연구성과 홍보

Mol Nutr Food Res. 2025 Jan;69(2):e2400494.

Title : Use of FISH-FLOW as a Method for the Identification and Quantification of Bacterial Populations

Authors : Jorge Enrique Vazquez Bucheli1,2, Yuri Lee1, Bobae Kim3, Nuno F Azevedo4,5,6, Andreia S Azevedo4,5,6, Svetoslav Dimitrov Todorov7,8, Yosep Ji3, Hyeji Kang1,3, Wilhelm H Holzapfel1,3*

Affiliations :

1Human Effective Microbes, Department of Advanced Convergence, Handong Global University, Pohang, Gyeongbuk, 37554, Republic of Korea.

2Facultad de medicina veterinaria y zootecnia, Universidad Autónoma de Chiapas, Chiapas, Calz. Emiliano Zapata Km. 8, Del Frigorífico, Chiapas, 29060, México.

3HEMPharma Inc., Pohang, Gyeongbuk, 37554, Republic of Korea.

4LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, rua Dr. Roberto Frias, Porto, 4200-465, Portugal.

5i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, 4200-135, Portugal.

6IPATIMUP - Instituto de Patologia e Imunologia Molecular, Universidade do Porto, Porto, 4200-135, Portugal.

7ProBacLab, Department of Advanced Convergence, Handong Global University, Pohang, Gyeongbuk, 37554, Republic of Korea.

8ProBacLab, Laboratório de Microbiologia de Alimentos, Departamento de Alimentos e Nutrição, Experimental, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, São Paulo (SP), 05501-000, Brazil.

DOI: 10.1002/mnfr.202400494.

Abstract :

The gastrointestinal tract (GIT) harbors the largest group of microbiotas among the microbial communities of the human host. The resident organisms typical of a healthy gut are well adapted to the gastrointestinal environment while alteration of these populations can trigger disorders that may affect the health and well-being of the host. Various investigations have applied different tools to study bacterial communities in the gut and their correlation with gastrointestinal disorders such as inflammatory bowel disease (IBD), obesity, and diabetes. This study proposes fluorescent in situ hybridization, combined with flow cytometry (FISH-FLOW), as an alternative approach for phylum level identification of Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria and quantification of target bacteria from the GIT based on analysis of fecal samples, where results are validated by quantitative polymerase chain reaction (qPCR) and 16S ribosomal ribonucleic acid (16s rRNA) sequencing. The results obtained via FISH-FLOW experimental approach show high specificity for the developed probes for hybridization with the target bacteria. The study, therefore, suggests the FISH-FLOW as a reliable method for studying bacterial communities in the gut with results correlating well with those of metagenomic investigations of the same fecal samples.