Molecular detection of Ehrlichia chaffeensis and a divergent Anaplasma lineage in free-ranging anteaters from southeastern Brazil

Molecular detection of Ehrlichia chaffeensis and a divergent Anaplasma lineage in free-ranging anteaters from southeastern Brazil | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Molecular detection of Ehrlichia chaffeensis and a divergent Anaplasma lineage in free-ranging anteaters from southeastern Brazil João Otávio Mochiuti, Mateus Oliveira Machado, Lígia Souza Lima Silveira da Mota This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9201768/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 4 You are reading this latest preprint version Abstract Bacteria of the family Anaplasmataceae are tick-borne pathogens of recognized veterinary and zoonotic relevance, widely distributed among domestic and wild vertebrate hosts. Although increasingly reported in Brazilian wildlife species, molecular data on these agents in xenarthrans remain scarce, particularly in regions undergoing intense environmental transformation. This study investigated the occurrence of Anaplasma spp. and Ehrlichia spp. in free-ranging giant anteaters ( Myrmecophaga tridactyla ), a threatened species in Brazil, and southern tamanduas ( Tamandua tetradactyla ) from the central-western region of São Paulo state, Brazil. Twenty-six blood samples (21 M. tridactyla and 5 T. tetradactyla ) were analyzed using conventional and nested PCR assays targeting the 23S rRNA gene ( Anaplasma spp.) and the dsb gene ( Ehrlichia spp.), followed by sequencing and phylogenetic inference. Ehrlichia chaffeensis DNA was detected in two M. tridactyla samples (7.69%), whereas Anaplasma DNA was detected in one T. tetradactyla sample (3.85%). The E. chaffeensis sequences showed high nucleotide identity (99.6–100%) with reference strains and clustered within well-supported clades in Maximum Likelihood analyses, supporting species-level identification. The Anaplasma sequence displayed high similarity to uncultured Anaplasma spp. clones and grouped phylogenetically near Anaplasma marginale and Anaplasma centrale . These findings provide molecular evidence of the circulation of Anaplasmataceae in free-ranging xenarthrans and suggest that these hosts may participate in sylvatic transmission networks of zoonotic tick-borne pathogens in anthropogenically altered landscapes in southeastern Brazil. Anaplasmataceae Anaplasma spp. Ehrlichia chaffeensis tick-borne pathogens Xenarthra One Health Figures Figure 1 Introduction Bacteria of the family Anaplasmataceae comprise obligate intracellular, tick-borne pathogens of recognized veterinary and zoonotic relevance. Species within the genera Anaplasma and Ehrlichia infect a wide range of domestic and wild vertebrate hosts, and several are associated with emerging infectious diseases of public health importance (Paddock and Childs 2003; Zanella 2016). In Brazil, molecular detection of Ehrlichia and Anaplasma species has been reported in cervids, carnivores, rodents, marsupials, birds, and captive wild mammals, highlighting the broad host spectrum of these bacteria in wildlife (Machado et al. 2006; André et al. 2010; André et al. 2012; Benevenute et al. 2017; Alabí Córdova et al. 2024). However, despite the increasing number of molecular investigations in wild mammals, data regarding Anaplasmataceae in xenarthrans remain limited. Giant anteaters ( Myrmecophaga tridactyla ) and southern tamanduas ( Tamandua tetradactyla ) are widely distributed in Brazil and frequently inhabit fragmented or anthropogenically altered landscapes, which may increase their exposure to tick vectors. Previous studies have documented infections in anteaters with hemoparasites and other vector-borne agents, including Trypanosoma cruzi , T. rangeli, and Leishmania infantum (De Araujo et al. 2013; Calchi et al. 2020). Nevertheless, molecular characterization of Anaplasma and Ehrlichia in these species remains scarce, particularly in São Paulo state. Given the ecological relevance of xenarthrans and the ongoing environmental transformation in southeastern Brazil, investigating the circulation of Anaplasmataceae in these hosts is epidemiologically relevant. Therefore, the present study aimed to investigate the occurrence of Anaplasma spp. and Ehrlichia spp. in free-ranging anteaters from the central-western region of São Paulo state, Brazil, using PCR-based detection and phylogenetic analysis. Materials and Methods Blood samples (n = 26) from giant anteaters ( Myrmecophaga tridactyla , n = 21) and southern tamanduas ( Tamandua tetradactyla , n = 5) were obtained from animals admitted to the Center for Wild Animal Medicine and Research (CEMPAS), UNESP, Botucatu, São Paulo, Brazil. Animals were rescued due to road accidents, illegal wildlife trade, or habitat disturbance. All procedures involving animals were conducted in accordance with institutional and national ethical standards. Sampling and handling protocols were approved by the Animal Use Ethics Committee of São Paulo State University (CEUA-UNESP; protocol no. 000.051/2024) and authorized by the Brazilian Biodiversity Authorization and Information System (SISBIO; permit no. 86964-1). Total genomic DNA was extracted using the DNeasy Blood & Tissue Kit (QIAGEN, Germany), according to the manufacturer’s instructions. DNA integrity and the absence of PCR inhibitors were verified by amplification of the endogenous GAPDH gene (Birkenheuer et al. 2003). Detection of Anaplasma spp. was performed by conventional PCR targeting a 515 bp fragment of the 23S rRNA gene (Dahmani et al. 2015). Detection of Ehrlichia spp. was conducted using nested PCR targeting the dsb gene (Doyle et al. 2005; Almeida et al. 2013). Sequencing was performed using the Sanger method. Consensus sequences were edited and assembled using Geneious and compared with reference sequences using BLASTn. The sequences obtained in this study were deposited in GenBank under accession numbers PZ060335–PZ060336 (dsb gene, Ehrlichia spp.) and PX496885.1 (23S rRNA gene, Anaplasma sp.). Phylogenetic analyses were conducted in MEGA11 using the Maximum Likelihood method under the Tamura–Nei model with 1,000 bootstrap replicates. The dsb gene was used for Ehrlichia , and the 23S rRNA gene for Anaplasma . For phylogenetic reconstruction, Anaplasma phagocytophilum was used as outgroup for the Ehrlichia dataset, whereas Ehrlichia ruminantium was used as outgroup for the Anaplasma dataset. Results and Discussion Detection of Anaplasmataceae DNA revealed the presence of Ehrlichia chaffeensis in two Myrmecophaga tridactyla individuals (7.69%) and Anaplasma spp. in one Tamandua tetradactyla individual (3.85%), as demonstrated by PCR amplification. Subsequent BLASTn analysis demonstrated 100% nucleotide identity (sample 8) and 99.61% identity (sample 14) with reference strains of E. chaffeensis , including the Wakulla and Arkansas. Phylogenetic inference placed both sequences within well-supported clades comprising E. chaffeensis , supporting species-level identification (Fig. 1 A). The high nucleotide identity in the dsb gene, combined with robust phylogenetic clustering, supports the assignment of the detected sequences to E. chaffeensis . The dsb sequences generated in this study were deposited in GenBank under accession numbers PZ060335 (sample 8) and PZ060336 (sample 14). The detection of E. chaffeensis in M. tridactyla is epidemiologically relevant given the zoonotic importance of this pathogen and its documented occurrence in wildlife and domestic hosts in Brazil (Paddock and Childs 2003; Machado et al. 2006; André et al. 2010). Although reports of Ehrlichia spp. in free-ranging mammals have increased in recent years (Sacchi et al. 2012; Benevenute et al. 2017), molecular evidence in xenarthrans remains limited. The Anaplasma -positive sample showed 97.35% nucleotide identity with uncultured Anaplasma sp. clone RHM5 and 96.57% identity with clone Sen3-1. In Maximum Likelihood phylogenetic analysis, the sequence clustered within a well-supported clade (bootstrap = 96%) comprising lineages related to Anaplasma marginale and Anaplasma centrale (Fig. 1 B). Despite its phylogenetic proximity to these species, the detected sequence did not group identically with any single reference strain, indicating genetic divergence from currently characterized taxa. The Anaplasma 23S rRNA sequence obtained in this study was deposited in GenBank under accession number PX496885.1. Similar findings of divergent or uncultured Anaplasma lineages in wildlife have been reported in Brazil and other regions (Dahmani et al. 2015; Calchi et al. 2020), suggesting the existence of sylvatic transmission cycles involving poorly characterized variants. However, because only a partial 23S rRNA fragment was analyzed, definitive species-level classification cannot be established, and additional molecular markers or multilocus approaches would be required for precise taxonomic resolution. From an ecological perspective, the detection of E. chaffeensis and a genetically divergent Anaplasma lineage in free-ranging xenarthrans suggests the potential involvement of these hosts in the maintenance of enzootic cycles of Anaplasmataceae in fragmented and human-modified ecosystems. Although the epidemiological significance of these findings remains to be fully elucidated, the occurrence of zoonotic and genetically distinct lineages in anthropogenically altered landscapes underscores the importance of continuous surveillance at the wildlife–livestock–human interface. The low detection frequency observed may reflect limited sample size, subclinical infection with low bacterial load, seasonal variation in vector activity, or intrinsic sensitivity limitations of conventional PCR assays (Paddock and Childs 2003). Additionally, the absence of concomitant tick collection and identification represents a limitation of the present study, as vector species potentially involved in local transmission cycles remain undetermined. The lack of bacterial isolation also precludes confirmation of pathogen viability. From a conservation and public health perspective, the detection of Anaplasmataceae in threatened species such as M. tridactyla underscores the need to incorporate vector-borne pathogens into wildlife health monitoring programs. Although clinical implications were not assessed in this study, the presence of zoonotic and genetically distinct lineages supports the relevance of integrating wildlife pathogen surveillance into broader One Health strategies, particularly in regions undergoing rapid environmental change. Future investigations including expanded geographic sampling, vector identification, and multilocus or genomic approaches will be essential to clarify the epidemiological role of xenarthrans in the maintenance and transmission of these agents. Conclusion This study provides molecular evidence of the circulation of Anaplasmataceae in free-ranging anteaters from central-western São Paulo State, southeastern Brazil. Ehrlichia chaffeensis was detected in Myrmecophaga tridactyla , while a genetically divergent Anaplasma lineage was identified in Tamandua tetradactyla . These findings expand current knowledge of the occurrence of Anaplasma and Ehrlichia in xenarthrans and highlight the importance of integrating wildlife pathogen surveillance into broader One Health frameworks in regions undergoing rapid environmental change. Further investigations incorporating expanded sampling, vector identification, and multilocus or genomic approaches are necessary to clarify the epidemiological role and taxonomic status of these lineages. Declarations Acknowledgements We acknowledge CEMPAS for their assistance in animal care and providing the samples for the study. Authors’ contributions JOM: conceptualization, methodology, formal analysis, and writing – original draft preparation. MOM: sample collection, laboratory procedures, and data curation. LSLM: supervision and writing – review and editing. All authors read and approved the final manuscript. Funding This research received no external funding. Availability of data and materials The nucleotide sequences generated in this study are available in GenBank under accession numbers PZ060335–PZ060336 (dsb gene, Ehrlichia chaffeensis ) and PX496885.1 (23S rRNA gene, Anaplasma sp.). Additional information supporting the findings of this study is available from the corresponding author upon reasonable request. Ethics approval and consent to participate All procedures involving animals were conducted in accordance with institutional and national ethical guidelines. Sampling and handling protocols were approved by the Animal Use Ethics Committee of São Paulo State University (CEUA-UNESP; protocol no. 000.051/2024) and authorized by the Brazilian Biodiversity Authorization and Information System (SISBIO; permit no. 86964-1). Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. References Alabí Córdova AS, Fecchio A, Calchi AC, Dias CM, Mongruel ACB, Neves LF, Lee DAB, Machado RZ, André MR (2024) Novel tick-borne Anaplasmataceae genotypes in tropical birds from the Brazilian Pantanal Wetland. Microorganisms. https://doi.org/10.3390/microorganisms12050962 Almeida AP, Souza TD, Marcili A, Labruna MB (2013) Novel Ehrlichia and Hepatozoon agents infecting the crab-eating fox ( Cerdocyon thous ) in southeastern Brazil. J Med Entomol. https://doi.org/10.1603/ME12272 André MR, Machado RZ, Labruna MB, Baldani CD, Duarte JMB (2010) Molecular and serologic detection of Ehrlichia spp. in endangered Brazilian wild captive felids. J Wildl Dis. https://doi.org/10.7589/0090-3558-46.3.1017 André MR, Adania AC, Machado RZ, Allegretti M, Felippe SS, Silva PC, Labruna MB (2012) Molecular detection of tick-borne bacterial agents in Brazilian and exotic captive carnivores. Ticks Tick Borne Dis. https://doi.org/10.1016/j.ttbdis.2012.04.002 Benevenute JL, Machado RZ, André MR, Labruna MB, Roque ALR (2017) Assessment of a quantitative 5′ nuclease real-time polymerase chain reaction using the groEL gene for Ehrlichia and Anaplasma species in rodents in Brazil. Ticks Tick Borne Dis. https://doi.org/10.1016/j.ttbdis.2017.04.005 Birkenheuer AJ, Levy MG, Breitschwerdt EB (2003) Development and evaluation of a seminested PCR for detection and differentiation of Babesia gibsoni (Asian genotype) and Babesia canis DNA in canine blood samples. J Clin Microbiol. https://doi.org/10.1128/JCM.41.9.4172-4177.2003 Calchi AC, André MR, Machado RZ, Labruna MB (2020) Ehrlichia spp. and Anaplasma spp. in Xenarthra mammals from Brazil, with evidence of novel “ Candidatus Anaplasma spp.” Sci Rep. https://doi.org/10.1038/s41598-020-69564-8 Dahmani M, Davoust B, Benterki MS, Fenollar F, Raoult D, Mediannikov O (2015) Development of a new PCR-based assay to detect Anaplasmataceae and the first report of Anaplasma phagocytophilum and Anaplasma platys in cattle from Algeria. Comp Immunol Microbiol Infect Dis. https://doi.org/10.1016/j.cimid.2015.02.002 De Araujo VAL, Boite MC, Cupolillo E, Jansen AM, Roque ALR (2013) Mixed infection in the anteater Tamandua tetradactyla (Mammalia: Pilosa) from Pará State, Brazil: Trypanosoma cruzi , T. rangeli , and Leishmania infantum . Parasitology. https://doi.org/10.1017/S0031182012001886 Doyle CK, Labruna MB, Breitschwerdt EB, Tang YW, Corstvet RE, Hegarty BC, Bloch KC, Li P, Walker DH (2005) Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene. J Mol Diagn. https://doi.org/10.1016/S1525-1578(10)60581-8 Machado RZ, André MR, Labruna MB (2006) Detection of Ehrlichia chaffeensis in Brazilian marsh deer ( Blastocerus dichotomus ). Vet Parasitol. https://doi.org/10.1016/j.vetpar.2006.02.038 Paddock CD, Childs JE (2003) Ehrlichia chaffeensis : a prototypical emerging pathogen. Clin Microbiol Rev. https://doi.org/10.1128/CMR.16.1.37-64.2003 Sacchi ABV, André MR, Machado RZ, Labruna MB, Duarte JMB (2012) Prevalence and molecular characterization of Anaplasmataceae agents in free-ranging Brazilian marsh deer ( Blastocerus dichotomus ). Comp Immunol Microbiol Infect Dis. https://doi.org/10.1016/j.cimid.2012.02.002 Zanella JRC (2016) Zoonoses emergentes e reemergentes e sua importância para saúde e produção animal. Pesq Agropec Bras. https://doi.org/10.1590/S0100-204X2016000500011 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 27 Mar, 2026 Editor assigned by journal 26 Mar, 2026 Submission checks completed at journal 26 Mar, 2026 First submitted to journal 23 Mar, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-9201768","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":613073342,"identity":"ed5c501e-b171-423f-a2c5-b3384ebed9cd","order_by":0,"name":"João Otávio Mochiuti","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxElEQVRIiWNgGAWjYBACxgYwIcHAwMx8ACIkQawWHma2BOK0wPXxMPAYEKeFuYE78cHPHRaJ+9l5vkkXVNTlMUj3PiBgAe9mw94zEok9zLzbpGecOVzMIHPcgJCWbdKMbVAtvG0HEhsk0gh5A66F55k077860rSwSfM2MBOhpRnklzYJ457DbMbWPMcOF7PJHMOvxbC9d+ODn211su39hx/e5qmpy+OXbiOgpRlNIIENvwYGBnl0gQRCOkbBKBgFo2DkAQCAKDxqnU7EGgAAAABJRU5ErkJggg==","orcid":"","institution":"Universidade Estadual Paulista (Unesp)","correspondingAuthor":true,"prefix":"","firstName":"João","middleName":"Otávio","lastName":"Mochiuti","suffix":""},{"id":613073343,"identity":"25881157-fb92-44a3-871d-1a7266bc1593","order_by":1,"name":"Mateus Oliveira Machado","email":"","orcid":"","institution":"Universidade Estadual Paulista (Unesp)","correspondingAuthor":false,"prefix":"","firstName":"Mateus","middleName":"Oliveira","lastName":"Machado","suffix":""},{"id":613073344,"identity":"a867a51d-2aa4-454e-89cf-0eb3b078f556","order_by":2,"name":"Lígia Souza Lima Silveira da Mota","email":"","orcid":"","institution":"Universidade Estadual Paulista (Unesp)","correspondingAuthor":false,"prefix":"","firstName":"Lígia","middleName":"Souza Lima Silveira da","lastName":"Mota","suffix":""}],"badges":[],"createdAt":"2026-03-23 14:24:16","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9201768/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9201768/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106537478,"identity":"7a780250-9b9c-4284-aceb-23c776fadb8a","added_by":"auto","created_at":"2026-04-09 15:26:43","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":219962,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic analysis of \u003cem\u003eEhrlichia\u003c/em\u003e and \u003cem\u003eAnaplasma \u003c/em\u003edetected in anteaters.\u003c/p\u003e\n\u003cp\u003eTrees were constructed using the Maximum Likelihood method under the Tamura–Nei model with 1,000 bootstrap replicates in MEGA11. Bootstrap values ≥50% are shown at nodes. Sequences generated in the present study are indicated in bold, and GenBank accession numbers are shown before species names. (A) The dsb gene dataset of \u003cem\u003eEhrlichia\u003c/em\u003espp. was rooted using \u003cem\u003eAnaplasma phagocytophilum\u003c/em\u003e as outgroup. (B) The 23S rRNA gene dataset of \u003cem\u003eAnaplasma\u003c/em\u003e spp. was rooted using \u003cem\u003eEhrlichia ruminantium\u003c/em\u003e as outgroup. Scale bars represent substitutions per site.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-9201768/v1/4421828dbed1d443353cdb9d.png"},{"id":106537535,"identity":"8b9385df-a99e-4991-ac4b-b9d3384a5947","added_by":"auto","created_at":"2026-04-09 15:27:01","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":578725,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9201768/v1/08fe6ea2-62ef-4b80-9db0-11dbb02c4953.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Molecular detection of Ehrlichia chaffeensis and a divergent Anaplasma lineage in free-ranging anteaters from southeastern Brazil","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBacteria of the family Anaplasmataceae comprise obligate intracellular, tick-borne pathogens of recognized veterinary and zoonotic relevance. Species within the genera \u003cem\u003eAnaplasma\u003c/em\u003e and \u003cem\u003eEhrlichia\u003c/em\u003e infect a wide range of domestic and wild vertebrate hosts, and several are associated with emerging infectious diseases of public health importance (Paddock and Childs 2003; Zanella 2016).\u003c/p\u003e \u003cp\u003eIn Brazil, molecular detection of \u003cem\u003eEhrlichia\u003c/em\u003e and \u003cem\u003eAnaplasma\u003c/em\u003e species has been reported in cervids, carnivores, rodents, marsupials, birds, and captive wild mammals, highlighting the broad host spectrum of these bacteria in wildlife (Machado et al. 2006; Andr\u0026eacute; et al. 2010; Andr\u0026eacute; et al. 2012; Benevenute et al. 2017; Alab\u0026iacute; C\u0026oacute;rdova et al. 2024). However, despite the increasing number of molecular investigations in wild mammals, data regarding Anaplasmataceae in xenarthrans remain limited.\u003c/p\u003e \u003cp\u003eGiant anteaters (\u003cem\u003eMyrmecophaga tridactyla\u003c/em\u003e) and southern tamanduas (\u003cem\u003eTamandua tetradactyla\u003c/em\u003e) are widely distributed in Brazil and frequently inhabit fragmented or anthropogenically altered landscapes, which may increase their exposure to tick vectors. Previous studies have documented infections in anteaters with hemoparasites and other vector-borne agents, including \u003cem\u003eTrypanosoma cruzi\u003c/em\u003e, \u003cem\u003eT.\u003c/em\u003e rangeli, and \u003cem\u003eLeishmania infantum\u003c/em\u003e (De Araujo et al. 2013; Calchi et al. 2020). Nevertheless, molecular characterization of \u003cem\u003eAnaplasma\u003c/em\u003e and \u003cem\u003eEhrlichia\u003c/em\u003e in these species remains scarce, particularly in S\u0026atilde;o Paulo state.\u003c/p\u003e \u003cp\u003eGiven the ecological relevance of xenarthrans and the ongoing environmental transformation in southeastern Brazil, investigating the circulation of Anaplasmataceae in these hosts is epidemiologically relevant. Therefore, the present study aimed to investigate the occurrence of \u003cem\u003eAnaplasma\u003c/em\u003e spp. and \u003cem\u003eEhrlichia\u003c/em\u003e spp. in free-ranging anteaters from the central-western region of S\u0026atilde;o Paulo state, Brazil, using PCR-based detection and phylogenetic analysis.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003eBlood samples (n\u0026thinsp;=\u0026thinsp;26) from giant anteaters (\u003cem\u003eMyrmecophaga tridactyla\u003c/em\u003e, n\u0026thinsp;=\u0026thinsp;21) and southern tamanduas (\u003cem\u003eTamandua tetradactyla\u003c/em\u003e, n\u0026thinsp;=\u0026thinsp;5) were obtained from animals admitted to the Center for Wild Animal Medicine and Research (CEMPAS), UNESP, Botucatu, S\u0026atilde;o Paulo, Brazil. Animals were rescued due to road accidents, illegal wildlife trade, or habitat disturbance.\u003c/p\u003e \u003cp\u003e All procedures involving animals were conducted in accordance with institutional and national ethical standards. Sampling and handling protocols were approved by the Animal Use Ethics Committee of S\u0026atilde;o Paulo State University (CEUA-UNESP; protocol no. 000.051/2024) and authorized by the Brazilian Biodiversity Authorization and Information System (SISBIO; permit no. 86964-1).\u003c/p\u003e \u003cp\u003eTotal genomic DNA was extracted using the DNeasy Blood \u0026amp; Tissue Kit (QIAGEN, Germany), according to the manufacturer\u0026rsquo;s instructions. DNA integrity and the absence of PCR inhibitors were verified by amplification of the endogenous GAPDH gene (Birkenheuer et al. 2003).\u003c/p\u003e \u003cp\u003eDetection of \u003cem\u003eAnaplasma\u003c/em\u003e spp. was performed by conventional PCR targeting a 515 bp fragment of the 23S rRNA gene (Dahmani et al. 2015). Detection of \u003cem\u003eEhrlichia\u003c/em\u003e spp. was conducted using nested PCR targeting the dsb gene (Doyle et al. 2005; Almeida et al. 2013).\u003c/p\u003e \u003cp\u003eSequencing was performed using the Sanger method. Consensus sequences were edited and assembled using Geneious and compared with reference sequences using BLASTn. The sequences obtained in this study were deposited in GenBank under accession numbers PZ060335\u0026ndash;PZ060336 (dsb gene, \u003cem\u003eEhrlichia\u003c/em\u003e spp.) and PX496885.1 (23S rRNA gene, \u003cem\u003eAnaplasma\u003c/em\u003e sp.).\u003c/p\u003e \u003cp\u003ePhylogenetic analyses were conducted in MEGA11 using the Maximum Likelihood method under the Tamura\u0026ndash;Nei model with 1,000 bootstrap replicates. The dsb gene was used for \u003cem\u003eEhrlichia\u003c/em\u003e, and the 23S rRNA gene for \u003cem\u003eAnaplasma\u003c/em\u003e. For phylogenetic reconstruction, \u003cem\u003eAnaplasma phagocytophilum\u003c/em\u003e was used as outgroup for the \u003cem\u003eEhrlichia\u003c/em\u003e dataset, whereas \u003cem\u003eEhrlichia ruminantium\u003c/em\u003e was used as outgroup for the \u003cem\u003eAnaplasma\u003c/em\u003e dataset.\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cp\u003eDetection of Anaplasmataceae DNA revealed the presence of \u003cem\u003eEhrlichia chaffeensis\u003c/em\u003e in two \u003cem\u003eMyrmecophaga tridactyla\u003c/em\u003e individuals (7.69%) and \u003cem\u003eAnaplasma\u003c/em\u003e spp. in one \u003cem\u003eTamandua tetradactyla\u003c/em\u003e individual (3.85%), as demonstrated by PCR amplification.\u003c/p\u003e \u003cp\u003eSubsequent BLASTn analysis demonstrated 100% nucleotide identity (sample 8) and 99.61% identity (sample 14) with reference strains of \u003cem\u003eE. chaffeensis\u003c/em\u003e, including the Wakulla and Arkansas. Phylogenetic inference placed both sequences within well-supported clades comprising \u003cem\u003eE. chaffeensis\u003c/em\u003e, supporting species-level identification (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). The high nucleotide identity in the dsb gene, combined with robust phylogenetic clustering, supports the assignment of the detected sequences to \u003cem\u003eE. chaffeensis\u003c/em\u003e. The dsb sequences generated in this study were deposited in GenBank under accession numbers PZ060335 (sample 8) and PZ060336 (sample 14).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe detection of \u003cem\u003eE. chaffeensis\u003c/em\u003e in \u003cem\u003eM. tridactyla\u003c/em\u003e is epidemiologically relevant given the zoonotic importance of this pathogen and its documented occurrence in wildlife and domestic hosts in Brazil (Paddock and Childs 2003; Machado et al. 2006; Andr\u0026eacute; et al. 2010). Although reports of \u003cem\u003eEhrlichia\u003c/em\u003e spp. in free-ranging mammals have increased in recent years (Sacchi et al. 2012; Benevenute et al. 2017), molecular evidence in xenarthrans remains limited.\u003c/p\u003e \u003cp\u003eThe \u003cem\u003eAnaplasma\u003c/em\u003e-positive sample showed 97.35% nucleotide identity with uncultured \u003cem\u003eAnaplasma\u003c/em\u003e sp. clone RHM5 and 96.57% identity with clone Sen3-1. In Maximum Likelihood phylogenetic analysis, the sequence clustered within a well-supported clade (bootstrap\u0026thinsp;=\u0026thinsp;96%) comprising lineages related to \u003cem\u003eAnaplasma marginale\u003c/em\u003e and \u003cem\u003eAnaplasma centrale\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eDespite its phylogenetic proximity to these species, the detected sequence did not group identically with any single reference strain, indicating genetic divergence from currently characterized taxa. The Anaplasma 23S rRNA sequence obtained in this study was deposited in GenBank under accession number PX496885.1.\u003c/p\u003e \u003cp\u003eSimilar findings of divergent or uncultured \u003cem\u003eAnaplasma\u003c/em\u003e lineages in wildlife have been reported in Brazil and other regions (Dahmani et al. 2015; Calchi et al. 2020), suggesting the existence of sylvatic transmission cycles involving poorly characterized variants. However, because only a partial 23S rRNA fragment was analyzed, definitive species-level classification cannot be established, and additional molecular markers or multilocus approaches would be required for precise taxonomic resolution.\u003c/p\u003e \u003cp\u003eFrom an ecological perspective, the detection of \u003cem\u003eE. chaffeensis\u003c/em\u003e and a genetically divergent \u003cem\u003eAnaplasma\u003c/em\u003e lineage in free-ranging xenarthrans suggests the potential involvement of these hosts in the maintenance of enzootic cycles of Anaplasmataceae in fragmented and human-modified ecosystems. Although the epidemiological significance of these findings remains to be fully elucidated, the occurrence of zoonotic and genetically distinct lineages in anthropogenically altered landscapes underscores the importance of continuous surveillance at the wildlife\u0026ndash;livestock\u0026ndash;human interface.\u003c/p\u003e \u003cp\u003eThe low detection frequency observed may reflect limited sample size, subclinical infection with low bacterial load, seasonal variation in vector activity, or intrinsic sensitivity limitations of conventional PCR assays (Paddock and Childs 2003). Additionally, the absence of concomitant tick collection and identification represents a limitation of the present study, as vector species potentially involved in local transmission cycles remain undetermined. The lack of bacterial isolation also precludes confirmation of pathogen viability.\u003c/p\u003e \u003cp\u003eFrom a conservation and public health perspective, the detection of Anaplasmataceae in threatened species such as \u003cem\u003eM. tridactyla\u003c/em\u003e underscores the need to incorporate vector-borne pathogens into wildlife health monitoring programs. Although clinical implications were not assessed in this study, the presence of zoonotic and genetically distinct lineages supports the relevance of integrating wildlife pathogen surveillance into broader One Health strategies, particularly in regions undergoing rapid environmental change. Future investigations including expanded geographic sampling, vector identification, and multilocus or genomic approaches will be essential to clarify the epidemiological role of xenarthrans in the maintenance and transmission of these agents.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study provides molecular evidence of the circulation of Anaplasmataceae in free-ranging anteaters from central-western S\u0026atilde;o Paulo State, southeastern Brazil. \u003cem\u003eEhrlichia chaffeensis\u003c/em\u003e was detected in \u003cem\u003eMyrmecophaga tridactyla\u003c/em\u003e, while a genetically divergent \u003cem\u003eAnaplasma\u003c/em\u003e lineage was identified in \u003cem\u003eTamandua tetradactyla\u003c/em\u003e. These findings expand current knowledge of the occurrence of \u003cem\u003eAnaplasma\u003c/em\u003e and \u003cem\u003eEhrlichia\u003c/em\u003e in xenarthrans and highlight the importance of integrating wildlife pathogen surveillance into broader One Health frameworks in regions undergoing rapid environmental change. Further investigations incorporating expanded sampling, vector identification, and multilocus or genomic approaches are necessary to clarify the epidemiological role and taxonomic status of these lineages.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe acknowledge CEMPAS for their assistance in animal care and providing the samples for the study.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eJOM: conceptualization, methodology, formal analysis, and writing \u0026ndash; original draft preparation.\u003c/p\u003e\n\u003cp\u003eMOM: sample collection, laboratory procedures, and data curation.\u003c/p\u003e\n\u003cp\u003eLSLM: supervision and writing \u0026ndash; review and editing.\u003c/p\u003e\n\u003cp\u003eAll authors read and approved the final manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research received no external funding.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe nucleotide sequences generated in this study are available in GenBank under accession numbers PZ060335\u0026ndash;PZ060336 (dsb gene, \u003cem\u003eEhrlichia chaffeensis\u003c/em\u003e) and PX496885.1 (23S rRNA gene, \u003cem\u003eAnaplasma\u003c/em\u003e sp.). Additional information supporting the findings of this study is available from the corresponding author upon reasonable request.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;All procedures involving animals were conducted in accordance with institutional and national ethical guidelines. Sampling and handling protocols were approved by the Animal Use Ethics Committee of S\u0026atilde;o Paulo State University (CEUA-UNESP; protocol no. 000.051/2024) and authorized by the Brazilian Biodiversity Authorization and Information System (SISBIO; permit no. 86964-1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAlab\u0026iacute; C\u0026oacute;rdova AS, Fecchio A, Calchi AC, Dias CM, Mongruel ACB, Neves LF, Lee DAB, Machado RZ, Andr\u0026eacute; MR (2024) Novel tick-borne Anaplasmataceae genotypes in tropical birds from the Brazilian Pantanal Wetland. Microorganisms. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.3390/microorganisms12050962\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAlmeida AP, Souza TD, Marcili A, Labruna MB (2013) Novel Ehrlichia and Hepatozoon agents infecting the crab-eating fox (\u003cem\u003eCerdocyon thous\u003c/em\u003e) in southeastern Brazil. J Med Entomol. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1603/ME12272\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndr\u0026eacute; MR, Machado RZ, Labruna MB, Baldani CD, Duarte JMB (2010) Molecular and serologic detection of \u003cem\u003eEhrlichia\u003c/em\u003e spp. in endangered Brazilian wild captive felids. J Wildl Dis. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.7589/0090-3558-46.3.1017\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAndr\u0026eacute; MR, Adania AC, Machado RZ, Allegretti M, Felippe SS, Silva PC, Labruna MB (2012) Molecular detection of tick-borne bacterial agents in Brazilian and exotic captive carnivores. Ticks Tick Borne Dis. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1016/j.ttbdis.2012.04.002\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBenevenute JL, Machado RZ, Andr\u0026eacute; MR, Labruna MB, Roque ALR (2017) Assessment of a quantitative 5\u0026prime; nuclease real-time polymerase chain reaction using the \u003cem\u003egroEL\u003c/em\u003e gene for \u003cem\u003eEhrlichia\u003c/em\u003e and \u003cem\u003eAnaplasma\u003c/em\u003e species in rodents in Brazil. Ticks Tick Borne Dis. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1016/j.ttbdis.2017.04.005\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBirkenheuer AJ, Levy MG, Breitschwerdt EB (2003) Development and evaluation of a seminested PCR for detection and differentiation of \u003cem\u003eBabesia gibsoni\u003c/em\u003e (Asian genotype) and \u003cem\u003eBabesia canis\u003c/em\u003e DNA in canine blood samples. J Clin Microbiol. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1128/JCM.41.9.4172-4177.2003\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCalchi AC, Andr\u0026eacute; MR, Machado RZ, Labruna MB (2020) \u003cem\u003eEhrlichia\u003c/em\u003e spp. and \u003cem\u003eAnaplasma\u003c/em\u003e spp. in Xenarthra mammals from Brazil, with evidence of novel \u0026ldquo;\u003cem\u003eCandidatus Anaplasma\u003c/em\u003e spp.\u0026rdquo; Sci Rep. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1038/s41598-020-69564-8\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDahmani M, Davoust B, Benterki MS, Fenollar F, Raoult D, Mediannikov O (2015) Development of a new PCR-based assay to detect Anaplasmataceae and the first report of \u003cem\u003eAnaplasma phagocytophilum\u003c/em\u003e and \u003cem\u003eAnaplasma platys\u003c/em\u003e in cattle from Algeria. Comp Immunol Microbiol Infect Dis. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1016/j.cimid.2015.02.002\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Araujo VAL, Boite MC, Cupolillo E, Jansen AM, Roque ALR (2013) Mixed infection in the anteater \u003cem\u003eTamandua tetradactyla\u003c/em\u003e (Mammalia: Pilosa) from Par\u0026aacute; State, Brazil: \u003cem\u003eTrypanosoma cruzi\u003c/em\u003e, \u003cem\u003eT. rangeli\u003c/em\u003e, and \u003cem\u003eLeishmania infantum\u003c/em\u003e. Parasitology. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1017/S0031182012001886\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDoyle CK, Labruna MB, Breitschwerdt EB, Tang YW, Corstvet RE, Hegarty BC, Bloch KC, Li P, Walker DH (2005) Detection of medically important \u003cem\u003eEhrlichia\u003c/em\u003e by quantitative multicolor TaqMan real-time polymerase chain reaction of the \u003cem\u003edsb\u003c/em\u003e gene. J Mol Diagn. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1016/S1525-1578(10)60581-8\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMachado RZ, Andr\u0026eacute; MR, Labruna MB (2006) Detection of \u003cem\u003eEhrlichia chaffeensis\u003c/em\u003e in Brazilian marsh deer (\u003cem\u003eBlastocerus dichotomus\u003c/em\u003e). Vet Parasitol. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1016/j.vetpar.2006.02.038\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePaddock CD, Childs JE (2003) \u003cem\u003eEhrlichia chaffeensis\u003c/em\u003e: a prototypical emerging pathogen. Clin Microbiol Rev. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1128/CMR.16.1.37-64.2003\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSacchi ABV, Andr\u0026eacute; MR, Machado RZ, Labruna MB, Duarte JMB (2012) Prevalence and molecular characterization of Anaplasmataceae agents in free-ranging Brazilian marsh deer (\u003cem\u003eBlastocerus dichotomus\u003c/em\u003e). Comp Immunol Microbiol Infect Dis. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1016/j.cimid.2012.02.002\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZanella JRC (2016) Zoonoses emergentes e reemergentes e sua import\u0026acirc;ncia para sa\u0026uacute;de e produ\u0026ccedil;\u0026atilde;o animal. Pesq Agropec Bras. \u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003ehttps://doi.org/10.1590/S0100-204X2016000500011\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"veterinary-research-communications","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"verc","sideBox":"Learn more about [Veterinary Research Communications](https://www.springer.com/journal/11259)","snPcode":"11259","submissionUrl":"https://submission.nature.com/new-submission/11259/3","title":"Veterinary Research Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Anaplasmataceae, Anaplasma spp., Ehrlichia chaffeensis, tick-borne pathogens, Xenarthra, One Health","lastPublishedDoi":"10.21203/rs.3.rs-9201768/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9201768/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBacteria of the family Anaplasmataceae are tick-borne pathogens of recognized veterinary and zoonotic relevance, widely distributed among domestic and wild vertebrate hosts. Although increasingly reported in Brazilian wildlife species, molecular data on these agents in xenarthrans remain scarce, particularly in regions undergoing intense environmental transformation. This study investigated the occurrence of \u003cem\u003eAnaplasma\u003c/em\u003e spp. and \u003cem\u003eEhrlichia\u003c/em\u003e spp. in free-ranging giant anteaters (\u003cem\u003eMyrmecophaga tridactyla\u003c/em\u003e), a threatened species in Brazil, and southern tamanduas (\u003cem\u003eTamandua tetradactyla\u003c/em\u003e) from the central-western region of S\u0026atilde;o Paulo state, Brazil. Twenty-six blood samples (21 \u003cem\u003eM. tridactyla\u003c/em\u003e and 5 \u003cem\u003eT. tetradactyla\u003c/em\u003e) were analyzed using conventional and nested PCR assays targeting the 23S rRNA gene (\u003cem\u003eAnaplasma\u003c/em\u003e spp.) and the \u003cem\u003edsb\u003c/em\u003e gene (\u003cem\u003eEhrlichia\u003c/em\u003e spp.), followed by sequencing and phylogenetic inference. \u003cem\u003eEhrlichia chaffeensis\u003c/em\u003e DNA was detected in two \u003cem\u003eM. tridactyla\u003c/em\u003e samples (7.69%), whereas \u003cem\u003eAnaplasma\u003c/em\u003e DNA was detected in one \u003cem\u003eT. tetradactyla\u003c/em\u003e sample (3.85%). The \u003cem\u003eE. chaffeensis\u003c/em\u003e sequences showed high nucleotide identity (99.6\u0026ndash;100%) with reference strains and clustered within well-supported clades in Maximum Likelihood analyses, supporting species-level identification. The \u003cem\u003eAnaplasma\u003c/em\u003e sequence displayed high similarity to uncultured \u003cem\u003eAnaplasma\u003c/em\u003e spp. clones and grouped phylogenetically near \u003cem\u003eAnaplasma marginale\u003c/em\u003e and \u003cem\u003eAnaplasma centrale\u003c/em\u003e. These findings provide molecular evidence of the circulation of Anaplasmataceae in free-ranging xenarthrans and suggest that these hosts may participate in sylvatic transmission networks of zoonotic tick-borne pathogens in anthropogenically altered landscapes in southeastern Brazil.\u003c/p\u003e","manuscriptTitle":"Molecular detection of Ehrlichia chaffeensis and a divergent Anaplasma lineage in free-ranging anteaters from southeastern Brazil","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-09 15:24:58","doi":"10.21203/rs.3.rs-9201768/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-27T06:28:27+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-03-27T03:50:44+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-03-27T03:50:22+00:00","index":"","fulltext":""},{"type":"submitted","content":"Veterinary Research Communications","date":"2026-03-23T14:14:18+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"veterinary-research-communications","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"verc","sideBox":"Learn more about [Veterinary Research Communications](https://www.springer.com/journal/11259)","snPcode":"11259","submissionUrl":"https://submission.nature.com/new-submission/11259/3","title":"Veterinary Research Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"bff0ebb3-e900-42d6-a463-2c7356c2937a","owner":[],"postedDate":"April 9th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-04-28T08:10:13+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-09 15:24:58","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9201768","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9201768","identity":"rs-9201768","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}