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Searching for Bacteria within Acute Cholecystitis Using Next-Generation Sequencers

19 Pages Posted: 24 Mar 2025 Publication Status: Under Review

See all articles by Tomohiro Otsuka

Tomohiro Otsuka

Juntendo University

Yoichi Ishizaki

Juntendo University

Jiro Yoshimoto

Juntendo University

Kenji Takamori

Juntendo University

Shin Watanabe

Juntendo University

Abstract

Introduction: A biliary microbiome comprising flora within normal gallbladders was recently uncovered through analyses targeting the bacterial 16S ribosomal RNA (16S rRNA) gene, despite the gallbladder previously being regarded as a sterile environment. In the present study, we subjected bile samples from patients with acute cholecystitis to gene analysis targeting bacterial flora. 


Methods: We targeted patients diagnosed as having Grade I or Grade II acute cholecystitis (in accordance with the Tokyo Guidelines 2018 established by the Japanese Society of Hepato-Biliary-Pancreatic Surgery) who underwent laparoscopic cholecystectomy within 24 h of diagnosis at Juntendo University Urayasu Hospital between July 2021 and January 2024 for evaluation. We drew bile samples from the gallbladder of each patient to confirm the presence of biliary bacterial flora, using both standard bacteriology (culture tests) and 16S rRNA gene sequencing. 

Results: Of the 29 samples, 15 yielded cultures positive for bacterial flora, and gene analysis revealed the presence of bacterial biliary flora in all 14 samples that had tested negative in standard bacteriology. Considering the bacterial flora of a normal gallbladder without lesions as “normal flora,” bacteria other than normal flora—Propionibacterium spp., Coprococcus spp., Prevotella spp., Sediminibacterium spp., and Collinesella spp.—were detected in 25 of the 29 cases (86%). 

Conclusions: Bacteria not detected in non-inflammatory gallbladders such as Propiobacterium spp., Coprococcus spp., Prevotella spp., Sediminibacterium spp., and Collinesella spp. may play a role in the mechanism underlying development of acute cholecystitis.

Note:
Funding declaration: This work was supported by Asahi Group Holdings, Ltd. The funder had no role in the study design, in the collection, analysis, or interpretation of data, in the writing of the report, or in the decision to submit the article for publication.

Conflict of Interests: This study received funding from Asahi Group Holdings, Ltd. The authors declare no remaining conflicts of interest.

Ethical Approval: Informed consent was obtained from all individual participants included in the study. The study was approved by the ethics committee of Juntendo University, Urayasu Hospital (Approval No. 3-009; 13 March 2021).

Keywords: Acute cholecystitis, Bacteria flora, Next-generation sequencer, 16S ribosomal RNA, 16S rRNA, Biliary microbiome

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Suggested Citation

Otsuka, Tomohiro and Ishizaki, Yoichi and Yoshimoto, Jiro and Takamori, Kenji and Watanabe, Shin, Searching for Bacteria within Acute Cholecystitis Using Next-Generation Sequencers. Available at SSRN: https://ssrn.com/abstract=5184653 or http://dx.doi.org/10.2139/ssrn.5184653

Tomohiro Otsuka (Contact Author)

Juntendo University ( email )

Yoichi Ishizaki

Juntendo University ( email )

2 Chome-1-1 Hongo
Tokyo
Japan

Jiro Yoshimoto

Juntendo University ( email )

2 Chome-1-1 Hongo
Tokyo
Japan

Kenji Takamori

Juntendo University ( email )

Shin Watanabe

Juntendo University ( email )

2 Chome-1-1 Hongo
Tokyo
Japan

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