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Due to the extensive use of green urban areas as recreation places, city residents are exposed to tick-borne pathogens. The objectives of our study were (i) to determine the occurrence of ticks in urban green areas, focussing on areas used by humans such as parks, schools and kindergartens, and urban forests, and (ii) to assess the prevalence of Borrelia infections in ticks in Zielona Góra, a medium-sized city in western Poland. A total of 161 ticks representing the two species Ixodes ricinus (34 males, 51 females, 30 nymphs) and Dermacentor reticulatus (20 males, 26 females) were collected from 29 of 72 (40.3%) study sites. In total, 26.1% of the ticks (85.7% of I. ricinus and 14.3% of D. reticulatus) yielded DNA of Borrelia. The difference in the infection rate between I. ricinus and D. reticulatus was significant. Among infected ticks, the most frequent spirochete species were B. lusitaniae (50.0%) and B. afzelii (26.2%), followed by B. spielmanii (9.5%), B. valaisiana (7.1%), B. burgdorferi sensu stricto, (4.8%) and B. miyamotoi (2.4%). No co-infections were found. We did not observe a correlation in the occurrence of Borrelia spirochetes in ticks found in individual study sites that differed in terms of habitat type and height of vegetation. Our findings demonstrate that the Borrelia transmission cycles are active within urban habitats, pointing the need for monitoring of tick-borne pathogens in public green areas. They could serve as guidelines for authorities for the proper management of urban green spaces in a way that may limit tick populations and the potential health risks posed by tick-borne pathogens. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the level of infection of Ixodes ricinus ticks with pathogens (Borrelia spp., Rickettsia spp., and Anaplasma spp.) collected from Lacerta agilis and Zootoca vivipara lizards in the urban areas of Wrocław (SW Poland). The study was carried out in July-August 2020. Lizards were caught by a noose attached to a pole or by bare hands, identified by species, and examined for the presence of ticks. Each lizard was then released at the site of capture. Ticks were removed with tweezers, identified by species using keys, and molecular tests were performed for the presence of pathogens. From 28 lizards (17 specimens of Z. vivipara and 11 specimens of L. agilis) a total of 445 ticks, including 321 larvae and 124 nymphs, identified as I. ricinus were collected. A larger number of ticks were obtained from L. agilis compared to Z. vivipara. Molecular tests for the presence of pathogens were performed on 445 specimens of I. ricinus. The nested PCR method for the fla gene allowed the detection of Borrelia spp. in 9.4% of ticks, and it was higher in ticks from L. agilis (12.0%) than from Z. vivipara (1.0%). The RFLP method showed the presence of three species, including two belonging to the B. burgdorferi s.l. complex (B. lusitaniae and B. afzelii), and B. miyamotoi. The overall level of infection of Rickettsia spp. was 19.3%, including 27.2% in ticks collected from Z. vivipara and 17.0% from L. agilis. Sequencing of randomly selected samples confirmed the presence of R. helvetica. DNA of Anaplasma spp. was detected only in one pool of larvae collected from L. agilis, and sample sequencing confirmed the presence of (A) phagocytophilum. The research results indicate the important role of lizards as hosts of ticks and their role in maintaining pathogens in the environment including urban agglomeration as evidenced by the first recorded presence of (B) miyamotoi and (A) phagocytophilum in I. ricinus ticks collected from L. agilis. However, confirmation of the role of sand lizards in maintaining (B) miyamotoi and A. phagocytophilum requires more studies and sampling of lizard tissue. [ABSTRACT FROM AUTHOR]

The invasive Asian longhorned tick Haemaphysalis longicornis that vectors and transmits several animal pathogens is significantly expanding in the United States. Recent studies report that these ticks also harbor human pathogens including Borrelia burgdorferi sensu lato, Babesia microti, and Anaplasma phagocytophilum. Therefore, studies that address the interactions of these ticks with human pathogens are important. In this study, we report the characterization of H. longicornis organic anion-transporting polypeptides (OATPs) in interactions of these ticks with A. phagocytophilum. Using OATP-signature sequence, we identified six OATPs in the H. longicornis genome. Bioinformatic analysis revealed that H. longicornis OATPs are closer to other tick orthologs rather than to mammalian counterparts. Quantitative real-time PCR analysis revealed that OATPs are highly expressed in immature stages when compared to mature stages of these ticks. In addition, we noted that the presence of A. phagocytophilum upregulates a specific OATP in these ticks. We also noted that exogenous treatment of H. longicornis with xanthurenic acid, a tryptophan metabolite, influenced OATP expression in these ticks. Immunoblotting analysis revealed that antibody generated against Ixodes scapularis OATP cross-reacted with H. longicornis OATP. Furthermore, treatment of H. longicornis with OATP antibody impaired colonization of A. phagocytophilum in these ticks. These results not only provide evidence that the OATP-tryptophan pathway is important for A. phagocytophilum survival in H. longicornis ticks but also indicate OATP as a promising candidate for the development of a universal anti-tick vaccine to target this bacterium and perhaps other rickettsial pathogens of medical importance. [ABSTRACT FROM AUTHOR]

Uncovering the complexity of systems in non-model organisms is critical for understanding arthropod immunology. Prior efforts have mostly focused on Dipteran insects, which only account for a subset of existing arthropod species in nature. Here we use and develop advanced techniques to describe immune cells (hemocytes) from the clinically relevant tick Ixodes scapularis at a single-cell resolution. We observe molecular alterations in hemocytes upon feeding and infection with either the Lyme disease spirochete Borrelia burgdorferi or the rickettsial agent Anaplasma phagocytophilum. We reveal hemocyte clusters exhibiting defined signatures related to immunity, metabolism, and proliferation. Depletion of phagocytic hemocytes affects hemocytin and astakine levels, two I. scapularis hemocyte markers, impacting blood-feeding, molting behavior, and bacterial acquisition. Mechanistically, astakine alters hemocyte proliferation, whereas hemocytin affects the c-Jun N-terminal kinase (JNK) signaling pathway in I. scapularis. Altogether, we discover a role for tick hemocytes in immunophysiology and provide a valuable resource for comparative biology in arthropods. Rolandelli et al. define how immune cells from the tick Ixodes scapularis are affected by infection and show that this cell population have distinct signatures and functions in immunity, metabolism and proliferation. [ABSTRACT FROM AUTHOR]

Background: A newly discovered zoonotic infection carried by ixodid ticks, Anaplasma capra, affects a wide variety of hosts, including numerous mammals. A. capra most likely infects erythrocytes or endothelial cells in mammals. This study aimed to investigate the A. capra pathogen in goats in Türkiye's Van province. Methods: A total of 200 goat blood samples were examined. Goat samples were subjected to partial amplification of the gltA gene fragment using a nested polymerase chain reaction. Results: A. capra DNA was detected in 0.5% of goat blood samples. Phylogenetic analysis of a partial gltA gene fragment showed that the Eastern Türkiye isolate, closely grouped with A. capra isolates reported from wild and domestic ruminants in France, Türkiye, and Kyrgyzstan, formed a distinct clade. Conclusions: This is the first report of A. capra in goats in Van province, Eastern Türkiye. [ABSTRACT FROM AUTHOR]

Dermacentor reticulatus is tick species with an expanding geographical range in Europe, which creates the possibility of spreading microorganisms of significant veterinary and medical importance. The study aimed to investigate the prevalence and genetic diversity of Rickettsia spp., Babesia spp., Borrelia spp. and Anaplasma phagocytophilum in adult D. reticulatus ticks from the Eastern European population in the urban and the natural biotopes of north-eastern Poland. Microorganisms were detected by PCR and identified by DNA sequencing. The overall infection rate of at least one of the pathogens was 29.6%. The predominantly was Rickettsia spp. (27.1%) (with R. raoultii—9.1%) followed by Babesia spp. (2.4%) with B. canis (1.5%) as the most frequent. Based on 18S rRNA gene sequence, three B. canis genotypes were revealed. The prevalence of R. raoultii and B. canis was significantly higher in ticks from natural biotopes. The infection rates of B. afzelii and A. phagocytophilum were determined at 0.9% and 0.3%, respectively. Co-infections were detected in 3.8% of infected ticks. In diagnosing tick-borne diseases in humans, tick-borne lymphadenopathy should not be excluded. The prevalence of different genotypes of B. canis suggests differences in the clinical picture of canine babesiosis in the area. [ABSTRACT FROM AUTHOR]

There is a knowledge gap in the study of Argasidae soft ticks and the pathogens they can transmit. These hematophagous arthropods are widely distributed and are often considered typical bird ectoparasites. Tick-parasitized birds can act not only as a reservoir of pathogens but also can carry these pathogen-infected arthropods to new areas. Seven griffon vulture nestlings were sampled in northeastern Spain, collecting ticks (n = 28) from two individuals and blood from each vulture (n = 7). Blood samples from griffon vultures tested PCR positive for Flavivirus (7/7), Anaplasma (6/7), piroplasms (4/7), and Rickettsia (1/7). A total of 27 of the 28 analyzed ticks were positive for Rickettsia, 9/28 for Anaplasma, 2/28 for piroplasms, and 5/28 for Crimean-Congo Hemorrhagic fever virus (CCHFv). Sequencing and phylogenetic analyses confirmed the presence of Rickettsia spp., Babesia ardeae, and zoonotic Anaplasma phagocytophilum in vultures and Rickettsia spp., B. ardeae, and CCHFv genotype V in ticks. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize the gene expression of host immune- and cellular responses to a Norwegian virulent strain of Anaplasma phagocytophilum, the cause of tick-borne fever in sheep. Ten sheep were intravenously inoculated with a live virulent strain of A. phagocytophilum. Clinical-, observational-, hematological data as well as bacterial load, flow cytometric cell count data from peripheral blood mononuclear cells and host's gene expression post infection was analysed. The transcriptomic data were assessed for pre-set time points over the course of 22 days following the inoculation. Briefly, all inoculated sheep responded with clinical signs of infection 3 days post inoculation and onwards with maximum bacterial load observed on day 6, consistent with tick-borne fever. On days, 3–8, the innate immune responses and effector processes such as IFN1 signaling pathways and cytokine mediated signaling pathways were observed. Several pathways associated with the adaptive immune responses, namely T-cell activation, humoral immune responses, B-cell activation, and T- and B-cell differentiation dominated on the days of 8, 10 and 14. Flow-cytometric analysis of the PBMCs showed a reduction in CD4+CD25+ cells on day 10 and 14 post-inoculation and a skewed CD4:CD8 ratio indicating a reduced activation and proliferation of CD4-T-cells. The genes of important co-stimulatory molecules such as CD28 and CD40LG, important in T- and B-cell activation and proliferation, did not significantly change or experienced downregulation throughout the study. The absence of upregulation of several co-stimulatory molecules might be one possible explanation for the low activation and proliferation of CD4-T-cells during A. phagocytophilum infection, indicating a suboptimal CD4-T-cell response. The upregulation of T-BET, EOMES and IFN-γ on days 8–14 post inoculation, indicates a favoured CD4 Th1- and CD8-response. The dynamics and interaction between CD4+CD25+ and co-stimulatory molecules such as CD28, CD80, CD40 and CD40LG during infection with A. phagocytophilum in sheep needs further investigation in the future. [ABSTRACT FROM AUTHOR]

We analysed both pooled and individual tick samples collected from four countries in Eastern Europe and the Black Sea region, using metagenome-based nanopore sequencing (NS) and targeted amplification. Initially, 1337 ticks, belonging to 11 species, were screened in 217 pools. Viruses (21 taxa) and human pathogens were detected in 46.5% and 7.3%, respectively. Tick-borne viral pathogens comprised Tacheng Tick Virus 2 (TTV2, 5.9%), Jingmen Tick Virus (JMTV, 0.9%) and Tacheng Tick Virus 1 (TTV1, 0.4%). An association of tick species with individual virus taxa was observed, with the exception of TTV2, which was observed in both Dermacentor and Haemaphysalis species. Individual ticks from pools with pathogen detection were then further screened by targeted amplification and then NS, which provided extensive genome data and revealed probable pathogen Haseki Tick Virus (HTV, 10.2%). Two distinct TTV2 clades were observed in phylogenetic analysis, one of which included closely related Dermacentor reticulatus Uukuviruses. JMTV detection indicated integrated virus sequences. Overall, we observed an expansion of newly documented pathogenic tick-borne viruses into Europe, with TTV1 being identified on the continent for the first time. These viruses should be included in the diagnostic assessment of symptomatic cases associated with tick bites and vector surveillance efforts. NS is shown as a useful tool for monitoring tick-associated pathogens in pooled or individual samples. [ABSTRACT FROM AUTHOR]

Human contact with wild animals in synanthropic habits is often mediated by arthropod vectors such as ticks. This is an important method of spreading infectious agents that pose a risk to human health. Thus, this study aimed to molecularly detect Ehrlichia spp., Anaplasma spp., Borrelia spp., and protozoa of the order Piroplasmida in ticks collected from coatis of Iguaçu National Park (PNI), Paraná, Brazil. This study involved 553 ticks DNA, including Amblyomma spp. larvae, Haemaphysalis juxtakochi nymphs, Amblyomma brasiliense, Amblyomma coelebs, and adults of Amblyomma ovale. The DNA extracted from each sample was subjected to polymerase chain reaction (PCR) targeting the genes 23S rRNA for the Anaplasmataceae family, 16S rRNA for Anaplasma spp., dsb for Ehrlichia spp., flaB, 16S rRNA, hpt, and glpQ for Borrelia spp., and 18S rRNA for Piroplasmid protozoans. DNA from Anaplasma sp. was detected in ticks of the species A. coelebs (4/553); Borrelia sp. DNA was detected in A. coelebs (3/553), A. ovale (1/553), and Amblyomma larvae (1/553); and Theileria sp. was detected in A. coelebs (2/553). All tested samples were negative for Ehrlichia spp. Our study constitutes the newest report in South America of these microorganisms, which remain poorly studied. [ABSTRACT FROM AUTHOR]

We evaluated the presence of Ehrlichia spp. in unfed capybara ticks, Amblyomma dubitatum, and explored its association with capybaras density, ticks density and environmental variables. We observed that in the Iberá wetlands ecoregion A. dubitatum is infected by "Candidatus Ehrlichia hydrochoerus" and in a lesser extent with an Ehrlichia species closely related to Ehrlichia chaffeensis. The frequency of "Ca. Ehrlichia hydrochoerus" presence in A. dubitatum was not associated with vector abundance, but the probability of finding "Ca. Ehrlichia hydrochoerus"-infected ticks increased when the density of capybaras was low two months before. We hypothesize that when the density of capybaras decreases, A. dubitatum immature stages may seek out alternative hosts one of which could exhibit high realized reservoir competence for "Ca. Ehrlichia hydrochoerus", leading to an increased prevalence of this ehrlichiae in questing A. dubitatum. High minimum temperatures and high cumulative rainfall in the time period previous to tick collection (15 to 60 days) were positively correlated with the prevalence of "Ca. Ehrlichia hydrochoerus" infection in A. dubitatum. Our results suggest that a combination of factors (both biological and abiotic) could raise the risk of human exposure to tick-borne Ehrlichia in the Iberá wetlands ecoregion. [ABSTRACT FROM AUTHOR]

Technological and computational advancements in the fields of genomics and bioinformatics are providing exciting new opportunities for pathogen discovery and genomic surveillance. In particular, single-molecule nucleotide sequence data originating from Oxford Nanopore Technologies (ONT) sequencing platforms can be bioinformatically leveraged, in real-time, for enhanced biosurveillance of a vast array of zoonoses. The recently released nanopore adaptive sampling (NAS) strategy facilitates immediate mapping of individual nucleotide molecules to a given reference as each molecule is being sequenced. User-defined thresholds then allow for the retention or rejection of specific molecules, informed by the real-time reference mapping results, as they are physically passing through a given sequencing nanopore. Here, we show how NAS can be used to selectively sequence DNA of multiple bacterial tick-borne pathogens circulating in wild populations of the blacklegged tick vector, Ixodes scapularis. [ABSTRACT FROM AUTHOR]

Cardio-pulmonary parasites, such as Angiostrongylus vasorum, Crenosoma vulpis, and Eucoleus aerophilus, pose a significant concern on account of pulmonary and cardiac problems they induce in dogs. While the red fox is known to be a key reservoir host for A. vasorum and may also play a role in transmitting C. vulpis and E. aerophilus, there has been no recent research on these parasites in foxes from Sardinia, with the most current studies dating back to 1986. A survey was conducted on red foxes in Sardinia, where a total of 51 foxes were collected, necropsied, and examined for adult worms in their hearts and lungs. The worms were identified using morphometric analysis and molecular methods. The results showed a 54.9% overall prevalence at dissection: 45.1% of the foxes were positive for E. aerophilus, 17.6% for C. vulpis, and 13.7% for A. vasorum. The molecular analyses validated the morphological characterization. In comparison to previous research, which found 13 out of 85 foxes to be positive for A. vasorum with a prevalence rate of 15.3% and 1 for E. aerophilus with a prevalence of 1.2%, this study showed an increased prevalence of E. aerophilus and C. vulpis, and a decrease in the prevalence of A. vasorum. These results indicate that the red foxes in Sardinia represent a reservoir host for cardio-pulmonary nematodes and it should be considered in the differential diagnosis of respiratory distress syndrome in dogs. [ABSTRACT FROM AUTHOR]

Human anaplasmosis caused by Anaplasma phagocytophilum is one of the most common tick-borne diseases in the United States. The black-legged ticks, Ixodes scapularis, vector and transmit this bacterium to humans. In this study, we provide evidence that targeting I. scapularis membrane-bound organic anion transporting polypeptide 4056 (IsOATP4056) with an anti-vector vaccine affects transmission of A. phagocytophilum from ticks to the vertebrate host. Anaplasma phagocytophilum induces expression of IsOATP4056 in ticks and tick cells. Increased membrane localization of IsOATP4056 was evident in A. phagocytophilum-infected tick cells. Treatment with high dose (10 µg/ml) but not low dose (5 µg/ml) of EL-6 antibody that targets the largest extracellular loop of IsOATP4056 showed cytotoxic effects in tick cells but not in human keratinocyte cell line (HaCaT). Passive immunization, tick-mediated transmission and in vitro studies performed with mice ordered from two commercial vendors and with tick cells showed that EL-6 antibody not only impairs A. phagocytophilum transmission from ticks to the murine host but also aids in the reduction in the bacterial loads within engorged ticks and in tick cells by activation of arthropod Toll pathway. Furthermore, reduced molting efficiency was noted in ticks fed on EL-6 antibody-immunized mice. Collectively, these results provide a good candidate for the development of anti-tick vaccine to target the transmission of A. phagocytophilum and perhaps other tick-borne pathogens of medical importance. [ABSTRACT FROM AUTHOR]

Tick-borne infections, including those of bacterial origin, are significant public health issues. Antimicrobial resistance (AMR), which is one of the most pressing health challenges of our time, is driven by specific genetic determinants, primarily by the antimicrobial resistance genes (ARGs) of bacteria. In our work, we investigated the occurrence of ARGs in the genomes of tick-borne bacterial species that can cause human infections. For this purpose, we processed short/long reads of 1550 bacterial isolates of the genera Anaplasma (n = 20), Bartonella (n = 131), Borrelia (n = 311), Coxiella (n = 73), Ehrlichia (n = 13), Francisella (n = 959) and Rickettsia (n = 43) generated by second/third generation sequencing that have been freely accessible at the NCBI SRA repository. From Francisella tularensis, 98.9% of the samples contained the FTU-1 beta-lactamase gene. However, it is part of the F. tularensis representative genome as well. Furthermore, 16.3% of them contained additional ARGs. Only 2.2% of isolates from other genera (Bartonella: 2, Coxiella: 8, Ehrlichia: 1, Rickettsia: 2) contained any ARG. We found that the odds of ARG occurrence in Coxiella samples were significantly higher in isolates related to farm animals than from other sources. Our results describe a surprising lack of ARGs in these bacteria and suggest that Coxiella species in farm animal settings could play a role in the spread of AMR. [ABSTRACT FROM AUTHOR]

This document is a letter to the editor of the journal International Orthopaedics. The authors of the letter are responding to a recent publication by Sanpera et al. that examines the outcomes of malrotated elbow supracondylar fractures in children. The authors of the letter raise concerns about biases in the study and question the measurement techniques used to detect loss of rotational reduction. They also suggest that the authors should have considered the Beighton hypermobility scale and its impact on shoulder rotation. Overall, the letter provides a critical analysis of the original study and raises important questions about its methodology and findings. [Extracted from the article]

Anaplasma marginale is transmitted biologically by infected ticks or mechanically by biting flies and contaminated fomites. In tick-free areas, such as southern Uruguay, horseflies could be the principal vectors of this pathogen for bovines, causing anaplasmosis. The objective of this work was to detect the presence of A. marginale by MSP-5 PCR and Sanger sequencing in the most prevalent species of horseflies obtained using different collection methods in Colonia, Tacuarembó and Paysandú, Uruguay. Eight horsefly species were tested (Dasybasis missionum, Poeciloderas lindneri, Tabanus campestris, T. claripennis, T. fuscofasciatus, T. platensis, T. tacuaremboensis and T. triangulum); four species were found to be positive for A. marginale, with D. missionum and P. lindneri having the most frequent infections, while only one individual each of T. fuscofasciatus and T. tacuaremboensis was positive. Both D. missionum and P. lindneri were positive for A. marginale in tick-free areas, and the implications are discussed in this report. [ABSTRACT FROM AUTHOR]

Tick-borne encephalitis (TBE) is an infectious viral disease, the pathogenesis of which is still not fully understood. Additionally, TBE can be complicated by co-infections with various bacteria that are also transmitted by ticks, which can affect the proper diagnosis and treatment. Therefore, the aim of the study was to evaluate changes in the plasma phospholipid (PL) and ceramide (CER) profile of patients with TBE and patients with bacterial co-infection (B. burgdorferi or A. phagocytophilum) in relation to healthy subjects. For this purpose, a high-resolution LC-QTOF-MS/MS platform as well as univariate and multivariate statistics were used. The results of this study showed that the levels of phosphatidylcholines (PC) and lysophosphatidylcholines (LPC) species were increased in the plasma of patients with TBE and patients with TBE co-infected with bacteria. On the other hand, observed differences in the content of phosphoethanolamines (PE) and sphingomyelins (SM) make it possible to distinguish TBE patients from patients with co-infections. The opposite direction of changes was also observed in the CER content. This study showed significant modifications to the metabolic pathways of linoleic (LA) and arachidonic acid (AA), as confirmed by the quantitative analysis of these fatty acids. The obtained results allow to distinguish the pathomechanism of TBE from TBE with bacterial co-infection, and consequently may improve the diagnostic process and enable more efficient pharmacotherapy against both pathogens. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum is a tick-borne Gram-negative intracellular bacterium that could infect a few leukocytes of a large range of mammalian hosts together with humans. The clinical symptoms of the disease (granulocytic anaplasmosis) can vary from subclinical to fatal. The study aimed to detect A. phagocytophilum among camels (Camelus dromedarius) in Iran through two distinctive diagnostic techniques. One hundred twenty-five blood samples of male camels (C. dromedarius) were taken. Thin blood films were stained by Giemsa and DNA was extracted from whole blood for molecular examination. In this study, A. phagocytophilum was detected by microscopy and molecular examination among 125 male camels (C. dromedarius) in Yazd, central Iran. The A. phagocytophilum morula was observed in one sample out of 125 (0.8%) in microscopic examination and was detected in 4 samples out of 125 (3.2%) in the molecular examination. According to the results, A. phagocytophilum infects camels (C. dromedarius) consequently, camels can play a role in the infection of ticks, other animals, or humans. Many aspects of this disease continue to be set up by way of further studies in Iran. [ABSTRACT FROM AUTHOR]

Purpose: Babesia canis infection occurs in many locations throughout Europe. However, various studies report different clinicopathological findings in affected dogs. This study was focused on changes in clinical and hematologic parameters in dogs with B. canis infection from eastern Slovakia. Methods: The study was prospective and included 45 dogs with suspected babesiosis. Babesia canis infection was confirmed by PCR in 34 cases and by blood smear microscopy in 24 (70.6%) of them. Hematology results, clinical examination from these dogs, and possible co-infection with other tick-borne pathogens by PCR were subsequently evaluated. Results: The major clinical signs found included lethargy (91%), fever (59%), anorexia (59%), pigmenturia (47%) and icterus (18%). Mortality rate was 6%. Thrombocytopenia was the most common hematologic change, observed in 100% of the dogs with B. canis infection. Other frequent findings were lymphopenia (82%) and anemia (68%). No co-infections were detected. Anaplasma phagocytophilum infection was diagnosed by PCR only in one dog, which was not infected with B. canis. Conclusions: This study showed that B. canis infection in eastern Slovakia should be diagnosed by PCR when there is clinical suspicion of the disease, as almost 30% of the infected sick dogs did not have demonstrable parasites in their blood smear by microscopy. Lymphopenia is a frequent hematologic finding in B. canis infection and observed even more often than anemia. However, in agreement with previous studies, thrombocytopenia remains the most common hematologic finding associated with B. canis infection. [ABSTRACT FROM AUTHOR]

Tortoises of the genus Testudo are the main hosts of Hyalomma aegyptium ticks. This species serves as a vector of several zoonotic pathogens. Therefore, the present study aimed to investigate the presence of four pathogens associated with H. aegyptium ticks obtained from tortoises from Tunisia. Conventional, multiplex and nested PCRs were used for Aanaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii and Babesia spp. screening. The molecular analyses revealed the presence of A. phagocytophilum and Babesia spp. None of the ticks, were infected by E. canis or C. burnetii species. Co-infection was detected in four ticks. As a conclusion, this is the first detection of A. phagocytophilum and Babesia spp. in H. aegyptium ticks collected from tortoises, in Tunisia. Thus, considering these results, the spur-thighed tortoise constitute a potential host of H. aegyptium which plays an important role in the transmission of pathogenic agents affecting both human and animals. In term of public health, a strict control and surveillance should be carried to reduce the circulation of such pathogens between different hosts. [ABSTRACT FROM AUTHOR]

Ticks are ectoparasitic arthropods of great medical and veterinary importance and are important vectors of several pathogens that affect livestock, humans, and pets. Nevertheless, the requirements of individual tick species for climatic factors for tick survival and seasonality and the temporal dynamics of tick-borne pathogens have still not been sufficiently explored, especially in areas with the simultaneous occurrence of several tick species. When the occurrence and ecology of a single tick species are studied at the model site, all the factors which determine their occurrence must be taken into account during the study. Research at the site with the sympatric occurrence of different tick species eliminates the need to study all other factors (biotic and abiotic) that affect the occurrence of a given species in a particular locality because these species are present at the same site under the same environmental conditions. These locations are therefore highly appropriate for the research of seasonal activity that is related to climatic factors. As the host range of tick species overlaps at these locations, this is highly useful also for studying pathogen flow between individual tick species. In the present study, we evaluated the influence of the most important climatic conditions, such as temperature, relative humidity, and saturation deficit for each tick species individually. Except for the coldest parts of the year, we observed a year-round occurrence of individual tick species, with species-specific differences in temporal distribution. Ixodes ricinus was the most common species, followed by Haemaphysalis inermis, Dermacentor marginatus and D. reticulatus. Altogether, 660 ticks were examined for the presence of zoonotic bacteria and protozoans. The widest spectrum of tick-borne pathogens was detected in I. ricinus. The highest was the prevalence of spirochetes from Borrelia burgdorferi s.l. complex (12.43 %), with a predominance of B. garinii and B. afzelii, followed by Babesia spp. (7.73 %) represented by B. microti and B. venatorum. Anaplasma phagocytophilum was confirmed in 2.76 % of tested ticks. The presence of co-infection with Borrelia spp. and Babesia spp. was recorded in I. ricinus (1.1 %). We also confirmed the presence of borreliae in D. reticulatus and H. inermis for the first time in Slovakia. The results confirm that I. ricinus is epidemiologically the most important tick species in Slovakia and Central Europe. [ABSTRACT FROM AUTHOR]

Ticks are medically important hematophagous ectoparasites transmitting pathogens such as Borrelia, Rickettsia, Babesia, Anaplasma, and Coxiella species that may seriously affect the health of animals and humans. More than a million cases of rickettsiosis, anaplasmosis, and Q fever occur annually worldwide, often in farmers or other occupationally exposed people. Despite the confirmed presence of the causative agents in endemic areas, the attention to these intriguing illnesses is frequently neglected in many countries, including Slovakia. We performed serological and molecular analyses of 403 human sera and 118 blood samples obtained from Slovak patients in the last decade. Herein, we describe seven particular cases to illustrate how clinicians and public health professionals are performing laboratory diagnostic work together to monitor the circulation of rickettsiae and rickettsia-like pathogens and solve health challenges associated with these infections. The results of laboratory testing were related to the symptoms of the diseases and possible tick infestations. Fever and erythema or rash were commonly reported in the cases of rickettsial infections, which were confirmed by the presence of IgM anti-SFGR antibodies and PCR assay positive for either Rickettsia helvetica, Rickettsia raoultii, or Rickettsia sp. On the other hand, an acute Q fever was commonly associated with fever and close contact with livestock or their products. Additionally, for the first time in Slovakia, we described a case with the presence of Anaplasma phagocytophilum DNA in human patient's blood for 47 days. In conclusion, we assume that effective surveillance of rickettsioses and Rickettsia-like infections is essential for public health. It certainly contributes to better prevention and management of these communicable diseases. [ABSTRACT FROM AUTHOR]

The black-legged tick, Ixodes scapularis, is a well-known vector for the Lyme disease-causing pathogen (Borrelia burgdorferi) but can also carry other disease-causing pathogens such as Rickettsia, Anaplasma, Bartonella, Ehrlichia, and Theileria. Hence, tick screening using highly specific protein signatures for specific pathogens will help assess the prevalence of infected ticks and understand the pathogen-tick interactions in a specific geographic area. In this study, we used data-dependent acquisition to key pathogen protein signatures in black-legged ticks collected from the Southern Tier New York. Bottom-up proteomic analysis of extract from five combined ticks identified 2,052 tick proteins and 41 pathogen proteins with high confidence (≥ 99% C.I.). Results show high peptide spectral match counts for Rickettsia species and Borrelia species and lower counts for other rarer pathogens such as Anaplasma phagocytophilum. Parallel reaction monitoring performed on protein extracts from individual ticks (n = 10) revealed that 8 out of the 10 screened ticks carried Rickettsia species, 5 carried Borrelia species, 3 carried both pathogens, and only 1 tick carried no detectable bacteria. Mass spectrometry-based proteomics is a highly specific way to define the expression of different types of pathogen proteins in infected ticks. This might bring insights into the tick-pathogen interactions at the molecular level and especially expression pathogen surface proteins in ticks. [ABSTRACT FROM AUTHOR]

In recent years, the distribution of Dermacentor reticulatus ticks has expanded into new territories in many European countries, including Poland, with increased population densities in areas of their regular occurrence. The spread of D. reticulatus enhances the risk of exposure of domestic animals and their owners to tick-borne diseases. The objective of this study was to assess the prevalence of infection of D. reticulatus ticks feeding on dogs with the pathogens Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. The study material comprised 152 D. reticulatus ticks collected from dogs in the northeastern part of Lublin Province (eastern Poland). A ready-made AmpliSens® TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis/E.muris-FRT PCR kit was used for qualitative detection and differentiation of tick-borne infections. The assessment of the degree of infection of the analyzed ticks with the two pathogens revealed that 9.2% (14/152) of the examined ticks were infected with one of the pathogens. No co-infections with the pathogens were detected in any of the ticks. The highest specific percentage of infections (8.6%, 13/152) was associated with A. phagocytophilum. The presence of B. burgdorferi s.l. was detected in only one of the examined ticks (0.7%). The spread of D. reticulatus to new territories and the increase in population density in areas of their regular occurrence implies the need for further studies of the prevalence of pathogens with medical and veterinary importance in order to assess the risk of tick-borne diseases. [ABSTRACT FROM AUTHOR]

Context: Pathogens can manipulate microbial interactions to ensure survival, potentially altering the functional patterns and microbiome assembly. The present study investigates how Anaplasma phagocytophilum infection affects the functional diversity, composition, and assembly of the Ixodes scapularis microbiome, with a focus on high central pathways-those characterized by elevated values in centrality metrics such as eigenvector, betweenness, and degree measures, in the microbial community., Methods: Using previously published data from nymphs' gut V4 region's amplicons of bacterial 16S rRNA, we predicted the functional diversity and composition in control and A. phagocytophilum-infected ticks and inferred co-occurrence networks of taxa and ubiquitous pathways in each condition to associate the high central pathways to the microbial community assembly., Results: Although no differences were observed concerning pathways richness and diversity, there was a significant impact on taxa and functional assembly when ubiquitous pathways in each condition were filtered. Moreover, a notable shift was observed in the microbiome's high central functions. Specifically, pathways related to the degradation of nucleosides and nucleotides emerged as the most central functions in response to A. phagocytophilum infection. This finding suggests a reconfiguration of functional relationships within the microbial community, potentially influenced by the pathogen's limited metabolic capacity. This limitation implies that the tick microbiome may provide additional metabolic resources to support the pathogen's functional needs., Conclusions: Understanding the metabolic interactions within the tick microbiome can enhance our knowledge of pathogen colonization mechanisms and uncover new disease control and prevention strategies. For example, certain pathways that were more abundant or highly central during infection may represent potential targets for microbiota-based vaccines., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Coinfections with the tick-borne pathogens Theileria luwenshuni and Anaplasma phagocytophilum can cause significant economic losses in sheep and goat farming. The difficulty in detecting these two pathogens by microscopic examination warrants the development of a rapid detection test to discriminate them. In this study, a duplex polymerase chain reaction (PCR) assay was developed to simultaneously detect T. luwenshuni and A. phagocytophilum. Alignment of the sequences from related pathogens allowed us to design a primer pair targeting the 18S ribosomal RNA gene in T. luwenshuni and generate a target product of 962 bp, whereas a previously reported species-specific primer (SSAP2f/SSAP2r) for A. phagocytophilum was used in the same reaction to generate a product of 641 bp. Genomic DNA from T. luwenshuni and A. phagocytophilum was 10-fold serially diluted for testing PCR sensitivity. Under the optimal PCR conditions we established, the lower limit of detection of the assay was 29.13 fg/μL for T. luwenshuni and 1.53 fg/μL for A. phagocytophilum, and PCR primers used in this study were confirmed to be 100% species-specific using other hemoparasites previously identified by other methods. No significant difference was found between conventional and duplex PCR protocols used to detect the two species. Our study provides an effective, sensitive, specific, and accurate tool for the diagnosis and epidemiological surveillance of mixed infections of the two pathogens in sheep and goats. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum is an important tick-borne zoonotic agent of human granulocytic anaplasmosis (HGA). In Europe, the Ixodes ticks are the main vector responsible for A. phagocytophilum transmission. A wide range of wild animals is involved in the circulation of this pathogen in the environment. Changes in populations of vertebrates living in different ecosystems impact the ecology of ticks and the epidemiology of tick-borne diseases. In this study, we investigated four species, Western European hedgehog (Erinaceus europaeus), northern white-breasted hedgehog (Erinaceus roumanicus), Eurasian red squirrel (Sciurus vulgaris), and the common blackbird (Turdus merula), to describe their role in the circulation of A. phagocytophilum in urban and periurban ecosystems. Ten different tissues were collected from cadavers of the four species, and blood and ear/skin samples from live blackbirds and hedgehogs. Using qPCR, we detected a high rate of A. phagocytophilum: Western European hedgehogs (96.4%), northern white-breasted hedgehogs (92.9%), Eurasian red squirrels (60%), and common blackbirds (33.8%). In the groEL gene, we found nine genotypes belonging to three ecotypes; seven of the genotypes are associated with HGA symptoms. Our findings underline the role of peridomestic animals in the ecology of A. phagocytophilum and indicate that cadavers are an important source of material for monitoring zoonotic pathogens. Concerning the high prevalence rate, all investigated species play an important role in the circulation of A. phagocytophilum in municipal areas; however, hedgehogs present the greatest anaplasmosis risk for humans. Common blackbirds and squirrels carry different A. phagocytophilum variants some of which are responsible for HGA. [ABSTRACT FROM AUTHOR]

Pathogens carried by ticks pose a threat to both human and animal health across the world. Typically associated with rural landscapes, ticks appear to adapt well to life in urban recreational areas. Although Dermacentor reticulatus is commonly found across Europe, data on the prevalence of pathogens in this tick species, in an urban environment, are very limited. PCR was used to examine 368 D. reticulatus individuals collected in the Zwierzyniecki Forest Nature Reserve in Białystok, Poland. In total, 10.3% of ticks were infected, with Babesia spp. (9.2%), Anaplasma phagocytophilum (0.8%) and Borrelia burgdorferi sensu lato (0.3%). Rickettsia spp., Bartonella spp., and Coxiella burnetii were not detected. Sequence analysis for Babesia-positive samples identified 79.4% of them as Babesia canis, 8.8% as Babesia microti, 5.9% as Babesia spp., 2.9% as Babesia venatorum, and 2.9% as Babesia vogeli. Results obtained in this study indicate that D. reticulatus ticks found within the urban premises of the study area are infected with at least three pathogens and therefore are an important factor in public health risk for tick-borne diseases. [ABSTRACT FROM AUTHOR]

Purpose: The Anaplasma genus includes a Gram-negative bacterium infecting the blood cells of wild and domestic mammals, causing tick-borne fever. Infection with pathogenic Anaplasma phagocytophilum strains may cause Human Granulocytic Anaplasmosis. Wild boars (Sus scrofa) may act as natural wild reservoir hosts for potentially zoonotic A. phagocytophilum strains; however, there is still little data to confirm this statement. The aim of this study was to verify whether wild boars can be classified as natural reservoirs of Anaplasma spp. and to compare the suitability of spleen and liver samples for such analysis. Methods: Liver and spleen samples were collected from 59 wild boars (2017–2019). The organs were tested for Anaplasma phagocytophilum using short (partial) fragments of three markers: 16S rRNA, groEL, ankA. Results: Anaplasma spp. DNA was detected in 12 out of 59 samples, with a prevalence of 20.34%. The presence of A. phagocytophilum was confirmed by sequencing of the partial 16S rRNA gene. Positive individuals were tested for the characteristic markers: groEL and ankA. The analysis of the nucleotide sequences of 16S rRNA, groEL and ankA, indicated that the strains of A. phagocytophilum detected in these studies are potentially zoonotic for humans. Conclusion: Wild boars from Poland can be classified as a natural reservoir of the zoonotic strain of Anaplasma phagocytophilum. Both the spleen and the liver tissues were found to be suitable materials for the detection of A. phagocytophilum. [ABSTRACT FROM AUTHOR]

Ticks may carry several pathogens as vectors and their pathogen load may vary due to differences in geography, climate and vegetation. In this study, we collected ticks from 39 districts of Istanbul (Turkey) between May and October, from 2013 to 2017, and identified them under stereo-microscope. In addition, we investigated the pathogens that the ticks carry (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia sp. and Babesia sp.) by using multiplex polymerase chain reaction (PCR) method. We collected a total of 875 ticks from the ground and from various animals and kept them at 4 °C until experiments. We identified 248 Rhipicephalus bursa (28.3% of the total), 205 (23.4%) Rhipicephalus annulatus, 197 (22.5%) Haemaphysalis concinna, 149 (17.0%) Rhipicephalus sanguineus, 24 (2.7%) Hyalomma marginatum, 21 (2.4%) Ixodes ricinus, 13 (1.5%) Rhipicephalus kohlsi, 5 (0.6%) Hyalomma anatolicum, 5 (0.6%) Hyalomma aegyptium, 5 (0.6%) Dermacentor niveus and 3 (0.3%) Ixodes hexagonus. We included a total of 328 questing ticks in the study: 63 R. bursa, 63 R. sanguineus, 63 R. annulatus, 63 H. concinna, 24 H. marginatum, 21 I. ricinus, 13 R. kohlsi, 5 H. anatolicum, 5 H. aegyptium, 5 D. niveus and 3 I. hexagonus. Multiplex PCR indicated that 80 (24.4%) ticks were infected with Rickettsia sp., 5 (1.5%) with B. burgdorferi and 1 (0.3%) with Babesia sp. Our study indicated that Rickettsia is more common in ticks collected around Istanbul. [ABSTRACT FROM AUTHOR]

This document is a correction notice for an article titled "Tick-Borne Pathogens Screening Using a Multiplex Real-Time Polymerase Chain Reaction-Based Method" published in Acta Parasitologica. The original article contained a mistake in reference 28, which has now been corrected. The correction provides the accurate reference and the original article can be accessed online. The publisher, Springer Nature, remains neutral and unbiased in terms of jurisdictional claims and institutional affiliations. The authors of the article are listed at the end of the correction notice. [Extracted from the article]

The importance of vectors and vector-borne diseases (VBDs) is increasing on a global scale. Many vectors and pathogens benefit from global warming and can spread to novel habitats where they were formerly not present, including higher altitudes. Various vector-borne pathogens (VBPs), such as Anaplasma phagocytophilum, have been reported in, for instance, red foxes and wild ungulates in the Western Austrian Alps. However, these animals are known to migrate to lower regions in the winter season, and therefore, it is of interest to investigate if VBPs are also present in mammals faithful to their higher altitude alpine habitat all year round. Blood parasites and other VBPs, namely. Trypanosomatidae, piroplasms, Hepatozoon spp., filarioid helminths, Anaplasmataceae, and Rickettisa spp., were thus analysed with PCR in 148 alpine marmots (Marmota marmota). None of the marmots' blood samples was positive for these VBPs, indicating a low abundance or absence of competent vectors in the alpine region. Alpine marmots seem to be naïve for VBPs (at least in our study area). An overview of VBD agents in other marmot species is given. [ABSTRACT FROM AUTHOR]

Ixodes ricinus (Acari: Ixodida) is the main vector in Europe of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. Wolinski National Park (WNP) is situated by the Baltic Sea and is frequently visited by tourists. The aim of the study was to determine the potential risk of exposure to tick borne infection with B. burgdorferi s.l., A. phagocytophilum and B. microti on the areas of WNP. In total, 394 I. ricinus were tested. The pathogens in ticks were detected by PCR, nested PCR, RFLP and sequencing. Altogether, pathogens were detected in 12.69% of the studied ticks. B. burgdorferi s.l., was shown in 0.25% of the studied I. ricinus, while A. phagocytophilum and B. microti were detected in 1.01% and 10.65% of studied ticks, respectively. Co-infection by A. phagocytophilum and B. microti was shown in only one I. ricinus nymph. Analysis of B. burgdorferi s.l., genospecies showed that 0.25% of the studied ticks were infected with Borrelia garinii. The obtained results show the potentially high human risk of exposure to tick-borne infection with B. microti, and the low potential risk of infection with B. garinii and A. phagocytophilum on the studied areas of WNP. [ABSTRACT FROM AUTHOR]

Tick-borne diseases constitute a problem for livestock and public health. Given the socio-economic and environmental conditions of the Colombian Caribbean, ticks are particularly abundant, in turn exposing domestic animals and people in contact with them to such diseases. This study evaluates the presence of Babesia spp., Anaplasma spp., Coxiella spp. and Borrelia spp. in domestic animal ticks (Amblyomma mixtum, A. dissimile, Dermacentor nitens, Rhipicephalus sanguineus and R. microplus) by conventional PCR. Findings show a prevalence of 12.5% of Babesia, 0% of Borrelia, 39.4% of Anaplasma and 52.9% of Coxiella, whereas 6.2% of a total sample of 104 tick pools presented coinfections between Babesia and Anaplasma. Among the molecularly identified species are Ba. vogeli, Ba. bigemina and A. marginale, in addition to two Coxiella species—one being C. mudrowiae and the other similar to an undescribed endosymbiont of Rhipicephalus sp. It is necessary to evaluate the vector capacity of ticks such as A. mixtum, D. nitens and R. sanguineus in the transmission of A. marginale. Moreover, it is necessary to explore the role that bacteria of the genus Coxiella might have both in the health of humans and animals, and in the metabolism and reproduction of ticks. This is the first report on Babesia vogeli and B. bigemina in ticks from the Colombian Caribbean, representing a risk to animal and human health. [ABSTRACT FROM AUTHOR]

Ticks are ectoparasites that feed on blood of a broad taxonomic range of terrestrial and flying vertebrates and are distributed across a wide range of environmental conditions. Here, we explore the biotic and abiotic factors on infestation probability of ticks of the genus Amblyomma and assess the degree of host specificity based on analysis of 1028 birds surveyed across Brazil. We show that tick infestation rates exhibited considerable variation across the 235 avian species analyzed and that the probability of an individual bird being parasitized by immature ticks (larvae and nymphs) increased with annual precipitation. Host phylogeny and two host ecological traits known to promote tick exposure (body mass and foraging behavior) did not predict infestation probability. Moreover, immature ticks displayed a low degree of host specificity at the family level. Lastly, tick occurrence in birds carrying infection with avian malaria and related parasites did not differ from those free of these haemosporidian parasites, indicating a lack of parasite avoidance by immature ticks. Our findings demonstrate that tick occurrence in birds across Brazilian biomes responds to environmental factors rather than ecological and evolutionary host attributes. [ABSTRACT FROM AUTHOR]

Southern Germany is known as tick-borne encephalitis (TBE) risk area; however, the north of the country is almost free of human TBE cases. Due to its location in the transition zone between TBE risk areas and areas with only sporadic cases, Saxony is of importance in the surveillance of TBE. Roe deer (Capreolus capreolus), showing high seroprevalence of TBE virus (TBEV) antibodies, are considered to be sentinels for TBE risk assessment. Thus, roe deer could be used as indicators helping to better understand the focality of the TBEV in nature and as a possible source to isolate TBEV. Therefore, the aims of this study were to examine roe deer coats for the presence of ticks to establish the tick burden and to detect the TBEV in attached ticks. One hundred thirty-four roe deer coats were provided by hunters from the Hunting Association in Saxony (August 2017–January 2019). The coats were frozen at − 80 °C and after de-freezing examined on both sides—inside and outside. Attached and nonattached ticks were collected, morphologically identified and tested using real-time RT-PCR for the presence of TBEV. In total, 1279 ticks were found on 48 coats. The predominant species was Ixodes ricinus (99.76%; n = 1276). Three remaining specimens were Ixodes spp. (0.16%, 1 female and 1 nymph) and Dermacentor reticulatus (0.08%, 1 male). The average infestation rate was 26.7 (SD = 69.5), with maximum of 439 ticks per animal. Females were the dominant life stage of ticks (n = 536; 42%), followed by nymphs (n = 397; n = 31.1%), males (n = 175; 13.7%), and larvae (n = 168; 13.2%). Only half of collected ticks were attached (n = 662; 51.8%). TBEV was detected only in one tick out of 1279 tested ticks. It was a female infesting a roe deer from Saxon Switzerland-East Ore Mountain. The results show that the method used in this study is not sufficient as a sentinel marker to predict TBEV spreading in nature. Although previous studies demonstrated the usefulness of serological testing of roe deer in order to trace TBE-endemic regions, using ticks attached to them to get virus isolates is not productive. [ABSTRACT FROM AUTHOR]

The roe deer (Capreolus capreolus) has been identified as an intermediate host for six known Sarcocystis species, S. capreolicanis, S. entzerothi, S. gracilis, S. linearis, S. oviformis, and S. silva. In this study, we identified Sarcocystis species in the diaphragm and tongue muscles from the Lithuanian and Spanish roe deer, respectively, on the basis of a microscopic examination and DNA analysis. A total of 43 and 27 sarcocysts were isolated and characterized from the Lithuanian and Spanish roe deer, respectively. Overall six Sarcocystis species were identified in roe deer from Lithuania, and only three of them, S. gracilis, S. linearis, and S. silva were found to have infecting animals from Spain. The current paper represents first molecular results of Sarcocystis species in the Spanish roe deer. Furthermore, transmission electron microscopy examination revealed specific wall structure of sarcocysts studied, S. linearis was characterized by ribbon-like villar protrusions (vp) (type 8a), and S. oviformis was distinguished by elongated vp resembling spades or mushroom-like structures (type 39). Based on 18S rDNA and cox1 sequences, Sarcocystis species from the roe deer showed considerable intraspecific genetic variability. However, similar values of intraspecific genetic variation were estimated at both genes analysed. The highest variability was observed for S. capreolicanis and S. linearis in both genes and for S. silva at cox1. Consequently, the level of genetic variability of Sarcocystis from the roe deer varied depending on species rather than on gene analysed or geographical area. [ABSTRACT FROM AUTHOR]

Ticks are important parasites from economic and public health points of view because of their ability to reduce farm animals' productivity and transmit zoonotic diseases. We conducted this cross-sectional study between January and March 2016 and between March and April 2017 to identify tick species in West Darfur, Al-Jazeera, and the River Nile states in the Sudan and to investigate whether these ticks carry Rickettsia spp. and Crimean-Congo hemorrhagic fever (CCHF) virus. In total, 1593 ticks were collected from 207 animals and identified based on morphology or 16S rRNA gene and tested for Rickettsia spp. and CCHF virus either individually or as pools containing 2 to 10 pooled ticks using molecular methods. Overall, 14 tick species belonging to three genera, namely Amblyomma, Hyalomma, and Rhipicephalus, were identified. Hyalomma anatolicum and Rhipicephalus evertsi evertsi were the most frequent ticks. A total of 561 tests comprised of individual or pooled ticks were conducted and 13.7% (77/561) were positive for Rickettsia spp. which were mostly Rickettsia aeschlimannii and R. africae. The highest positivity was noticed among H. rufipes collected from cattle and camels in West Darfur. However, none of the screened Hyalomma ticks harbored CCHF viral RNA. These findings suggest that there might be a risk of zoonotic transmission of Rickettsia spp. by ticks but zoonotic transmission of CCHF virus is apparently doubtful. An in-depth and a country-wide epidemiological study is needed to better understand the dynamic of Rickettsia spp. and CCHF virus in the Sudan. [ABSTRACT FROM AUTHOR]

Purpose: The aim of the study is to assess anti-Coxiella burnetii antibodies presence in inhabitants of north-eastern Poland, to assess the risk of Q fever after tick bite and to assess the percentage of co-infection with other pathogens. Methods: The serological study included 164 foresters and farmers with a history of tick bite. The molecular study included 540 patients, hospitalized because of various symptoms after tick bite. The control group consisted of 20 honorary blood donors. Anti-Coxiella burnetii antibodies titers were determined by Coxiella burnetii (Q fever) Phase 1 IgG ELISA (DRG International Inc. USA). PCR was performed to detect DNA of C. burnetii, Borrelia burgdorferi and Anaplasma phagocytophilum. Results: Anti-C. burnetii IgG was detected in six foresters (7.3%). All foresters with the anti-C. burnetii IgG presence were positive toward anti-B. burgdorferi IgG and anti-TBE (tick-borne encephalitis). Anti-C. burnetii IgG was detected in five farmers (6%). Four farmers with anti-C. burnetii IgG presence were positive toward anti-B. burgdorferi IgG and two with anti-TBE. Among them one was co-infected with B. burgdorferi and TBEV. Correlations between anti-C. burnetii IgG and anti-B. burgdorferi IgG presence and between anti-C. burnetii IgG presence and symptoms of Lyme disease were observed. C. burnetii DNA was not detected in any of the 540 (0%) patients. Conclusions: C. burnetii is rarely transmitted by ticks, but we proved that it is present in the environment, so it may be a danger to humans. The most common co-occurrence after tick bite concerns C. burnetii and B. burgdorferi. [ABSTRACT FROM AUTHOR]

Context: By modifying ecosystems, land cover changes influence the emergence, the spread and the incidence of vector-borne diseases. Objective: This study aimed at identifying associations between landscape structure and the prevalence of two tick-borne infectious agents, Anaplasma phagocytophilum and Borrelia burgdorferi s.l., in small mammal communities. Methods: Small mammals were sampled in 24 sites along a gradient of woodland fragmentation and hedgerow network density, and screened for infectious agents with real-time PCR techniques. For each site, structural variables (composition and configuration) of the surrounding landscape at various scales (0–500 m) and variables of wooded habitats connectivity based on graph theory and least cost path distances for the two dominant species, bank voles (Myodes glareolus) and wood mice (Apodemus sylvaticus), were computed. Results: The A. phagocytophilum prevalence increased with wooded habitats cover (0–500 m), likely through host population size, and increased slightly with bank vole abundance, which has a higher reservoir competence than wood mouse. The B. burgdorferi s.l. prevalence increased with wooded ecotones only at local scales (50–100 m). Wooded habitats connectivity measures did not improve models built with simple land cover variables. A more marked spatial pattern was observed for the prevalence of A. phagocytophilum than B.burgdorferi s.l. Conclusions: This study highlights the interest of considering together the ecology of infectious agents (e.g. host specificity) and the host species community ecology to better understand the influence of the landscape structure on the spatial distribution of vector-borne infectious agents. [ABSTRACT FROM AUTHOR]

PCR-based single-strand conformation polymorphism (SSCP) analyses combined with DNA sequencing of the prokaryotic 16S ribosomal (r) RNA gene encompassing the hypervariable V4 region was used to determine the bacterial composition of Rocky Mountain wood ticks (Dermacentor andersoni) attached to Richardson's ground squirrels (Urocitellus richardsonii) and questing on vegetation in southern Saskatchewan, Canada. The bacteria present in questing adult ticks from Saskatchewan Landing Provincial Park included Rickettsia peacockii, a Francisella-like endosymbiont (FLE) and an Arsenophonus-like endosymbiont. Bacteria in the adult and nymphal ticks attached to U. richardsonii collected from Beechy included R. peacockii, a FLE, and several other genera (e.g., Ralstonia, Sphingobium, Comamonas and Pseudomonas). The bacteria detected in D. andersoni in the present study are consistent with the findings of other studies that have characterized the microbiome of this tick species in the USA using next generation sequencing. This result demonstrates that the SSCP-based approach used in this study is cost- and time-effective for examining bacterial composition in ticks. [ABSTRACT FROM AUTHOR]

The capability of imidacloprid 10% + flumethrin 4.5% (Seresto®) collars to prevent transmission of Borrelia burgdorferi sensu lato (Bbsl) and Anaplasma phagocytophilum (Ap) by naturally infected ticks was evaluated in two studies with 44 dogs. In each study, one group served as non-treated control, whereas the other groups were treated with the Seresto® collar. All dogs were exposed to naturally Bbsl- and Ap-infected hard ticks (Ixodes ricinus, Ixodes scapularis). In study 1, tick infestation was performed on study day (SD) 63 (2 months post-treatment [p.t.]); in study 2, it was performed on SD 32 (one month p.t.) respectively SD 219 (seven months p.t.). In situ tick counts were performed 2 days after infestation. Tick counts and removals followed 6 (study 1) or 5 days (study 2) later. Blood sampling was performed for the detection of specific Bbsl and Ap antibodies and, in study 1, for the documentation of Ap DNA by PCR. Skin biopsies were examined for Bbsl by PCR and culture (only study 1). The efficacy against Ixodes spp. was 100% at all time points. In study 1, two of six non-treated dogs became infected with Bbsl, and four of six tested positive for Ap; none of the treated dogs tested positive for Bbsl or Ap. In study 2, ten of ten non-treated dogs became infected with Bbsl and Ap; none of the treated dogs tested positive for Bbsl or Ap; 100% acaricidal efficacy was shown in both studies. Transmission of Bbsl and Ap was successfully blocked for up to 7 months. [ABSTRACT FROM AUTHOR]

We investigated the effectiveness of integrated tick management (ITM) approaches in reducing the burden of infection with Borrelia burgdorferi, Babesia microti, and Anaplasma phagocytophilum in Ixodes scapularis. We found a 52% reduction in encountering a questing nymph in the Metarhizium anisopliae (Met52) and fipronil rodent bait box treatment combination as well as a 51% reduction in the combined white-tailed deer (Odocoileus virginianus) removal, Met52, and fipronil rodent bait box treatment compared to the control treatment. The Met52 and fipronil rodent bait box treatment combination reduced the encounter potential with a questing nymph infected with any pathogen by 53%. Compared to the control treatment, the odds of collecting a parasitizing I. scapularis infected with any pathogen from a white-footed mouse (Peromyscus leucopus) was reduced by 90% in the combined deer removal, Met52, and fipronil rodent bait box treatment and by 93% in the Met52 and fipronil rodent bait box treatment combination. Our study highlights the utility of these ITM measures in reducing both the abundance of juvenile I. scapularis and infection with the aforementioned pathogens. [ABSTRACT FROM AUTHOR]

In this report, concurrent infection of Leptospira interrogans serovar icterohaemorrhagiae, Hepatozoon canis and Anaplasma phagocytophilum in a Labrador Retriever dog and its successful therapeutic management have been described. A 3-year-old female dog was presented with a history of anorexia, fever, lethargy, weakness, vomiting, black-coloured faeces and yellowish urine. Clinical and laboratory examination revealed pyrexia, icteric mucous membranes, hepatomegaly, splenomegaly, anaemia, leukocytosis, thrombocytopaenia, hypoalbuminemia and hyperbilirubinemia. Hepatozoon canis and Anaplasma phagocytophilum infections were confirmed by blood smear examination. Leptospira icterohaemorrhagiae was confirmed by microscopic agglutination test (MAT). Animal was stabilised with polyionic fluids, and therapy was started with imidocarb dipropionate (6.6 mg/Kg, SC, single dose), doxycycline (5 mg/kg, PO, bid) for 28 days, N-acetyl cysteine (30 mg/Kg, IV, SID) for 5 days and fipronil (0.25%) spray for topical use. Supportive therapy was done with plasma volume expanders, multivitamins, haematinics and hepatoprotectants. The animal showed complete recovery after 2 months of therapy. [ABSTRACT FROM AUTHOR]

In 2014, a new tick species, Ixodes inopinatus, was described, which is closely related to Ixodes ricinus. So far, I. inopinatus has been found in Tunisia, Morocco, Spain, Portugal, Romania, Austria, and southern Germany. No data is yet available regarding occurrence of I. inopinatus in northern Germany and the potential role of I. inopinatus as a vector for tick-borne pathogens. Therefore, 3845 DNA samples from Ixodes ticks collected for prevalence studies on Borrelia spp., Rickettsia spp., and Anaplasma phagocytophilum during the years 2010–2015 in the northern German cities of Hamburg and Hanover were differentiated into I. ricinus or I. inopinatus by sequencing a part of the 16S rRNA gene. In total, 4% (137/3845) of the sequenced ticks were assigned to the species I. inopinatus and 96% (3708/3845) to I. ricinus. The prevalence of Borrelia spp., Rickettsia spp., and A. phagocytophilum DNA in I. inopinatus was 34% (46/137), 46% (63/137), and 3% (4/137), respectively, whereas the prevalence of these bacteria in I. ricinus was 25% (919/3708), 47% (1729/3708), and 4% (135/3708), respectively. Compared with I. ricinus, significantly more I. inopinatus ticks tested positive for Borrelia. To the best of our knowledge, this is the first report of I. inopinatus in northern Germany. Detection of the DNA of Borrelia spp., Rickettsia spp., and A. phagocytophilum in questing I. inopinatus indicates a potential role of this tick species as a vector of these pathogens, which needs to be confirmed by transmission experiments. [ABSTRACT FROM AUTHOR]

Purpose: There are more than 500,000 employees in Germany alone who are at risk of being bitten by a tick at their workplace and thus also at risk of being infected with Borrelia burgdorferi s.l. or the tick-borne meningoencephalitis virus. So far, there are only a small number of studies on the risk of tick bites in Central Europe, in particular, for military personnel during relevant training activities. Methods: During a total of 36 months of training in 2008/2009 and from 2012 to 2014, the number of tick bites and any resulting diseases of 1156 recruits under comparable conditions of exposure and prevention were documented based on their medical records. The incidence of tick bites was calculated with each recruit's individual exposure time for field training. Results: There were a total of 66 tick bites during an exposure time of 317,059 h of field training (0.21 tick bites per 1000 h of training). The risk of tick bites was found to have a seasonal dependency. In 6 out of the 66 cases in which someone was bitten, the patients consulted a physician for a follow-up examination, and in one of these cases the bite resulted in an infection requiring treatment. Conclusions: It turns out that there is a rather low but relevant risk of being exposed to tick bites for military personnel during their field training. Under the given study conditions, months with a high risk of tick bites can be distinguished from months with a low risk of tick bites, which is of vital importance, in particular, for guidance and prevention. [ABSTRACT FROM AUTHOR]

Context: Tick-borne diseases are becoming increasingly prevalent world-wide. This is likely due in part to land-cover change, particularly forest fragmentation, but this evidence is largely limited to Lyme disease. It is unknown whether this is generalizable to other, emergent tick-borne pathogens. Objectives: Motivated by hypotheses regarding landscape context and vertebrate hosts, we asked how landscape context, namely habitat amount, quality, and fragmentation, is related to the distribution of Ehrlichia chaffeensis, a tick-borne pathogen of increasing concern, and the interannual occupancy dynamics of its vector, the lone star tick (Amblyomma americanum). Methods: We collected nymphal ticks from 130 plots in southeastern Virginia, U.S., for 5 years and tested for E. chaffeensis via targeted PCR. We derived metrics of landscape context from Landsat data and related these to pathogen prevalence and tick turnover using hierarchical Bayesian models. Results: Landscape context was associated with both pathogen prevalence and tick turnover. Pathogen prevalence was negatively associated with total forest landcover, coniferous forest landcover, and forest edge density. Tick turnover was positively associated with coniferous landcover and with an interaction between total forest landcover and edge. This interaction was such that turnover was predicted to be lowest in small contiguous forests, and highest in small fragmented forests. Conclusions: Landscape context affects E. chaffeensis prevalence and occupancy dynamics of its tick host, though these processes appear decoupled. We hypothesize that pathogen prevalence may be more driven by reservoir host movement and social behavior and tick dynamics are more driven by host population density. [ABSTRACT FROM AUTHOR]

Tick-borne rickettsioses, most of them belonging to the spotted fever group (SFG), have been recognized as important emerging vector-borne zoonotic diseases. In order to determine the presence of Rickettsia spp. in questing ticks from north-western Spain, 1056 Ixodes ricinus, 19 Dermacentor marginatus, 17 Dermacentor reticulatus and one Ixodes acuminatus were processed. Rickettsia DNA was detected by PCR targeting rOmpA and rOmpB genes. A total of 219 (20.7%) I. ricinus, 19 (100%) D. marginatus and four D. reticulatus (23.5%) were positive. The prevalence was significantly higher in I. ricinus from coastal areas and in winter. Five species were identified: Rickettsia felis, Rickettsia monacensis, Rickettsia raoultii, Rickettsia slovaca and "Candidatus Rickettsia rioja". Our results reveal a significant presence of some pathogenic Rickettsia species in questing tick populations from this area which involves a noticeable risk of rickettsiosis. As R. raoultii, R. slovaca and "Ca. R. rioja" DNA were identified in I. ricinus, considered an unusual vector for these Rickettsia species, further studies are needed to unravel the role of that tick species in the maintenance and transmission of these three Rickettsia species in north-western Spain. [ABSTRACT FROM AUTHOR]