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Background: Bovine anaplasmosis caused by Anaplasma marginale is highly endemic in tropical and sub-tropical regions of the world where it accounts for huge economic losses to the livestock sector. Approximately 30% of water buffalo (Bubalus bubalis) herds in India are located in Uttar Pradesh (U.P). However in India, most of the research studies on Anaplasma are focused on cattle whereas studies on this pathogen in water buffaloes have been neglected. Therefore, present investigation was undertaken for determining the prevalence of A. marginale infection in water buffaloes in U.P, North India. Methods: In the current investigation during 2017-2019, the prevalence of A. marginale infection in 228 water buffaloes in three districts of North-West U.P was studied by microscopy and by nested-PCR amplification of major surface protein 5 (msp5) gene. Result: Microscopic examination of blood smears of 228 water buffaloes revealed 7.89% animals positive for A. marginale. Conventional PCR of full length major surface protein 5 (msp5) gene detected 29.39% and nested-PCR amplification of this gene detected 64.0% of these animals as positive for A. marginale infection. A high incidence of the infection detected in this study area suggests that water buffaloes may serve as reservoir hosts of A. marginale and play a vital role in the spread of infection to other susceptible animals. However further studies are required on larger buffalo populations for determining the epidemiological status of anaplasmosis in these animals in India. [ABSTRACT FROM AUTHOR]

Since tick-borne diseases (TBDs) incidence, both in human and animal populations, is increasing worldwide, there is the need to assess the presence, distribution and prevalence of tick-borne pathogens. Reliable estimates on tick-borne pathogens (TBPs) prevalence represent the public health foundation to create risk maps and take effective prevention and control actions against TBDs. Tick surveillance consists of collecting and testing (usually in pools) thousands of specimens. Construction and analysis of tick pools represent a challenge due to the complexity of tick-borne pathogens and tick-borne diseases ecology. The aim of this study is to provide a practical guideline on appropriate pooling strategies and statistical analysis of infection prevalence through: (i) reporting the different pooling strategies and statistical methodologies commonly used to calculate pathogen prevalence in tick populations and (ii) practical comparison between statistical methods utilising a real dataset of infection prevalence in ticks collected in Northern Italy. Reporting detailed information on tick pool composition and size is as important as the correct TBPs prevalence estimation. Among the prevalence indexes, we suggest using maximum-likelihood estimates of pooled prevalence instead of minimum infection rate or pool positivity rate given the merits of the method and availability of software. [ABSTRACT FROM AUTHOR]

Birds are one of the most species-diverse vertebrate groups and are susceptible to numerous hematophagous ectoparasites. Migratory birds likely contribute to the circulation of these ectoparasites and their associated pathogens. One of the many migration paths crosses the Mediterranean islands including Corsica and its wetlands, which are migration stopovers. In our study, we collected blood samples and hematophagous ectoparasites in migratory and sedentary bird populations in two coastal lagoons: Biguglia and Gradugine. A total of 1377 birds were captured from which 762 blood samples, 37 louse flies, and 44 ticks were collected. All the louse flies were identified as Ornithomya biloba and all the ticks were from the Ixodes genus: Ixodes sp. (8.5%), I. accuminatus/ventalloi (2.9%), I. arboricola/lividus (14.3%), I. frontalis (5.7%) and I. ricinus (68.6%). Five pathogens were detected: Anaplasma phagocytophilum, Erhlichia chaffeensis, and Rickettsia helvetica in ticks, and Trypanosoma sp. in louse flies. Ehrlichia chaffeensis and the West Nile virus were both detected in bird blood samples in Corsica. This is the first report of these tick, louse fly and pathogen species isolated on the bird population in Corsica. Our finding highlights the importance of bird populations in the presence of arthropod-borne pathogens in Corsican wetlands. [ABSTRACT FROM AUTHOR]

The present study was performed to survey the dominant tick populations and molecularly determine the pathogenic agents of anaplasmosis in ticks from Gyeongsang, Republic of Korea. A total of 3825 questing ticks were collected by the flagging method from 12 sites near animal farms in Gyeongsang from March to October 2021. A molecular genomic study was performed with ticks stored in 70% ethanol to detect Anaplasma genes by the previously described method. The monthly incidence of ticks varied by developmental stages, i.e., nymphs, adults, and larvae, and each of their populations peaked in May, March, and October, respectively. The predominant tick species were Haemaphysalis longicornis, Haemaphysalis sp., Haemaphysalis flava, Ixodes nipponensis, and Amblyomma testudinarium in order. To determine the Anaplasma infection rate, collected ticks were pooled into 395 groups. The minimum infection rate (MIR) of Anaplasma was 0.7% (27 pools). That of A. phagocytophilum was highest (23 pools, MIR 0.6%), followed by A. phagocytophilum-like Anaplasma spp. clade B (2 pools, MIR 0.1%), A. bovis (1 pool, MIR 0.1%), and A. capra (1 pool, MIR 0.1%), respectively. In this study, five species of ticks, including unidentified Haemaphysalis species, were collected in 12 survey sites in Gyeongsang, but their prevalence was somewhat different according to the tick species and survey sites. Further, the incidence rate (6.8%) of 4 Anaplasma spp. was not as high in tick pools. However, the results of this study may offer a basis for future epidemiological research and risk assessment of tick-borne diseases. [ABSTRACT FROM AUTHOR]

We report 2 cases of Spiroplasma ixodetis infection in an immunocompetent patient and an immunocompromised patient who had frequent tick exposure. Fever, thrombocytopenia, and increased liver aminotransferase levels raised the suspicion of anaplasmosis, but 16S rRNA PCR and Sanger sequencing yielded a diagnosis of spiroplasmosis. Both patients recovered after doxycycline treatment.

According to the worldwide growing number of examined tick-borne diseases, the aim of this study was to evaluate the changes in proteomic profile of human serum induced by the development of Erythema migrans (EM), human granulocytic anaplasmosis (AP) and co-infection before/after antibiotic therapy. A proteomics approach based on SDS-PAGE/LC-MS/MS analysis was used to determine the proteins expression and 15d-PGE2 adducts level with albumin isolated from the serum of patients and sex/age-matched healthy donors. In the serum proteome of the patients with EM or/and AP, significant changes occurred in the expression of the same top 15 modified proteins; however, each protein level was modified in the different way in terms of comparing B. burgdorferi and A. phagocytophilum infections, as well as before and after therapy. In the case of co-infection, the differences in protein expression before and after therapy were significantly lower than in the monoinfection. Moreover, both EM and AP infections/co-infection significantly increased the albumin ability to create adducts with 15d-PGE2. The therapy partially reversed this property only in the case of a single infection, but this effect was not observed for therapy of co-infections. The results demonstrate how challenging is the treatment of the tick-borne co-infections and how important is further analysis of this subject. Individual differences are also observed in each of examined in this study diseases, which makes it more difficult to develop a common biomarker for each of the tick-borne diseases. • Proteins are modified differently in B. burgdorferi and A. phagocytophilum infection. • Protein expression changes more after therapy in co-infection than monoinfection. • EM and AP infections significantly increase the albumin-15d-PGE2 adducts creation. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum is an obligatory intracellular bacterium that causes tick-borne zoonosis called human granulocytic anaplasmosis. Mechanisms by which Anaplasma replicates inside of the membrane-bound compartment called "inclusion" in neutrophils are incompletely understood. A small GTPase Rab27a is found in the secretory granules and multivesicular endosomes. In this study we found Rab27a-containing granules were localized to Anaplasma inclusions in guanine nucleotide-dependent manner, and constitutively active Rab27a enhanced Anaplasma infection and dominant-negative Rab27a inhibited Anaplasma infection. Rab27a effector, JFC1 is known to mediate docking/fusion of Rab27a-bearing granules for exocytosis in leukocytes. shRNA stable knockdown of Rab27a or JFC1 inhibited Anaplasma infection in HL-60 cells. Similar to Rab27a, both endogenous and transfected JFC1 were localized to Anaplasma inclusions by immunostaining or live cell imaging. The JFC1 C2A domain that binds 3′-phosphoinositides, was sufficient and required for JFC1 and Rab27a localization to Anaplasma inclusions which were enriched with phosphatidylinositol 3-phosphate. Nexinhib20, the small molecule inhibitor specific to Rab27a and JFC1 binding, inhibited Anaplasma infection. Taken together, these results imply elevated phosphatidylinositol 3-phosphate in the inclusion membrane recruits JFC1 to mediate Rab27a-bearing granules/vesicles to dock/fuse with Anaplasma inclusions, the lumen of which is topologically equivalent to the exterior of the cell to benefit Anaplasma proliferation. [ABSTRACT FROM AUTHOR]

The incidence of tick-borne infections other than Lyme borreliosis and tick-borne encephalitis is rising in Europe, including the Netherlands. Nature management workers, being highly exposed to ticks, serve as valuable sentinels for seroprevalence studies on tick-borne pathogens (TBPs). This study assessed nature management workers' seropositivity to TBPs including Anaplasma phagocytophilum, Babesia divergens, B. microti, Borrelia burgdorferi s.l., Rickettsia conorii and R. typhi in the Netherlands. In addition, the study examined coexposure to multiple TBPs and identified risk factors for B. burgdorferi s.l.- and A. phagocytophilum -seropositivity. The study included 525 nature management workers who donated serum and completed a questionnaire. Sera were analysed for exposure to A. phagocytophilum, B. divergens, B. microti, R. conorii and R. typhi using immunofluorescence assays. For B. burgdorferi s.l. antibody detection, the recommended two-tier testing strategy was used. Risk factor analysis was performed using logistic regression modelling. Seropositivity was 30.9 % for B. burgdorferi s.l.; 16.4 % for A. phagocytophilum ; 6.5 % for R. conorii ; 2.3 % for R. typhi ; 4.2 % for B. divergens ; and 0.4 % for B. microti. Almost half (49.3 %) of the participants demonstrated seropositivity for one or more pathogens. Risk factors for B. burgdorferi s.l.-seropositivity included being male, increasing age and tick bite frequency. For A. phagocytophilum -seropositivity, increasing age and working in North Holland province were significant risk factors. This study illustrates the exposure to TBPs in the Netherlands, emphasizing the need for ongoing vigilance and international collaborations to better understand and address the growing threat of TBPs in regions with demonstrated environmental TBP circulation. [ABSTRACT FROM AUTHOR]

Background: The Gram-negative bacterium Anaplasma phagocytophilum is an intracellular pathogen and an etiological agent of human and animal anaplasmosis. Its natural reservoir comprises free-ranging ungulates, including roe deer (Capreolus capreolus) and red deer (Cervus elaphus). These two species of deer also constitute the largest group of game animals in Poland. The aim of the study was to genotype and perform a phylogenetic analysis of A. phagocytophilum strains from roe deer and red deer. Methods: Samples were subjected to PCR amplification, sequencing, and phylogenetic analysis of strain-specific genetic markers (groEL, ankA). Results: Five haplotypes of the groEL gene from A. phagocytophilum and seven haplotypes of ankA were obtained. The phylogenetic analysis classified the groEL into ecotypes I and II. Sequences of the ankA gene were classified into clusters I, II, and III. Conclusions: Strains of A. phagocytophilum from red deer were in the same ecotype and cluster as strains isolated from humans. Strains of A. phagocytophilum from roe deer represented ecotypes (I, II) and clusters (II, III) that were different from those isolated from red deer, and these strains did not show similarity to bacteria from humans. However, roe deer can harbor nonspecific strains of A. phagocytophilum more characteristic to red deer. It appears that the genetic variants from red deer can be pathogenic to humans, but the significance of the variants from roe deer requires more study.

Abstract Background In Mongolia, the taiga tick Ixodes persulcatus is the major vector of tick-borne pathogens. Knowledge about co-infections of these pathogens in ticks is necessary both for understanding their persistence in nature and for diagnosing and treating tick-borne diseases. Methods The prevalence of seven tick-borne infections in 346 I. persulcatus collected from the Selenge and Bulgan provinces of Mongolia was evaluated using real-time PCR. Quantification of Borrelia spp. was performed using multiplex quantitative PCR targeting the 16S rRNA gene. Genetic analysis of Borrelia spp. in 11 ticks infected with Borrelia miyamotoi, including six ticks co-infected with Borrelia burgdorferi sensu lato (s.l.), was performed by high-throughput sequencing of the flaB gene fragment. Results Six ticks (1.7%) were infected with tick-borne encephalitis virus (TBEV); 171 (49.4%), with B. burgdorferi sensu lato; 17 (4.9%), with B. miyamotoi; 47 (13.6%), with Anaplasma phagocytophilum; and 56 (16.2%), with Ehrlichia sp. Neither Rickettsia sibirica nor R. heilongjiangensis were detected. Borrelia burgdorferi s.l. occurred as co-infection in 55 (32.2%) of all infected ticks. The other pathogens co-infected ticks in 58.8–70.2% of cases. No pairwise associations between co-infecting pathogens were observed, with the exception of a positive association between A. phagocytophilum and Ehrlichia sp. infections. The spirochete loads of B. miyamotoi were significantly higher than those of B. burgdorferi s.l. (mean: 5.2 vs 4.0 log10 genome copies/tick, respectively). Ten isolates of B. miyamotoi belonged to the Siberian lineage. Borrelia burgdorferi s.l was represented by nine isolates of B. afzelii, B. bavariensis and B. garinii. Conclusions In populations of I. persulcatus inhabiting the Selenge and Bulgan provinces of Mongolia, five vector-borne pathogens, i.e. TBEV, B. burgdorferi s.l., B. miyamotoi, A. phagocytophilum and Ehrlichia sp., persist independently from each other, with the exception of A. phagocytophilum and Ehrlichia sp. which seem to share the circulation mode. The discrepancies in B. burgdorferi s.l. and B. miyamotoi prevalence and spirochete load per tick suggest that different ecological niches are occupied by Lyme disease and relapsing fever agents. High-throughput sequencing allows genetic identification of borreliae species in co-infected ticks. Graphical Abstract

Abstract Background Human granulocytic anaplasmosis is a tick-borne zoonotic disease caused by Anaplasma phagocytophilum. Coinfections with A. phagocytophilum and other tick-borne pathogens are reported frequently, whereas the relationship between A. phagocytophilum and flea-borne Yersnia pestis is rarely concerned. Results A. phagocytophilum and Yersnia pestis were discovered within a Marmota himalayana found dead in the environment, as determined by 16S ribosomal rRNA sequencing. Comparative genomic analyses of marmot-derived A. phagocytophilum isolate demonstrated its similarities and a geographic isolation from other global strains. The 16S rRNA gene and GroEL amino acid sequence identity rates between marmot-derived A. phagocytophilum (JAHLEX000000000) and reference strain HZ (CP000235.1) are 99.73% (1490/1494) and 99.82% (549/550), respectively. 16S rRNA and groESL gene screenings show that A. phagocytophilum is widely distributed in marmots; the bacterium was more common in marmots found dead (24.59%, 15/61) than in captured marmots (19.21%, 29/151). We found a higher Y. pestis isolation rate in dead marmots harboring A. phagocytophilum than in those without it (2 = 4.047, p

Background: The Ixodes ricinus tick is the main vector of Borrelia burgdorferi and tick-borne encephalitis virus in Switzerland. Spotted fever group Rickettsiae (SFG) and Anaplasma phagocytophilum have been detected in Swiss ticks, however, information about the extent and clinical presentation of these infections in humans is scant. Methods: Indirect fluorescent antibody tests for SFG rickettsiae and Anaplasma phagocytophilum were performed on serum samples of 121 Borrelia burgdorferi seropositive patients with and without Lyme disease and 43 negative controls. Results: Out of 121 Borrelia burgdorferi seropositive individuals, 65 (53.7%) were seropositive for IgG and 15 (12.4%) for IgM antibodies to SFG rickettsiae. IgM antibodies were detected more frequently in early-than in late-stage of Lyme disease (12 out of 51 and 2 out of 49; respectively; p ​= ​0.0078). Significantly higher IgG antibody titers against SFG rickettsiae were found in patients with late-stage compared to patients with early-stage Lyme disease (mean titer 1:261 and 1:129, respectively; p ​= ​0.038). This difference was even more pronounced in patients with acrodermatitis chronica atrophicans compared to patients with early stage of Lyme disease (mean titer 1:337 and 1:129, respectively; p ​= ​0.009).In patients presenting with fatigue, headache and myalgia, the prevalence of IgG antibodies against SFG rickettsiae was significantly higher (7 out of 11; 63.6%) than in Borrelia burgdorferi seropositive individuals without clinical illness (1 out of 10; 10%; p ​= ​0.024). IgG antibodies to Anaplasma phagocytophilum were detected in 12 out of 121 individuals (9.9%), no IgM antibodies were found. Conclusion: Infections with SFG rickettsiae and Anaplasma phagocytophilum are underdiagnosed and should be ruled out after a tick bite. Further studies are needed to elucidate the possible causative role of SFG rickettsiae for myalgia, headache and long-lasting fatigue after a tick bite and to determine the necessity for an antibiotic treatment.

Tick-borne infectious diseases pose a serious health threat in certain regions of the world. Emerging infectious diseases caused by novel tick-borne pathogens have been reported that are causing particular concern. Several tick-borne diseases often coexist in the same foci, and a single vector tick can transmit two or more pathogens at the same time, which greatly increases the probability of co-infection in host animals and humans and can lead to an epidemic of tick-borne disease. The lack of epidemiological data and information on the specific clinical symptoms related to co-infection with tick-borne pathogens means that it is not currently possible to accurately and rapidly distinguish between a single pathogen infection and co-infection with multiple pathogens, which can have serious consequences. Inner Mongolia in the north of China is endemic for tick-borne infectious diseases, especially in the eastern forest region. Previous studies have found that more than 10% of co-infections were in host-seeking ticks. However, the lack of data on the specific types of co-infection with pathogens makes clinical treatment difficult. In our study, we present data on the co-infection types and the differences in co-infection among different ecological regions through genetic analysis of tick samples collected throughout Inner Mongolia. Our findings may aid clinicians in the diagnosis of concomitant tick-borne infectious diseases. Author summary: Ticks carry and transmit a variety of pathogens, and their host animals are widely distributed in nature with diverse epidemic links, so that several tick-borne diseases can coexist in the same foci, and even one type of tick or one tick can carry a variety of pathogens at the same time, these factors pose a serious public health threat. In our study, we summarized the regional differences in tick infection and co-infection rates outcomes in all Inner Mongolia of China, and found that pathogen diversity varied with ecogeographic differences such as gobi desert, grassland and forest. A variety of tick-borne bacterial pathogens were identified, with an overall high prevalence rate (61.4% of ticks infected), among infected ticks, 24.3% were co-infected. An unexpected high infection and co-infection rates of ticks collected from the Forest region of eastern Inner Mongolia (sample area 1) was detected. Geographical differences affect tick species and tick-borne pathogens, so the potential threat to humans or animals is also different. In particular, that that knowledge of co-infections is important as they do not just present diagnostic challenges, but the pathogens might play different roles within their respective hosts, thus modulating disease severity. [ABSTRACT FROM AUTHOR]

Background: The objective of this field study was to assess the effect of treating a considerable portion of a dog population naturally exposed to canine vector-borne pathogens (CVBPs) in endemic areas with a 10% w/w imidacloprid/4.5% w/w flumethrin collar (Seresto®) on the transmission of CVBPs and the resulting incidence of infection. Methods: A total of 479 dogs from two sites were enrolled in the study. Collars were placed on all dogs continuously for 21 months, with replacement of the collar every 7 months. All dogs were examined, including body weight and blood/conjunctival swab collections, every 7 months. Serum samples were analysed for the presence of antibodies against Leishmania infantum, Ehrlichia canis and Anaplasma phagocytophilum. PCR assays were also performed on blood samples and conjunctival swab collected from the dogs for the presence of L. infantum, and on blood samples only for the presence of Ehrlichia spp. and Anaplasma spp. Sand flies were collected, identified to species level and molecularly tested for L. infantum throughout two vector activity seasons. Results: The results showed that the Seresto collar was safe with continuous use. At study inclusion, 419, 370 and 453 dogs tested negative for L. infantum, Ehrlichia spp. and Anaplasma spp., respectively (353 dogs tested negative for any pathogen). Overall, 90.2% of the dogs were protected from L. infantum infection on both sites combined. The entomological survey confirmed the presence of competent vectors of L. infantum at all monitored locations, namely the sand flies Phlebotomus neglectus and Phlebotomus tobbi, both of which are regarded as the most important competent vectors in the Mediterranean basin. All captured sand flies tested negative for L. infantum. Protection against ticks and fleas was high, with only two dogs showing a low number of ticks and seven dogs having low numbers of fleas at single evaluation time points. Across the entire study population, a number of dogs became infected with tick-transmitted pathogens, but prevention of transmission was 93% for E. canis and 87.2% for Anaplasma spp. when all cases from both sites were combined. Conclusions: The Seresto® (10% w/w imidacloprid/4.5% w/w flumethrin) collar significantly reduced the risk of CVBP transmission when compared to previously observed incidences of CVBP infections in two highly endemic areas under field conditions. [ABSTRACT FROM AUTHOR]

Human granulocytic anaplasmosis (HGA) is a tick-borne infection caused by the bacterium Anaplasma phagocytophilum. In this study, we report an indigenous case of clinically diagnosed HGA. The patient was a 41-year-old man who experienced a tick bite and later developed fever, chills, myalgia, malaise, thrombocytopenia, leukocytosis with a left shift, elevated hepatic transaminase levels, and splenomegaly upon admission to the hospital. Immunofluorescence assays detected seroconversion against A. phagocytophilum , whereas tests for spotted fever group rickettsiae, murine typhus, scrub typhus, Q fever, and ehrlichiosis were negative. ELISA and Western blot analysis using recombinant MSP2 protein confirmed the exposure to A. phagocytophilum. Oral doxycycline and intravenous ceftriaxone were prescribed, and the patient made a full recovery. Our findings indicate the presence of HGA on the main island of Taiwan. Precautions against tick bites should be taken when engaging in outdoor activities, and HGA should be considered by physicians in the differential diagnosis. [ABSTRACT FROM AUTHOR]

Equine and canine anaplasmosis and borreliosis are major tick-borne zoonotic diseases caused by Anaplasma phagocytophilum and various species of Borrelia (the most important being Borrelia burgdorferi s.l.), respectively. This study evaluated the seroexposure to Anaplasma and Borrelia in dogs and horses used in Animal-Assisted Interventions or living in contact with children, elderly people or immunocompromised persons. A total of 150 horses and 150 dogs living in Italy were equally divided into clinically healthy animals and animals with at least one clinical sign compatible with borreliosis and/or anaplasmosis (present at clinical examination or reported in the medical history). Serum samples were tested with ELISA and immunoblot for the presence of antibodies against A. phagocytophilum and B. burgdorferi s.l., and the association between seropositivity and possible risk factors was analyzed using multivariate and univariate tests. Overall, 13 dogs (8.7%) and 19 horses (12.7%) were positive for at least one of the two pathogens. In addition, 1 dog (0.7%) and 12 horses (8%) were positive for antibodies against A. phagocytophilum, while 12 dogs (8.0%) and 10 horses (6.7%) had antibodies against B. burgdorferi s.l. Tick infestation in the medical history of the dogs was significantly associated with seropositivity to at least one pathogen (p = 0.027; OR 7.398). These results indicate that, in Italy, ticks infected with A. phagocytophilum and/or B. burgdorferi circulate in places where horses and dogs are in contact with people at risk of developing severe diseases. Awareness should be increased, and adequate control plans need to be developed to protect human and animal health, especially where vulnerable, at-risk individuals are concerned. [ABSTRACT FROM AUTHOR]

In Europe, raccoons are invasive neozoons with their largest population in Germany. Globally, this mesocarnivore acts as a wildlife reservoir for many (non-)zoonotic (re-)emerging pathogens, but very little epidemiological data is available for southwest Germany. This exploratory study aimed to screen free-ranging raccoons in Baden-Wuerttemberg (BW, Germany) for the occurrence of selected pathogens with One Health relevance. Organ tissue and blood samples collected from 102 animals, obtained by hunters in 2019 and 2020, were subsequently analysed for two bacterial and four viral pathogens using a qPCR approach. Single samples were positive for the carnivore protoparvovirus-1 (7.8%, n = 8), canine distemper virus (6.9%, n = 7), pathogenic Leptospira spp. (3.9%, n = 4) and Anaplasma phagocytophilum (15.7%, n = 16). West Nile virus and influenza A virus were not detected. Due to their invasive behaviour and synanthropic habit, raccoons may increase the risk of infections for wildlife, domestic animals, zoo animals and humans by acting as a link between them. Therefore, further studies should be initiated to evaluate these risks. [ABSTRACT FROM AUTHOR]

The aim of the study was to determine prevalence of Anaplasmataceae-infected ticks in the Black Sea Coast and the Pleven regions of Bulgaria. A total of 350 ticks from different tick species were collected. Two hundred fifty-five ticks were removed from dogs in the Black Sea Coast region, and 95 Ixodes ricinus ticks were collected by flagging vegetation with a white flannel cloth in two areas in the region of Pleven. After the DNA isolation of the ticks, a genus-specific polymerase chain reaction (PCR) was performed to identify Anaplasmataceae. Second PCRs were performed with species-specific primers to identify Ehrlichia canis (E. canis) and Anaplasma phagocytophilum (A. phagocytophilum). The results showed that 26.9% of the Ixodes ricinus ticks were infected with Anaplasmataceae in the Black Sea Coast region and 36.8% in the Pleven region. The infection with E. canis was detected in 35.7% and A. phagocytophilum in 25.0% of positive ticks from the Black Sea Coast region. In the Pleven region, 22.9% of ticks were positive for E. canis, while 42.9% were positive for A. phagocytophilum. The molecular identification of E. canis in ticks collected from Bulgaria was performed for the first time. In conclusion, the present study revealed a higher prevalence of ticks infected with Anaplasmataceae, particularly A. phagocytophilum, in the Pleven region than in the Black Sea Coast region. [ABSTRACT FROM AUTHOR]

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Anaplasmosis, caused by the tickborne bacterium Anaplasma phagocytophilum, is an emerging public health threat in the United States. In the northeastern United States, the blacklegged tick (Ixodes scapularis) transmits the human pathogenic genetic variant of A. phagocytophilum (Ap-ha) and a nonpathogenic variant (Ap-V1). New York has recently experienced a rapid and geographically focused increase in cases of anaplasmosis. We analyzed A. phagocytophilum–infected I. scapularis ticks collected across New York during 2008–2020 to differentiate between variants and calculate an entomological risk index (ERI) for each. Ap-ha ERI varied between regions and increased in all regions during the final years of the study. Space-time scan analyses detected expanding clusters of Ap-ha located within documented anaplasmosis hotspots. Ap-ha ERI was more positively correlated with anaplasmosis incidence than non-genotyped A. phagocytophilum ERI. Our findings help elucidate the relationship between the spatial ecology of A. phagocytophilum variants and anaplasmosis. [ABSTRACT FROM AUTHOR]

Background: Although the tick-borne pathogen Anaplasma phagocytophilum is currently described as a single species, studies using genetic markers can distinguish groups of variants associated with different hosts, pathogenicity, zoonotic potential and biotic and geographic niches. The objective of our study was to investigate the genetic diversity of A. phagocytophilum and Ixodes ricinus ticks attached to people. Methods: In collaboration with a commercial diagnostic company, a total of 52 DNA samples were obtained from ticks that tested positive for A. phagocytophilum by quantitative PCR. The genetic profile of each sample was determined using the groEL and ankA genes. Identification of the tick species was confirmed by partial sequencing of the COI subunit and a portion of the TROSPA gene. Results: All 52 ticks were identified as I. ricinus. Two protocols of nested PCR amplifying 1293- and 407-bp fragments of groEL of A. phagocytophilum yielded amplicons of the expected size for all 52 samples. Among all sequences, we identified 10 unique genetic variants of groEL belonging to ecotype I and ecotype II. The analysis targeting ankA was successful in 46 of 52 ticks. Among all sequences, we identified 21 unique genetic variants phylogenetically belonging to three clusters. Conclusions: Our results indicate that ticks attached to people harbor distant genetic variants of A. phagocytophilum, some of which are not recognized as zoonotic. Further studies are needed to determine the risk of human infection by genetic variants other than those designated as zoonotic. [ABSTRACT FROM AUTHOR]

Borrelia miyamotoi is a relapsing fever spirochete that shares the same vector [fas Lyme disease causing Borrelia. This epidemiological study of B. miyamotoi was conducted in rodent reservoirs, tick vectors and human populations simultaneously. A total of 640 rodents and 43 ticks were collected from Phop Phra district, Tak province, Thailand. The prevalence rate for all Borrelia species was 2.3% and for B. miyamotoi was 1.1% in the rodent population, while the prevalence rate was quite high in ticks collected from rodents with an infection rate of 14.5% (95% CI: 6.3–27.6%). Borrelia miyamotoi was detected in Ixodes granulatus collected from Mus caroli and Berylmys bowersi, and was also detected in several rodent species (Bandicota indica, Mus spp., and Leopoldamys sabanus) that live in a cultivated land, increasing the risk of human exposure. Phylogenetic analysis revealed that the B. miyamotoi isolates detected in rodents and I. granulatus ticks in this study were similar to isolates detected in European countries. Further investigation was conducted to determine the serological reactivity to B. miyamotoi in human samples received from Phop Phra hospital, Tak province and in rodents captured from Phop Phra district using an in-house, direct enzyme-linked immunosorbent assay (ELISA) assay with B. miyamotoi recombinant glycerophosphodiester-phosphodiesterase (rGlpQ) protein as coated antigen. The results showed that 17.9% (15/84) of human patients and 9.0% (41/456) of captured rodents had serological reactivity to B. miyamotoi rGlpQ protein in the study area. While a low level of IgG antibody titers (100–200) was observed in the majority of seroreactive samples, higher titers (400–1,600) were also detected in both humans and rodents. This study provides the first evidence of B. miyamotoi exposure in human and rodent populations in Thailand and the possible roles of local rodent species and Ixodes granulatus tick in its enzootic transmission cycle in nature. Author summary: Borrelia miyamotoi, a causative agent of relapsing fever, has been recognized as a human pathogen worldwide. In Thailand, the spirochete bacteria was first detected in rodents collected from the far north in the country. The further investigation revealed its detection in several rodent species and Ixodes granulatus tick, indicating they could play a role in maintaining the enzootic transmission cycle of B. miyamotoi in nature. Epidemiological surveillance was conducted in Tak province where B. miyamotoi was previously detected in rodents. Although B. miyamotoi was not detected in human samples by molecular methods, the serological reactivity to B. miyamotoi recombinant protein in humans and rodents along with the detection of spirochete bacteria in rodents and ticks is a strong indication of B. miyamotoi transmission in this area. This is also the first report of B. miyamotoi serological reactivity in rodents and humans in Thailand; therefore, the possibility of B. miyamotoi infection should not be neglected. Understanding their epidemiological factors regarding their enzootic transmission, the risk of human exposure, and the awareness of disease prevalence will help control and prevent the disease. [ABSTRACT FROM AUTHOR]

Simple Summary: The bacterium Anaplasma phagocytophilum can cause granulocytic anaplasmosis in domestic animals, wildlife, and humans. The pathogen is transmitted by ticks and is predominantly endemic in Central and Northern Europe. We discuss the percentages of dogs that tested positive for A. phagocytophilum by direct detection of the pathogen (4.9%) and by detecting antibodies (23.3%). We found a statistically significant impact of seasonality and years of testing on PCR results as well as sex, age, years of testing and seasonality on antibody results. The dynamics of infections with A. phagocytophilum in dogs in Germany are consistent with peaks in vector activity. There is a notable increase in canine granulocytic anaplasmosis in Germany over the time of this study, which calls for increased attention and demonstrates its rising importance in clinical practice. Anaplasma phagocytophilum is an obligate intracellular bacterium that causes granulocytic anaplasmosis in domestic animals, wildlife, and humans and is primarily transmitted by ticks of the Ixodes persulcatus complex. This retrospective study aims to determine the percentages of dogs that tested positive for A. phagocytophilum in Germany. It included the results of direct (polymerase chain reaction [PCR]) and indirect (immunofluorescence antibody test [IFAT], antibody-enzyme-linked immunosorbent assay [ELISA]) detection methods performed in the laboratory LABOKLIN on canine samples provided by German veterinarians from 2008 to 2020. Out of a total of 27,368 dogs tested by PCR, 1332 (4.9%) tested positive, while 24,720 (27.4%) of the 90,376 dogs tested by IFAT/ELISA had positive serology. High rates of positive PCR results were observed in months with known peaks in vector activity, showing that the dynamics of A. phagocytophilum infections in dogs in Germany are consistent with vector activity. In dogs with a positive PCR result, peaks in serology could be observed four weeks after initial testing. Male and senior dogs had higher rates of positive serology. A possible impact of environmental factors such as changes in climate should be investigated further. Overall, the upward trend in positive test results over the years indicates that canine granulocytic anaplasmosis will continue to become increasingly important for veterinary medicine. [ABSTRACT FROM AUTHOR]

The intracellular bacteria Anaplasma spp. and Coxiella burnetii and the tick-borne encephalitis virus (TBEV) are tick-transmitted pathogens circulating in the southern German sheep population. Knowledge of interaction among Anaplasma spp., C. burnetii and TBEV in sheep is lacking, but together they might promote and reinforce disease progression. The current study aimed to identify co-exposure of sheep to Anaplasma spp., C. burnetii and TBEV. For this purpose, 1,406 serum samples from 36 sheep flocks located in both southern German federal states, Baden-Wuerttemberg and Bavaria, were analysed by ELISAs to determine the antibody levels of the three pathogens. Inconclusive and positive results from the TBEV ELISA were additionally confirmed by a serum neutralisation assay. The proportion of sheep with antibodies against Anaplasma spp. (47.2%), C. burnetii (3.7%) and TBEV (4.7%) differed significantly. Significantly more flocks with Anaplasma spp. seropositive sheep (91.7%) were detected than flocks with antibodies against TBEV (58.3%) and C. burnetii (41.7%), but there was no significant difference between the number of flocks which contained TBEV and C. burnetii seropositive sheep. Seropositivity against at least two pathogens was detected in 4.7% of sheep from 20 flocks. Most co-exposed sheep had antibodies against Anaplasma spp./TBEV (n = 36), followed by Anaplasma spp./C. burnetii (n = 27) and Anaplasma spp./C. burnetii/TBEV (n = 2). Only one sheep showed an immune response against C. burnetii and TBEV. Flocks with sheep being positive against more than one pathogen were widely distributed throughout southern Germany. The descriptive analysis revealed no association between the antibody response of the three pathogens at animal level. Taking the flocks as a cluster variable into account, the exposure to TBEV reduced the probability of identifying C. burnetii antibodies in sheep significantly (odds ratio 0.46; 95% confidence interval 0.24–0.85), but the reason for this is unknown. The presence of Anaplasma spp. antibodies did not influence the detection of antibodies against C. burnetii and TBEV. Studies under controlled conditions are necessary to evaluate any possible adverse impact of co-exposure to tick-borne pathogens on sheep health. This can help to clarify rare disease patterns. Research in this field may also support the One Health approach due to the zoonotic potential of Anaplasma spp., C. burnetii and TBEV. [ABSTRACT FROM AUTHOR]

The occurrence of co-infected hosts and questing ticks with more than one tick-borne pathogen—as in the case of Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato—is expected in endemic regions. Their synergy—in terms of pathogenesis and disease severity—has been suggested previously in humans. Limited data exist on the clinicopathological alterations in co-infected sheep. In this study, we investigated the impact of A. phagocytophilum and B. burgdorferi s.l. seropositivity, alone and in combination, on the hematological parameters of naturally infected sheep. A complete blood count was performed, and indirect immunofluorescence assays were used to detect IgG antibodies against A. phagocytophilum and IgG and IgM antibodies against B. burgdorferi s.l. Single natural exposure to B. burgdorferi s.l. was characterized by low Packed Cell Volume (PCV) values and platelet (PLT) counts, while single exposure to A. phagocytophilum was characterized by low PCV values, low white blood cell (WBC) counts, and an increased risk for leukopenia and neutropenia. Co-exposure resulted in the most severe blood abnormalities; all the blood parameters decreased, and the sheep presented an increased risk for anemia. Our study showed that natural co-exposure to A. phagocytophilum and B. burgdorferi s.l. in sheep leads to more severe blood abnormalities and enhances the pathogenic processes. More studies are needed to clarify the possible background mechanisms. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum, the aetiologic agent of human granulocytic anaplasmosis (HGA), is an obligate intracellular Gram-negative bacterium. During infection, A. phagocytophilum enhances the adhesion of neutrophils to the infected endothelial cells. However, the bacterial factors contributing to this phenomenon remain unknown. In this study, we characterized a type IV secretion system substrate of A. phagocytophilum, AFAP (an actin filament-associated Anaplasma phagocytophilum protein) and found that it dynamically changed its pattern and subcellular location in cells and enhanced cell adhesion. Tandem affinity purification combined with mass spectrometry identified host nucleolin as an AFAP-interacting protein. Further study showed the disruption of nucleolin by RNA interference, and the treatment of a nucleolin-binding DNA aptamer AS1411 attenuated AFAP-mediated cell adhesion, indicating that AFAP enhanced cell adhesion in a nucleolin-dependent manner. The characterization of cell adhesion-enhancing AFAP and the identification of host nucleolin as its interaction partner may help understand the mechanism underlying A. phagocytophilum-promoting cell adhesion, facilitating the elucidation of HGA pathogenesis. [ABSTRACT FROM AUTHOR]

Simple Summary: Anaplasmosis is a bacterial disease that has a severe impact on livestock production, such as reduced milk yield, poor growth and increased susceptibility to other diseases. This pilot study investigated the presence of Anaplasma bacteria and associated antibodies in blood samples collected from a subset of the Swedish goat population. The samples were analysed using polymerase chain reaction and an enzyme-linked immunosorbent assay. One of 40 goats tested positive for the presence of genetic material from the bacteria, while 33% of the serum samples contained antibodies for anaplasmosis. These results indicate that anaplasmosis is widespread in the goat population in certain areas of Sweden. More research is needed to understand the impact of anaplasmosis on Swedish goats and goat farmers. Anaplasmosis is a tick-borne disease that has a severe impact on livestock production and welfare. The aim of this pilot study was to investigate the presence of Anaplasma spp. and associated antibodies in a subset of the Swedish goat population. In 2020, six goat herds located in different parts of Sweden were visited and whole blood and serum samples were collected. The whole blood samples (n = 40) were analysed for the presence of Anaplasma phagocytophilum, A. ovis and A. capra using quantitative and conventional polymerase chain reaction (PCR). The serum samples (n = 59) were analysed for the presence of antibodies to Anaplasma spp. using a commercial competitive enzyme-linked immunosorbent assay, and the same analysis was carried out on additional serum samples previously collected in 2018, 2019 and 2020 (n = 166). One goat (2.5%) tested positive for the presence of A. phagocytophilum genetic material, while the seropositivity rate ranged from 20 to 71%, depending on the surveyed year and area. These results indicate widespread exposure to Anaplasma spp. in the Swedish goat population. To inform future risk assessments and control efforts, further research is warranted to determine the prevalence of anaplasmosis and its impact on goat farming in Sweden. [ABSTRACT FROM AUTHOR]

Objectives: Aeromonas spp. often cause life‐threatening diseases, including necrotizing fasciitis, which may lead to septic shock and ultimately death. Aeromonas infections are believed to be transmitted via minor wounds or the consumption of fresh fish. However, after the detection of Aeromonas hydrophila in ticks in areas endemic to Japanese‐spotted fever (JSF), a novel transmission route of A. hydrophila (i.e., via tick bites) has been proposed. We investigated the prevalence of A. hydrophila in ticks in areas endemic and not endemic to JSF in the Mie Prefecture, Japan. Methods: We collected ticks from endemic and nonendemic areas in summer and winter and assessed them for presence of A. hydrophila using polymerase chain reaction. Results: Six A. hydrophila isolates were obtained from 95 ticks in endemic areas, whereas one A. hydrophila isolate was obtained from 142 ticks in non‐endemic areas, in summer. All ticks that harboured A. hydrophila were Haemaphysalis longicornis (H.L); these ticks were almost at the larval stage and also carried Rickettsia spp. in the endemic area. In contrast, 51 and 41 ticks in the endemic and non‐endemic areas were captured in winter, respectively; A. hydrophila was not detected in these. Conclusions: This study revealed the prevalence of tick‐borne A. hydrophila. Therefore, the risk of transmission of A. hydrophila via a tick bite should be considered in the following conditions: areas abundant in H. L. harbouring Rickettsia spp., in areas endemic for JSF, presence of ticks in the larval stage and during the summer season. [ABSTRACT FROM AUTHOR]

Urban fauna is defined as animal species that can live in urban environments. Several species, including the western-European hedgehog (Erinaceus europaeus), have now been identified as part of this urban fauna, becoming permanent residents of parks and gardens in different cities across Europe. Due to the importance that this phenomenon represents for zoonotic disease surveillance, several authors have been conducting zoonotic agents' surveys on hedgehog. The aim of this study is to compare zoonotic diseases' prevalence in hedgehogs in urban environments with those from more rural areas. A systematic review with meta-analysis of twelve studied of zoonotic diseases' (in urban and rural areas of Europe) was therefore conducted for this purpose. Fifteen different zoonoses have been assessed in urban environments and six in rural areas. Anaplasma phagocytophilum was the most prevalent zoonotic agent found in urban habitats (96%). Dermatophytosis shows statistically significant differences between locations (p-value < 0.001), with a higher prevalence in urban Poland (55%; n = 182). Our results suggest further research and a standardized monitoring of different hedgehog populations are essential to understanding the epidemiology of several zoonotic pathogens in different habitat types (urban, rural, natural, industrial, etc.) and preventing possible disease outbreaks. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum is the causative agent of tick-borne fever in sheep, pasture fever in cattle, and granulocytic anaplasmosis in humans. The increasing prevalence and transboundary spread of A. phagocytophilum in livestock, ticks, and wildlife in the UK poses a potential zoonotic risk that has yet to be estimated. Several ecotypes of A. phagocytophilum show variable zoonotic potential. To evaluate the possible risk associated with the transmission of A. phagocytophilum from ruminants to humans, the ecotype was determined by sequencing the groEL gene from 71 positive blood and tissue samples from UK ruminants. Thirty-four groEL sequences were obtained, fourteen of which were identified in multiple samples. Of the 13 nucleotide polymorphisms identified through pairwise comparison, all corresponded to synonymous substitutions. The subsequent phylogenetic estimation of the relationship with other European/world isolates indicated that all the groEL sequences clustered with other ecotype I sequences. The presence of ecotype I closely reflects that observed in ruminants in continental Europe and suggests a lower risk of zoonotic transmission from this reservoir. [ABSTRACT FROM AUTHOR]

Acaricides are hypothesized to reduce human risk of exposure to tick-borne pathogens by decreasing the abundance and/or infection prevalence of the ticks that serve as vectors for the pathogens. Acaricides targeted at reservoir hosts such as small mammals are expected to reduce infection prevalence in ticks by preventing their acquisition of zoonotic pathogens. By reducing tick abundance, reservoir-targeted or broadcast acaricides could reduce tick infection prevalence by interrupting transmission cycles between ticks and their hosts. Using an acaricide targeted at small-mammal hosts (TCS bait boxes) and one sprayed on low vegetation (Met52 fungal biocide), we tested the hypotheses that infection prevalence of blacklegged ticks with zoonotic pathogens would be more strongly diminished by TCS bait boxes, and that any effects of both acaricidal treatments would increase during the four years of deployment. We used a masked, placebo-controlled design in 24 residential neighborhoods in Dutchess County, New York. Analyzing prevalence of infection with Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in 5380 nymphal Ixodes scapularis ticks, we found little support for either hypothesis. TCS bait boxes did not reduce infection prevalence with any of the three pathogens compared to placebo controls. Met52 was associated with lower infection prevalence with B. burgdorferi compared to placebo controls but had no effect on prevalence of infection with the other two pathogens. Although significant effects of year on infection prevalence of all three pathogens were detected, hypothesized cumulative reductions in prevalence were observed only for B. burgdorferi. We conclude that reservoir-targeted and broadcast acaricides might not generally disrupt pathogen transmission between reservoir hosts and tick vectors or reduce human risk of exposure to tick-borne pathogens. [ABSTRACT FROM AUTHOR]

The Soprano pipistrelle bat, Pipistrellus pygmaeus, is a common species in large parts of Sweden. Many of its natural habitats are near human habitations. This creates opportunities for ticks infesting these bats to encounter humans and possibly transmit zoonotic pathogens by tick bites. The bats are often infested with Carios vespertilionis, a tick species that, in addition to bats, has been recorded to bite humans on occasion. This study aimed to investigate if C. vespertilionis acts as a reservoir for Anaplasma phagocytophilum, Neoehrlichia mikurensis, Tick-borne encephalitis virus, and species of Babesia and Rickettsia and to improve currently used conventional PCR protocols for molecular species determination of Rickettsia spp. Ninety-two C. vespertilionis ticks were collected from underneath a bat-box harbouring P. pygmaeus. Pathogen-specific PCR assays showed that 58.4% were positive for Rickettsia spp. and negative for the other pathogens analysed. Phylogenetic analyses indicate that the species belong to R. parkeri, R. conorii, R. slovaca, R. sibirica subsp. mongolotimonae, R. rickettsii, and a hitherto uncultured Rickettsia sp. Several of these species are considered pathogenic to humans. Given the ecology and behaviour of C. vespertilionis, it may be a vector of these rickettsiae among bats and occasionally humans. To determine the Rickettsia species with certainty, and to determine if C. vespertilionis may be a reservoir and vector of the Rickettsia spp., further studies are needed. [ABSTRACT FROM AUTHOR]

Plasmids that carry antibiotic resistance genes occur frequently in Aeromonas salmonicida subsp. salmonicida, an aquatic pathogen with severe consequences in salmonid farming. Here, we describe a 67 kb plasmid found in the A. salmonicida subsp. salmonicida Strain SHY15-2939 from Quebec, Canada. This new plasmid, named pAsa-2939 and identified by high throughput sequencing, displays features never found before in this bacterial species. It contains a transposon related to the Tn21 family, but with an unusual organization. This transposon bears a catB3 gene (chloramphenicol resistance) that has not been detected yet in A. salmonicida subsp. salmonicida. The plasmid is transferable by conjugation into Aeromonas hydrophila, but not into Escherichia coli. Based on PCR analysis and genomic sequencing (Illumina and PacBio), we determined that the transposon is unstable in A. salmonicida subsp. salmonicida Strain SHY15-2939, but it is stable in A. hydrophila trans-conjugants, which explains the chloramphenicol resistance variability observed in SHY15-2939. These results suggest that this bacterium is likely not the most appropriate host for this plasmid. The presence of pAsa-2939 in A. salmonicida subsp. salmonicida also strengthens the reservoir role of this bacterium for antibiotic resistance genes, even those that resist antibiotics not used in aquaculture in Québec, such as chloramphenicol. [ABSTRACT FROM AUTHOR]

Background: The composition of the microbial flora associated with ixodid ticks has been studied in several species, revealing the importance of geographical origin, developmental stage(s) and feeding status of the tick, as well as substantial differences between tissues and organs. Studying the microbiome in the correct context and scale is therefore necessary for understanding the interactions between tick-borne pathogens and other microorganisms as well as other aspects of tick biology. Methods: In the present study the microbial flora of whole Ixodes ricinus, I. persulcatus and I. trianguliceps ticks were analyzed with 16S rRNA amplicon sequencing. Additionally, tick organs (midguts, Malpighian tubules, ovaries, salivary glands) from flat and engorged I. ricinus female ticks were examined with the same methodology. Results: The most abundant bacteria belonged to the group of Proteobacteria (Cand. Midichloria mitochondrii and Cand. Lariskella). 16S amplicon sequencing of dissected tick organs provided more information on the diversity of I. ricinus-associated microbial flora, especially when organs were collected from engorged ticks. Bacterial genera significantly associated with tick feeding status as well as genera associated with the presence of tick-borne pathogens were identified. Conclusions: These results contribute to the knowledge of microbial flora associated with ixodid ticks in their northernmost distribution limit in Europe and opens new perspectives for other investigations on the function of these bacteria, including those using other approaches like in vitro cultivation and in vitro models. [ABSTRACT FROM AUTHOR]

Ixodes ricinus and Ixodes persulcatus ticks are the main vectors of tick-borne encephalitis virus (TBEV), which has three main subtypes connected with certain tick species: the European subtype, associated with I. ricinus, and the Siberian and Far-Eastern subtypes, associated with I. persulcatus. Distribution ranges of these species overlap and form large sympatric areas in the East European Plain and Baltic countries. It has previously been shown that crossing of I. ricinus and I. persulcatus is possible, with the appearance of sterile hybrids. Hybridization of ticks can affect not only the spread of ticks but also the properties of natural foci of arbovirus infections, in particular TBEV. In the present study, we analyzed the effectiveness of virus transmission from infected mice to larvae and nymphs and trans-stadial transmission (from larvae to nymph and adult) in I. ricinus, I. persulcatus, and hybrids. For this purpose, we bred a hybrid generation from the crossing of I. persulcatus females and I. ricinus males, and we used the Siberian and European subtypes of TBEV. We showed that after feeding on infected mice, virus prevalence in engorged ticks decreased over time, and after molting, the opposite was true. In hybrids we observed the highest acquisition effectiveness and RNA copy numbers during Siberian TBEV subtype transmission. The efficiency of trans-stadial transmission of both TBEV subtypes was similar in hybrids and parental species. After the second trans-stadial TBEV transmission, a significant increase in ticks' infection rates was observed only in specific subtype-tick combination. Our data demonstrate the possible features of TBEV circulation in the I. ricinus and I. persulcatus sympatry area. [ABSTRACT FROM AUTHOR]

The ectoparasite Ixodes ricinus is an important vector for many tick-borne diseases (TBD) in the northern hemisphere, such as Lyme borreliosis, rickettsiosis, human granulocytic anaplasmosis, or tick-borne encephalitis virus. As climate change will lead to rising temperatures in the next years, we expect an increase in tick activity, tick population, and thus in the spread of TBD. Consequently, it has never been more critical to understand relationships within the microbial communities in ticks that might contribute to the tick's fitness and the occurrence of TBD. Therefore, we analyzed the microbiota in different tick tissues such as midgut, salivary glands, and residual tick material, as well as the microbiota in complete Ixodes ricinus ticks using 16S rRNA gene amplicon sequencing. By using a newly developed DNA extraction protocol for tick tissue samples and a self-designed mock community, we were able to detect endosymbionts and pathogens that have been described in the literature previously. Further, this study displayed the usefulness of including a mock community during bioinformatic analysis to identify essential bacteria within the tick. [ABSTRACT FROM AUTHOR]

Simple Summary: A disease outbreak in a Swedish herd of beef cattle which may be related to a tick-borne infection initiated an in-depth study to investigate the presence of bacteria and viruses in clinically healthy and diseased cattle, as well as in wild deer and boars from the region and in ticks collected from the cattle and deer. The results indicated unusual types of Mycoplasma as possible causative factors, probably in combination with immune suppression due to granulocytic anaplasmosis caused by Anaplasma phagocytophilum. The latter bacterium was widespread in the herd and was also found in wild deer in the region, and in ticks collected from the cattle and deer. Genetic comparisons indicated that Anaplasma phagocytophilum was circulating among the cattle population, while circulation between cattle and deer occurred infrequently. An outbreak of disease in a Swedish beef cattle herd initiated an in-depth study to investigate the presence of bacteria and viruses in the blood of clinically healthy (n = 10) and clinically diseased cattle (n = 20) using whole-genome shotgun sequencing (WGSS). The occurrence of infectious agents was also investigated in ticks found attached to healthy cattle (n = 61) and wild deer (n = 23), and in spleen samples from wild deer (n = 30) and wild boars (n = 10). Moreover, blood samples from 84 clinically healthy young stock were analysed for antibodies against Anaplasma phagocytophilum and Babesia divergens. The WGSS revealed the presence of at least three distinct Mycoplasma variants that were most closely related to Mycoplasma wenyonii. Two of these were very similar to a divergent M. wenyonii variant previously only detected in Mexico. These variants tended to be more common in the diseased cattle than in the healthy cattle but were not detected in the ticks or wild animals. The DNA of A. phagocytophilum was detected in similar proportions in diseased (33%) and healthy (40%) cattle, while 70% of the deer, 8% of ticks collected from the cattle and 19% of the ticks collected from deer were positive. Almost all the isolates from the cattle, deer and ticks belonged to Ecotype 1. Based on sequencing of the groEL-gene, most isolates of A. phagocytophilum from cattle were similar and belonged to a different cluster than the isolates from wild deer. Antibodies against A. phagocytophilum were detected in all the analysed samples. In conclusion, uncommon variants of Mycoplasma were detected, probably associated with the disease outbreak in combination with immune suppression due to granulocytic anaplasmosis. Moreover, A. phagocytophilum was found to be circulating within this cattle population, while circulation between cattle and deer occurred infrequently. [ABSTRACT FROM AUTHOR]

Tick-transmitted bacterial pathogens thrive in enzootic infection cycles, colonizing disparate vertebrate and arthropod tissues, often establishing persistent infections. Therefore, the evolution of robust immune evasion strategies is central to their successful persistence or transmission between hosts. To survive in nature, these pathogens must counteract a broad range of microbicidal host responses that can be localized, tissue-specific, or systemic, including a mix of these responses at the host–vector interface. Herein, we review microbial immune evasion strategies focusing on Lyme disease spirochetes and rickettsial or tularemia agents as models for extracellular and intracellular tick-borne pathogens, respectively. A better understanding of these adaptive strategies could enrich our knowledge of the infection biology of relevant tick-borne diseases, contributing to the development of future preventions. Tick-transmitted bacterial pathogens have evolved a plethora of immune evasion strategies that support their successful persistence through a complex enzootic infection cycle involving a variety of vertebrate hosts and arthropods. A range of major immune evasion strategies employed by widely-studied bacterial pathogens – such as the avoidance of host recognition or immune offense via the modulation of surface components, inhibition of complement, blocking of antimicrobial molecules, and interference with cellular defense effector pathways or cytokine production – are also reflected by major tick-borne pathogens like Borrelia burgdorferi sensu lato, relapsing fever spirochetes, and several intracellular bacterial pathogens from the genera Anaplasma , Francisella , and Ehrlichia. The immune evasion strategies of tick-borne pathogens are relatively more well-studied in mammalian hosts, as compared to vector ticks. A better understanding of the basic mechanisms of microbial immune evasion in both mammals and arthropods, focusing on contributions toward pathogen fitness, could contribute to the development of novel control measures against tick-borne infections. [ABSTRACT FROM AUTHOR]

Background Diagnosis of undifferentiated non-malaria fevers (NMF) in returning travellers is a great challenge. Currently, there is no consensus about the use of empirical antibiotics in returning travellers with undifferentiated NMF. Although studies in endemic areas showed that a wide range of pathogens implicated in undifferentiated NMF are treatable with doxycycline, the role of doxycycline in returning travellers with fever still has to be explored. Methods Prospective European multicentre cohort study of febrile international travellers (November 2017—November 2019). Immunological and molecular diagnostic techniques for doxycycline responding illnesses (DRI) agents such as Anaplasma phagocytophilum , spotted fever group Rickettsia spp. typhus group Rickettsia spp. Coxiella burnetii , Bartonella spp. Orientia tsutsugamushi, Borrelia miyamotoi, Borrelia recurrentis and Leptospira spp. were systematically performed in all patients with undifferentiated NMF. We estimated the prevalence and predictive factors of DRI in returning travellers with undifferentiated NMF. Results Among 347 travellers with undifferentiated NMF, 106 (30·5%) were finally diagnosed with DRI. Only 57 (53·8%) of the 106 DRI infections were diagnosed by the standard of care. The main causes of DRI were: 55 (51·9%) Rickettsia spp. 16 (15·1%) C. burnetii ; 15 (14·2%) Bartonella spp.; 13 (12·3%) Leptospira spp. and 10 (9·5%) A. phagocytophilum. The only predictive factor associated with DRI was presenting an eschar (aOR 39·52, 95%CI 4·85–322·18). Features of dengue such as retro-orbital pain (aOR 0·40, 95%CI 0·21–0·76) and neutropenia (aOR 0·41, 95%CI 0·21–0·79) were negatively associated with DRI. Conclusions Although DRI are responsible for 30% of undifferentiated NMF cases in travellers, those are seldom recognized during the first clinical encounter. Empirical treatment with doxycycline should be considered in returning travellers with undifferentiated fever and negative tests for malaria and dengue, particularly when presenting severe illness, predictive factors for rickettsiosis or no features of dengue. [ABSTRACT FROM AUTHOR]

The genus Anaplasma (Anaplasmataceae, Rickettsiales) includes tick-transmitted bacterial species of importance to both veterinary and human medicine. Apart from the traditionally recognized six Anaplasma species (A. phagocytophilum, A. platys, A. bovis, A. ovis, A. centrale, A. marginale), novel strains and candidate species, also of relevance to veterinary and human medicine, are emerging worldwide. Although species related to the zoonotic A. platys and A. phagocytophilum have been reported in several African and European Mediterranean countries, data on the presence of these species in sub-Saharan countries are still lacking. This manuscript reports the investigation of Anaplasma strains related to zoonotic species in ruminants in Senegal by combining different molecular tests and phylogenetic approaches. The results demonstrated a recent introduction of Candidatus (Ca) Anaplasma turritanum, a species related to the pathogenic A. platys, possibly originating by founder effect. Further, novel undetected strains related to Candidatus (Ca) Anaplasma cinensis were detected in cattle. Based on groEL and gltA molecular comparisons, we propose including these latter strains into the Candidatus (Ca) Anaplasma africanum species. Finally, we also report the emergence of Candidatus (Ca) A. boleense in Senegal. Collectively, results confirm that Anaplasma species diversity is greater than expected and should be further investigated, and that Anaplasma routine diagnostic procedures and epidemiological surveillance should take into account specificity issues raised by the presence of these novel strains, suggesting the use of a One Health approach for the management of Anaplasmataceae in sub-Saharan Africa. [ABSTRACT FROM AUTHOR]

Ovine and caprine anaplasmosis is an endemic tick-borne disease in Europe, the Middle East and some African and Asian countries, including Turkey. This study was conducted to determine the prevalence of Anaplasma ovis and Anaplasma phagocytophilum in sheep and goats in the Southeastern Anatolia region of Turkey. For each animal group, 384 individuals were included in the study. Microscopy and Rt-PCR were used to determine the prevalence of anaplasmosis in sheep and goats. Microscopic examination revealed 21.88% (n:84) and 39.58% (n:152) Anaplasma sp. positivity in sheep and goats, respectively. The study revealed that the molecular prevalence values were higher than the microscopic prevalence values for each microorganism. The molecular prevalence of A. ovis was 89.32% (n:343) in sheep and 78.91% (n:303) in goats and the difference was statistically significant (P<0.05). On the other hand, A. phagocytophilum was detected at 42.97% (n:165) in sheep and 15.89% (n:61) in goats. This is the first study in Turkey in which the Rt-PCR method was used for the detection of Anaplasma species in small ruminants. This study demonstrated that Rt-PCR provides more accurate and specific results than microscopic examination. The present study is expected to have contributed in determining the prevalence of Anaplasma species, one of the tick-borne pathogens, in Turkey and provide data on future control strategies for small ruminant anaplasmosis. [ABSTRACT FROM AUTHOR]

Background: Ticks are obligate hematophagous arthropods capable of transmitting a great variety of endemic and emerging pathogens causing diseases in animals and humans. Objectives: The aim of this study was to investigate the presence of Bartonella spp., Rickettsia spp., Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum in ticks collected from cattle in Benin and Togo. Methods: Overall, 396 (148 males, 205 females and 43 nymphs) ticks were collected from cattle in 17 districts (Benin and Togo) between 2019 and 2020. Ticks were pooled into groups of 2–6 ticks per pool according to individual host, location, species and developmental stage. The DNA of each pool was extracted for molecular screening. Results: PCR results revealed that 20 tick pools were positive for Bartonella spp. (Benin and Togo) and 23 tick pools positive for Rickettsia spp. (Benin), while all pools were negative for A. phagocytophilum and B. burgdorferi s.l. Sequence analysis of positive Rickettsia samples revealed the presence of Rickettsia aeschlimannii. Conclusions: The present study highlights the presence of zoonotic agents in ticks collected from cattle in Benin and Togo. This information will raise awareness of tick‐borne diseases among physicians and veterinarians, stimulate further studies to monitor these pathogens, and advise on necessary measures to control the spread of these zoonoses. [ABSTRACT FROM AUTHOR]

Identifying the species that act as hosts, vectors, and vehicles of vector-borne pathogens is vital for revealing the transmission cycles, dispersal mechanisms, and establishment of vector-borne pathogens in nature. Ticks are common vectors for pathogens causing human and animal diseases, and they transmit a greater variety of pathogenic agents than any other arthropod vector group. Ticks depend on the movements by their vertebrate hosts for their dispersal, and tick species with long feeding periods are more likely to be transported over long distances. Wild birds are commonly parasitized by ticks, and their migration patterns enable the long-distance range expansion of ticks. The African–Palearctic migration system is one of the world's largest migrations systems. African–Western Palearctic birds create natural links between the African, European, and Asian continents when they migrate biannually between breeding grounds in the Palearctic and wintering grounds in Africa and thereby connect different biomes. Climate is an important geographical determinant of ticks, and with global warming, the distribution range and abundance of ticks in the Western Palearctic may increase. The introduction of exotic ticks and their microorganisms into the Western Palearctic via avian vehicles might therefore pose a greater risk for the public and animal health in the future. [ABSTRACT FROM AUTHOR]

The prevalence of potential human pathogenic members of the order Rickettsiales differs between Borrelia burgdorferi sensu lato-positive and -negative tick microbiomes. Here, co-infection of members of the order Rickettsiales, such as Rickettsia spp., Anaplasma phagocytophilum, Wolbachia pipientis, and Neoehrlichia mikurensis as well as B. burgdorferi s.l. in the tick microbiome was addressed. This study used conventional PCRs to investigate the diversity and prevalence of the before-mentioned bacteria in 760 nucleic acid extracts of I. ricinus ticks detached from humans, which were previously tested for B. burgdorferi s.l.. A gltA gene-based amplicon sequencing approach was performed to identify Rickettsia species. The prevalence of Rickettsia spp. (16.7%, n = 127) and W. pipientis (15.9%, n = 121) were similar, while A. phagocytophilum was found in 2.8% (n = 21) and N. mikurensis in 0.1% (n = 1) of all ticks. Co-infection of B. burgdorferi s. l. with Rickettsia spp. was most frequent. The gltA gene sequencing indicated that Rickettsia helvetica was the dominant Rickettsia species in tick microbiomes. Moreover, R, monacensis and R. raoultii were correlated with autumn and area south, respectively, and a negative B. burgdorferi s. l. finding. Almost every fifth tick carried DNA of at least two of the human pathogenic bacteria studied here. [ABSTRACT FROM AUTHOR]

The European mole (Talpa europaea) has a widespread distribution throughout Europe. However, little is known about the presence of zoonotic pathogens in European moles. We therefore tested 180 moles from the middle and the south of the Netherlands by (q)PCR for the presence of multiple (tick-borne) zoonotic pathogens. Spotted fever Rickettsia was found in one (0.6%), Leptospira spp. in three (1.7%), Bartonella spp. in 69 (38.3%) and Hantaviridae in 89 (49.4%) of the 180 moles. Infections with Anaplasma phagocytophilum, Babesia spp., Neoehrlichia mikurensis, Borrelia spp., Spiroplasma spp. and Francisella tularensis were not found. In addition, in a subset of 35 moles no antibodies against Tick-borne encephalitis virus were found. The obtained sequences of Bartonella spp. were closely related to Bartonella spp. sequences from moles in Spain and Hungary. The Hantaviridae were identified as the mole-borne Nova virus, with high sequence similarity to sequences from other European countries, and Bruges virus. Though the zoonotic risk from moles appears limited, our results indicate that these animals do play a role in multiple host-pathogen cycles. [ABSTRACT FROM AUTHOR]

Jingmen tick virus (JMTV) is a novel tick‐borne virus first identified from Jingmen city, Hubei Province of China in 2010. It has been proved that JMTV can cause human diseases and is widely distributed both inside and outside of China. However, the survival mode and transmission characteristics of JMTV still need further research, particularly in terms of transovarial transmission. In this study, an investigation was conducted to explore the presence of JMTV from engorged female ticks to their offspring. All engorged female adult ticks were collected from domestic cattle and allowed to lay eggs in appropriate humidity and temperature conditions. Maternal ticks, eggs and larvae were screened for JMTV RNA through real‐time polymerase chain reaction (RT‐PCR) and nested PCR methods. The results revealed the positive rate of 10.53% (10/95) in engorged ticks, 9.09% (2/22) in eggs and 8% (4/50) in larvae pools, respectively. Phylogenetic analysis confirmed that sequences from eggs and larvae had closer relationship with those isolates from maternal engorged ticks with more than 99.7% homology and JMTV manifested with evolutional conservatism. Our study has identified for the first time that JMTV could be transmitted from mother generation to offspring of Haemaphysalis Longicornis. Nonetheless, the efficiency of transovarial transmission in JMTV and the significance of ticks as amplification hosts still need to be further illustrated. [ABSTRACT FROM AUTHOR]

Query fever (Q fever) is a disease caused by Coxiella burnetii, an obligate intracellular bacterium that affects humans and other animals. This organism is uncommon, but may be found in ruminant, and other domestic mammals. This study aimed to detect Coxiella burnetii (C. burnetii) in cattle and record the risk assessment associated with coxiellosis. Total of 100 bulk tank milk (BTM) samples were collected from dairy cattle located in different governorates in Egypt. C. burnetii antibodies were detected in the collected milk using ELISA. In addition, 15 samples were subjected to PCR targeting ISIIII gene specific to C. burnetii. All farms in the investigated area had a positive case for C. burnetii infection. The overall seroprevalence of C. burnetii was 51% among the examined farms. However, significant higher seropositive samples were observed in Alexandria, Menoufia, Giza, Faiyum, Ismailia, Beheira, followed by Gharbia, Sharqia, then Dakahlia, and BeniSuef. Using questionnaire some risk factors such as region, breed type, age, season herd size, water sources, feeding management, abortion, mastitis, presence of arthropods or rodents were investigated among the study area. In conclusion, the study revealed a wide distribution of infections over the study area and further epidemiological study is recommended to prevent and control of coxiellosis. [ABSTRACT FROM AUTHOR]

Anaplasma phagocytophilum is an emerging tick-borne zoonotic bacterium, which is considered a significant risk to the health and industry of cattle in tropical and sub-tropical regions worldwide. This research focuses on examining the worldwide occurrence of A. phagocytophilum in cattle. Several databases, including Scopus, PubMed, ProQuest, Web of Science, ScienceDirect, and Google Scholar, were searched for publications spanning October 2004 to November 2024. The pooled prevalence was calculated with a 95% confidence interval (CI) using a random-effects model based on the Freeman-Tukey double arcsine transformation. A total of 72 studies satisfied the inclusion criteria, revealing a global prevalence of A. phagocytophilum in cattle estimated at 8.5% (5.9%-11.5%). Mongolia (51.9%, 45.9%-56.2%) and Guatemala (51%, 41.2%-60.7%) were countries that accounted for the highest prevalence. Moreover, the infection was most prevalent in African region with prevalence of 11.3% (3.9%-21.5%). The highest prevalence rate was observed in hot-summer Mediterranean climate (13.7%, 4.7%-26.2%). The analysis indicated that immunological techniques were associated with the highest prevalence rate (14.2%, 6.5%-24.3%). The findings of the present research highlighted important geographical and environmental factors that affect the prevalence of disease. In the fields of veterinary medicine and public health, these findings enhance disease management plans and preventative initiatives., (© 2025 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

Background: Anaplasma spp., zoonotic tick-borne pathogens affecting livestock, companion animals, and humans, exhibits 15-18% seropositivity among hunting dogs in the Republic of Korea (South Korea). The dominant tick species in South Korea, Haemaphysalis longicornis, can transmit these pathogens to both humans and animals. Given the limited understanding of transovarial transmission of Anaplasma spp., our study aimed to assess the prevalence of questing larval ticks containing Anaplasma DNA. Additionally, we aimed to gather data for establishing a nationwide forecasting and alert system on seasonal variation of tick developmental stages and tick-borne zoonotic pathogens., Methods: From March to October 2021 and again from March to October 2022, we collected a total of 36,912 unfed, questing ticks of Haemaphysalis spp. from 149 sites in South Korea. Ticks were collected from herbaceous vegetation using the flagging method using a white flannel cloth. After species identification, one-third of collected ticks underwent analysis for Anaplasma DNA. Nymph ticks were pooled in groups of 1-10 and larvae in groups of 1-50, while adults were examined individually. Nested polymerase chain reaction (PCR) was performed to detect the genus Anaplasma by amplifying the 16S rRNA gene, followed by sequencing for species identification and phylogenetic analysis., Results: Of the 36,912 questing ticks collected, 13,082 (35.4%) were identified as nymphs and adults of H. longicornis and 3850 (10.4%) as those of Haemaphysalis flava. The morphologically indistinguishable larval stage of Haemaphysalis spp. predominated, with 19,980 (54.1%) collected primarily from July to October. From the 939 tick pools, 24 pools (2.6%) tested positive for Anaplasma, with the larval stage exhibiting the highest number of positive pools (16, 1.7%). Phylogenetic analysis revealed that 21 of the 24 Anaplasma-positive pools contained A. phagocytophilum-specific genes, with 1 identified as Anaplasma sp. and the remaining 2 as A. bovis., Conclusions: Our study provides evidence of transovarial transmission of A. phagocytophilum in Haemaphysalis spp. larvae under field conditions, showing that the bacteria are transmitted from mother ticks to unengorged, questing larvae. Additionally, our findings contribute significant data for establishing a nationwide forecasting and alert system on seasonal variation of tick developmental stages and tick-borne zoonotic pathogens., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Approval from Chonnam National University’s Institutional Animal Care and Use Committee was not required for the present study, because the collected samples from questing ticks in the environment in this study did not cause hazard to any animals. Ticks are not included in the endangered species of South Korea. Specific approval for each collection site was not needed because the sites were not located within national parks or protected regions. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)