Construction and evaluation of a O139 Vibrio cholerae vaccine candidate based on a hemA gene mutation

Ravichandran, Manickam; Ali, Syed Atif; Rashid, Nur Haslindawaty Abdul; Kurunathan, S.; Yean, Chan Yean; Ting, Lai Chin; Bakar, Afifi Sheikh Abu; Lalitha, P.; Zafarina, Zainuddin

doi:10.1016/j.vaccine.2005.07.016

Cited by 15 publications

(14 citation statements)

References 34 publications

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“…Gross serosal hemorrhage (Figure 1) was also found to be similar with previous reports [6,[79][80][81][82][83]. The histopathological sections reflected trans mural congestion and excessive ulceration in mucosa and submucosa which also support the previous report [6,[84][85][86][87][88]. The detection of Vibrio cholerae was confirmed with all possible microbiological and molecular techniques.…”

Section: Discussionsupporting

confidence: 89%

“…The PCR was carried out by the method described previously [85] For the PCR confirmation of Vibrio cholerae, two reported primers (VHMF: 5' TGG GAG CAG CGT CCA TTG TG 3 and VHA-AS5: 5' CAA TCA CAC CAA GTC ACT C 3') specific for Vibrio cholerae were used. The PCR conformation was done using 1.2% agarose gel electrophoresis in TBE buffer.…”

Section: Molecular Confirmation Of Vibrio Choleraementioning

confidence: 99%

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Isolation of Vibrio cholerae in Homogenized Tissues of Liver, Gall Bladder and Bile in Rabbit Model

Abdullah

Aamenah³

et al. 2014

Interdiscip J Microinflammation

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Vibrio cholerae O139 is well known as the causative agent of cholera. It is noninvasive but many reports suggested it to cause bacteremia which brings in a little controversy with the previously reported old literature especially after reported histopathological invasion pattern of Vibrio cholerae O139 and Vibrio cholerae O1 ElTor. The rabbits were processes for ileac loops inoculation of Vibrio cholerae O139 followed by biochemical and molecular analysis of the presence of Vibrio cholerae O139 in liver, gall bladder and bile. We concluded the presence of Vibrio cholerae in liver, bile and gall bladder homogenized tissue. Retrograde spreading of bacteria to the liver via the common bile duct was excluded by complete closure of the intestinal lumen distant to the duct in the intestinal lumen. However, the bacteria might traffic, probably via macrophages, to distant area in the body including the liver. Another study in future is a need to identify the lesions of Vibrio cholerae invasions within liver and gall bladder by immunohistochemistry and using GFP visibility for Vibrio cholerae passage.

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Section: Discussionsupporting

confidence: 89%

Section: Molecular Confirmation Of Vibrio Choleraementioning

confidence: 99%

Isolation of Vibrio cholerae in Homogenized Tissues of Liver, Gall Bladder and Bile in Rabbit Model

Abdullah

Aamenah³

et al. 2014

Interdiscip J Microinflammation

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“…All together, these findings offer a comprehensive information related to kinetics and quality of immune response in this model and parallel our experience to that one from strain 638 [34]. Immunological profiles, however, were different from those reported by Ravichandran et al which could be related to the inoculation route used in each study [48].…”

Section: Discussionsupporting

confidence: 88%

TLP01, an mshA mutant of Vibrio cholerae O139 as vaccine candidate against cholera

Ledón

Ferrán

Pérez

et al. 2012

Microbes and Infection

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“…Such availability of ALA does not seem to be universal in host-associated bacteria. For example, a hemA mutant of Vibrio cholerae was significantly (10-fold) attenuated in colonizing the infant mouse gut (57), and a Staphylococcus aureus hemA mutant was similarly reduced in virulence and colonization of hearts and livers in a mouse model (58). In the case of V. fischeri, symbionts in the light organ are provisioned with many amino acids and peptides from the host (59), which may prime cells for uptake of ALA via peptide transport systems (60).…”

Section: Resultsmentioning

confidence: 99%

An Expanded Transposon Mutant Library Reveals that Vibrio fischeri δ-Aminolevulinate Auxotrophs Can Colonize Euprymna scolopes

Lyell

Septer

Dunn

et al. 2017

Appl Environ Microbiol

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Libraries of defined mutants are valuable research tools but necessarily lack gene knockouts that are lethal under the conditions used in library construction. In this study, we augmented a Vibrio fischeri mutant library generated on a rich medium (LBS, which contains [per liter] 10 g of tryptone, 5 g of yeast extract, 20 g of NaCl, and 50 mM Tris [pH 7.5]) by selecting transposon insertion mutants on supplemented LBS and screening for those unable to grow on LBS. We isolated strains with insertions in alr, glr (murI), glmS, several heme biosynthesis genes, and ftsA, as well as a mutant disrupted 14 bp upstream of ftsQ. Mutants with insertions in ftsA or upstream of ftsQ were recovered by addition of Mg 2ϩ to LBS, but their cell morphology and motility were affected. The ftsA mutant was more strongly affected and formed cells or chains of cells that appeared to wind back on themselves helically. Growth of mutants with insertions in glmS, alr, or glr was recovered with N-acetylglucosamine (NAG), D-alanine, or D-glutamate, respectively. We hypothesized that NAG, D-alanine, or D-glutamate might be available to V. fischeri in the Euprymna scolopes light organ; however, none of these mutants colonized the host effectively. In contrast, hemA and hemL mutants, which are auxotrophic for ␦-aminolevulinate (ALA), colonized at wild-type levels, although mutants later in the heme biosynthetic pathway were severely impaired or unable to colonize. Our findings parallel observations that legume hosts provide Bradyrhizobium symbionts with ALA, but they contrast with virulence phenotypes of hemA mutants in some pathogens. The results further inform our understanding of the symbiotic light organ environment.IMPORTANCE By supplementing a rich yeast-based medium, we were able to recover V. fischeri mutants with insertions in conditionally essential genes, and further characterization of these mutants provided new insights into this bacterium's symbiotic environment. Most notably, we show evidence that the squid host can provide V. fischeri with enough ALA to support its growth in the light organ, paralleling the finding that legumes provide Bradyrhizobium ALA in symbiotic nodules. Taken together, our results show how a simple method of augmenting already rich media can expand the reach and utility of defined mutant libraries.KEYWORDS Photobacterium, Aliivibrio, symbiosis, aminolevulinic acid, hemin, photobacteria D efined knockout mutant libraries are useful resources that have been generated to promote research in several bacterial experimental models and pathogens (1-10). In such studies, researchers have often listed essential genes, the knockouts of which were apparently lethal under the conditions of library construction and therefore not represented by mutants in the library. In order to recover insertion mutants collectively

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Construction and evaluation of a O139 Vibrio cholerae vaccine candidate based on a hemA gene mutation

Cited by 15 publications

References 34 publications

Isolation of Vibrio cholerae in Homogenized Tissues of Liver, Gall Bladder and Bile in Rabbit Model

Isolation of Vibrio cholerae in Homogenized Tissues of Liver, Gall Bladder and Bile in Rabbit Model

TLP01, an mshA mutant of Vibrio cholerae O139 as vaccine candidate against cholera

An Expanded Transposon Mutant Library Reveals that Vibrio fischeri δ-Aminolevulinate Auxotrophs Can Colonize Euprymna scolopes

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