandfly-associated phleboviruses Bogoria virus, Perkerra virus and Ntepes virus recently detected in the same region. Further studies are needed to investigate the transmission and maintenance cycles of these viruses, as well as to assess their potential to infect vertebrates., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest. The funders had no role in the study design, data collection and interpretation, or the decision to submit the work for publication., (Copyright © 2022. Published by Elsevier GmbH.)

Ticks are primary vectors for many tick-borne pathogens (TBPs) and pose a serious threat to veterinary and public health. Information on the presence of TBPs in Chinese Milu deer (Elaphurus davidianus) is limited. In this study, a total of 102 Chinese Milu deer blood samples were examined for Anaplasma spp., Theileria spp., Babesia spp., Rickettsia spp., and Borrelia spp., and three TBPs were identified: Anaplasma phagocytophilum (48; 47.1 %), Candidatus Anaplasma boleense (47; 46.1%), and Theileria capreoli (8; 7.8 %). Genetic and phylogenetic analysis of the 16S rRNA and 18S rRNA confirmed their identity with corresponding TBPs. To our knowledge, this is the first report on Candidatus A. boleense and T. capreoli detection in Chinese Milu deer. A high prevalence of A. phagocytophilum with veterinary and medical significance was identified in endangered Chinese Milu deer, which could act as potential zoonotic reservoirs. The identification of the TBPs in Chinese Milu deer provides useful information for the prevention and control of tick-borne diseases., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Ticks are blood-sucking ectoparasites that secrete immunomodulatory substances in saliva to hosts during engorging. Cystatins, a tick salivary protein and natural inhibitor of Cathepsins, are attracting growing interest globally because of the immunosuppressive activities and the feasibility as an antigen for developing anti-tick vaccines. This review outlines the classification and the structure of tick Cystatins, and focuses on the anti-inflammatory effects and molecular mechanisms. Tick Cystatins can be divided into four families based on structures and cystatin 1 and cystatin 2 are the most abundant. They are injected into hosts during blood feeding and effectively mitigate the host inflammatory response. Mechanically, tick Cystatins exert anti-inflammatory properties through the inhibition of TLR-NF-κb, JAK-STAT and p38 MAPK signaling pathways. Further investigations are crucial to confirm the reduction of inflammation in other cell types like neutrophils and mast cells, and fully elucidate the underlying mechanism (like the structural mechanism) to make Cystatin a potential candidate for the development of novel anti-inflammation agents., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Vector-borne diseases pose a significant threat to human and animal health worldwide. Their emergence is influenced by various factors such as environmental changes, host characteristics, and human behavior. The One Health approach is necessary to thoroughly investigate tick-borne diseases and understand the complex interactions between environmental, animal, and human health. Anthropogenic changes have impacted predators, leading to cascading effects on wildlife prey species and the emergence of vector-borne diseases. The increase in global trade and travel has led to the introduction of several invasive vector species, increasing the risk of zoonotic pathogen spillover. Tick and tick-borne disease research requires an interdisciplinary approach to address challenges in a One Health paradigm., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

In Central and Eastern Europe, wolf populations have been increasing over the last two decades, recolonizing areas from which the species had been previously exterminated. As wolves are still recovering after years of persecution by humans, recognizing pathogens infecting this species, including tick-borne infections, is crucial for its conservation. On the other hand the high mobility of wolves and their frequent contacts with humans, dogs, and other domestic species make them a potentially important zoonotic reservoir. In this paper, we used molecular methods to determine the prevalence of tick-borne pathogens in the following genera: Anaplasma, Babesia, Bartonella, Borrelia, and Rickettsia in 50 free-ranging wolves from Poland. We detected Babesia canis in the blood of nine individuals (prevalence 9/50=18 %). The obtained sequence showed the highest similarity to B. canis isolated from dogs and ticks, and all infected individuals originated from regions endemic to the ornate tick, Dermacentor reticulatus. Anaplasma phagocytophilum was found in tissue from one individual (1/50=2 %), and the sequence was assigned to the zoonotic ecotype I., (Copyright © 2023. Published by Elsevier GmbH.)

In this report we described a case of aural hematomas in three lambs associated with Otobius megnini (Ixodida: Argasidae) infestation. From April to May 2021, five 3-month-old Hampshire cross lambs presented with unilateral aural hematomas. Upon otoscopic examination, engorged soft ticks (O. megnini) were observed in the external ear canals of three of the five lambs. The remaining two lambs had lesions consistent with infestation and were in a shared environment and deemed likely to have been infected. The treatment of all animals was based on the drainage of the serosanguinous fluid through an incision in the internal space of the ear pinna. Upon physical inspection of the entire flock (n = 310), O. megnini infestation was observed in one additional animal that did not have a hematoma. Following animal and environmental ectoparasiticide treatment with permethrin, no recurrences or additional cases of aural hematomas were observed in the flock in the following two-year period. To the authors' knowledge, this is the first description of aural hematomas in lambs associated with O. megnini infestation with successful recovery after surgery and off-label acaricide treatment., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In large parts of the northern hemisphere, multiple deer species coexist, and management actions can strongly influence wild deer communities. Such changes may also indirectly influence other species in the community, such as small mammals and birds, because deer can have strong effects on their habitats and resources. Deer, small mammals and birds play an important role in the dynamics of tick-borne zoonotic diseases. It is, however, relatively underexplored how the abundance and composition of vertebrate communities may affect the outbreak potential, maintenance and circulation of tick-borne pathogens. In this study we focus on the outbreak potential by exploring how the basic reproduction number R 0 for different tick-borne pathogens depends on host community composition. We used published data on co-varying roe deer (Capreolus capreolus) and fallow deer (Dama dama) densities following a hunting ban, and different small mammal and bird densities, to investigate how the change in host community influences the R 0 of four tick-borne pathogens: one non-zoonotic, namely Anaplasma phagocytophilum ecotype 2, and three zoonotic, namely A. phagocytophilum ecotype 1, Borrelia afzelii and Borrelia garinii. We calculated R 0 using a next generation matrix approach, and used elasticities to quantify the contributions to R 0 of the different groups of host species. The value of R 0 for A. phagocytophilum ecotype 1 was higher with high fallow deer density and low roe deer density, while it was the other way round for A. phagocytophilum ecotype 2. For B. afzelii, R 0 was mostly related to the density of small mammals and for B. garinii it was mostly determined by bird density. Our results show that the effect of species composition is substantial in the outbreak potential of tick-borne pathogens. This implies that also management actions that change this composition, can (indirectly and unintentionally) affect the outbreak potential of tick-borne diseases., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2023. Published by Elsevier GmbH.)

This paper describes a new tick-host record of Haemaphysalis wellingtoni from the helmeted guineafowl (Numida meleagris) in Peninsular Malaysia. This record is also the first report of a tick infestation from N. meleagris in Asia. A checklist of the tick-guineafowl associations has been designed, resulting in 32 tick species including H. wellingtoni recorded as infesting N. meleagris worldwide. A list of pathogens harboured or transmitted by H. wellingtoni is provided. The status of H. wellingtoni as a true parasite of N. meleagris, and the health threat posed to the guineafowls are discussed in this paper., Competing Interests: Declaration of Competing Interest All authors declare no competing interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Ticks act as vectors and hosts of numerous arboviruses. Examples of medically important arboviruses include the tick-borne encephalitis virus, Crimean Congo hemorrhagic fever, and severe fever with thrombocytopenia syndrome. Recently, some novel arboviruses have been identified in blood specimens of patients with unexplained fever and a history of tick bites in Inner Mongolia. Consequently, tick-borne viruses are a major focus of infectious disease research. However, the spectrum of tick-borne viruses in subtropical areas of China has yet to be sufficiently characterized. In this study, we collected 855 ticks from canine and bovine hosts in four locations in Hainan Province. The ticks were combined into 18 pools according to genus and location. Viral RNA-sequence libraries were subjected to transcriptome sequencing analysis. Molecular clues from metagenomic analyses were used to classify sequence reads into virus species, genera, or families. The diverse viral reads closely associated with mammals were assigned to 12 viral families and important tick-borne viruses, such as Jingmen, Beiji nairovirus, and Colorado tick fever. Our virome and phylogenetic analyses of the arbovirus strains provide basic data for preventing and controlling human infectious diseases caused by tick-borne viruses in the subtropical areas of China., Competing Interests: Declaration of competing interest None., (Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Lyme disease, or also known as Lyme borreliosis, is caused by the spirochetes belonging to the Borrelia burgdorferi sensu lato complex, which can enter the human body following the bite of an infected tick. Many membrane lipid-bound proteins, also known as lipoproteins, are located on the surface of B. burgdorferi sensu lato and play a crucial role in the spirochete to interact with its environment, whether in ticks or mammals. Since the spirochete needs to perform various tasks, such as resisting the host's immune system or spreading throughout the organism, it is not surprising that numerous surface proteins have been found to be essential for B. burgdorferi sensu lato complex bacteria in causing Lyme disease. In this study, we have determined (at 2.4 Å resolution) and characterized the 3D structure of BB0158, one of the few chromosomally encoded outer surface proteins from B. burgdorferi sensu stricto. BB0158 belongs to the paralogous gene family 44 (PFam44), consisting of four other members (BB0159, BBA04, BBE09 and BBK52). The characterization of BB0158, which appears to form a domain-swapped dimer, in conjunction with the characterization of the corresponding PFam44 members, certainly contribute to our understanding of B. burgdorferi sensu stricto proteins., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest in this study., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Hemotropic mycoplasmas (hemoplasmas) are small Gram-negative bacteria that parasitize red blood cells and can cause mild to severe anemia in a wide range of vertebrates, including ruminants. Cattle population in Somalia is around 3.9 million heads, with animals more concentrated around the river areas, mainly in the Juba River and Shabelle River Valleys. Information on hemoplasmas in Sub-Saharan Africa are scarce, mainly in Somalia, where no studies have been performed to date. Accordingly, this study aimed to assess the molecular occurrence of hemoplasmas in 131 cattle blood samples from Somalia. Thirty out of 131 (22.90%; 95% CI: 16.54-30.81%) cattle were infested by ticks: Rhipicephalus pulchellus (68.18%), Amblyomma gemma (18.18%), Amblyomma lepidum (9.09%), Hyalomma marginatum (1.51%), Hyalmomma rufipes (1.51%), and Rhipicephalus pravus (1.51%). A total of 74/131 (56.48%; 95% CI: 47.93-64.67%) cattle were positive for hemotropic Mycoplasma spp. by real-time PCR (qPCR) based on the 16S rRNA gene. Hemoplasma-positive samples were later subjected to species-specific PCR assays for Mycoplasma wenyonii and 'Candidatus Mycoplasma haematobovis' based on the 16S rRNA gene. A total of 34/74 (45.94%; 95% CI: 35.07-57.22%) animals were coinfected by both species; 31/74 (41.89%; 95% CI: 31.32-53.26%) and 3/74 (4.05%; 95% CI: 01.39-11.25%) cattle were solely positive to M. wenyonii and 'Ca. M. haematobovis', respectively. Six out of 74 (8.1%; 95% CI: 03.77-16.58%) cattle were negative on species-specific conventional PCR assays but tested positive by a semi-nested PCR assay based on the 16S rRNA gene of hemoplasmas. Sequencing of the detected hemotropic Mycoplasma sp. 16S rRNA gene confirmed that animals were infected by M. wenyonii and 'Ca. M. haematobovis'. To the best of our knowledge, this is the first study on the detection of hemoplasmas in cattle from Somalia., Competing Interests: Declaration of Competing Interest The authors have no competing financial interests to declare., (Copyright © 2022. Published by Elsevier B.V.)

The Brazilian Atlantic Forest helds one of the most diverse and unique avifauna in the world. Many vertebrate species are reservoirs of tick-borne pathogens, and birds are an important group among them due to their mobility which facilitates the dispersion of ticks and the infectious agents they carry. This study brings data on the tick diversity parasitizing birds and the molecular detection of Rickettsia spp. in these arthropods. Birds (n = 773) were captured, identified, and banded at Mata do Paraíso Research, Training, and Environmental Education Center located in Viçosa, Minas Gerais, Brazil. Birds were checked for the presence of ticks, which were individually collected, identified, and molecularly processed through Polymerase Chain Reaction (PCR) for the detection of Rickettsia spp. A total of 130 individuals were infested by ticks, and 479 tick specimens were collected, showing a seasonal distribution of the life stages throughout the year. Ticks were identified as Amblyomma longirostre (59/479); Amblyomma calcaratum (20/479); Amblyomma varium (3/479); Amblyomma sculptum (2/479) and Amblyomma spp. larvae (395/479). Seasonal distribution of the life stages of ticks was observed along the year and significant negative correlations were found between temperature and collected ticks and temperature and infested birds. From the evaluated samples of ticks, 25.44% (n = 43/169) scored positive for Rickettsia spp., and sequence analysis indicated high nucleotide identity with Rickettsia rhipicephali, R. massiliae, R. africae and R. honei marmionii. The potential for dispersal of ticks by birds added to the aggressiveness of species of the genus Amblyomma and the zoonotic potential of some species of Rickettsia are quite worrying when we consider that the study area is widely attended by students, researchers, people from the city and neighboring municipalities., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Records of accidental parasitism by ticks in humans from Brazil are scarce, with most being reported by researchers who are parasitized during their research and by professionals who work with animals. In order to compile these records, an extensive literature review was carried out. Our revision includes studies published between 1909 and 2022, including nine species of the Argasidae family and 32 species of the Ixodidae family that were reported biting humans in the six biomes of the Brazilian territory. The species with the highest number of records of human parasitism was Amblyomma sculptum, followed by Amblyomma coelebs, Amblyomma cajennense sensu stricto, and Amblyomma brasiliense. The Atlantic Forest was the most frequent biome where human parasitism occurred, probably due to the greater number of inhabitants, universities, and researchers in the region; however, this does not mean that this biome is more conducive to the development of ticks and their parasitism in humans. In addition to Amblyomma ovale, a vector of Rickettsia parkeri in the country, two of the main species that act as vectors of Rickettsia rickettsii, A. sculptum, and Amblyomma aureolatum, have been reported, which is quite worrying considering that the wide distribution of the species and life stages most frequently mentioned in parasitism (i.e., nymphs and adults) are the ones that favour pathogen transmission. This research provides a significant contribution to the knowledge of tick species associated with human parasitism in Brazil; however, due to environmental change potentiated by deforestation and fires, it is expected that there will be a geographic expansion of some tick species and the pathogens that use them as a vector and an increase in human parasitism., (Copyright © 2022. Published by Elsevier B.V.)

Crimean-Congo haemorrhagic fever virus is a tick-borne zoonotic agent of the genus Orthonairovirus, and the causal agent of Crimean-Congo haemorrhagic fever in humans. The study is aimed at investigating the presence of CCHFV RNA in ticks and cattle in Bosnia and Herzegovina (B&H), as well as the exposure of cattle to CCHFV in this country. Only one pool of three male Hyalomma marginatum ticks tested positive for viral RNA (3/760, 0.39%). No viral RNA was detected in 206 bovine blood samples using real-time reverse-transcriptase PCR (RT-qPCR). The overall seroprevalence of CCHFV in the cattle population was estimated as 14.97% (72/481). The results indicate the presence and the established circulation of CCHFV in B&H. Considering the endemic character of CCHFV in the Balkans as well as our results, the possibility that CCHFV is endemic in the territory of Bosnia and Herzegovina is considered to be very high., (Copyright © 2022. Published by Elsevier GmbH.)

Ticks are obligatory parasites and transmit several pathogens to animals and humans. This study aimed to report the parasitism of ticks on carcasses of animals hunted by hunters from communities near Mapinguari National Park, at the shared border of Amazonas and Rondonia states, northern Brazil. The ticks were collected from heads of carcasses of the following animals: South American tapir (Tapirus terrestris), collared peccary (Pecari tajacu) and long-nosed armadillo (Dasypus beniensis). Six Amblyomma tick species were identified: Amblyomma coelebs, Amblyomma naponense, Amblyomma latepunctatum, Amblyomma oblongoguttatum, Amblyomma scalpturatum, Amblyomma triste and Amblyomma spp. The most abundant tick species were A. oblongoguttatum and A. scalpturatum. DNA extraction and polymerase chain reaction (PCR) were performed on all ticks to test for the presence of rickettsial gltA gene fragments, but no amplification was observed. This study reports, for the first time, the parasitism of A. coelebs in the armadillo D. beniensis and the presence of A. triste in the states of Amazonas and Rondonia, updating the tick fauna to 24 and 39 species for these states, respectively., (Copyright © 2022. Published by Elsevier GmbH.)

Objective of this study is to find the optimal conditions for preparing the samples, resulting in quality, reproducible and specific MS spectra of the ticks, with a shelf life in 70% ethanol of more than ten years. Amblyomma (Am.) variegatum species which had been stored in alcohol for more than twenty years and for which numerous specimens were available were used to compare the performance of four protocols tested. Spectra of insufficient quality were obtained from Am. variegatum legs preserved in alcohol for long periods with the reference protocol, named DO that we had set up years ago. The same observation was made on the spectra from Am. variegatum legs from dry (evaporated alcohol, DO-mod protocol). With new protocols named ReDO and PReDO the spectra were of good quality with high intensities (> 3000 a.u.). Blind testing showed that 94%, and 93% of the spectra were correctly identified with relevant log score values (LSVs ≥1.8), respectively for ReDO and PReDO protocols. All soft ticks treated in this study by PReDO protocol exhibited low intensity spectra with background noise. This study revealed that MALDI-TOF MS is able to identify hard ticks stored during decades in alcohol or dry (evaporated alcohol). SIGNIFICANCE OF THE STUDY: The correct identification of ticks, including vectors responsible for the transmission of infectious diseases in humans and animals is essential for their control. MALDI-TOF MS, a proteomic tool that has emerged in recent years, has become an innovative, accurate and alternative tool for the identification of arthropods, including ticks. However, previous studies reported that preservation of arthropods in alcohol modified the MS spectra obtained from specimens of the same species freshly collected or frozenly stored. In this study, a standard protocol was established for the identification of tick collections which had been stored for more than ten years in alcohol. Four different protocols were assessed. The analysis of the results showed that among the four protocols tested, two protocols named ReDO (Rehydration and incubation of the legs in 40 μl of HPLC water for 12 h in a dry bath at 37°) and PreDO (Drying of the legs for 12 h in a dry bath at 37 °C followed by rehydration and incubation in 40 μl of HPLC water for 12 h.) seem to be more appropriate for the MALDI-TOF MS identification of ticks from old collections preserved in alcohol or dry. This study is promising for the future, as it will make it possible to create a MALDI-TOF MS database from a wide range of ticks which have been stored for a long time in alcohol or which are dry stored in laboratories and museums around the world., (Copyright © 2022 Elsevier B.V. All rights reserved.)