{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T09:26:35Z","timestamp":1783416395759,"version":"3.54.6"},"reference-count":25,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T00:00:00Z","timestamp":1783382400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000774","name":"DTRA","doi-asserted-by":"publisher","award":["HDTRA11710050"],"award-info":[{"award-number":["HDTRA11710050"]}],"id":[{"id":"10.13039\/100000774","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Anthrax is an endemic and undercharacterized zoonotic disease in the Middle East, including Jordan. Between 2018 and 2020, we conducted a comprehensive investigation of 13 confirmed anthrax outbreaks in Jordan, analyzing 822 samples from animal farm environments, including carcasses, asymptomatic livestock, and abiotic environmental surfaces. All samples were tested by qPCR targeting the chromosomal marker (ba177) and the plasmid marker pXO1 (pag). Among carcass samples, 75\/195 (38.5%) were ba177-positive, of which 81% harbored the pXO1. Of specimens from live-animals and from environmental surfaces, 218\/627 (35%) were positive by qPCR, likely reflecting environmental contamination during active outbreak periods. Serological analysis using anti-protective antigen (PA) ELISA revealed a high seroprevalence of 53% (75\/141) among asymptomatic animals, indicating widespread sub-clinical exposure to B. anthracis antigens and previously undocumented endemicity. An integrated approach combining qPCR with ELISA demonstrated that 11% of seropositive animals with paired swab testing also yielded swab samples that were positive by qPCR, suggesting environment-to-host transition. Molecular strain typing using canonical SNP (canSNP) analysis identified the rare sublineage C.USA.A1055 within lineage C.Br.A1005 across two distinct outbreaks, suggesting the environmental persistence of this endemic lineage. Overall, our findings provide the first systematic molecular and serological surveillance baseline data for Jordan, demonstrating a complex genomic persistence and subclinical exposure landscape. This study suggests the need for enhanced surveillance strategies under the One Health framework to mitigate the risk of anthrax endemicity.<\/jats:p>","DOI":"10.3390\/microorganisms14071483","type":"journal-article","created":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T08:04:49Z","timestamp":1783411489000},"page":"1483","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Molecular Characterization, SNP-Based Strain Profiling, and Seroprevalence of Bacillus anthracis in Ruminants in Jordan"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5142-6699","authenticated-orcid":false,"given":"Amin A.","family":"Aqel","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Medicine, Mutah University, Al-Karak 61710, Jordan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3658-5675","authenticated-orcid":false,"given":"Mohammad Abu","family":"Lubad","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Medicine, Mutah University, Al-Karak 61710, Jordan"},{"name":"Faculty of Medicine, Aqaba Medical Sciences University, Aqaba 77110, Jordan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8617-2476","authenticated-orcid":false,"given":"Hamed","family":"Alzoubi","sequence":"additional","affiliation":[{"name":"Department of Pathology and Microbiology, Faculty of Medicine, Jordan University of Science and Technology, Irbid 22110, Jordan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-2970-8188","authenticated-orcid":false,"given":"Daniel S.","family":"Schabacker","sequence":"additional","affiliation":[{"name":"Argonne National Laboratory, Lemont, IL 60439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Forrester","sequence":"additional","affiliation":[{"name":"Argonne National Laboratory, Lemont, IL 60439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scott","family":"Schlueter","sequence":"additional","affiliation":[{"name":"Argonne National Laboratory, Lemont, IL 60439, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Khemmani","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Loyola University Chicago, Chicago, IL 60660, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4532-0545","authenticated-orcid":false,"given":"Alan J.","family":"Wolfe","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Loyola University Chicago, Chicago, IL 60660, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tahir","family":"Yaqub","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3447-5629","authenticated-orcid":false,"given":"Muhammad Waqar","family":"Aziz","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan"},{"name":"Department of Microbiology, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7065-5059","authenticated-orcid":false,"given":"Mohammed","family":"Alsbou","sequence":"additional","affiliation":[{"name":"Department of Pathological Sciences, College of Medicine, Ajman University, Ajman P.O. Box 346, United Arab Emirates"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2244-4406","authenticated-orcid":false,"given":"Yasser","family":"Gaber","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef 62511, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,7]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (WHO) (2008). Anthrax in Humans and Animals, WHO. [4th ed.]. Available online: https:\/\/www.who.int\/publications\/b\/31250."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2236","DOI":"10.1001\/jama.287.17.2236","article-title":"Anthrax as a biological weapon, 2002: Updated recommendations for management","volume":"287","author":"Inglesby","year":"2002","journal-title":"JAMA"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1164\/rccm.201102-0209CI","article-title":"Anthrax infection","volume":"184","author":"Sweeney","year":"2011","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Timofeev, V., Bakhteeva, I., Khlopova, K., Mironova, R., Titareva, G., Goncharova, Y., Solomentsev, V., Kravchenko, T., Dyatlov, I., and Vergnaud, G. (2023). New Research on the Bacillus anthracis Genetic Diversity in Siberia. Pathogens, 12.","DOI":"10.3390\/pathogens12101257"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"920","DOI":"10.36899\/JAPS.2023.4.0685","article-title":"Endemic Seroprevalence and Farmer Control Measures of Anthrax among Ruminate Animals in Pakistan","volume":"33","author":"Sardar","year":"2023","journal-title":"J. Anim. Plant Sci."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Sardar, N., Aziz, M.W., Mukhtar, N., Yaqub, T., Anjum, A.A., Javed, M., Ashraf, M.A., Tanvir, R., Wolfe, A.J., and Schabacker, D.S. (2023). One Health Assessment of Bacillus anthracis Incidence and Detection in Anthrax-Endemic Areas of Pakistan. Microorganisms, 11.","DOI":"10.3390\/microorganisms11102462"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1016\/j.ijid.2020.07.083","article-title":"Palpebral anthrax, a rare though important condition in villagers: A case report and literature review","volume":"99","author":"Eshraghi","year":"2020","journal-title":"Int. J. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"196","DOI":"10.4274\/balkanmedj.galenos.2019.2018.12.19","article-title":"Cutaneous Anthrax Outbreak in the Trakya Region of Turkey","volume":"36","author":"Kuloglu","year":"2019","journal-title":"Balk. Med. J."},{"key":"ref_9","first-page":"269","article-title":"Lessons to be learned from recent biosafety incidents in the United States","volume":"17","author":"Weiss","year":"2015","journal-title":"Isr. Med. Assoc. J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e187428582205250","DOI":"10.2174\/18742858-v16-e2205250","article-title":"Cutaneous Anthrax in a Farmer Man: A Case Report","volume":"16","author":"Mahmoudi","year":"2022","journal-title":"Open Microbiol. J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1475","DOI":"10.3906\/sag-1504-41","article-title":"Evaluation of cutaneous anthrax cases during an outbreak in the east region of Turkey","volume":"46","year":"2016","journal-title":"Turk. J. Med. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Ochai, S.O., Hassim, A., Dekker, E.H., Magome, T., Lekota, K.E., Makgabo, S.M., de Klerk-Loris, L.-M., van Schalkwyk, L.O., Kamath, P.L., and Turner, W.C. (2024). Comparing microbiological and molecular diagnostic tools for the surveillance of anthrax. PLoS Negl. Trop. Dis., 18.","DOI":"10.1101\/2024.04.02.24305203"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Ganz, H.H., Turner, W.C., Brodie, E.L., Kusters, M., Shi, Y., Sibanda, H., Torok, T., and Getz, W.M. (2014). Interactions between Bacillus anthracis and plants may promote anthrax transmission. PLoS Negl. Trop. Dis., 8.","DOI":"10.1371\/journal.pntd.0002903"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"896","DOI":"10.1128\/JCM.41.2.896-899.2003","article-title":"PCR assay to detect Bacillus anthracis spores in heat-treated specimens","volume":"41","author":"Fasanella","year":"2003","journal-title":"J. Clin. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Van Ert, M.N., Easterday, W.R., Huynh, L.Y., Okinaka, R.T., Hugh-Jones, M.E., Ravel, J., Zanecki, S.R., Pearson, T., Simonson, T.S., and U\u2019REn, J.M. (2007). Global genetic population structure of Bacillus anthracis. PLoS ONE, 2.","DOI":"10.1371\/journal.pone.0000461"},{"key":"ref_16","first-page":"295","article-title":"The ecology of anthrax spores: Tough but not invincible","volume":"36","author":"Dragon","year":"1995","journal-title":"Can. Vet. J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"104029","DOI":"10.1016\/j.resmic.2023.104029","article-title":"The persistence of time: The lifespan of Bacillus anthracis spores in environmental reservoirs","volume":"174","author":"Barandongo","year":"2023","journal-title":"Res. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Ogunleye, S.C., Olorunshola, M.M., Fasina, K.A., Aborode, A.T., Akinsulie, O.C., Amoo, A., Olatoye, B.J., Bakare, A., Lawal, M.A., and Adekanye, O. (2024). Anthrax outbreak: Exploring its biological agents and public health implications. Front. Trop. Dis., 4.","DOI":"10.3389\/fitd.2023.1297896"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1103","DOI":"10.4269\/ajtmh.2007.77.1103","article-title":"Modeling the geographic distribution of Bacillus anthracis, the causative agent of anthrax disease, for the contiguous United States using predictive ecologic niche modeling","volume":"77","author":"Blackburn","year":"2007","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Aminu, O.R., Lembo, T., Zadoks, R.N., Biek, R., Lewis, S., Kiwelu, I., Mmbaga, B.T., Mshanga, D., Shirima, G., and Denwood, M. (2020). Practical and effective diagnosis of animal anthrax in endemic low-resource settings. PLoS Negl. Trop. Dis., 14.","DOI":"10.1101\/2020.03.24.004382"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1586\/14760584.2015.981533","article-title":"Host immunity to Bacillus anthracis lethal factor and other immunogens: Implications for vaccine design","volume":"14","author":"Altmann","year":"2015","journal-title":"Expert Rev. Vaccines"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1128\/9781555819323.ch14","article-title":"Animal models for the pathogenesis, treatment, and prevention of infection by Bacillus anthracis","volume":"3","author":"Welkos","year":"2016","journal-title":"Bact. Spore Mol. Syst."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"101095","DOI":"10.1016\/j.padiff.2025.101095","article-title":"A Study of the anthrax transmission model in herbivorous animals involving vaccination and harvesting","volume":"13","author":"Triska","year":"2025","journal-title":"Partial. Differ. Equ. Appl. Math."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"16912","DOI":"10.1038\/s41598-026-48459-6","article-title":"Hair and wool of ruminants and canines as environmental vectors of Bacillus anthracis and potential human exposure in anthrax-endemic regions","volume":"16","author":"Aziz","year":"2026","journal-title":"Sci. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.1007\/s11250-015-0882-8","article-title":"Molecular characterization of the circulating Bacillus anthracis in Jordan","volume":"47","author":"Aqel","year":"2015","journal-title":"Trop. Anim. Health Prod."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/7\/1483\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T09:06:23Z","timestamp":1783415183000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/7\/1483"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,7]]},"references-count":25,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["microorganisms14071483"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14071483","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,7]]}}}