Vibrio cholerae is a globally important pathogen that is endemic in many areas of the world and causes 3–5 million reported cases of cholera every year. Historically, there have been seven acknowledged cholera pandemics; recent outbreaks in Zimbabwe and Haiti are included in the seventh and ongoing pandemic1. Only isolates in serogroup O1 (consisting of two biotypes known as ‘classical’ and ‘El Tor’) and the derivative O139 (refs 2, 3) can cause epidemic cholera2. It is believed that the first six cholera pandemics were caused by the classical biotype, but El Tor has subsequently spread globally and replaced the classical biotype in the current pandemic1. Detailed molecular epidemiological mapping of cholera has been compromised by a reliance on sub-genomic regions such as mobile elements to infer relationships, making El Tor isolates associated with the seventh pandemic seem superficially diverse. To understand the underlying phylogeny of the lineage responsible for the current pandemic, we identified high-resolution markers (single nucleotide polymorphisms; SNPs) in 154 whole-genome sequences of globally and temporally representative V. cholerae isolates. Using this phylogeny, we show here that the seventh pandemic has spread from the Bay of Bengal in at least three independent but overlapping waves with a common ancestor in the 1950s, and identify several transcontinental transmission events. Additionally, we show how the acquisition of the SXT family of antibiotic resistance elements has shaped pandemic spread, and show that this family was first acquired at least ten years before its discovery in V. cholerae.

Evidence for several waves of global transmission in the seventh cholera pandemic

As the causative agent of cholera, the bacterium Vibrio cholerae represents an enormous public health burden, especially in developing countries around the world. Cholera is a self-limiting illness; however, antibiotics are commonly administered as part of the treatment regimen. Here we review the initial identification and subsequent evolution of antibiotic-resistant strains of V. cholerae. Antibiotic resistance mechanisms, including efflux pumps, spontaneous chromosomal mutation, conjugative plasmids, SXT elements and integrons, are also discussed. Numerous multidrug-resistant strains of V. cholerae have been isolated from both clinical and environmental settings, indicating that antibiotic use has to be restricted and alternative methods for treating cholera have to be implemented.

https://www.ualberta.ca/~pukatzki/labpage/Publications_files/Maya_antibiotics_2011.pdf

Antibiotic resistance mechanisms of Vibrio cholerae.

The presence of SXT/R391-related integrating conjugative elements (ICEs) in bacterial strains isolated from fish obtained from marine aquaculture environments in 2001 to 2010 in the northwestern Iberian Peninsula was studied. ICEs were detected in 12 strains taxonomically related to Vibrio scophthalmi (3 strains), Vibrio splendidus (5 strains), Vibrio alginolyticus (1 strain), Shewanella haliotis (1 strain), and Enterovibrio nigricans (2 strains), broadening the known host range able to harbor SXT/R391-like ICEs. Variable DNA regions, which confer element-specific properties to ICEs of this family, were characterized. One of the ICEs encoded antibiotic resistance functions in variable region III, consisting of a tetracycline resistance locus. Interestingly, hot spot 4 included genes providing resistance to rifampin (ICEVspPor2 and ICEValPor1) and quaternary ammonium compounds (QACs) (ICEEniSpa1), and variable region IV included a mercury resistance operon (ICEVspSpa1 and ICEEniSpa1). The S exclusion group was more represented than the R exclusion group, accounting for two-thirds of the total ICEs. Mating experiments allowed ICE mobilization to Escherichia coli strains, showing the corresponding transconjugants' rifampin, mercury, and QAC resistance. These results show the first evidence of ICEs providing rifampin and QAC resistances, suggesting that these mobile genetic elements contribute to the dissemination of antimicrobial, heavy metal, and QAC resistance determinants in aquaculture environments.

Integrating Conjugative Elements as Vectors of Antibiotic, Mercury, and Quaternary Ammonium Compound Resistance in Marine Aquaculture Environments

A conjugative plasmid, pMRV150, which mediated multiple-drug resistance (MDR) to at least six antibiotics, including ampicillin, streptomycin, gentamicin, tetracycline, chloramphenicol, and trimethoprim-sulfamethoxazole, was identified in a Vibrio cholerae O139 isolate from Hangzhou, eastern China, in 2004. According to partial pMRV150 DNA sequences covering 15 backbone regions, the plasmid is most similar to pIP1202, an IncA/C plasmid in an MDR Yersinia pestis isolate from a Madagascar bubonic plague patient, at an identity of 99.99% (22,180/22,183 nucleotides). pMRV150-like plasmids were found in only 7.69% (1/13) of the O139 isolates tested during the early period of the O139 epidemic in Hangzhou (1994, 1996, and 1997); then the frequency increased gradually from 60.00% (3/5) during 1998 and 1999 to 92.16% (47/51) during 2000 to 2006. Most (42/51) of the O139 isolates bearing pMRV150-like plasmids were resistant to five to six antibiotics, whereas the plasmid-negative isolates were resistant only to one to three antibiotics. In 12 plasmid-bearing O139 isolates tested, the pMRV150-like plasmids ranged from approximately 140 kb to 170 kb and remained at approximately 1 or 2 copies per cell. High (4.50 × 10−2 and 3.08 × 10−2) and low (0.88 × 10−8 to 3.29 × 10−5) plasmid transfer frequencies, as well as no plasmid transfer (under the detection limit), from these O139 isolates to the Escherichia coli recipient were observed. The emergence of pMRV150-like or pIP1202-like plasmids in many bacterial pathogens and nonpathogens occupying diverse niches with global geographical distribution indicates an increasing risk to public health worldwide. Careful tracking of these plasmids in the microbial ecosystem is warranted.

Vibrio cholerae O139 Multiple-Drug Resistance Mediated by Yersinia pestis pIP1202-Like Conjugative Plasmids

Vibrio cholerae O139 Multidrug Resistance Mediated by Yersinia pestis pIP1202-like Conjugative Plasmids

We analyzed 28 epidemic Vibrio cholerae O1 strains isolated in the region of Thua Thien Hue (Vietnam) in 2003. Ubiquitous amoxicillin, prevalent aminoglycosides and sporadic erythromycin resistances were observed. All were devoid of plasmids, class 1 integrons and ICEs and showed the same BglI ribotype, irrespective of their site of isolation and resistance pattern. A strain isolated in 1990 in the same area was resistant to amoxicillin and aminoglycosides but characterized by a different ribotype. This strain contained ICEVchVie0, belonging to the SXT/R391 ICE family, devoid of any resistance cluster. The molecular analysis of three conserved and six variable regions outlined an original genetic profile. ICEs not coding for resistance to drugs seem to be more frequent than supposed, and this finding reinforces the idea that the SXT/R391 family of genetic elements is wide and composite. The clearance of ICEVchVie0 in the 2003 epidemic may be explained by the lack of any resistance determinant as a favorable selective marker.

Molecular characterization of ICEVchVie0 and its disappearance in Vibrio cholerae O1 strains isolated in 2003 in Vietnam

The genus Chlamydia comprises obligate intracellular bacteria that infect a wide variety of hosts, with infection leading to a range of diseases in humans and animals; they thus constitute a major public health threat. Among the members of the Chlamydiaceae family, Chlamydia suis, C. abortus, C. pecorum, and C. psittaci represent the most important pathogenic species infecting a large range of hosts and are a well-established threat to livestock. Information regarding the circulation of Chlamydia species in ruminants from Vietnam is lacking. In this study, DNA extracted from 60 blood samples collected from goats in Hue province was used for Chlamydia spp. identification by classic PCR and Sanger sequencing. Chlamydia spp. were detected in eleven samples (18.3%) and C. abortus and C. psittaci were molecularly identified by sequencing. Despite the limited sample size in this study, findings point out the relevance of ruminants as hosts of chlamydial species in Central Vietnam and the importance of monitoring chlamydial strains through the activation of surveillance programs in this country. The need for a deeper evaluation of human and animal health risk analysis in terms of chlamydiosis should be also considered.

/pdf/first-molecular-detection-of-zoonotic-chlamydia-species-in-1rhbj8ig.pdf

First Molecular Detection of Zoonotic Chlamydia Species in Vietnamese Goats

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Molecular characterization of ICEVchVie0 and its disappearance in Vibrio cholerae O1 strains isolated in 2003 in Vietnam

First Molecular Detection of Zoonotic Chlamydia Species in Vietnamese Goats