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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.

The tickborne bacterial pathogen Neoehrlichia mikurensis has been detected in <1% of blood donors in Sweden. N. mikurensis can give rise to asymptomatic persistent infections. Up to 25% of Ixodes ricinus ticks in southern Norway are infected with N. mikurensis. We investigated the incidence of N. mikurensis infection among blood donors in this region. We detected N. mikurensis in the blood of 45/499 (9%) blood donors by independent PCR methods; 69% of those were repeatedly positive 1-7 months after the first detection and tested negative after doxycycline treatment. We tested 8/19 adult recipients of potentially infected blood; none tested positive for N. mikurensis at the time of testing (191-301 days after transfusion). Our study identified a very high rate of infection with N. mikurensis in blood donors in Norway; whether infection can be transmitted by transfusion of blood products, however, remains unclear.

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.

PurposeTo increase knowledge about the varied clinical manifestations of human infection with the emerging tick-borne pathogen Neoehrlichia mikurensis.MethodsAll patients diagnosed in Sweden with N. mikurensis infection during a 10-year period (2013–2023) were investigated regarding their demographic factors, risk factors, comorbidities, clinical signs and symptoms, and laboratory results. Multivariate models were generated using “Orthogonal Projections to Latent Structures-Discriminant Analysis” to identify clinical and immune parameters associated with N. mikurensis infection.ResultsDuring the 10-year period, 134 patients were diagnosed with N. mikurensis infection, 102 of whom were included in this study. Most of the patients (79%) were immunosuppressed. The main comorbidities were malignant B-cell lymphomas, multiple sclerosis, and rheumatoid arthritis. Rituximab therapy (59%) and splenectomy (14%) featured prominently. All patients resided in the southern tick-endemic part of Sweden, yet one-third of them were diagnosed in wintertime when ticks are inactive. Two asymptomatically infected blood donors were identified but transfusion-transmitted infection was not confirmed. Increased levels of C-reactive protein, orosomucoid, and total IgM in serum were associated with neoehrlichiosis. Previously unreported symptoms such as ankle edema, neck pain, numbness, and sudden deafness were detected in some patients. One case of aplastic anemia partially improved after eradication of the infection.ConclusionsNeoehrlichiosis is a multi-faceted emerging infectious disease.Key pointsHuman infections with tick-borne Neoehrlichia mikurensis can result in asymptomatic carriage, severe disease with fever and vascular events, and a possibly causal association with a variety of hematologic conditions as well as with sudden deafness.

Emerging tick-borne infections pose public health challenges and may complicate treatment decisions. The EMBio study, a multicenter observational study, aims to describe erythema migrans (EM), an early localized manifestation of Borrelia burgdorferi sensu lato (s.l.) infection, and investigate the occurrence of tick-borne co-infections among patients presenting with this skin lesion. Additionally, the study seeks to determine relations between EM morphology, other clinical manifestations, specific pathogens, and disease prognosis. Clinical characteristics, skin biopsies, and blood samples were analyzed from 26 patients to assess co-infections, quantity, Borrelia species, and spirochete load. Borrelia DNA was detected in 88% of EM skin lesions, with Borrelia afzelii as the predominant species. Two cases of co-infections were identified, one involving two Borrelia species and one involving Borrelia afzelii and the intracellular bacterium Neoehrlichia mikurensis . Notably, homogeneous EM lesions harbored significantly higher spirochete quantities in the central zone compared to annular lesions, suggesting that lesion morphology reflects local bacterial density. This supports the value of molecular diagnostics in detecting mixed infections and supports morphology-guided biopsy strategies in the clinical assessment of cutaneous infections. This study contributes to a better understanding of co-infection dynamics and may improve diagnostic accuracy and patient management in endemic settings.

Piroplasmosis, a disease of domestic and wild animals, is caused by tick-borne protozoa of the genera Babesia and Theileria, while anaplasmosis is caused by tick-borne bacteria of genera Anaplasma. Hyalomma dromedarii is the most dominant tick species infesting camels in Egypt and act as a vector of piroplasms, Anaplasma, Rickettsia and Ehrlichia spp. The available information concerning the detection of these pathogens in H. dromedarii infesting camels is limited. The present study aimed to evaluate the status of these pathogens in H. dromedarii ticks over four seasons of a year, in addition to investigate the infections of piroplasms and Anaplasmataceae besides their genetic diversity starting from June 2021 till April 2022. A total of 275 semi-engorged females of H. dromedarii were collected from different slaughtered camels, Toukh city slaughterhouse then investigated by Polymerase Chain Reaction (PCR) to detect piroplasms (Babesia spp., Theileria spp.) and Anaplasmataceae DNA targeting 18 S rRNA and 16 S rRNA genes, respectively followed by sequencing and phylogenetic analyses. Overall, piroplasms were detected in 38 ticks (13.8%), Babesia spp. was detected in 35 ticks (12.7%), while Theileria spp. was detected in one tick (0.4%). Anaplasmataceae was detected in 57 ticks (20.7%). Mixed infections of piroplasms and Anaplasmataceae were detected in 13 ticks (5%). Single infection either with piroplasms or Anaplasmataceae was detected in 25 (9%) and 44 (16%) ticks, respectively. The highest monthly rate of piroplasms was in April (spring) and Anaplasmataceae was in July (summer). Sequence analysis revealed that Babesia bigemina, Wolbachia spp. and Anaplasma marginale are the most dominant species in the examined tick samples. To the best of our knowledge, this study confirms the presence of B. bigemina, Wolbachia spp. and A. marginale in H. dromedarii in Egypt by sequencing.

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.