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By using PCR testing, we found Neoehrlichia mikurensis DNA in 1.1% of ticks removed from persons in Sweden and Finland. Symptoms developed in 2 immunocompetent persons. Despite low transmission risk, infection can occur after short tick attachment. Our findings highlight the need to consider N. mikurensis in patients with unexplained symptoms after tick bite.

Background Anaplasma marginale is an obligate intracellular bacterium and the main cause of bovine anaplasmosis in tropical and subtropical regions. In Egypt, data regarding the prevalence of A. marginale in ruminant hosts and of the circulating genotypes is lacking. This study therefore aimed to (i) investigate the presence, epidemiology and genotypes of A. marginale in cattle and buffaloes in Egypt, (ii) to evaluate suitable diagnostic tools and (iii) to identify co-infections of A. marginale with other selected tick-borne pathogens. Methods Blood samples were collected from 394 animals (309 cattle and 85 buffaloes) from three different areas in Egypt. For the detection of A. marginale infection, several tests were compared for their sensitivity and specificity: blood smear analysis, enzyme-linked immunosorbent assay (ELISA), PCR, real-time PCR and reverse line blot (RLB) assay. Co-infections with A. marginale, piroplasms and other Anaplasmataceae were surveyed by RLB while A. marginale genotypes were identified by amplifying and sequencing the partial msp1α gene. Results Anaplasma marginale DNA was amplified by qPCR in 68.3% of cattle and 29.4% of buffaloes. RLB showed infection with A. marginale in 50.2% of cattle and 42.5% of buffaloes. Blood smear analysis detected this agent in 16.2% of cattle and 2.4% of buffaloes. ELISA showed specific antibodies against A. marginale in 54.9% of cattle. Anaplasma marginale was associated, in cattle and buffaloes, with several tick-borne pathogens ( Theileria annulata , Babesia bovis , Babesia bigemina , Babesia occultans and Anaplasma platys ). A significant difference of A. marginale infection level was noticed in cattle, where animals between 3–5-years-old had a higher prevalence (79.2%) compared to those older than 5 years (36.4%) and younger than 3 years (59.7%) and one year (64.5%), respectively ( P  = 0.002281). Microsatellite analysis identified 15 different genotypes. Conclusions The epidemiological findings revealed high prevalence of A. marginale in cattle and buffaloes in all the investigated areas. The circulation of diverse genotypes was observed, most of these A. marginale genotypes being specific for Egypt. The qPCR assay was confirmed to be the most sensitive tool for detection of A. marginale in cattle and buffaloes even in the carrier state, highlighting the importance of using suitable diagnostic tests.

Neoehrlichia mikurensis is vector-borne gram-negative intracellular bacteria that has a wide range of hosts, and are recognized as emerging human pathogens. This study aimed to investigate the prevalence and genetic characteristics of N. mikurensis infections in rodents from central and southern of Shanxi Province, China. The rodents were captured, and the liver, spleen and kidney were collected for N . mikurensis detection and identification by 16 S rRNA and groEL genes sequencing. N . mikurensis was detected in Apodemus agrarius from Yangquan and Eothenomys inez from Yuncheng, with a positivity rate of 0.66% (2/301). Phylogenetic analysis based on 16 S rRNA and groEL genes revealed that the sequences in this study formed distinct branches separate from reported clusters (Clusters I-IV), clustered near Clusters III and IV, which the pathogenicity was unclear. Analysis of groEL gene polymorphism identified 12 haplotypes (Hap-1 to Hap-12) among 64 sequences (one novel sequence from this study and 63 from GenBank), with high haplotype diversity (Hd = 0.770 ± 0.030). Dominant haplotypes (Hap-2, Hap-3, Hap-4; 82.8% of sequences) exhibited distinct geographic specific patterns. In addition, Asian sequences exhibited greater diversity than European sequences, and human-derived sequences showed the higher genetic diversity than rodent- and tick-derived sequences. Overall, the detection of N . mikurensis in rodents in Shanxi Province expands its known distribution and provides novel insights into the genetic diversity in different regions and hosts.

Background Globally, the disease ecology of reptiles remains understudied, even for threatened and iconic species such as the Galápagos marine iguana ( Amblyrhynchus cristatus ). Although marine iguanas are parasitized by distinct species of ticks and mites, research on vector-borne diseases for this species is limited. Methods In this study, we detected 16S ribosomal RNA (rRNA) sequences of Candidatus Allocryptoplasma in transcriptomic data from marine iguana blood samples. These 16S rRNA sequences were further characterized through phylogenetic analysis and a haplotype network. Results Our analysis revealed the first molecular evidence for the infection of marine iguanas with Candidatus Allocryptoplasma, a candidate genus in the family Anaplasmataceae with unknown pathogenic potential, likely transmitted by ticks. Phylogenetic analysis of the novel 16S rRNA sequences together with available Anaplasmataceae sequences confirmed their assignment to this candidate genus. A haplotype network analysis indicated that the agent infecting the marine iguana represents a distinct lineage within the known Ca . Allocryptoplasma diversity. Conclusions Candidatus Allocryptoplasma had a high prevalence within marine iguanas, infecting individuals across most of the geographical range of this species. To elucidate the transmission dynamics of this bacterium in the Galápagos ecosystem, ectoparasites of the marine iguana and shared vertebrate hosts should be screened for infection with Ca . Allocryptoplasma. Graphical Abstract

Anaplasmataceae agents comprise obligate intracellular bacteria that can cause disease in humans and animals. Between August 2013 and March 2015, 31 Nasua nasua (coati), 78 Cerdocyon thous (crab-eating fox), seven Leopardus pardalis (ocelot), 110 wild rodents, 30 marsupials, and 42 dogs were sampled in the Pantanal wetland, Brazil. In addition, ectoparasites found parasitizing the animals were collected and identified. The present work aimed to investigate the occurrence of Anaplasmataceae agents in wild mammals, domestic dogs and ectoparasites, by molecular and serological techniques. Overall, 14 (17·9%) C. thous, seven (16·6%) dogs and one (3·2%) N. nasua were seroreactive to Ehrlichia canis. Nine dogs, two C. thous, one N. nasua, eight wild rodents, five marsupials, eight Amblyomma sculptum, four Amblyomma parvum, 13 A. sculptum nymphal pools, two Amblyomma larvae pools and one Polygenis (Polygenis) bohlsi bohlsi flea pool were positive for Ehrlichia spp. closely related to E. canis. Seven N. nasua, two dogs, one C. thous, one L. pardalis, four wild rodents, three marsupials, 15 A. sculptum, two Amblyomma ovale, two A. parvum and one Amblyomma spp. larval pools were positive for Anaplasma spp. closely related to A. phagocytophilum or A. bovis. The present study provided evidence that wild animals from Brazilian Pantanal are exposed to Anaplasmataceae agents.

Raccoons ( Procyon lotor ) originated in North America and have been introduced to Europe. Due to their close contact with human settlements, they are important reservoirs for zoonotic pathogens, such as Baylisascaris procyonis . The relevance and prevalence of vector-borne pathogens have not yet been fully elucidated. In this study, we screened 285 spleen samples of raccoons, collected between 2019 and 2022 in Germany. The samples were analysed by PCR to detect Mycoplasma spp., Anaplasmataceae, Bartonella spp. , Babesia spp., Rickettsia spp., Filarioidea, Trypanosomatida and Hepatozoon spp., and positive PCR products were sequenced. In total, 104 samples were positive for Mycoplasma spp. (36.49%), making this the first study to detect Mycoplasma spp. in raccoons outside of North America. Three samples were positive for Babesia spp. (1.05%) and two for Anaplasma phagocytophilum (0.7%). Phylogenetic analysis revealed that the Mycoplasma spp. detected all belong to the haemotrophic mycoplasmas cluster and were grouped within a single phylogenetic clade. Two different Babesia spp. were detected, one of which was closely related to Babesia canis , while the other was more closely related to Babesia sp. from ruminants. It is unclear whether the pathogens detected have an impact on the health of raccoons or whether they may serve as a reservoir for other animals.

Backgrounds Corsica is a French island situated in the Mediterranean Sea. The island provides suitable natural conditions to study disease ecology, especially tick-borne diseases and emerging diseases in animals and ticks. The family Anaplasmataceae is a member of the order Rickettsiales ; it includes the genera Anaplasma , Ehrlichia , Neorickettsia and Wolbachia . Anaplasmosis and ehrlichiosis traditionally refer to diseases caused by obligate intracellular bacteria of the genera Anaplasma and Ehrlichia . The aim of this study was to identify and estimate the prevalence of Anaplasmataceae species infecting domestic animals and ticks in Corsica. Methods In this study, 458 blood samples from sheep, cattle, horses, goats, dogs, and 123 ticks removed from cattle, were collected in Corsica. Quantitative real-time PCR screening and genetic characterisation of Anaplasmataceae bacteria were based on the 23S rRNA, rpoB and groEl genes. Results Two tick species were collected in the present study: Rhipicephalus bursa (118) and Hyalomma marginatum marginatum (5). Molecular investigation showed that 32.1% (147/458) of blood samples were positive for Anaplasmataceae infection. Anaplasma ovis was identified in 42.3% (93/220) of sheep. Anaplasma marginale was amplified from 100% (12/12) of cattle and two R. bursa (2/123). Several potentially new species were also identified: Anaplasma cf. ovis , “ Candidatus Anaplasma corsicanum”, “ Candidatus Anaplasma mediterraneum” were amplified from 17.3% (38/220) of sheep, and Anaplasma sp. marginale-like was amplified from 80% (4/5) of goats. Finally, one R. bursa tick was found to harbour the DNA of E. canis . All samples from horses and dogs were negative for Anaplasmataceae infection. Conclusions To our knowledge, this study is the first epidemiological survey on Anaplasmataceae species infecting animals and ticks in Corsica and contributes toward the identification of current Anaplasmataceae species circulating in Corsica.

Background Our study aimed to assess the diversity of the species of Anaplasmataceae in Senegal that infect animals and ticks in three areas: near Keur Momar Sarr (northern region), Dielmo and Diop (Sine Saloum, central region of Senegal), and in Casamance (southern region of Senegal). Methods A total of 204 ticks and 433 blood samples were collected from ruminants, horses, donkeys and dogs. Ticks were identified morphologically and by molecular characterization targeting the 12S rRNA gene. Molecular characterization of species of Anaplasmataceae infecting Senegalese ticks and animals was conducted using the 23S rRNA, 16S rRNA, rpoB and groEL genes. Results Ticks were identified as Rhipicephalus evertsi evertsi (84.3%), Hyalomma rufipes (8.3%), Hyalomma impeltatum (4.9%), R. bursa (1.5%) and R. muhsamae (0.9%). The overall prevalence of Anaplasmataceae infection in ticks was 0.9%, whereas 41.1% of the sampled animals were found infected by one of the species belonging to this family. We identified the pathogen Anaplasma ovis in 55.9% of sheep, A. marginale and A. centrale in 19.4% and 8.1%, respectively, of cattle, as well as a putative new species of Anaplasmataceae. Two Anaplasma species commonly infecting ruminants were identified. Anaplasma cf. platys , closely related to A. platys was identified in 19.8% of sheep, 27.7% of goats and 22.6% of cattle, whereas a putative new species, named here provisionally “ Candidatus Anaplasma africae”, was identified in 3.7% of sheep, 10.3% of goats and 8.1% of cattle. Ehrlichia canis and Anaplasma platys were identified only from dogs sampled in the Keur Momar Sarr area. Ehrlichia canis was identified in 18.8% of dogs and two R. e. evertsi ticks removed from the same sheep. Anaplasma platys was identified in 15.6% of dogs. Neither of the dogs sampled from Casamance region nor the horses and donkeys sampled from Keur Momar Sarr area were found infected by an Anaplasmataceae species. Conclusions This study presents a summary of Anaplasmataceae species that infect animals and ticks in three areas from the northern, central and southern regions of Senegal. To our knowledge, our findings demonstrate for the first time the presence of multiple Anaplasmataceae species that infect ticks and domestic animals in Senegal. We recorded two potentially new species commonly infecting ruminants named here provisionally as Anaplasma cf . platys and “ Candidatus Anaplasma africae”. However, E. canis was the only species identified and amplified from ticks. None of the other Anaplasmataceae species identified in animals were identified in the tick species collected from animals.

Raccoons (Procyon lotor) are reservoirs for pathogens of other wildlife species, domestic animals, and humans, including several tick‐borne pathogens. A relatively understudied organism in raccoons is Candidatus Neoehrlichia procyonis which has been detected in raccoons from the southeastern United States. A related species in Europe and Asia, Neoehrlichia mikurensis, uses rodents as reservoirs and Ixodes spp. as vectors; however, studies on rodents suggest they are not susceptible to Ca. N. procyonis. N. mikurensis has been associated with cases of neoehrlichiosis in people and dogs, which emphasizes the need to better understand the natural history of Ca. N. procyonis. We conducted a molecular survey of raccoons from selected regions of the United States and Canada. Of 394 raccoons tested, 167 (42.4%) were confirmed to be positive for Ca. N. procyonis based on sequence analysis. There was spatial variation in prevalence with significantly higher prevalence (68%, 268/394) being detected in the Southeast region of the United States compared with all other regions, although a high prevalence (55.1%, 217/394) was detected in California. Lower prevalence was detected in the Midwest (3.8%, 15/394) and none of the raccoons from Canada were positive. These data suggest that Ca. N. procyonis is widespread in raccoon populations in the United States but there is spatial variation which may be related to vector distribution or some other factor. Although not known to infect hosts other than raccoons, neoehrlichiosis should be considered in cases of suspected ehrlichiosis in immunocompromised dogs or people that have no known etiologic agent. A total of 167/394 (42.4%) raccoons were positive for Candidatus Neoehrlichia procyonis. Spatial variation was noted with higher prevalences in the Southeast region of the United States and in California. Ca. N. procyonis is widespread in raccoon populations, but the spatial variation in prevalence may be related to vector distribution or another factor.

Background Neoehrlichia mikurensis s an emerging and vector-borne zoonosis: The first human disease cases were reported in 2010. Limited information is available about the prevalence and distribution of Neoehrlichia mikurensis in Europe, its natural life cycle and reservoir hosts. An Ehrlichia -like schotti variant has been described in questing Ixodes ricinus ticks, which could be identical to Neoehrlichia mikurensis. Methods Three genetic markers, 16S rDNA, gltA and GroEL, of Ehrlichia schotti -positive tick lysates were amplified, sequenced and compared to sequences from Neoehrlichia mikurensis . Based on these DNA sequences, a multiplex real-time PCR was developed to specifically detect Neoehrlichia mikurensis in combination with Anaplasma phagocytophilum in tick lysates. Various tick species from different life-stages, particularly Ixodes ricinus nymphs , were collected from the vegetation or wildlife. Tick lysates and DNA derived from organs of wild rodents were tested by PCR-based methods for the presence of Neoehrlichia mikurensis. Prevalence of Neoehrlichia mikurensis was calculated together with confidence intervals using Fisher's exact test. Results The three genetic markers of Ehrlichia schotti -positive field isolates were similar or identical to Neoehrlichia mikurensis . Neoehrlichia mikurensis was found to be ubiquitously spread in the Netherlands and Belgium, but was not detected in the 401 tick samples from the UK. Neoehrlichia mikurensis was found in nymphs and adult Ixodes ricinus ticks, but neither in their larvae, nor in any other tick species tested. Neoehrlichia mikurensis was detected in diverse organs of some rodent species. Engorging ticks from red deer, European mouflon, wild boar and sheep were found positive for Neoehrlichia mikurensis . Conclusions Ehrlichia schotti is similar, if not identical, to Neoehrlichia mikurensis. Neoehrlichia mikurensis is present in questing Ixodes ricinus ticks throughout the Netherlands and Belgium. We propose that Ixodes ricinus can transstadially, but not transovarially, transmit this microorganism, and that different rodent species may act as reservoir hosts. These data further imply that wildlife and humans are frequently exposed to Neoehrlichia mikurensis- infected ticks through tick bites. Future studies should aim to investigate to what extent Neoehrlichia mikurensis poses a risk to public health.