{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T20:29:39Z","timestamp":1787862579824,"version":"build-2784847793"},"reference-count":42,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,7,15]],"date-time":"2025-07-15T00:00:00Z","timestamp":1752537600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Botulism is a severe muscle paralysis disease mediated by the botulinum toxin. Here, we reported a foodborne botulism case caused by Clostridium botulinum subtype A5(b3) from self-packaged vacuum spicy rabbit heads. Treatment for this case was delayed due to misdiagnosis and insufficient diagnostic capacity in three hospitals, which resulted in progressive clinical deterioration, and eventually, the patient was transferred to Shandong Public Health Clinical Center for specialized therapy. The case was suspected as foodborne botulism by the Qilu Medical-Prevention Innovation Integration pathway and multi-disciplinary consultation. An epidemiological investigation and laboratory confirmation revealed that the botulinum neurotoxin originated from vacuum-packaged spicy rabbit heads distributed via interprovincial cold chain logistics. After treatment with botulism antiserum, the patient\u2019s condition significantly improved, and they were discharged after recovery. We revealed that this foodborne botulism outbreak was caused by the Clostridium botulinum A5(b3) subtype from food by whole-genome sequencing and SNP typing. All the strains belonged to Group I carrying the botulinum neurotoxin gene classified as the ha cluster. Toxin A was confirmed by MBA and other methods, while toxin B was non-functional due to the truncated bont\/B gene. Other virulence genes and antibiotic resistance genes were also detected. Our findings indicate that self-packaged vacuum meat products represent an emerging risk factor for botulism transmission when stored improperly. Importantly, the recurrent misdiagnosis in this case underscored the urgent need to enhance the training of healthcare professionals in medical institutions to improve the diagnostic accuracy and clinical management of botulism.<\/jats:p>","DOI":"10.3390\/microorganisms13071662","type":"journal-article","created":{"date-parts":[[2025,7,15]],"date-time":"2025-07-15T08:04:41Z","timestamp":1752566681000},"page":"1662","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Foodborne Botulism Caused by Clostridium botulinum Subtype A5(b3) by Self-Packaged Vacuum Spicy Rabbit Heads"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3729-777X","authenticated-orcid":false,"given":"Wen","family":"Cui","sequence":"first","affiliation":[{"name":"Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-1125-2808","authenticated-orcid":false,"given":"Chuanmin","family":"Ma","sequence":"additional","affiliation":[{"name":"Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5955-0109","authenticated-orcid":false,"given":"Ming","family":"Liu","sequence":"additional","affiliation":[{"name":"Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Li","sequence":"additional","affiliation":[{"name":"Huaiyin District Center for Disease Control and Prevention, Huaiyin District, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Zhou","sequence":"additional","affiliation":[{"name":"Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuwen","family":"Shi","sequence":"additional","affiliation":[{"name":"Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuefang","family":"Xu","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases, National Institute for Communicable Diseases Control and Prevention, Chinese Center for Disease Control and Prevention, Changping, Beijing 102206, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui","family":"Liu","sequence":"additional","affiliation":[{"name":"Jinan Center for Disease Control and Prevention Affiliated to Shandong University, Jinan 250021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rasetti-Escargueil, C., Lemichez, E., and Popoff, M.R. (2019). Public health risk associated with botulism as foodborne zoonoses. Toxins, 12.","DOI":"10.3390\/toxins12010017"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Centurioni, D.A., Egan, C.T., and Perry, M.J. (2022). Current developments in diagnostic assays for laboratory confirmation and investigation of botulism. J. Clin. Microbiol., 60.","DOI":"10.1128\/jcm.00139-20"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"S2","DOI":"10.1002\/mds.20003","article-title":"Historical notes on botulism, Clostridium botulinum, botulinum toxin, and the idea of the therapeutic use of the toxin","volume":"19","author":"Erbguth","year":"2004","journal-title":"Mov. Disord."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2205251","DOI":"10.1080\/21505594.2023.2205251","article-title":"Pathogenicity and virulence of Clostridium botulinum","volume":"14","author":"Rawson","year":"2023","journal-title":"Virulence"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bowe, B.K., Wentz, T.G., Gregg, B.M., Tepp, W.H., Schill, K.M., Sharma, S., and Pellett, S. (2022). Genomic Diversity, Competition, and toxin production by group I and II Clostridium botulinum strains used in food challenge studies. Microorganisms, 10.","DOI":"10.3390\/microorganisms10101895"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1016\/j.resmic.2014.10.010","article-title":"Genomes, neurotoxins and biology of Clostridium botulinum Group I and Group II","volume":"166","author":"Carter","year":"2015","journal-title":"Res. Microbiol."},{"key":"ref_7","first-page":"1","article-title":"Genetic diversity within Clostridium botulinum serotypes, botulinum neurotoxin gene clusters and toxin subtypes","volume":"364","author":"Hill","year":"2013","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1038\/nrmicro3295","article-title":"Botulinum neurotoxins: Genetic, structural and mechanistic insights","volume":"12","author":"Rossetto","year":"2014","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.toxicon.2015.09.009","article-title":"The long journey of botulinum neurotoxins into the synapse","volume":"107","author":"Rummel","year":"2015","journal-title":"Toxicon Off. J. Int. Soc. Toxinol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Peck, M.W., Smith, T.J., Anniballi, F., Austin, J.W., Bano, L., Bradshaw, M., Cuervo, P., Cheng, L.W., Derman, Y., and Dorner, B.G. (2017). Historical perspectives and guidelines for botulinum neurotoxin subtype nomenclature. Toxins, 9.","DOI":"10.3390\/toxins9010038"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/978-1-0716-0430-4_20","article-title":"Functional analysis of botulinum hemagglutinin (HA)","volume":"2132","author":"Matsumura","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2125","DOI":"10.1128\/AEM.03934-13","article-title":"Whole-genome single-nucleotide-polymorphism analysis for discrimination of Clostridium botulinum group I strains","volume":"80","author":"Timme","year":"2014","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1167","DOI":"10.1086\/444507","article-title":"Botulism","volume":"41","author":"Sobel","year":"2005","journal-title":"Clin. Infect. Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/S0065-2911(09)05503-9","article-title":"Biology and genomic analysis of Clostridium botulinum","volume":"55","author":"Peck","year":"2009","journal-title":"Adv. Microb. Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Le Bouquin, S., Lucas, C., Souillard, R., Le Mar\u00e9chal, C., Petit, K., Kooh, P., Jourdan-Da Silva, N., Meurens, F., Guillier, L., and Mazuet, C. (2022). Human and animal botulism surveillance in France from 2008 to 2019. Front. Public Health, 24.","DOI":"10.3389\/fpubh.2022.1003917"},{"key":"ref_16","first-page":"1730","article-title":"Foodborne Botulism, Canada, 2006\u20132021","volume":"29","author":"Harris","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"ref_17","first-page":"788","article-title":"Epidemiological analysis of foodborne botulism outbreaks-China, 2004\u20132020","volume":"4","author":"Li","year":"2022","journal-title":"China CDC Wkly."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Ma, X., Li, K., Li, F., Su, J., Meng, W., Sun, Y., Sun, H., Sun, J., Yuan, Y., and Lin, Y. (2022). Tracing Foodborne botulism events caused by Clostridium botulinum in Xinjiang province, China, using a core genome sequence typing scheme. Microbiol. Spectr., 10.","DOI":"10.1128\/spectrum.01164-22"},{"key":"ref_19","unstructured":"(2017). Microbiological Examination of Food Hygiene\u2014Examination of Clostridium Botulinum and Botulinus Toxin. Standard No. GB 4789.12-2016."},{"key":"ref_20","first-page":"235","article-title":"Rapid quantitative detection of botulinum toxin type A by using lanthanide fluorescence technology","volume":"43","author":"Wang","year":"2020","journal-title":"Chin. Front. Health Quar."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3935","DOI":"10.1128\/AEM.71.7.3935-3941.2005","article-title":"Evaluation of lateral-flow Clostridium botulinum neurotoxin detection kits for food analysis","volume":"71","author":"Sharma","year":"2005","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"i884","DOI":"10.1093\/bioinformatics\/bty560","article-title":"Fastp: An ultra-fast all-in-one FASTQ preprocessor","volume":"34","author":"Chen","year":"2018","journal-title":"Bioinformatics"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"760","DOI":"10.1016\/j.jemermed.2020.12.006","article-title":"An outbreak of botulinum types A, B, and E associated with vacuum-packaged salted fish and ham","volume":"60","author":"Min","year":"2021","journal-title":"J. Emerg. Med."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Pernu, N., Keto-Timonen, R., Lindstr\u00f6m, M., and Korkeala, H. (2020). High prevalence of Clostridium botulinum in vegetarian sausages. Food Microbiol., 91.","DOI":"10.1016\/j.fm.2020.103512"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1017\/S0950268800066991","article-title":"A food-poisoning incident caused by Clostridium botulinum toxin A in Japan","volume":"99","author":"Otofuji","year":"1987","journal-title":"Epidemiol. Infect."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lonati, D., Schicchi, A., Crevani, M., Buscaglia, E., Scaravaggi, G., Maida, F., Cirronis, M., Petrolini, V.M., and Locatelli, C.A. (2020). Foodborne botulism: Clinical diagnosis and medical treatment. Toxins, 12.","DOI":"10.3390\/toxins12080509"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1082","DOI":"10.1101\/gr.6282807","article-title":"Genome sequence of a proteolytic (Group I) Clostridium botulinum strain Hall A and comparative analysis of the clostridial genomes","volume":"17","author":"Sebaihia","year":"2007","journal-title":"Genome Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Brunt, J., van Vliet, A.H., Carter, A.T., Stringer, S.C., Amar, C., Grant, K.A., Godbole, G., and Peck, M.W. (2020). Diversity of the genomes and neurotoxins of strains of Clostridium botulinum group I and Clostridium sporogenes associated with foodborne, infant and wound botulism. Toxins, 12.","DOI":"10.3390\/toxins12090586"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"10786","DOI":"10.1074\/jbc.271.18.10786","article-title":"Genetic characterization of Clostridium botulinum type A containing silent type B neurotoxin gene sequences","volume":"271","author":"Hutson","year":"1996","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Dover, N., Barash, J.R., Hill, K.K., Davenport, K.W., Teshima, H., Xie, G., and Arnon, S.S. (2013). Clostridium botulinum strain Af84 contains three neurotoxin gene clusters: Bont\/A2, bont\/F4 and bont\/F5. PLoS ONE, 8.","DOI":"10.1371\/annotation\/b482f80f-c5b6-4b9c-8e9b-8b7139dc37f1"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Nawrocki, E.M., Bradshaw, M., and Johnson, E.A. (2018). Botulinum neurotoxin-encoding plasmids can be conjugatively transferred to diverse clostridial strains. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-21342-9"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Benoit, R.M. (2018). Botulinum neurotoxin diversity from a gene-centered view. Toxins, 10.","DOI":"10.3390\/toxins10080310"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1904","DOI":"10.1128\/AEM.02422-10","article-title":"Novel structural elements within the nonproteolytic Clostridium botulinum type F toxin gene cluster","volume":"77","author":"Dover","year":"2011","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"213","DOI":"10.4161\/mge.1.3.17617","article-title":"Horizontal gene transfer of toxin genes in Clostridium botulinum: Involvement of mobile elements and plasmids","volume":"1","author":"Skarin","year":"2011","journal-title":"Mob. Genet. Elements."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1012","DOI":"10.1128\/JCM.01774-09","article-title":"Further characterization of proteolytic Clostridium botulinum type A5 reveals that neurotoxin formation is unaffected by loss of the cntR (botR) promoter sigma factor binding site","volume":"48","author":"Carter","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.fm.2010.03.005","article-title":"Clostridium botulinum in the post-genomic era","volume":"28","author":"Michael","year":"2011","journal-title":"Food Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2351","DOI":"10.1128\/JB.00072-11","article-title":"Complete genome sequence of the proteolytic Clostridium botulinum type A5 (B3) strain H04402065","volume":"193","author":"Carter","year":"2011","journal-title":"J. Bacteriol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2349","DOI":"10.1128\/JCM.00799-09","article-title":"Novel Clostridium botulinum toxin gene arrangement with subtype A5 and partial subtype B3 botulinum neurotoxin genes","volume":"47","author":"Dover","year":"2009","journal-title":"J. Clin. Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1093\/gbe\/evw101","article-title":"Diversity of group I and II Clostridium botulinum strains from France including recently identified subtypes","volume":"8","author":"Mazuet","year":"2016","journal-title":"Genome Biol. Evol."},{"key":"ref_40","first-page":"43","article-title":"Research progress of cfr gene mediated bacteria resistance","volume":"21","author":"Xuan","year":"2019","journal-title":"J. Agric. Sci. Technol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.cofs.2016.09.006","article-title":"Impact of Clostridium botulinum genomic diversity on food safety","volume":"10","author":"Peck","year":"2016","journal-title":"Curr. Opin. Food Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15585\/mmwr.rr7002a1","article-title":"Clinical guidelines for diagnosis and treatment of botulism","volume":"70","author":"Rao","year":"2021","journal-title":"MMWR Recomm. Rep."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/7\/1662\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:09:57Z","timestamp":1760033397000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/7\/1662"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,15]]},"references-count":42,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["microorganisms13071662"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13071662","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,15]]}}}