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Use of Fecal Slurry Cultures to Study In Vitro Effects of Bacteriocins on the Gut Bacterial Populations of Infants

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Abstract

Bacteriocins are antimicrobial peptides produced by bacteria to compete with other bacteria for nutrients and ecological niches. The antimicrobial effect of these peptides on the bacterial populations in the gut is likely dynamic as the survival of the microbes in this environment depends on both competition and cooperation. In this study, we evaluated four different bacteriocins from lactic acid bacteria (LAB): nisin, enterocin A (EntA), enterocin K1 (EntK1), and garvicin ML (GarML), which have different inhibition spectra and physicochemical properties. The bacteriocins were tested in vitro using fecal slurry batch cultures from infants. The abundances of some bacterial populations in the cultures were determined using quantitative PCR (qPCR) and the metabolic activity of the gut microbiota was assessed by measuring the production of short-chain fatty acids (SCFA) using gas chromatography. The effects of the bacteriocins correlated well with their antimicrobial spectra and the administered concentrations. Nisin and GarML, with broad antimicrobial spectra, shifted the abundance of several intestinal bacterial groups, while EntA and EntK1, with relative narrower inhibition spectra, showed no or little effect. Moreover, the results from the SCFA analysis were consistent with changes obtained in the bacterial composition. In particular, a reduction in acetate concentration was observed in the samples with low abundance of Bifidobacterium, which is a well-known acetate producer. The variability imposed on the intestinal bacterial populations by the different bacteriocins tested suggests that this type of antimicrobials have great potential to modulate the gut microbiota for medical purposes.

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Acknowledgements

We thank Lars-Gustav Snipen for his advices on statistics.

Funding

OCOU was supported by a strategic scholarship program to food science research, from Norwegian University of Life Sciences (NMBU) (project 1205051025), and she has currently a postdoctoral position funded by Research Council of Norway. NS was the recipient of a “Clarín” postdoctoral contract (Marie Curie European CoFund Program) cofounded by the “Plan Regional de Investigación” of Principado de Asturias and she is currently the recipient of a postdoctoral contract awarded by the Fundación para la Investigación Biosanitaria de Asturias (FINBA). Travels and stays in Spain and Norway for this study were supported by EEA Coordinated Mobility of Researchers NILS Science and Sustainability Project 017-CM-01-2013.

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Authors and Affiliations

  1. Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), Ås, Norway

    Özgün C. O. Umu, Marije Oostindjer, Kirill V. Ovchinnikov & Dzung B. Diep

  2. Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain

    Miguel Gueimonde, Clara G. de los Reyes-Gavilán & Nuria Salazar

  3. Sección Departamental de Nutrición y Ciencia de los Alimentos (Nutrición, Bromatología, Higiene y Seguridad Alimentaria), Facultad de Veterinaria, Universidad Complutense de Madrid (UCM), Madrid, Spain

    Sara Arbulu & Pablo E. Hernández

  4. Department of Technology and Biotechnology of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain

    Beatriz Martínez

  5. Diet, Microbiota, and Health Group, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), 33011, Avda de Roma s/n, Oviedo, Spain

    Miguel Gueimonde, Clara G. de los Reyes-Gavilán & Nuria Salazar

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  1. Özgün C. O. Umu
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  2. Miguel Gueimonde
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Correspondence to Özgün C. O. Umu.

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Ethical Approval

The study was approved by the Regional Ethical Committee of Asturias Public Health Service (SESPA).

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Informed consent was obtained from the parents of all individual participant infants included in the study.

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Fig. S1

Molar concentrations of butyrate and isobutyrate in the fecal slurry cultures. Panels represents butyrate (A) and isobutyrate (B). Time points were shown with 0, reference time point zero after stabilization (shown in pink); 24 h, after 24-hour incubation; and 48 h, after 48-hour incubation. Concentrations of the bacteriocins in the treatments were indicated with 0, no bacteriocin addition; 10, 10 μg/mL of bacteriocin added; and 50, 50 μg/mL of bacteriocin added. The color of the data points represents donors: red rectangle, Donor-1; green rectangle, Donor-2; and blue rectangle, Donor-3. The data points from donor-2 and donor-3 overlap at the values of zero. CON, positive control; EntA, Enterocin A; EntK1, Enterocin K1; and GarML, Garvicin (PNG 4383 kb)

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Umu, Ö.C.O., Gueimonde, M., Oostindjer, M. et al. Use of Fecal Slurry Cultures to Study In Vitro Effects of Bacteriocins on the Gut Bacterial Populations of Infants. Probiotics & Antimicro. Prot. 12, 1218–1225 (2020). https://doi.org/10.1007/s12602-019-09614-w

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