{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T03:58:38Z","timestamp":1790135918365,"version":"4.0.1"},"reference-count":35,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,6,10]],"date-time":"2025-06-10T00:00:00Z","timestamp":1749513600000},"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>Probiotics are often advised to be taken separately from antibiotics due to their sensitivity to antibiotic activity and gastrointestinal (GI) stress. However, Clostridium butyricum spores, as next-generation probiotics, may withstand concurrent use with antibiotics due to their unique structural adaptations. This study systematically evaluated the survival rates and morphological changes of C. butyricum spores exposed to 10 clinically relevant antibiotics in simulated gastric\/intestinal fluids, exploring their feasibility for co-administration with antibiotics. Survival rates of C. butyricum spores were tested against 10 antibiotics across four classes (\u03b2-lactams, macrolides, aminoglycosides, and tetracyclines) in simulated GI fluids. Time\u2013kill curves analyzed spore survival over 0\u20134 h, while scanning electron microscopy (SEM) observed spore wall integrity and morphological changes under different conditions. The spore survival rates remained &gt;89% in intestinal fluid and &gt;60% in gastric fluid across all antibiotics. SEM revealed gastric acid and proteolysis increased spore wall fragmentation, reducing resistance, whereas the intestinal environment preserved spore integrity. This study was the first to demonstrate that C. butyricum spores can survive simultaneous exposure to antibiotics in the gastrointestinal tract, challenging traditional probiotic usage guidelines. The findings support their co-administration with antibiotics to simplify dosing regimens and improve medication adherence. Such an approach advances antimicrobial stewardship by optimizing therapeutic strategies for antibiotic\u2013probiotic combinations.<\/jats:p>","DOI":"10.3390\/microorganisms13061347","type":"journal-article","created":{"date-parts":[[2025,6,10]],"date-time":"2025-06-10T09:59:14Z","timestamp":1749549554000},"page":"1347","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Survival and Morphological Changes of Clostridium butyricum Spores Co-Exposed to Antibiotics and Simulated Gastrointestinal Fluids: Implications for Antibiotic Stewardship"],"prefix":"10.3390","volume":"13","author":[{"given":"Yi-Meng","family":"Yang","sequence":"first","affiliation":[{"name":"School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meng-Yue","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying-Ying","family":"Wu","sequence":"additional","affiliation":[{"name":"School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Hangzhou Grand Biologic Pharmaceutical Inc., Hangzhou 310030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4849-3660","authenticated-orcid":false,"given":"Yi-Xuan","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2019","DOI":"10.1007\/s10482-020-01474-7","article-title":"Human Gut Microbiota\/Microbiome in Health and Diseases: A Review","volume":"113","author":"Gomaa","year":"2020","journal-title":"Antonie Leeuwenhoek"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Ramirez, J., Guarner, F., Bustos Fernandez, L., Maruy, A., Sdepanian, V.L., and Cohen, H. (2020). Antibiotics as Major Disruptors of Gut Microbiota. Front. Cell. Infect. Microbiol., 10.","DOI":"10.3389\/fcimb.2020.572912"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1038\/s41564-018-0257-9","article-title":"Recovery of Gut Microbiota of Healthy Adults Following Antibiotic Exposure","volume":"3","author":"Palleja","year":"2018","journal-title":"Nat. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e09725","DOI":"10.1016\/j.heliyon.2022.e09725","article-title":"Probiotics, Their Prophylactic and Therapeutic Applications in Human Health Development: A Review of the Literature","volume":"8","author":"Tegegne","year":"2022","journal-title":"Heliyon"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1046\/j.1472-765x.2000.00764.x","article-title":"Antimicrobial Susceptibility of Bifidobacteria: Antimicrobial Susceptibility of Bifidobacteria","volume":"31","author":"Yazid","year":"2000","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"q1170","DOI":"10.1136\/bmj.q1170","article-title":"Antimicrobial Stewardship","volume":"385","author":"Powers","year":"2024","journal-title":"BMJ"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/j.jiac.2015.02.001","article-title":"Applications of the Pharmacokinetic\/Pharmacodynamic (PK\/PD) Analysis of Antimicrobial Agents","volume":"21","author":"Isla","year":"2015","journal-title":"J. Infect. Chemother."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.cmi.2015.10.014","article-title":"Clostridium butyricum: From Beneficial to a New Emerging Pathogen","volume":"22","author":"Cassir","year":"2016","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Ariyoshi, T., Hagihara, M., Takahashi, M., and Mikamo, H. (2022). Effect of Clostridium Butyricum on Gastrointestinal Infections. Biomedicines, 10.","DOI":"10.3390\/biomedicines10020483"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1907272","DOI":"10.1080\/19490976.2021.1907272","article-title":"Butyrate-Producing Human Gut Symbiont, Clostridium butyricum, and Its Role in Health and Disease","volume":"13","author":"Stoeva","year":"2021","journal-title":"Gut Microbes"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.4103\/jcrt.jcrt_1565_21","article-title":"The Relationship between Clostridium Butyricum and Colorectal Cancer","volume":"18","author":"Liu","year":"2022","journal-title":"J. Cancer Res. Ther."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Shin, Y., Han, S., Kwon, J., Ju, S., Choi, T., Kang, I., and Kim, S. (2023). Roles of Short-Chain Fatty Acids in Inflammatory Bowel Disease. Nutrients, 15.","DOI":"10.3390\/nu15204466"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1016\/j.chom.2013.05.013","article-title":"A Single Strain of Clostridium butyricum Induces Intestinal IL-10-Producing Macrophages to Suppress Acute Experimental Colitis in Mice","volume":"13","author":"Hayashi","year":"2013","journal-title":"Cell Host Microbe"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1007\/s00535-015-1084-x","article-title":"A Breakthrough in Probiotics: Clostridium Butyricum Regulates Gut Homeostasis and Anti-Inflammatory Response in Inflammatory Bowel Disease","volume":"50","author":"Kanai","year":"2015","journal-title":"J. Gastroenterol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.4049\/jimmunol.2000353","article-title":"The Butyrate-Producing Bacterium Clostridium Butyricum Suppresses Clostridioides difficile Infection via Neutrophil- and Antimicrobial Cytokine\u2013Dependent but GPR43\/109a-Independent Mechanisms","volume":"206","author":"Hayashi","year":"2021","journal-title":"J. Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"111755","DOI":"10.1016\/j.celrep.2022.111755","article-title":"Clostridium butyricum-Induced \u03c9-3 Fatty Acid 18-HEPE Elicits Anti-Influenza Virus Pneumonia Effects through Interferon-\u03bb Upregulation","volume":"41","author":"Hagihara","year":"2022","journal-title":"Cell Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/j.bbi.2023.11.016","article-title":"Probiotic Clostridium butyricum Ameliorates Cognitive Impairment in Obesity via the Microbiota-Gut-Brain Axis","volume":"115","author":"Zheng","year":"2024","journal-title":"Brain Behav. Immun."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Mun, C., Cai, J., Hu, X., Zhang, W., Zhang, N., and Cao, Y. (2022). Clostridium butyricum and Its Culture Supernatant Alleviate the Escherichia Coli-Induced Endometritis in Mice. Animals, 12.","DOI":"10.3390\/ani12192719"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.resmic.2014.12.001","article-title":"Updates on the Sporulation Process in Clostridium Species","volume":"166","author":"Talukdar","year":"2015","journal-title":"Res. Microbiol."},{"key":"ref_20","first-page":"TBS-0010-2012","article-title":"Physiology and Sporulation in Clostridium","volume":"2","year":"2014","journal-title":"Microbiol. Spectr."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"100772","DOI":"10.1016\/j.isci.2019.100772","article-title":"Clostridium butyricum Modulates the Microbiome to Protect Intestinal Barrier Function in Mice with Antibiotic-Induced Dysbiosis","volume":"23","author":"Hagihara","year":"2020","journal-title":"iScience"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1016\/j.jiac.2019.04.018","article-title":"Stability of Probiotics with Antibiotics via Gastric Tube by Simple Suspension Method: An in Vitro Study","volume":"25","author":"Mitsuboshi","year":"2019","journal-title":"J. Infect. Chemother."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"741590","DOI":"10.1016\/j.aquaculture.2024.741590","article-title":"Isolation and Characterization of a Strain Clostridium butyricum B3 from the Intestine of Pelteobagrus Fulvidraco and Its Potential Probiotic Roles","volume":"595","author":"Zhang","year":"2025","journal-title":"Aquaculture"},{"key":"ref_24","first-page":"195","article-title":"Research Progress on the Application of Clostridium butyricum in Shrimp Aquaculture","volume":"76","author":"Li","year":"2024","journal-title":"Isr. J. Aquac.-Bamidgeh"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1128\/AAC.00261-06","article-title":"Antibiotic Susceptibility Profile of Bifidobacteria as Affected by Oxgall, Acid, and Hydrogen Peroxide Stress","volume":"51","author":"Kheadr","year":"2007","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1046\/j.1442-200X.2003.01671.x","article-title":"Prevention of Antibiotic-associated Diarrhea in Children by Clostridium butyricum MIYAIRI","volume":"45","author":"Seki","year":"2003","journal-title":"Pediatr. Int."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Tomita, Y., Sakata, S., Imamura, K., Iyama, S., Jodai, T., Saruwatari, K., Hamada, S., Akaike, K., Anai, M., and Fukusima, K. (2023). Association of Clostridium Butyricum Therapy Using the Live Bacterial Product CBM588 with the Survival of Patients with Lung Cancer Receiving Chemoimmunotherapy Combinations. Cancers, 16.","DOI":"10.3390\/cancers16010047"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1139\/cjm-2016-0186","article-title":"Exploring the Relationship between Exposure to Technological and Gastrointestinal Stress and Probiotic Functional Properties of Lactobacilli and Bifidobacteria","volume":"62","author":"Amund","year":"2016","journal-title":"Can. J. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1007\/s10529-019-02729-8","article-title":"Autochthonous Lactic Acid Bacteria\u2014Presentation of Potential Probiotics Application","volume":"41","year":"2019","journal-title":"Biotechnol. Lett."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e00170-21","DOI":"10.1128\/mmbr.00170-21","article-title":"Stress Response in Bifidobacteria","volume":"86","author":"Zeidan","year":"2022","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"665","DOI":"10.3201\/eid3004.231633","article-title":"Clostridium Butyricum Bacteremia Associated with Probiotic Use, Japan","volume":"30","author":"Sada","year":"2024","journal-title":"Emerg. Infect. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2172666","DOI":"10.1080\/19490976.2023.2172666","article-title":"Neonatal Necrotizing Enterocolitis: Clostridium butyricum and Clostridium neonatale Fermentation Metabolism and Enteropathogenicity","volume":"15","author":"Ferraris","year":"2023","journal-title":"Gut Microbes"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Ashammakhi, N., Nasiri, R., Barros, N.R.D., Tebon, P., Thakor, J., Goudie, M., Shamloo, A., Martin, M.G., and Khademhosseini, A. (2020). Gut-on-a-Chip: Current Progress and Future Opportunities. Biomaterials, 255.","DOI":"10.1016\/j.biomaterials.2020.120196"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4130","DOI":"10.1038\/s41467-023-39429-x","article-title":"Decoding a Cryptic Mechanism of Metronidazole Resistance among Globally Disseminated Fluoroquinolone-Resistant Clostridioides Difficile","volume":"14","author":"Olaitan","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e00997-24","DOI":"10.1128\/aac.00997-24","article-title":"Insights into Antibiotic Resistance Promoted by Quinolone Exposure","volume":"69","author":"Bush","year":"2025","journal-title":"Antimicrob. Agents Chemother."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/6\/1347\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:49:34Z","timestamp":1760032174000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/6\/1347"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,10]]},"references-count":35,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["microorganisms13061347"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13061347","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,10]]}}}