Drug-Resistance Mechanisms in Vibrio cholerae O1 Outbreak Strain, Haiti, 2010

Sjölund-Karlsson, Maria; Reimer, Aleisha; Folster, Jason P.; Walker, Matthew; Dahourou, Georges; Batra, Dhwani; Martín, Irene; Joyce, Kevin; Parsons, Michele B.; Boncy, Jacques; Whichard, Jean M.; Gilmour, Matthew W.

doi:10.3201/eid1711.110720

Cited by 78 publications

(89 citation statements)

References 10 publications

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“…The primary target of fluoroquinolones appears to be gyrase rather than topoisomerase (11), so the role of the multiple mutations identified in the parC gene in the present investigation in fluoroquinolone susceptibility requires careful study. The resistance pattern of our isolates is similar to that of the Haitian outbreak strain (12), but the amino acid change in position 85 and the four additional mutations detected are hitherto unknown mutations. To our knowledge, the five mutations in the parC gene described here have not been reported previously.…”

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confidence: 55%

Novel Multiple Mutations in the Topoisomerase Gene of Haitian Variant Vibrio cholerae O1

Divya¹,

Sivakumar²,

Devi³

et al. 2014

Antimicrob Agents Chemother

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c C holera has reinstated its deadly grip on the planet with recent epidemics in Zimbabwe and Haiti (1, 2), along with the emergence of Vibrio cholerae having reduced susceptibility to fluoroquinolones. India also experiences frequent outbreaks caused by multidrug-resistant V. cholerae (3). We report reduced sensitivity to ciprofloxacin in V. cholerae O1 El Tor Ogawa and novel multiple mutations in the DNA topoisomerase parC gene.Twelve V. cholerae strains were obtained from the Government Medical College Hospital, Kozhikode, Kerala, India, and serotyped as V. cholerae O1 El Tor Ogawa. Detection of the ctxA, ctxB, and tcpA genes (encoding cholera toxin and toxin-coregulated pili) (4, 5) was done by PCR, and the ctxB gene was sequenced to determine the cholera toxin genotype. Antimicrobial susceptibility testing was performed by disc diffusion (6), and the MIC of ciprofloxacin was determined by Etest (AB bioMérieux, Solna, Sweden). To check for mutations in the quinolone resistancedetermining region (QRDR), the genes encoding DNA gyrase (gyrA, gyrB) and DNA topoisomerase (parC, parE) were amplified and subsequently sequenced (7).All of the isolates possessed the ctxA and ctxB genes and the El Tor allele of tcpA. The ctxB gene sequences were similar to those of the Haitian variant of V. cholerae O1 (5). This variant has DNA sequences identical to those of the classical type of ctxB with an additional mutation at position 58 resulting in the substitution of asparagine for histidine. Antimicrobial susceptibility testing showed that the isolates were sensitive to tetracycline, erythromycin, and chloramphenicol and resistant to ampicillin, furazolidone, nalidixic acid, streptomycin, trimethoprim, cotrimoxazole, and polymyxin B. All of the strains showed reduced susceptibility to ciprofloxacin (MICs, 0.25 to 0.5 mg/liter). The QRDRs of gyrA, gyrB, parC, and parE of all of the isolates were amplified. Since all of the isolates had the same antibiotic susceptibility pattern, four were selected for sequencing of the gyrase and topoisomerase genes. Sequence analysis revealed five mutations in the parC gene, i.e., TCG to AAT (Ser-60¡Asn), TAC to TTT (Tyr-65¡Phe), TCG to ATC (Ser-85¡Ile), GCC to TCC (Ala-128¡Ser), and AAA to CGG (Lys-129¡Arg) (Fig. 1A), and a single mutation in gyrA (Ser-83¡Ile), while no changes were detected in the gyrB and parE genes.From the available literature, the most common amino acid substitution observed in the parC gene is the replacement of serine with leucine at position 85 (TCG ¡TTG) (8, 9), while in this study, serine was replaced with isoleucine in the same position. To understand the effect of this mutation on ciprofloxacin resistance, molecular docking analysis of ciprofloxacin on wild-type (native) and mutant ParC was performed with Autodock 4.0 as shown in Fig. 1B. In the native ParC protein, the optimal binding energy in the QRDR was Ϫ4.95 kcal/mol, whereas in the mutant protein, no ciprofloxacin binding was observed. To explain the lack of ciprofloxacin interaction affinity in the QRDR...

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confidence: 55%

Novel Multiple Mutations in the Topoisomerase Gene of Haitian Variant Vibrio cholerae O1

Divya¹,

Sivakumar²,

Devi³

et al. 2014

Antimicrob Agents Chemother

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“…Clearly V. cholera was found resistant to a commonly used antibiotics such as tetracycline, erythromycin and sensitive to ceftriaxone, cefixim; such behavior may threat through a higher secondary infection rate and by causing illness of longer duration [33]. In general, it was found that the resistance genes are located on large conjugative elements (SXT constins) that are integrated into prfC on the V. cholera chromosome [100], and this finding have opened space for more investigations and discussion for studying the integrating conjugative elements as well as the mobile genetic elements that can potentially mediate transfer of antimicrobial drug resistance [61] [101].…”

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confidence: 99%

The Isolation of <i>Vibrio cholera</i> and Other Enteric Bacteria with Molecular Characterization of <i>Vibrio cholera</i> during the Outbreak of Baghdad/Iraq in 2015

Jameel¹,

Shafek²,

Abdulmohsen³

et al. 2016

AiM

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Vibrio cholera, causing acute watery diarrhea known as cholera disease, affects all ages and both genders. Cholera infection outbreaks in Iraq have been reported for several years. The recent cholera outbreak, emerged throughout 2015, was investigated using bacteriological laboratory tests, singleplex and multiplex PCR technique for the detection of V. cholera from stool samples. Furthermore the toxigenic potential coupled with the antibiotic susceptibility test for cholera and other bacteria were also investigated. The stool samples were collected from 5698 patients admitted to Al-Yarmouk Teaching hospital and health care centers in Baghdad/Al-Karkh, Iraq, from the 1 st of August to the 30 th of December 2015. The V. cholera was isolated from 194 cases (3.4% of the cases age between 21 -50 years). In addition, other enteric infections: Salmonellosis and Shigellosis 7 and 21 respectively, protozoan parasite Giardia lamblia and Entamoeba histolytica 2 and 43 cases respectively were also reported. High percentage of V. cholera infection was detected in October (122 cases, 62.8%), compared with other enteric infections that show high percentage of diarrheal disease in September and November. The results have confirmed that the cholera outbreak was caused by V. cholera O1, biotype El Tor, and serotype Inaba. Seven virulence genes were identified ctxA, toxR, zot, ace, rfbO1, tcpA and ompW. Moreover, the cholera isolated strains were found sensitive to most antibiotic but resistant to nalidixic acid.

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“…Integrated conjugated elements (ICEs) found in HCO1 strains were compared with ICEVchBan5 and ICEVchMoz in V. cholerae O1 CIRS101 and B33, respectively. Analysis of their genetic organization revealed HCO1 ICE is >99% similar to ICEVchBan5, differing only by 5SNPS (31). In contrast, the majority of the Haitian V. cholerae non-O1/O139 strains carry an ICE sequence variant; only three strains lacked an insertion at this site (Table S3).…”

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Genomic diversity of 2010 Haitian cholera outbreak strains

Hasan

Choi

Eppinger

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

171

212

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The millions of deaths from cholera during the past 200 y, coupled with the morbidity and mortality of cholera in Haiti since October 2010, are grim reminders that Vibrio cholerae , the etiologic agent of cholera, remains a scourge. We report the isolation of both V . cholerae O1 and non-O1/O139 early in the Haiti cholera epidemic from samples collected from victims in 18 towns across eight Arrondissements of Haiti. The results showed two distinct populations of V. cholerae coexisted in Haiti early in the epidemic. As non-O1/O139 V . cholerae was the sole pathogen isolated from 21% of the clinical specimens, its role in this epidemic, either alone or in concert with V . cholerae O1, cannot be dismissed. A genomic approach was used to examine similarities and differences among the Haitian V . cholerae O1 and V . cholerae non-O1/O139 strains. A total of 47 V . cholerae O1 and 29 V . cholerae non-O1/O139 isolates from patients and the environment were sequenced. Comparative genome analyses of the 76 genomes and eight reference strains of V . cholerae isolated in concurrent epidemics outside Haiti and 27 V . cholerae genomes available in the public database demonstrated substantial diversity of V. cholerae and ongoing flux within its genome.

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Drug-Resistance Mechanisms in Vibrio cholerae O1 Outbreak Strain, Haiti, 2010

Cited by 78 publications

References 10 publications

Novel Multiple Mutations in the Topoisomerase Gene of Haitian Variant Vibrio cholerae O1

Novel Multiple Mutations in the Topoisomerase Gene of Haitian Variant Vibrio cholerae O1

The Isolation of <i>Vibrio cholera</i> and Other Enteric Bacteria with Molecular Characterization of <i>Vibrio cholera</i> during the Outbreak of Baghdad/Iraq in 2015

Genomic diversity of 2010 Haitian cholera outbreak strains

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Drug-Resistance Mechanisms in Vibrio cholerae O1 Outbreak Strain, Haiti, 2010

Cited by 78 publications

References 10 publications

Novel Multiple Mutations in the Topoisomerase Gene of Haitian Variant Vibrio cholerae O1

Novel Multiple Mutations in the Topoisomerase Gene of Haitian Variant Vibrio cholerae O1

The Isolation of &lt;i&gt;Vibrio cholera&lt;/i&gt; and Other Enteric Bacteria with Molecular Characterization of &lt;i&gt;Vibrio cholera&lt;/i&gt; during the Outbreak of Baghdad/Iraq in 2015

Genomic diversity of 2010 Haitian cholera outbreak strains

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The Isolation of <i>Vibrio cholera</i> and Other Enteric Bacteria with Molecular Characterization of <i>Vibrio cholera</i> during the Outbreak of Baghdad/Iraq in 2015