{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T12:46:52Z","timestamp":1787662012282,"version":"build-2736575974"},"reference-count":48,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,5,23]],"date-time":"2023-05-23T00:00:00Z","timestamp":1684800000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Streptococcus equi sub. zooepidemicus (SEZ) is described as a commensal bacterium of several animal species, including humans. Growing evidence supports the potential role of SEZ in the onset and progression of severe clinical manifestations of diseases in horses and other animals. In the present communication, we describe the diagnostic procedure applied to characterize the streptococcal infections caused by a novel SEZ sequence type (ST525) in donkeys raised on a farm in Abruzzo, Italy. The diagnostic process began with anamnesis and anatomopathological analysis, which revealed a severe bacterial suppurative bronchopneumonia associated with systemic vascular damage and haemorrhages. Then, SEZ infection was confirmed by applying an integrative diagnostic strategy that included standard bacterial isolation techniques, analytical tools for bacteria identification (MALDI-TOF MS), and molecular analysis (qPCR). Furthermore, the application of the whole-genome sequencing approach helped us to identify the bacterial strains and the virulence factors involved in animal diseases. The novel SEZ-ST525 was identified in two cases of the disease. This new sequence type was isolated from the lung, liver, and spleen in Case 1, and from retropharyngeal lymph nodes in Case 2. Moreover, the presence of the virulence gene mf2, a virulence factor carried by prophages in Streptococcus pyogenes, was also found for the first time in an SEZ strain. The results of the present study highlight the need to apply an integrated diagnostic approach for the identification and tracking of pathogenic strains of SEZ, shedding new light on the re-evaluation of these bacteria as a causative agent of disease in animals and humans.<\/jats:p>","DOI":"10.3390\/pathogens12060750","type":"journal-article","created":{"date-parts":[[2023,5,23]],"date-time":"2023-05-23T09:13:50Z","timestamp":1684833230000},"page":"750","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Identification of the Novel Streptococcus equi subsp. zooepidemicus Sequence Type 525 in Donkeys of Abruzzo Region, Italy"],"prefix":"10.3390","volume":"12","author":[{"given":"Maria Chiara","family":"Cantelmi","sequence":"first","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"},{"name":"Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmine","family":"Merola","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"},{"name":"Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9225-9727","authenticated-orcid":false,"given":"Daniela","family":"Averaimo","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7412-2164","authenticated-orcid":false,"given":"Alexandra","family":"Chiaverini","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4982-476X","authenticated-orcid":false,"given":"Francesca","family":"Cito","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"},{"name":"Department of Veterinary Medicine, University of Teramo, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4411-618X","authenticated-orcid":false,"given":"Antonio","family":"Cocco","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9595-782X","authenticated-orcid":false,"given":"Giovanni","family":"Di Teodoro","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0668-0532","authenticated-orcid":false,"given":"Maria Elisabetta","family":"De Angelis","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Di Bernardo","sequence":"additional","affiliation":[{"name":"Freelance Veterinary Practitioner, 65019 Pescara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Davide","family":"Auzino","sequence":"additional","affiliation":[{"name":"Freelance Veterinary Practitioner, 65019 Pescara, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2089-4123","authenticated-orcid":false,"given":"Antonio","family":"Petrini","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale dell\u2019Abruzzo e Molise \u201cG. Caporale\u201d, Campo Boario, 64100 Teramo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,5,23]]},"reference":[{"key":"ref_1","unstructured":"Sellon, D.C., and Long, M.T. (2013). Equine Infectious Diseases E-Book, Elsevier Health Sciences."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.cimid.2019.03.012","article-title":"Genetic Diversity of Streptococcus Equi Subsp. Zooepidemicus Isolated from Horses","volume":"65","author":"Preziuso","year":"2019","journal-title":"Comp. Immunol. Microbiol. Infect Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/j.vetmic.2007.04.020","article-title":"Real-Time PCR for Detection and Differentiation of Streptococcus Equi Subsp. Equi and Streptococcus Equi Subsp. Zooepidemicus","volume":"124","author":"Johansson","year":"2007","journal-title":"Vet. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/j.syapm.2015.02.010","article-title":"Rapid and Accurate Identification of Streptococcus Equi Subspecies by MALDI-TOF MS","volume":"38","author":"Kudirkiene","year":"2015","journal-title":"Syst. Appl. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1177\/1040638717702687","article-title":"Discrimination of Streptococcus Equi Subsp. Equi and Streptococcus Equi Subsp. Zooepidemicus Using Matrix-Assisted Laser Desorption\/Ionization Time-of-Flight Mass Spectrometry","volume":"29","author":"Mani","year":"2017","journal-title":"J. Vet. Diagn. Investig."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1128\/JCM.40.3.1106-1108.2002","article-title":"Unusual Outbreak of Clinical Mastitis in Dairy Sheep Caused by Streptococcus Equi Subsp. Zooepidemicus","volume":"40","author":"Vela","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1136\/vr.161.15.528","article-title":"Outbreak and Control of Haemorrhagic Pneumonia Due to Streptococcus Equi Subspecies Zooepidemicus in Dogs","volume":"161","author":"Kim","year":"2007","journal-title":"Vet. Rec."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.1007\/s10096-010-1037-5","article-title":"Streptococcus Equi Subspecies Zooepidemicus Meningitis--a Case Report and Review of the Literature","volume":"29","author":"Eyre","year":"2010","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"142","DOI":"10.1016\/j.tvjl.2010.04.028","article-title":"Streptococcus Zooepidemicus: An Emerging Canine Pathogen","volume":"188","author":"Priestnall","year":"2011","journal-title":"Vet. J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1017\/S0950268810001184","article-title":"Human Meningitis from Streptococcus equi subsp. Zooepidemicus Acquired as Zoonoses","volume":"139","author":"Minces","year":"2011","journal-title":"Epidemiol. Infect."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"156","DOI":"10.3201\/eid2101.140640","article-title":"Streptococcus Equi Subsp. Zooepidemicus Infections Associated with Guinea Pigs","volume":"21","author":"Gruszynski","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1041","DOI":"10.3201\/eid1907.121365","article-title":"Transmission of Streptococcus Equi Subspecies Zooepidemicus Infection from Horses to Humans","volume":"19","author":"Pelkonen","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5","DOI":"10.3201\/eid2905.221338","article-title":"Early Release\u2014Severe Streptococcus Equi Subspecies Zooepidemicus Outbreak from Unpasteurized Dairy Product Consumption, Italy","volume":"29","author":"Bosica","year":"2023","journal-title":"Emerg. Infect. Dis. J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"537","DOI":"10.29261\/pakvetj\/2020.081","article-title":"Endometritis in Donkeys Associated with Streptococcus Equi Subspecies Zooepidemicus Infection","volume":"40","author":"Gao","year":"2020","journal-title":"PVJ"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.jevs.2019.04.003","article-title":"Selenium and Vitamin E Concentrations in a Healthy Donkey Population in Central Italy","volume":"78","author":"Bazzano","year":"2019","journal-title":"J. Equine Vet. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"228","DOI":"10.3168\/jdsc.2021-0175","article-title":"Valorization of Donkey Milk: Technology, Functionality, and Future Prospects","volume":"3","author":"Papademas","year":"2022","journal-title":"JDS Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.jevs.2018.02.026","article-title":"Infectious Diseases in Donkeys and Mules: An Overview and Update","volume":"65","author":"Barrandeguy","year":"2018","journal-title":"J. Equine Vet. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"C\u00e2mara, R.J.F., Bueno, B.L., Resende, C.F., Balasuriya, U.B.R., Sakamoto, S.M., and dos Reis, J.K.P. (2020). Viral Diseases That Affect Donkeys and Mules. Animals, 10.","DOI":"10.3390\/ani10122203"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1016\/j.cveq.2021.04.012","article-title":"Antimicrobial Selection for the Equine Practitioner","volume":"37","author":"Wilson","year":"2021","journal-title":"Vet. Clin. N. Am. Equine Pract."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"446","DOI":"10.3389\/fmicb.2018.00446","article-title":"A Validation Approach of an End-to-End Whole Genome Sequencing Workflow for Source Tracking of Listeria Monocytogenes and Salmonella Enterica","volume":"9","author":"Portmann","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.ijid.2018.04.4090","article-title":"The Italian Information System for the Collection and Analysis of Complete Genome Sequence of Pathogens Isolated from Animal, Food and Environment","volume":"73","author":"Cito","year":"2018","journal-title":"Int. J. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1529","DOI":"10.1128\/JCM.02981-13","article-title":"Benchmarking of Methods for Genomic Taxonomy","volume":"52","author":"Larsen","year":"2014","journal-title":"J. Clin. Microbiol."},{"key":"ref_23","first-page":"000748","article-title":"ResFinder\u2014An Open Online Resource for Identification of Antimicrobial Resistance Genes in next-Generation Sequencing Data and Prediction of Phenotypes from Genotypes","volume":"8","author":"Florensa","year":"2022","journal-title":"Microb. Genom."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"W16","DOI":"10.1093\/nar\/gkw387","article-title":"PHASTER: A Better, Faster Version of the PHAST Phage Search Tool","volume":"44","author":"Arndt","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e20","DOI":"10.7717\/peerj-cs.20","article-title":"CFSAN SNP Pipeline: An automated method for constructing SNP matrices from next-generation sequence data","volume":"1","author":"Davis","year":"2015","journal-title":"PeerJ Comput. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1111\/j.1751-0813.2000.tb11901.x","article-title":"Bacterial Infection of the Lower Respiratory Tract in 34 Horses","volume":"78","author":"Racklyeft","year":"2000","journal-title":"Aust. Vet. J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1111\/evj.13339","article-title":"Pathology, Infectious Agents and Horse- and Management-Level Risk Factors Associated with Signs of Respiratory Disease in Ethiopian Working Horses","volume":"53","author":"Laing","year":"2021","journal-title":"Equine Vet. J."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"604675","DOI":"10.3389\/fvets.2021.604675","article-title":"A Novel Real-Time PCR Assay for the Rapid Detection of Virulent Streptococcus Equi Subspecies Zooepidemicus\u2014An Emerging Pathogen of Swine","volume":"8","author":"Kuchipudi","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.vetmic.2017.06.020","article-title":"Diversity of Streptococcus Equi Subsp. Zooepidemicus Strains Isolated from the Spanish Sheep and Goat Population and the Identification, Function and Prevalence of a Novel Arbutin Utilisation System","volume":"207","author":"Steward","year":"2017","journal-title":"Vet. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"943","DOI":"10.3168\/jds.2008-1548","article-title":"Epidemiological Investigation of Streptococcus Equi Subspecies Zooepidemicus Involved in Clinical Mastitis in Dairy Goats","volume":"92","author":"Pisoni","year":"2009","journal-title":"J. Dairy Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1007\/s00203-002-0412-8","article-title":"Cloning and Characterization of Two Novel DNases from Streptococcus pyogenes","volume":"177","author":"Hasegawa","year":"2002","journal-title":"Arch. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Meurer, M., \u00d6hlmann, S., Bonilla, M.C., Valentin-Weigand, P., Beineke, A., Hennig-Pauka, I., Schwerk, C., Schroten, H., Baums, C.G., and von K\u00f6ckritz-Blickwede, M. (2020). Role of Bacterial and Host DNases on Host-Pathogen Interaction during Streptococcus Suis Meningitis. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21155289"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1042\/BST20130165","article-title":"Streptococcus Zooepidemicus and Streptococcus Equi Evolution: The Role of CRISPRs","volume":"41","author":"Waller","year":"2013","journal-title":"Biochem. Soc. Trans."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1016\/j.ics.2005.11.086","article-title":"P9, a Temperate Bacteriophage of Streptococcus Equi","volume":"1289","author":"Tiwari","year":"2006","journal-title":"Int. Congr. Ser."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1128\/microbiolspec.GPP3-0059-2018","article-title":"The Bacteriophages of Streptococcus pyogenes","volume":"7","author":"McShan","year":"2019","journal-title":"Microbiol. Spectr."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Nocera, F.P., D\u2019Eletto, E., Ambrosio, M., Fiorito, F., Pagnini, U., and De Martino, L. (2021). Occurrence and Antimicrobial Susceptibility Profiles of Streptococcus Equi Subsp. Zooepidemicus Strains Isolated from Mares with Fertility Problems. Antibiotics, 11.","DOI":"10.3390\/antibiotics11010025"},{"key":"ref_37","first-page":"64","article-title":"Clinical and Microbiological Observations on Strangles in Donkeys","volume":"54","author":"Firew","year":"2015","journal-title":"Haryana Vet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1111\/evj.13114","article-title":"An Outbreak of Strangles Associated with a Novel Genotype of Streptococcus Equi Subspecies Equi in Donkeys in China during 2018","volume":"51","author":"Dong","year":"2019","journal-title":"Equine Vet. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"645627","DOI":"10.3389\/fvets.2021.645627","article-title":"Nasopharyngeal Microbiomes in Donkeys Shedding Streptococcus Equi Subspecies Equi in Comparison to Healthy Donkeys","volume":"8","author":"Zhu","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_40","unstructured":"Allen, G.P., Kydd, J.H., Slater, J.D., and Smith, K.C. (1999). Infectious Diseases VIII, Proceedings of the Eighth International Conference, Dubai, United Arab Emirates, 23\u201326 March 1998, R & W Publications."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1016\/j.cveq.2019.08.009","article-title":"Respiratory Disorders of the Donkey","volume":"35","author":"Rickards","year":"2019","journal-title":"Vet. Clin. N. Am. Equine Pract."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"376","DOI":"10.1016\/j.jevs.2015.03.005","article-title":"Donkeys Are Different","volume":"35","author":"Burden","year":"2015","journal-title":"J. Equine Vet. Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1136\/inp.f5262","article-title":"Clinical Approach to the Dull Donkey","volume":"35","author":"Thiemann","year":"2013","journal-title":"Practice"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.prevetmed.2017.02.015","article-title":"Equine Infectious Anemia Prevalence in Feral Donkeys from Northeast Brazil","volume":"140","author":"Oliveira","year":"2017","journal-title":"Prev. Vet. Med."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Jerele, S., Davis, E., Mapes, S., Pusterla, N., Navas Gonz\u00e1lez, F.J., Iglesias Pastrana, C., Abdelfattah, E.M., and McLean, A. (2020). Survey of Serum Amyloid A and Bacterial and Viral Frequency Using QPCR Levels in Recently Captured Feral Donkeys from Death Valley National Park (California). Animals, 10.","DOI":"10.3390\/ani10061086"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1111\/evj.12775","article-title":"Hair Coat Properties of Donkeys, Mules and Horses in a Temperate Climate","volume":"50","author":"Osthaus","year":"2018","journal-title":"Equine Vet. J."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1629","DOI":"10.1128\/JVI.80.4.1629-1636.2006","article-title":"Respiratory Viruses Augment the Adhesion of Bacterial Pathogens to Respiratory Epithelium in a Viral Species- and Cell Type-Dependent Manner","volume":"80","author":"Avadhanula","year":"2006","journal-title":"J. Virol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1051\/vetres:2004025","article-title":"The Pathogenic Equine Streptococci","volume":"35","author":"Timoney","year":"2004","journal-title":"Vet. Res."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/12\/6\/750\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:40:26Z","timestamp":1760125226000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/12\/6\/750"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,5,23]]},"references-count":48,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,6]]}},"alternative-id":["pathogens12060750"],"URL":"https:\/\/doi.org\/10.3390\/pathogens12060750","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,5,23]]}}}