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ORIGINAL RESEARCH article

Front. Microbiol.

Sec. Food Microbiology

This article is part of the Research TopicRapid Pathogen Detection in Food Supply ChainsView all 4 articles

Detection and characterization of Clostridium botulinum isolated from powdered infant formula

Provisionally accepted
Cesar NadalaCesar Nadala1Eni ThemeliEni Themeli1Fereidoun ForghaniFereidoun Forghani1*Youngsil HaYoungsil Ha1Sukkyun HanSukkyun Han1Anna ShapovalovaAnna Shapovalova1John RoachJohn Roach1Seong Hong KimSeong Hong Kim1Jennifer LM ThorsonJennifer LM Thorson1Joo-Young LeeJoo-Young Lee1Matthew SpearMatthew Spear1Benjamin BuhrmanBenjamin Buhrman1Kevin ChiuKevin Chiu1Niall MullaneNiall Mullane2Mansour SamadpourMansour Samadpour1*
  • 1IEH Laboratories and Consulting Group, Lake Forest Park, United States
  • 2ByHeart, 131 Varick St, 11th Floor, New York, NY 10013, United States

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Introduction: As a part of an ongoing investigation of the 2025 multistate outbreak of infant botulism linked to consumption of powdered infant formula (PIF), various samples – including unopened containers of PIF and base powder (bulk PIF before packaging) – were analyzed for the presence of Clostridium botulinum. Methods: Samples were screened using a tiered analytical approach: anaerobic enrichment followed by real-time PCR targeting botulinum neurotoxin-associated genes, confirmatory PCR, amplicon sequencing, colony isolation, and whole-genome sequencing (WGS). C. botulinum was detected in both the finished product and base powder. Results: Genomic analysis revealed genetic identity between isolates from one finished product lot and a base powder, while an isolate from another lot was distinct. Importantly, detection occurred in samples with non-detectable sulfite-reducing clostridia (SRC). Conclusion: Results of this study demonstrate that indicator-based screening (such as SRC enumeration) even if it had been in place before the outbreak, would not have prevented its occurrance. Positive linkage of an infant botulism outbreak to PIF should compel manufacturers to recognize C. botulinum as a hazard reasonably likely to occur in certain ingredients, necessitating the design and implementation of specific preventative controls.

Keywords: Clostridium botulinum, Food Safety, Infant botulism, outbreak investigation, Powdered infant formula (PIF), sulfite-reducingclostridia (SRC), Whole-genome sequencing (WGS)

Received: 31 Jan 2026; Accepted: 16 Feb 2026.

Copyright: © 2026 Nadala, Themeli, Forghani, Ha, Han, Shapovalova, Roach, Kim, Thorson, Lee, Spear, Buhrman, Chiu, Mullane and Samadpour. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Fereidoun Forghani
Mansour Samadpour

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Supplementary Material