Evaluation of Genotypic and Phenotypic Methods To Distinguish Clinical from Environmental
            <i>Vibrio vulnificus</i>
            Strains

Sanjuán, Eva; Fouz, Belén; Oliver, James D.; Amaro, Carmen

doi:10.1128/aem.01594-08

Cited by 54 publications

(49 citation statements)

References 37 publications

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“…Previous studies linking clinical strains to genetic sequences including 16S rRNA type, vvhA, vcg, and vvpdh ultimately have concluded that variation in these factors is not exclusive to clinical strains (Table S1) (28). There is a strong correlation between a lineage I MLST and human strains, but human isolates also derive from within lineage II (Fig.…”

Section: Discussionmentioning

confidence: 86%

Vibrio vulnificus rtxA1 gene recombination generates toxin variants with altered potency during intestinal infection

Kwak

Jeong

Satchell

2011

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

143

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Vibrio vulnificus is a food-borne bacterial pathogen associated with 1% of all food-related deaths, predominantly because of consumption of contaminated seafood. The ability of V. vulnificus to cause disease is linked to the production of a large cytotoxin called the "multifunctional-autoprocessing RTX" (MARTX Vv ) toxin, a factor shown here to be an important virulence factor by the intragastric route of infection in mice. In this study, we examined genetic variation of the rtxA1 gene that encodes MARTX Vv in 40 V. vulnificus Biotype 1 strains and found four distinct variants of rtxA1 that encode toxins with different arrangements of effector domains. We provide evidence that these variants arose by recombination either with rtxA genes carried on plasmids or with the rtxA gene of Vibrio anguillarum. Contrary to expected results, the most common rtxA1 gene variant in clinical-type V. vulnificus encodes a toxin with reduced potency and is distinct from the toxin produced by strains isolated from market oysters. These results indicate that an important virulence factor of V. vulnificus is undergoing significant genetic rearrangement and may be subject to selection for reduced virulence in the environment. This finding would imply further that in the future on-going genetic variation of the MARTX Vv toxins could result in the emergence of novel strains with altered virulence in humans.phylogeny | RTX | oyster T he gram-negative bacterial pathogen Vibrio vulnificus inhabits coastal waters including the US Gulf Coast. The pathogen causes gastroenteritis, primary septicemia, and necrotizing fasciitis and can be difficult to treat. The bacteria are particularly rapid growers in vivo and are highly invasive; thus death can occur as quickly as 24-48 h after ingestion (1, 2). In the United States from 1998-2008, 985 cases of V. vulnificus infection resulting in 91 deaths were reported to the Centers for Disease Control. Of these cases, 60% were caused by food-borne illness, particularly associated with the consumption of raw seafood (3). In fact, V. vulnificus accounts for 1% of all food-related deaths in the United States (4). Numerous studies recently have described a V. vulnificus multifunctional-autoprocessing RTX toxin (MARTX Vv ) as having a major impact on virulence in mice (5-8). Toxins of the MARTX family are very large composite toxins. The 5,206-amino acid MARTX Vv toxin as found in the two sequenced strains of V. vulnificus, Korean isolate CMCP6 and Taiwanese isolate YJ016 (9, 10), are 99% identical. Similar to all MARTX toxins, the majority of MARTX Vv is comprised of conserved repeat regions at the N and C termini that are proposed to form a pore in the eukaryotic cell plasma membrane for translocation of the central portion of the secreted toxin to the cytosol (11). The central region includes a conserved cysteine protease domain (CPD) that is required for inositol hexakisphosphate-induced autoprocessing. After translocation, autoprocessing at leucine residues in unstructured regions between the effector do...

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Section: Discussionmentioning

confidence: 86%

Vibrio vulnificus rtxA1 gene recombination generates toxin variants with altered potency during intestinal infection

Kwak

Jeong

Satchell

2011

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

143

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“…Until now, biotype 1 has been considered non-pathogenic to eels . A great variety of biotype 1 strains was also found by Sanjuán et al (2009) based on many isolates of V. vulnificus. They recommended using molecular typing to study the epidemiology of V. vulnificus infections (Sanjuán & Amaro 2007, Sanjuán et al 2009).…”

Section: Discussionmentioning

confidence: 96%

“…A great variety of biotype 1 strains was also found by Sanjuán et al (2009) based on many isolates of V. vulnificus. They recommended using molecular typing to study the epidemiology of V. vulnificus infections (Sanjuán & Amaro 2007, Sanjuán et al 2009). Although biotype 1 was considered to be primarily pathogenic to humans and mainly foodborne (Tison et al 1982, Hsueh et al 2004, based on our results of hiMLST, we found no identical published human strains in the database (http://pubmlst.org/ vvulnificus).…”

Section: Discussionmentioning

confidence: 96%

“…Rapid, sensitive and accurate identification methods for zoonotic V. vulnificus strains were needed (Sanjuán & Amaro 2007, Sanjuán et al 2009). Sanjuán et al (2011) reported that sequence-based typing of 115 Vibrio vulnificus strains of biotypes 1, 2 and 3 subdivided them into 3 phylogenetic lineages which did not correspond to biotypes, with biotype 2 being polyphyletic.…”

Section: Introductionmentioning

confidence: 99%

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Vibrio vulnificus outbreaks in Dutch eel farms since 1996: strain diversity and impact

Haenen

Zanten²,

Jansen³

et al. 2014

Dis. Aquat. Org.

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Vibrio vulnificus is a potentially zoonotic bacterial pathogen of fish, which can infect humans (causing necrotic fasciitis). We analysed 24 V. vulnificus isolates (from 23 severe eel disease outbreaks in 8 Dutch eel farms during 1996 to 2009, and 1 clinical strain from an eel farmer) for genetic correlation and zoonotic potential. Strains were typed using biotyping and molecular typing by high-throughput multilocus sequence typing (hiMLST) and REP-PCR (Diversilab ® ). We identified 19 strains of biotype 1 and 5 of biotype 2 (4 from eels, 1 from the eel farmer), that were subdivided into 8 MLST types (ST) according to the international standard method. This is the first report of V. vulnificus biotype 1 outbreaks in Dutch eel farms. Seven of the 8 STs, of unknown zoonotic potential, were newly identified and were deposited in the MLST database. The REP-PCR and the MLST were highly concordant, indicating that the REP-PCR is a useful alternative for MLST. The strains isolated from the farmer and his eels were ST 112, a known potential zoonotic strain. Antimicrobial resistance to cefoxitin was found in most of the V. vulnificus strains, and an increasing resistance to quinolones, trimethoprim + sulphonamide and tetracycline was found over time in strain ST 140. Virulence testing of isolates from diseased eels is recommended, and medical practitioners should be informed about the potential risk of zoonotic infections by V. vulnificus from eels for the prevention of infection especially among high-risk individuals. Additional use of molecular typing methods such as hiMLST and Diversilab ® is recommended for epidemiological purposes during V. vulnificus outbreaks.

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“…When performing health status evaluations, considering correct identification and treatment of bacterial diseases and conditions, of (primarily) farmed crayfish, speed is always an issue. Rapid identification of environmental bacteria via commercial phenotypic tests allows for a wide choice of tests selection, and API 20E (Biomerieux, Marcy l'Etoile, France), an identification system for Enterobacteriaceae and other non-fastidious Gram-negative rods developed for clinical specimens, seems to be increasingly used for the identification of aquatic pathogens (Topić Popović et al, 2007;Sanjuán et al, 2009;Bastardo et al, 2012;Esteve et al, 2012;Soto et al, 2012). However, due to several shortcomings of this system, such as the wrong identifications and the need of comparison with the diagnostic schemes based on reactions in conventional phenotypic tests, more advanced methods for identification are sought after.…”

Section: Introductionmentioning

confidence: 99%

High-throughput discrimination of bacteria isolated fromAstacus astacusandA. leptodactylus

Popović

Klobučar

Maguire

et al. 2014

Knowl. Managt. Aquatic Ecosyst.

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Key-words: crayfish, bacteria, MALDI-TOF MS, API 20EBacterial diseases and pathogens of crayfish are common, widespread, and occasionally causing serious mortalities. In order to take rapid measures for correct treatment of crayfish diseases, the turnover time and accuracy in bacterial identification is an issue. Bacteria isolated from tissues of apparently healthy Astacus astacus and A. leptodactylus were identified by the commercial phenotypic tests (API 20E) and by the matrix assisted laser induced desorption ionization connected to the time of flight mass spectrometry (MALDI-TOF MS). For Gram-negative rods, API 20E resulted in fewer species identifications than MALDI-TOF MS (5.2% versus 52.61%). The most frequently identified genus from A. astacus and A. leptodactylus was Pseudomonas spp.: API 20E (47.82%) and MALDI-TOF MS (52.17%). Both systems identified 60.86% of total isolates identically to the genus. Hafnia alvei was the only isolate for which API 20E and MALDI-TOF MS had a concordant reading to the species. MALDI-TOF MS proved to be a powerful, low-cost, rapid tool in bacterial genus identification. This is the first report of a direct comparison between the two systems for the identification of bacteria in crayfish, and also the first report on using MALDI-TOF MS for discrimination of freshwater crayfish bacterial isolates. ont identifié 60,86 % des isolats totaux au même genre. Hafnia alvei était le seul isolat dont API 20E et MALDI-TOF MS ont une lecture concordante à l'espèce. MALDI-TOF MS s'est avéré être un outil rapide, puissant, à faible coût, pour l'identification au niveau du genre bactérien. Ce travail est le premier d'une comparaison directe entre les deux systèmes pour l'identification des bactéries chez les écrevisses, et aussi le premier travail sur l'utilisation de MALDI-TOF MS pour la discrimination des isolats bactériens d'écrevisses d'eau douce.

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Evaluation of Genotypic and Phenotypic Methods To Distinguish Clinical from Environmental Vibrio vulnificus Strains

Cited by 54 publications

References 37 publications

Vibrio vulnificus rtxA1 gene recombination generates toxin variants with altered potency during intestinal infection

Vibrio vulnificus rtxA1 gene recombination generates toxin variants with altered potency during intestinal infection

Vibrio vulnificus outbreaks in Dutch eel farms since 1996: strain diversity and impact

High-throughput discrimination of bacteria isolated fromAstacus astacusandA. leptodactylus

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