Clinical and In Vitro Safety of Heyndrickxia coagulans AO 1167B: A Double-Blind, Placebo-Controlled Trial

Gissel García¹, Josanne Soto², Antonio Díaz³, Jesús Barreto⁴, Carmen Soto⁵, Ana Beatriz Pérez⁶, Suselys Boffill⁴, Ángela Gutiérrez³, Raúl de Jesús Cano⁷

Affiliations

¹ Pathology Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
² Clinical Laboratory Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
³ Statistical Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
⁴ Nutrition Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
⁵ Biochemistry Department, Biology Faculty, Havana University Cuba, Calle 25 Esquina J Vedado, La Habana 10200, Cuba.
⁶ Cellular Immunology Laboratory, Virology Department, Tropical Medicine Institute "Pedro Kourí", Autopista Novia del Medio Día Km 6 ½, La Habana 11400, Cuba.
⁷ Biological Sciences Department, California Polytechnic State University, San Luis Obispo, CA 93407, USA.

PMID: 39770785
PMCID: PMC11677179
DOI: 10.3390/microorganisms12122584

Clinical and In Vitro Safety of Heyndrickxia coagulans AO 1167B: A Double-Blind, Placebo-Controlled Trial

Gissel García et al. Microorganisms. 2024.

. 2024 Dec 13;12(12):2584.

doi: 10.3390/microorganisms12122584.

Authors

Gissel García¹, Josanne Soto², Antonio Díaz³, Jesús Barreto⁴, Carmen Soto⁵, Ana Beatriz Pérez⁶, Suselys Boffill⁴, Ángela Gutiérrez³, Raúl de Jesús Cano⁷

Affiliations

¹ Pathology Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
² Clinical Laboratory Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
³ Statistical Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
⁴ Nutrition Department, Clinical Hospital "Hermanos Ameijeiras", Calle San Lázaro No 701, Esq. a Belascoaín, La Habana 10400, Cuba.
⁵ Biochemistry Department, Biology Faculty, Havana University Cuba, Calle 25 Esquina J Vedado, La Habana 10200, Cuba.
⁶ Cellular Immunology Laboratory, Virology Department, Tropical Medicine Institute "Pedro Kourí", Autopista Novia del Medio Día Km 6 ½, La Habana 11400, Cuba.
⁷ Biological Sciences Department, California Polytechnic State University, San Luis Obispo, CA 93407, USA.

PMID: 39770785
PMCID: PMC11677179
DOI: 10.3390/microorganisms12122584

Abstract

(1) Background: Heyndrickxia coagulans, a lactic acid-producing bacterium, displays characteristics of both Lactobacillus and Bacillus genera. Clinical evidence suggests its potential health benefits. This study evaluated the safety of H. coagulans AO1167B as a candidate probiotic supplement. (2) Methods: Strain identification was confirmed through morphological, cultural, and genomic analyses, including 16S RNA and whole genome sequencing to assess antimicrobial resistance and virulence factors. Phenotypic tests, such as disk diffusion for antimicrobial resistance, and safety assays for cytotoxicity and hemolytic activity, were conducted. In a phase I, double-blind, placebo-controlled clinical trial, healthy adults were randomized into H. coagulans AO1167B and placebo groups for 60 days. Daily capsule consumption was monitored through clinical and hematological evaluations, adverse event tracking, and health surveys. (3) Results: The genome of H. coagulans AO1167B revealed no concerning features. Disk diffusion tests showed no antimicrobial resistance. The strain exhibited no cytotoxic or hemolytic activity, indicating in vitro safety. No significant differences in clinical or hematological parameters were observed between groups. The most common adverse event, gas, diminished over time. (4) Conclusions: H. coagulans AO1167B demonstrates a suitable safety profile, genetic stability, and probiotic potential for gastrointestinal health, justifying further clinical research.

Keywords: Heyndrickxia coagulans; clinical trial; humans; probiotic; randomized; safety.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

**Figure 1**
CONSORT Flow Diagram of Recruitment and Retention Throughout the Study [35].

**Figure 2**
Whole genome phylogenetic analysis of *H. coagulans* AO1167B and related strains. The Neighbor-Joining tree represents the phylogenetic relationship of *H. coagulans* AO1167B with closely related strains known for probiotic properties. The tree was constructed based on whole genome sequences, with bootstrap values indicated at each node to demonstrate clustering robustness. The scale bar represents the number of substitutions per site. The tree was visualized using FigTree v1.4.4 (http://tree.bio.ed.ac.uk/software/figtree/ accessed on 4 October 2024).

**Figure 3**
Cytotoxicity of H. coagulans AO1167B. (A) Hemolysis induced by H. coagulans AO1167B on 1% of human erythrocytes solution for 1 h. The hemolytic activity was expressed as % of hemoglobin (hemolysis %). (B) Percent of viability induced by H. coagulans AO1167B in Vero cells. The cytotoxic activity was expressed as % of viability. Error bars represent the standard error (SE) of the mean, calculated from three independent replicates.

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Clinical and In Vitro Safety of Heyndrickxia coagulans AO 1167B: A Double-Blind, Placebo-Controlled Trial

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Clinical and In Vitro Safety of Heyndrickxia coagulans AO 1167B: A Double-Blind, Placebo-Controlled Trial

Authors

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Abstract

Conflict of interest statement

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