Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya

BackgroundCholera remains a public health challenge in Kenya. To better understand its dynamics, we analyzed Vibrio cholerae genomes from clinical and environmental samples collected during the 2022–2023 outbreak. These strains were compared with historical genomes from Kenya, Uganda, Tanzania, and...

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Main Authors: Lydia M. Mageto, Gabriel Oluga Aboge, Zelalem H. Mekuria, Peter Gathura, John Juma, Michael Mugo, Collins Kipkorir Kebenei, Diana Imoli, Beatrice Atieno Ongadi, Kelvin Kering, Cecilia Kathure Mbae, Samuel Kariuki
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Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1603736/full
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author Lydia M. Mageto
Lydia M. Mageto
Gabriel Oluga Aboge
Zelalem H. Mekuria
Peter Gathura
John Juma
Michael Mugo
Collins Kipkorir Kebenei
Diana Imoli
Beatrice Atieno Ongadi
Kelvin Kering
Cecilia Kathure Mbae
Samuel Kariuki
author_facet Lydia M. Mageto
Lydia M. Mageto
Gabriel Oluga Aboge
Zelalem H. Mekuria
Peter Gathura
John Juma
Michael Mugo
Collins Kipkorir Kebenei
Diana Imoli
Beatrice Atieno Ongadi
Kelvin Kering
Cecilia Kathure Mbae
Samuel Kariuki
author_sort Lydia M. Mageto
collection DOAJ
description BackgroundCholera remains a public health challenge in Kenya. To better understand its dynamics, we analyzed Vibrio cholerae genomes from clinical and environmental samples collected during the 2022–2023 outbreak. These strains were compared with historical genomes from Kenya, Uganda, Tanzania, and Haiti to inform strategies for cholera prevention, control, and elimination in Kenya.MethodsClinical (stool) and environmental (wastewater, drinking water, and household effluent) samples were collected from Nairobi county. Samples were analyzed for V. cholerae using culture and real time PCR. The environmental (n = 17) and clinical (n = 70) isolates were then subjected to phenotypic antimicrobial susceptibility testing using the Kirby-Bauer disk diffusion method. Whole genome sequencing was employed to characterize the genome, detect antimicrobial resistance genes, virulence factors, and mobile genetic elements. Phylogenetic analysis was performed to assess the genetic relationship and diversity of isolates from 2022 to 2023 outbreak, comparing them with isolates from historical outbreaks.ResultsClinical isolates carried key virulence genes (ctxA, ctxB7, zot, and hlyA) and were 100% resistant to multiple antibiotics, including ampicillin, cefotaxime, ceftriaxone, and cefpodoxime, but remained susceptible to gentamicin and chloramphenicol. In contrast, environmental isolates lacked ctxB gene but harbored toxR, als, and hlyA, showing variable antibiotic resistance (59% to ampicillin, 41% to trimethoprim-sulfamethoxazole, and 47% to nalidixic acid). All clinical isolates from 2022 to 2023 outbreak harbored IncA/C2 plasmids and several antimicrobial resistance genes including blaPER–7. Phylogenetic analysis revealed high genetic diversity in environmental strains, clustering outside the 7th pandemic El Tor lineage, while clinical isolates were highly clonal. Genomes from 2022 to 2023 outbreak were closely related to Kenyan cholera outbreak genomes from 2016 (15 single nucleotide polymorphisms, T13 lineage).ConclusionThe 2022–2023 outbreak likely resulted from re-emergence of previously circulating strains rather than a new introduction. While the role of environmental reservoirs as a source of human infection remains unclear in our study, environmental isolates possess virulent and antimicrobial resistance genes that may spread via horizontal gene transfer. This highlights the need for continuous genomic surveillance to monitor V. cholerae evolution, track transmission patterns, and mitigate the spread of antimicrobial resistance.
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spelling doaj-art-22c51fec34a84db4ba2df5f1a1b69e3b2025-08-20T03:33:18ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-07-011610.3389/fmicb.2025.16037361603736Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in KenyaLydia M. Mageto0Lydia M. Mageto1Gabriel Oluga Aboge2Zelalem H. Mekuria3Peter Gathura4John Juma5Michael Mugo6Collins Kipkorir Kebenei7Diana Imoli8Beatrice Atieno Ongadi9Kelvin Kering10Cecilia Kathure Mbae11Samuel Kariuki12Department of Public Health, Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Nairobi, Nairobi, KenyaWashington State University, Global Health Kenya, Nairobi, KenyaDepartment of Public Health, Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Nairobi, Nairobi, KenyaDepartment of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH, United StatesDepartment of Public Health, Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Nairobi, Nairobi, KenyaInternational Livestock Research Institute (ILRI), Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaCentre for Microbiology Research, Kenya Medical Research Institute, Nairobi, KenyaBackgroundCholera remains a public health challenge in Kenya. To better understand its dynamics, we analyzed Vibrio cholerae genomes from clinical and environmental samples collected during the 2022–2023 outbreak. These strains were compared with historical genomes from Kenya, Uganda, Tanzania, and Haiti to inform strategies for cholera prevention, control, and elimination in Kenya.MethodsClinical (stool) and environmental (wastewater, drinking water, and household effluent) samples were collected from Nairobi county. Samples were analyzed for V. cholerae using culture and real time PCR. The environmental (n = 17) and clinical (n = 70) isolates were then subjected to phenotypic antimicrobial susceptibility testing using the Kirby-Bauer disk diffusion method. Whole genome sequencing was employed to characterize the genome, detect antimicrobial resistance genes, virulence factors, and mobile genetic elements. Phylogenetic analysis was performed to assess the genetic relationship and diversity of isolates from 2022 to 2023 outbreak, comparing them with isolates from historical outbreaks.ResultsClinical isolates carried key virulence genes (ctxA, ctxB7, zot, and hlyA) and were 100% resistant to multiple antibiotics, including ampicillin, cefotaxime, ceftriaxone, and cefpodoxime, but remained susceptible to gentamicin and chloramphenicol. In contrast, environmental isolates lacked ctxB gene but harbored toxR, als, and hlyA, showing variable antibiotic resistance (59% to ampicillin, 41% to trimethoprim-sulfamethoxazole, and 47% to nalidixic acid). All clinical isolates from 2022 to 2023 outbreak harbored IncA/C2 plasmids and several antimicrobial resistance genes including blaPER–7. Phylogenetic analysis revealed high genetic diversity in environmental strains, clustering outside the 7th pandemic El Tor lineage, while clinical isolates were highly clonal. Genomes from 2022 to 2023 outbreak were closely related to Kenyan cholera outbreak genomes from 2016 (15 single nucleotide polymorphisms, T13 lineage).ConclusionThe 2022–2023 outbreak likely resulted from re-emergence of previously circulating strains rather than a new introduction. While the role of environmental reservoirs as a source of human infection remains unclear in our study, environmental isolates possess virulent and antimicrobial resistance genes that may spread via horizontal gene transfer. This highlights the need for continuous genomic surveillance to monitor V. cholerae evolution, track transmission patterns, and mitigate the spread of antimicrobial resistance.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1603736/fullcholeraantimicrobial resistancewhole genome sequencingphylogenetic analysisvirulence
spellingShingle Lydia M. Mageto
Lydia M. Mageto
Gabriel Oluga Aboge
Zelalem H. Mekuria
Peter Gathura
John Juma
Michael Mugo
Collins Kipkorir Kebenei
Diana Imoli
Beatrice Atieno Ongadi
Kelvin Kering
Cecilia Kathure Mbae
Samuel Kariuki
Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya
Frontiers in Microbiology
cholera
antimicrobial resistance
whole genome sequencing
phylogenetic analysis
virulence
title Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya
title_full Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya
title_fullStr Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya
title_full_unstemmed Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya
title_short Genomic characterization of Vibrio cholerae isolated from clinical and environmental sources during the 2022–2023 cholera outbreak in Kenya
title_sort genomic characterization of vibrio cholerae isolated from clinical and environmental sources during the 2022 2023 cholera outbreak in kenya
topic cholera
antimicrobial resistance
whole genome sequencing
phylogenetic analysis
virulence
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1603736/full
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