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Cholera is a life-threatening gastrointestinal infection caused by a toxigenic bacterium, Vibrio cholerae . After a lull of almost 30 years, a first case of cholera was detected in Lebanon in October 2022. The outbreak lasted three months, with 8007 suspected cases (671 laboratory-confirmed) and 23 deaths. In this study, we use phenotypic methods and microbial genomics to study 34 clinical and environmental Vibrio cholerae isolates collected throughout this outbreak. All isolates are identified as V. cholerae O1, serotype Ogawa strains from wave 3 of the seventh pandemic El Tor (7PET) lineage. Phylogenomic analysis unexpectedly reveals the presence of two different strains of the seventh pandemic El Tor (7PET) lineage. The dominant strain has a narrow antibiotic resistance profile and is phylogenetically related to South Asian V. cholerae isolates and derived African isolates from the AFR15 sublineage. The second strain is geographically restricted and extensively drug-resistant. It belongs to the AFR13 sublineage and clusters with V. cholerae isolates collected in Yemen. In conclusion, the 2022-2023 Lebanese cholera outbreak is caused by the simultaneous introduction of two different 7PET strains. Genomic surveillance with cross-border collaboration is therefore crucial for the identification of new introductions and routes of circulation of cholera, improving our understanding of cholera epidemiology. Here the authors provide results from a genomic epidemiology study of a cholera outbreak in Lebanon, showing that it was caused by two Vibrio cholerae strains of serogroup O1 (El Tor biotype), namely an AFR15 sublineage related to South Asian isolates and extensively drug-resistant Yemeni AFR13.

Analysis of clinical and environmental Vibrio cholerae O1 strains obtained during 2008-2015 in Nigeria showed that lineages Afr9 and Afr12 carrying cholera toxin and Vibrio pathogenicity island 1 can be isolated from water. Our findings raise concerns about the role of the environment in maintenance and emergence of cholera outbreaks in Nigeria.

Northern Cameroon in Central Africa has experienced recurring cholera outbreaks despite ongoing efforts to control the disease. While most cholera studies focus on O1 pandemic Vibrio cholerae strains, non-O1/O139 strains are increasingly recognized for their infection potential and dynamic relationships with O1 strains during outbreaks. Here we explore the genetic diversity and phylogenetic relationships of non-O1/O139 V. cholerae (NOVC) isolated from environmental water sources in Northern Cameroon. These NOVC strains show significant genetic diversity and virulence potential. They are closely related to environmental strains from Kenya and clinical strains from Argentina and Haiti, suggesting transmissions across countries and continents, likely facilitated by human carriers. The highly conserved tcpA gene found in some strains from Cameroon is closely related to the tcpA O1 Classical type, suggesting direct or indirect genetic interactions between these environmental NOVC strains and pandemic strains. Our findings underscore the importance of environmental surveillance and further studies of NOVC strains to better understand cholera outbreaks.

In October of 2010, an outbreak of cholera was confirmed in Haiti for the first time in more than a century. A single clone of toxigenic Vibrio cholerae O1 biotype El Tor serotype Ogawa strain was implicated as the cause. Five years after the onset of cholera, in October, 2015, we have discovered a major switch (ranging from 7 to 100%) from Ogawa serotype to Inaba serotype. Furthermore, using wbeT gene sequencing and comparative sequence analysis, we now demonstrate that, among 2013 and 2015 Inaba isolates, the wbeT gene, responsible for switching Ogawa to Inaba serotype, sustained a unique nucleotide mutation not found in isolates obtained from Haiti in 2012. Moreover, we show that, environmental Inaba isolates collected in 2015 have the identical mutations found in the 2015 clinical isolates. Our data indicate that toxigenic V. cholerae O1 serotype Ogawa can rapidly change its serotype to Inaba, and has the potential to cause disease in individuals who have acquired immunity against Ogawa serotype. Our findings highlight the importance of monitoring of toxigenic V. cholerae O1 and cholera in countries with established endemic disease.

Cholera has been endemic in Douala since 1971. Most outbreaks start from Bepanda, an overcrowded neighbourhood with poor hygiene and sanitary conditions. We investigated water sources in Bepanda as reservoirs of Vibrio cholerae, the causative agent of cholera, determined its antibiotic susceptibility and some physico-chemical characteristics that could maintain the endemicity of this organism in Bepanda. Three hundred and eighteen water samples collected from 45 wells, 8 taps and 1 stream from February to July 2009 were analyzed for V. cholerae using standard methods. Isolates were characterized morphologically, biochemically and serologically. The disc diffusion technique was employed to investigate antibiotic susceptibility. Differences in prevalence of organism between seasons were analysed. Correlation strength and direction of association between physico-chemical parameters and occurrence of V. cholerae was analyzed using the Kendall tau_b non-parametric correlation. This was further confirmed with the forward-stepwise binary logistic regression. Eighty-seven (27.4%) samples were positive for V. cholerae. Isolation was highest from wells. The organism was isolated in the rainy season and dry season but the frequency of isolation was significantly higher (χ2 = 7.009, df = 1, P = 0.008) in the rainy season. Of the 96 confirmed V. cholerae isolates, 32 (33.3%) belonged to serogroup O1 and 64 (66.6%) were serogroup non-O1/non-O139. Isolates from tap (municipal water) were non-O1/non-O139 strains. Salinity had a significant positive correlation with isolation in the dry season (+0.267, P = 0.015) and rainy season (+0.223, P = 0.028). The forward-stepwise method of binary logistic regression indicated that as pH (Wald = 11.753, df = 1), P = 0.001) increased, odds of isolation of V. cholerae also increased (B = 1.297, S.E = 0.378, Exp(B) = 3.657). All isolates were sensitive to ciprofloxacin and ofloxacin. Multi-drug resistance was predominant among the non-O1/non-O139 isolates. V. cholerae was found in wells and stream in both seasons. Cholera will continue to be a health threat in Bepanda if intervention measures to prevent outbreak are not implemented. Continuous monitoring of water sources in this and other cholera high-risk areas in Cameroon is necessary, for a better preparedness and control of cholera.

Vibrio cholerae causes cholera, a severe diarrheal disease. Understanding the local genetic diversity and transmission of V. cholerae will improve our ability to control cholera. Vibrio cholerae isolates clustered in genetically related groups (clonal complexes, CC) by multilocus variable tandem-repeat analysis (MLVA) were compared by whole genome sequencing (WGS). Isolates in CC1 had been isolated from two geographical locations. Isolates in a second genetically distinct group, CC2, were isolated only at one location. Using WGS, CC1 isolates from both locations revealed, on average, 43.8 nucleotide differences, while those strains comprising CC2 averaged 19.7 differences. Strains from both MLVA-CCs had an average difference of 106.6. Thus, isolates comprising CC1 were more closely related (P < 10−6) to each other than to isolates in CC2. Within a MLVA-CC, after removing all paralogs, alternative alleles were found in all possible combinations on separate chromosomes indicative of recombination within the core genome. Including recombination did not affect the distinctiveness of the MLVA-CCs when measured by WGS. We found that WGS generally reflected the same genetic relatedness of isolates as MLVA, indicating that isolates from the same MLVA-CC shared a more recent common ancestor than isolates from the same location that clustered in a distinct MLVA-CC. Genetic relatedness of Vibrio cholerae isolates from distinct locations was similar when measured by multilocus variable tandem-repeat analysis or whole genome sequencing.

Phylodynamic analysis of genome-wide single-nucleotide polymorphism (SNP) data is a powerful tool to investigate underlying evolutionary processes of bacterial epidemics. The method was applied to investigate a collection of 65 clinical and environmental isolates of Vibrio cholerae from Haiti collected between 2010 and 2012. Characterization of isolates recovered from environmental samples identified a total of four toxigenic V. cholerae O1 isolates, four non-O1/O139 isolates, and a novel nontoxigenic V. cholerae O1 isolate with the classical tcpA gene. Phylogenies of strains were inferred from genome-wide SNPs using coalescent-based demographic models within a Bayesian framework. A close phylogenetic relationship between clinical and environmental toxigenic V. cholerae O1 strains was observed. As cholera spread throughout Haiti between October 2010 and August 2012, the population size initially increased and then fluctuated over time. Selection analysis along internal branches of the phylogeny showed a steady accumulation of synonymous substitutions and a progressive increase of nonsynonymous substitutions over time, suggesting diversification likely was driven by positive selection. Short-term accumulation of nonsynonymous substitutions driven by selection may have significant implications for virulence, transmission dynamics, and even vaccine efficacy. IMPORTANCE Cholera, a dehydrating diarrheal disease caused by toxigenic strains of the bacterium Vibrio cholerae , emerged in 2010 in Haiti, a country where there were no available records on cholera over the past 100 years. While devastating in terms of morbidity and mortality, the outbreak provided a unique opportunity to study the evolutionary dynamics of V. cholerae and its environmental presence. The present study expands on previous work and provides an in-depth phylodynamic analysis inferred from genome-wide single nucleotide polymorphisms of clinical and environmental strains from dispersed geographic settings in Haiti over a 2-year period. Our results indicate that even during such a short time scale, V. cholerae in Haiti has undergone evolution and diversification driven by positive selection, which may have implications for understanding the global clinical and epidemiological patterns of the disease. Furthermore, the continued presence of the epidemic strain in Haitian aquatic environments has implications for transmission. Cholera, a dehydrating diarrheal disease caused by toxigenic strains of the bacterium Vibrio cholerae , emerged in 2010 in Haiti, a country where there were no available records on cholera over the past 100 years. While devastating in terms of morbidity and mortality, the outbreak provided a unique opportunity to study the evolutionary dynamics of V. cholerae and its environmental presence. The present study expands on previous work and provides an in-depth phylodynamic analysis inferred from genome-wide single nucleotide polymorphisms of clinical and environmental strains from dispersed geographic settings in Haiti over a 2-year period. Our results indicate that even during such a short time scale, V. cholerae in Haiti has undergone evolution and diversification driven by positive selection, which may have implications for understanding the global clinical and epidemiological patterns of the disease. Furthermore, the continued presence of the epidemic strain in Haitian aquatic environments has implications for transmission.

Two methods were compared for their ability to accurately identify Vibrio species of interest: whole genome sequencing as the reference method and MALDI-TOF MS (matrix-assisted laser desorption/ionization-time of flight mass spectrometry) proteome fingerprinting. The accuracy of mass spectrometry–based identification method was evaluated for its ability to accurately identify isolates of Vibrio cholerae and Vibrio parahaemolyticus . Identification result of each isolate obtained by mass spectrometry was compared to identification by whole genome sequencing (WGS). The MALDI-TOF MS system had excellent performance for identification of V. cholerae and V. parahaemolyticus isolates grown on a non-selective solid agar media. Unlike the biochemical characterization performed by API20E. In this study, 161 isolates ( V. cholerae, n  = 33; V. parahaemolyticus, n  = 102; V. spp., n  = 23; other enteropathogens, Salmonella and E. coli , n  = 3) were used to assess accuracy. The MALDI-TOF MS system was able to accurately identify 100% (33/33) of the V. cholerae isolates and 99.9% (101/102) of V. parahaemolyticus isolates, with 100% for both sensitivity and specificity for V. cholerae and 99% sensitivity and 98% specificity for V. parahaemolyticus . Thus, mass spectrometry for bacterial identification is comparable to the WGS. Furthermore, in comparison to a biochemical characterization, the use of MALDI-TOF MS system shortens the analysis time from over 72 h to less than 24 h. Key points • V. cholerae and V. parahaemolyticus were successfully ID-ed by MALDI-TOF • MALDI-TOF sensitivity and specificity parallels the WGS method of identification • MALDI-TOF is several days faster than the battery of culture-dependent methods

Latin America has experienced two of the largest cholera epidemics in modern history; one in 1991 and the other in 2010. However, confusion still surrounds the relationships between globally circulating pandemic Vibrio cholerae clones and local bacterial populations. We used whole-genome sequencing to characterize cholera across the Americas over a 40-year time span. We found that both epidemics were the result of intercontinental introductions of seventh pandemic El Tor V. cholerae and that at least seven lineages local to the Americas are associated with disease that differs epidemiologically from epidemic cholera. Our results consolidate historical accounts of pandemic cholera with data to show the importance of local lineages, presenting an integrated view of cholera that is important to the design of future disease control strategies.

A comparison of the genomes of two “outbreak” Vibrio cholerae isolates from Haiti with those of other isolates indicates that this outbreak strain is distinct from circulating Latin American isolates and bears striking similarity to recent isolates from South Asia. The outbreak of cholera that began in Haiti in late October 2010 illustrates the continued public health threat of this ancient scourge. 1 Cholera, an acutely dehydrating diarrheal disease that can rapidly kill its victims, is caused by Vibrio cholerae, a gram-negative bacterium. 2 This disease, which is usually transmitted through contaminated water, can and has spread in an explosive fashion. In the weeks since cases were first confirmed in the Artibonite province of Haiti on October 19, 2010, the disease has reached all 10 provinces in Haiti and has spread to the neighboring Dominican Republic on the island of Hispaniola. Of . . .