Selection and evaluation of lactic acid bacteria from chicken feces in Thailand as potential probiotics

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Microbiology

Main article text

 

Introduction

Materials and Methods

Sample collections

Lactic acid bacteria isolation

Identification of lactic acid bacteria

Evaluation of probiotic functional properties of LAB isolates

Survival of LAB at low pH and varying bile salt concentrations

Pre-screening for resistance at pH 2.5 and with 0.3% bile salts
Resistance to acid at various pH levels and bile salt concentrations at a temperature of 42 °C

Resistance to 0.4% phenol

Cell surface properties

Cell surface hydrophobicity
Auto-aggregation
Co-aggregation

Antimicrobial activity against Salmonella enterica

Evaluation of probiotic safety

Antimicrobial resistance detection

Antimicrobial susceptibility
Genotyping antimicrobial resistance assay using a real-time PCR with specific probes
Whole-genome search for antimicrobial resistance genes

Hemolytic activity

Statistical analysis

Results

Isolation and identification of lactic acid bacteria

Survival of LAB under low pH and at different concentrations of bile

Phenol tolerance

Cell surface properties

Antimicrobial activity against Salmonella enterica

Antimicrobial resistance detection

Hemolytic activity

Discussion

Conclusions

Supplemental Information

LAB species recovered from the different chicken types.

DOI: 10.7717/peerj.16637/supp-1

The acid, bile salts and phenol tolerance of LAB isolates.

DOI: 10.7717/peerj.16637/supp-2

Raw data to determine the correlation.

DOI: 10.7717/peerj.16637/supp-3

Antimicrobial activity of 5 LAB isolates against 23 Salmonella enterica.

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Antimicrobial susceptibility of 5 LAB isolates.

DOI: 10.7717/peerj.16637/supp-5

Disc diffusion agar presenting susceptibility of Salmonella to CFS from the 5 selected strains.

S1: Antimicrobial activity of CFS (a) and NCFS (b) from 5 selected LAB isolates including; L. salivarius BF12 (1); P. acidilactici BF9 (2); P. acidilactici BF14 (3); P. acidilactici BYF20 (4); P. acidilactici BYF26 (5) against some isolates of 9 serovars of Salmonella enterica including; S. Typhimurium 3 (A); S. Agona 1 (B); S. Kentucky 2 (C); S. Virchow 2 (D); S. Albany (E); S. Braenderup (F); S. Hadar (G); S. Enteritidis 2 (H); S. Give (I).

DOI: 10.7717/peerj.16637/supp-6

List of antimicrobial resistance genes detected in the real-time PCR with specific probes assay.

DOI: 10.7717/peerj.16637/supp-7

Real-time PCR resulting AMR genes contained in the 5 selected strains.

Use of the real-time PCR assay only identified ermB, an erythromycin resistance gene, and only in P. acidilactici isolates BF9 and BF14.

DOI: 10.7717/peerj.16637/supp-8

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Benjamas Khurajog conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Yuda Disastra performed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Lum Dau Lawwyne performed the experiments, prepared figures and/or tables, and approved the final draft.

Wandee Sirichokchatchawan conceived and designed the experiments, performed the experiments, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Waree Niyomtham performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Jitrapa Yindee performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

David John Hampson analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Nuvee Prapasarakul conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

All isolates were obtained from freshly passed feces at the chicken farms. The Faculty of Veterinary Science Institutional Biosafety Committee approved this experimental work (agreement number IBC20310148).

Data Availability

The following information was supplied regarding data availability:

All oligonucleotide sequence primers are available in the Supplemental File.

Funding

The present scientific research was financially supported by the 2022-Fundamental Fund, Thailand Science Research and Innovation (TSRI), Chulalongkorn University (FOOD66310012), the Secondary Century Fund (C2F) for Doctoral Scholarship and the 90th Anniversary of Chulalongkorn University Scholarship (Ratchadaphiseksomphot Endowment Fund) and the ThaiFoods Group public company limited, Bangkok, Thailand. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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