---
title: "Citrobacter freundii Identified as Pathogen Causing Disease in Farmed Singhi (Heteropneustes fossi"
id: "pubmed-42424302"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42424302"
content_type: "clinical_feed_article"
specialty: "Infectious Disease"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42424302/"
doi: "10.1093/jahafs/vsag006"
published_at: "2026-08-27T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# Citrobacter freundii Identified as Pathogen Causing Disease in Farmed Singhi (Heteropneustes fossi
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42424302
- **Specialty:** [Infectious Disease](https://medichelpline.com/clinical-feed/infectious-disease.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42424302/)
- **DOI:** [10.1093/jahafs/vsag006](https://doi.org/10.1093%2Fjahafs%2Fvsag006)
- **Published At:** 2026-08-27T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- An outbreak in intensively farmed Singhi (Stinging Catfish, Heteropneustes fossilis) in Tripura, India, was linked to a bacterial isolate identified as **Citrobacter freundii** (strain COF_AHE25; GenBank ON798627). - Diseased fish displayed white patches, skin ulcers, hemorrhages, fin rot and mortality; an isolate was recovered and characterized by phenotypic/biochemical tests and 16S rRNA sequencing with phylogenetic analysis. - Experimental infection reproduced natural signs; mortality began at day 4 postinfection and increased in a dose-dependent manner, confirming virulence in Singhi. - The median lethal dose (LD50) for strain COF_AHE25 was estimated at 2.5 × 10^7 CFU per fish, establishing pathogenicity quantitatively. - Infected fish showed significant hematological, biochemical and immunological changes consistent with systemic bacterial septicemia, indicating systemic host response to infection. - Antimicrobial susceptibility testing (Kirby-Bauer, CLSI guidelines) showed the isolate was multidrug resistant, including resistance to **tetracycline**, **erythromycin**, β-lactams (penicillin, oxacillin, cefoxitin) and **vancomycin**. - The study highlights the first reported association of **C. freundii** with disease in Singhi, raising concerns for intensive freshwater aquaculture and the importance of surveillance, stewardship and improved health management. - Funding sources listed include the National Fisheries Development Board (NFDB) and the Department of Fisheries under PMMSY; authors declared no conflicts of interest.
## Clinical Analysis & Structured Key Points
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Affiliations Expand ### Affiliation * 1 Department of Aquatic Health and Environment, College of Fisheries, Central Agricultural University (Imphal), Lembucherra, Tripura,India. * PMID: **42424302** * DOI: [ 10.1093/jahafs/vsag006 ](https://doi.org/10.1093/jahafs/vsag006) Item in Clipboard # Discovery of Citrobacter freundii as a disease-causing agent in Singhi Heteropneustes fossilis: A first report Lija Ghosh et al. J Aquat Anim Health. 2026. Show details Display options Display options Format Abstract PubMed PMID J Aquat Anim Health Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22J+Aquat+Anim+Health%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22J+Aquat+Anim+Health%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42424302/) . 2026 Aug 27;38(3):154-168. doi: 10.1093/jahafs/vsag006. ### Authors [Lija Ghosh](https://pubmed.ncbi.nlm.nih.gov/?term=Ghosh+L&cauthor_id=42424302)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42424302/#short-view-affiliation-1 "Department of Aquatic Health and Environment, College of Fisheries, Central Agricultural University \(Imphal\), Lembucherra, Tripura,India."), [C Laltlanmawia](https://pubmed.ncbi.nlm.nih.gov/?term=Laltlanmawia+C&cauthor_id=42424302)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42424302/#short-view-affiliation-1 "Department of Aquatic Health and Environment, College of Fisheries, Central Agricultural University \(Imphal\), Lembucherra, Tripura,India."), [Himadri Saha](https://pubmed.ncbi.nlm.nih.gov/?term=Saha+H&cauthor_id=42424302)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42424302/#short-view-affiliation-1 "Department of Aquatic Health and Environment, College of Fisheries, Central Agricultural University \(Imphal\), Lembucherra, Tripura,India.") ### Affiliation * 1 Department of Aquatic Health and Environment, College of Fisheries, Central Agricultural University (Imphal), Lembucherra, Tripura,India. * PMID: **42424302** * DOI: [ 10.1093/jahafs/vsag006 ](https://doi.org/10.1093/jahafs/vsag006) Item in Clipboard Full text links Cite Display options Display options Format Abstract PubMed PMID ## Abstract **Objective:** The purpose of this study was to identify and describe Citrobacter freundii, which was associated with a disease outbreak in Singhi (also known as Stinging Catfish) Heteropneustes fossilis that are cultured under intensive freshwater farming conditions in Tripura, India, and to evaluate its pathogenicity and antimicrobial resistance profile. **Methods:** A bacterial isolate (COF_AHE25; GenBank accession no. ON798627) was recovered from diseased fish that were exhibiting white patches, skin ulcers, hemorrhages, fin rot, and mortality. The isolate was identified using phenotypic and biochemical evaluation, followed by molecular confirmation based on 16S ribosomal RNA gene sequencing and phylogenetic analysis. Pathogenicity was assessed through experimental infection trials, -daily monitoring of cumulative mortality, and estimation of the median lethal dose for the species. Host responses were evaluated through hematological, biochemical, and immunological analyses. Antimicrobial susceptibility was determined using the Kirby-Bauer disk diffusion method in accordance with the Clinical and Laboratory Standards Institute guidelines. **Results:** Experimental infection confirmed the virulence of the isolate, with infected Singhi developing clinical signs that were comparable to those observed during the natural outbreak. Mortality commenced at 4 d postinfection and progressed in a dose-dependent manner. Significant alterations in hematological, biochemical, and immunological parameters were observed, consistent with systemic bacterial septicemia. The median lethal dose of C. freundii strain COF_AHE25 was estimated at 2.5 × 107 CFU/fish. The isolate exhibited multidrug resistance, including resistance to tetracycline and erythromycin as well as to β-lactam antibiotics (penicillin, oxacillin, and cefoxitin) and vancomycin. **Conclusions:** The findings establish C. freundii as a pathogenic bacterium capable of causing significant disease and mortality in intensively farmed Singhi. The detection of resistance to antimicrobials commonly used in freshwater aquaculture highlights the need for strengthened disease surveillance, responsible antimicrobial stewardship, and improved health management practices in intensive fish farming systems. **Keywords:** Citrobacter freundii; Heteropneustes fossilis; LD50; antimicrobial resistance; pathogenicity. ## Plain language summary A bacterial outbreak in farmed Singhi in Tripura, India, was linked to Citrobacter freundii. The pathogen caused ulcers and mortality and showed multidrug resistance, highlighting risks to aquaculture health and sustainability. © American Fisheries Society 2026. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com. 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