Purification and Characterization of Enterotoxigenic El Tor-Like Hemolysin Produced by<i>Vibrio fluvialis</i>

Kothary, Mahendra H.; Lowman, Heather; McCardell, Barbara A.; Tall, Ben D.

doi:10.1128/iai.71.6.3213-3220.2003

Cited by 52 publications

(64 citation statements)

References 31 publications

(16 reference statements)

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“…In this study, we showed that the crude toxin (culture supernate) has both cytotoxic and vacuolating effects on HeLa cells. Our study also supports the fact that the nucleotide sequence of the V. fluvialis haemolysin gene shares significant homology with that of the El Tor haemolysin of V. cholerae (Kothary et al, 2003). However, it is still not clear whether the El Tor-like haemolysin of V. fluvialis has any correlation with vacuolation effects on HeLa cells similar to V. cholerae.…”

Section: R Chakraborty and Otherssupporting

confidence: 69%

“…The effect of culture supernate and cell lysate of all the V. fluvialis strains was also tested with CHO cells. The cytotoxic effect that was detected in the CHO cells treated with the culture supernate was possibly due to the presence of V. fluvialis haemolysin (Kothary et al, 2003). None of our strains was found to contain the cell elongation factor in their cell lysate as reported earlier (Lockwood et al, 1982).…”

supporting

confidence: 57%

“…Primer pairs targeting ctxA and tcpA (classical and El Tor biotypes of V. cholerae O1) were used in a multiplex PCR format as described elsewhere (Keasler & Hall, 1993). PCR assays were performed to detect the presence of V. cholerae regulatory genes toxR (Miller et al, 1987) and toxT (Carroll et al, 1997), and other genes like aldA, tagA, int (Kovach et al, 1996;Karaolis et al, 1998;Mitra et al, 2001), hlyA (El Tor haemolysin) specific for V. cholerae (Manning et al, 1984;Alm et al, 1988; Mitra et al, 2000), V. fluvialis haemolysin gene (Kothary et al, 2003) and stn (Ogawa et al, 1990). The species-specific ompW gene of V. cholerae was used in a PCR assay to confirm that the strains identified as V. fluvialis were not V. cholerae.…”

Section: Methodsmentioning

confidence: 99%

“…Since the culture supernate of V. fluvialis strains failed to evoke fluid accumulation in the suckling mice assay, it can be assumed that none of the strains produced heatstable toxin. Interestingly, the V. fluvialis haemolysin, a heatlabile toxin, can induce fluid accumulation in suckling mice in a dose-dependent manner (Kothary et al, 2003). For this reason, with the tested strains we consider that the amount of haemolysin present in the culture supernate was not sufficient to elicit fluid accumulation.…”

mentioning

confidence: 98%

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Cytotoxic and cell vacuolating activity of Vibrio fluvialis isolated from paediatric patients with diarrhoea

Chakraborty

et al. 2005

Journal of Medical Microbiology

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Vibrio fluvialis is a halophilic Vibrio species associated with acute diarrhoeal illness in humans. It has the potential to cause outbreaks and has an association with paediatric diarrhoea. In this study, 11 V. fluvialis strains isolated from hospitalized patients with acute diarrhoea at the Infectious Diseases Hospital, Kolkata were extensively characterized. All the strains showed growth in peptone broth containing 7 % NaCl. The strains showed variable results in Voges-Proskauer test and to a vibriostatic agent. There was also variation in their antibiograms, and some of the strains were multidrug resistant. Among the 11 strains, two showed only a single band difference in their PFGE profile and the remaining strains showed nine different PFGE patterns. However, unlike PFGE, the strains exhibited close matches and clustering in their ribotype patterns. The haemolytic effect on sheep red blood cells varied with strains. Partial sequence analysis revealed that the V. fluvialis haemolysin gene has 81 % homology with that of the El Tor haemolysin of Vibrio cholerae. A striking finding was the capability of all the strains to evoke distinct cytotoxic and vacuolation effects on HeLa cells. INTRODUCTIONDuring the late 1970s, a previously unidentified group of vibrios was isolated from diarrhoeic stools of infants, children and young adults in Bangladesh (Huq et al., 1980). These vibrios were designated 'group F' vibrios in England (Furniss et al., 1977) and 'CDC group EF6' in the USA (Huq et al., 1980). Taxonomic studies concluded that these organisms represented a new species, and they were designated as Vibrio fluvialis (Lee et al., 1981).The distribution of V. fluvialis is worldwide (McNicol et al., 1980;Thekdi et al., 1982) and this organism is not only isolated from human diarrhoeal cases (Huq et al., 1980;Tacket et al., 1982;Thekdi et al., 1982;Hickman-Brenner et al., 1984;Klontz et al., 1994;Hodge et al., 1995) but also from marine and estuarine environments (Seidler et al., 1980;Lee et al., 1981;Morris & Black, 1985). There are reports of food poisoning caused by this organism (Kobayashi & Ohnaka, 1989;Thekdi et al., 1990), especially due to consumption of raw shellfish (Levine & Griffin, 1993). V. fluvialis is also associated with extraintestinal infections (Yoshii et al., 1987;Albert et al., 1991). The clinical symptoms of the disease include mild to moderate dehydration, vomiting, fever, abdominal pain and diarrhoea (Seidler et al., 1980). The halophilic V. fluvialis (Lee et al., 1981;Lockwood et al., 1982) phenotypically resembles Aeromonas species (Seidler et al., 1980) and taxonomically lies between Aeromonas and Vibrio species (Thekdi et al., 1990). Among the halophilic vibrios, it has close similarity to V. furnissii, but, unlike V. fluvialis, V. furnissii is aerogenic in nature (Brenner et al., 1983).Since V. fluvialis enteritis is reported infrequently, the epidemiology of this infection is not adequately understood. There is very little information available on the virulence factors associated wit...

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Section: R Chakraborty and Otherssupporting

confidence: 69%

supporting

confidence: 57%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 98%

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Cytotoxic and cell vacuolating activity of Vibrio fluvialis isolated from paediatric patients with diarrhoea

Chakraborty

et al. 2005

Journal of Medical Microbiology

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“…Vibrio species have been incriminated in cases of diarrhoea, accounting for a substantial degree of morbidity and mortality in different age groups worldwide (Obi et al 2004). The most notable of Vibrio pathogens are V. cholerae, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio fluvialis (CDC Centers for Disease Control and Prevention 1999; Finkelstein et al 2002;Kothary et al 2003;Chakraborty et al 2006) which are mainly transmitted via water and food. They all cause diarrhoea, but in entirely different ways; V. vulnificus and V. parahaemolyticus are invasive organisms, affecting primarily the colon, while V. cholerae is non-invasive, affecting the small intestine through secretion of an enterotoxin (Todar 2005), and is the etiologic agent of cholera.…”

Section: Introductionmentioning

confidence: 99%

Prevalence and Characterisation of Non-Cholerae Vibrio spp. in Final Effluents of Wastewater Treatment Facilities in Two Districts of the Eastern Cape Province of South Africa: Implications for Public Health

Okoh¹,

Sibanda²,

Nongogo³

et al. 2015

Wastewater and Public Health

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Vibrios and other enteric pathogens can be found in wastewater effluents of a healthy population. We assessed the prevalence of three non-cholerae vibrios in wastewater effluents of 14 wastewater treatment plants (WWTP) in Chris Hani and Amathole district municipalities in the Eastern Cape Province of South Africa for a period of 12 months. With the exception of WWTP10 where presumptive vibrios were not detected in summer and spring, presumptive vibrios were detected in all seasons in other WWTP effluents. When a sample of 1,000 presumptive Vibrio isolates taken from across all sampling sites were subjected to molecular confirmation for Vibrio, 668 were confirmed to belong to the genus Vibrio, giving a prevalence rate of 66.8 %. Further, molecular characterisation of 300 confirmed Vibrio isolates revealed that 11.6 % (35) were Vibrio parahaemolyticus, 28.6 % (86) were Vibrio fluvialis and 28 % (84) were Vibrio vulnificus while 31.8 % (95) belonged to other Vibrio spp. not assayed for in this study. Antibiogram profiling of the three Vibrio species showed that V. parahaemolyticus was ≥50 % susceptible to 8 of the test antibiotics and ≥50 % resistant to only 5 of the 13 test antibiotics, while V. vulnificus showed a susceptibility profile of ≥50 % to 7 of the test antibiotics and a resistance profile of ≥50 % to 6 of the 13 test antibiotics. V. fluvialis showed ≥50 % resistance to 8 of the 13 antibiotics used while showing ≥50 % susceptibility to only 4 antibiotics used. All three Vibrio species were susceptible to gentamycin, cefuroxime, meropenem and imipenem. Multiple antibiotic resistance patterns were also evident especially against such antibiotics as tetracyclin, polymixin B, penicillin G, sulfamethazole and erythromycin against which all Vibrio species were resistant. These results indicate a significant threat to public health, more so in the Eastern Cape Province of South Africa which is characterised by widespread poverty, with more than a third of the population directly relying on surface water sources for drinking and daily use.

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Bacterial Foodborne Disease

Green

Furuno

Horneman

et al. 2009

Bacterial Infections of Humans

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Purification and Characterization of Enterotoxigenic El Tor-Like Hemolysin Produced byVibrio fluvialis

Cited by 52 publications

References 31 publications

Cytotoxic and cell vacuolating activity of Vibrio fluvialis isolated from paediatric patients with diarrhoea

Cytotoxic and cell vacuolating activity of Vibrio fluvialis isolated from paediatric patients with diarrhoea

Prevalence and Characterisation of Non-Cholerae Vibrio spp. in Final Effluents of Wastewater Treatment Facilities in Two Districts of the Eastern Cape Province of South Africa: Implications for Public Health

Bacterial Foodborne Disease

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