Characterisation of a Listeria monocytogenes mutant deficient in D-arabitol fermentation

Saklani-Jusforgues, Hélène; Fontan, Élisabeth; Goossens, Pierre L.

doi:10.1016/s0923-2508(01)01189-5

Cited by 10 publications

(15 citation statements)

References 5 publications

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“…Despite the fact that the genome sequence indicated that this operon was important for dulcitol transport, L. monocytogenes could not grow on dulcitol. Rather, our data as well as those of Saklani-Jusforgues et al (49) suggest that this operon is important for arabitol transport. The growth of this mutant in an aqueous radish suspension resulted in a generation time equivalent to that of the wild type at 30°C, so we concluded that the defect in attachment at 30°C and the slight defect at 10°C were not the result of a growth defect during the course of the assay.…”

Section: Discussionsupporting

confidence: 75%

“…However, we determined that even wild-type L. monocytogenes does not grow on dulcitol, either as a sole carbon and energy source or cometabolized in TSYE medium. Saklani-Jusforgues et al (49) reported that an L. monocytogenes Tn917lac insertion mutant was incapable of growth on the sugar arabitol and had a disruption in the dulcitol transport operon, specifically in a gene three ORFs downstream from the gene disrupted by ⍀003 (25, 41). The ⍀003 strain was incapable of utilizing arabitol as a sole carbon and energy source, nor could it cometabolize arabitol as a supplement to TSYE medium.…”

Section: Attachment Of L Monocytogenes Strains To Radish Tissuementioning

confidence: 99%

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Attachment of Listeria monocytogenes to Radish Tissue Is Dependent upon Temperature and Flagellar Motility

Gorski

Palumbo

Mandrell

2003

Appl Environ Microbiol

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Outbreaks of listeriosis and febrile gastroenteritis have been linked to produce contamination by Listeria monocytogenes. In order to begin to understand the physiology of the organism in a produce habitat, the ability of L. monocytogenes to attach to freshly cut radish tissue was examined. All strains tested had the capacity to attach sufficiently well such that they could not be removed during washing of the radish slices. A screen was developed to identify Tn917-LTV3 mutants that were defective in attachment to radish tissue, and three were characterized. Two of the three mutations were in genes with unknown functions. Both of the unknown genes mapped to a region predicted to contain genes necessary for flagellar export; however, only one of the two insertions caused a motility defect. The third insertion was found to be in an operon encoding a phosphoenolpyruvate-sugar phosphotransferase system. All three mutants were defective in attachment when tested at 30°C; the motility mutant had the most severe phenotype. However, not all of the mutants were defective when tested at other temperatures. These results indicate that L. monocytogenes may use different attachment factors at different temperatures and that temperature should be considered an important variable in studies of the molecular mechanisms of Listeria fitness in complex environments.Listeria monocytogenes is a gram-positive pathogen that causes a multitude of symptoms, including septicemia, abortion, liver failure, and meningitis (50). When L. monocytogenes is not living as an intracellular invader, it can be isolated from the feces of animals as well as humans and from the soil, where it can survive as a saprophyte for 10 to 12 years. From these locations, it can contaminate the food supply in many ways. It can survive in sewage and enter the soil, where it may become associated with plants and/or farm animals before entering food-processing plants, grocery stores, and home refrigerators (2, 10). In all of these niches, L. monocytogenes can survive and even thrive, leading to contamination of foods and ingestion of the pathogen. L. monocytogenes is quite hardy under a variety of environmental stresses. It has been reported that it can grow at temperatures of between Ϫ0.4 and 50°C and withstand extremes of osmotic pressure, as evidenced by growth in 10% NaCl, and it displays a pH range of 4.3 to 9 (18, 21).While food contamination with L. monocytogenes is most often associated with smoked meats and dairy products, fresh produce has also been implicated in outbreaks and sporadic cases of listeriosis. Specific produce-related outbreaks of listeriosis and febrile gastroenteritis caused by L. monocytogenes have been associated with contaminated cabbage, corn, and lettuce and/or celery (8,30,51). In addition, this organism has caused the recall of vegetables, including red peppers, sprouts, and lettuce, and potato salad (4-7, 13). L. monocytogenes has been isolated from radishes, potatoes, cucumbers, and cabbage in surveys of produce from U.S. grocery s...

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Section: Discussionsupporting

confidence: 75%

Section: Attachment Of L Monocytogenes Strains To Radish Tissuementioning

confidence: 99%

Attachment of Listeria monocytogenes to Radish Tissue Is Dependent upon Temperature and Flagellar Motility

Gorski

Palumbo

Mandrell

2003

Appl Environ Microbiol

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“…Auxotrophic mutations successfully applied for virulence attenuation in other bacterial carrier systems were also used for obtaining attenuated L. monocytogenes strains (1,29,45,47). However, these mutations either led to insufficient virulence attenuation (1,29,45) or required coinjection of the impaired metabolite for the induction of an efficient immune response against L. monocytogenes (47).…”

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confidence: 99%

Growth, Virulence, and Immunogenicity ofListeria monocytogenes aroMutants

et al. 2004

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Mutants of Listeria monocytogenes with deletions in genes of the common branch of the biosynthesis pathway leading to aromatic compounds were constructed as possible virulence-attenuated carrier strains for protein antigens or vaccine DNA. aroA, aroB, and in particular aroE mutants showed strongly reduced growth rates in epithelial cells and even in rich culture media. The metabolism of the aro mutants under these conditions was predominantly anaerobic. Aerobic metabolism and a wild-type growth rate were, however, regained upon the addition of vitamin K 2 , suggesting that the aro mutants are deficient in oxidative respiration due to the lack of menaquinone. Replication of the aro mutants in the host cell's cytosol and cell-to-cell spread were drastically slowed down, and all aro mutants showed high virulence attenuation in mice, i.e., the 50% lethal dose in BALB/c mice was increased at least 10 4 -fold for the aroA, aroB, and aroA/B mutants and >10 5 -fold for the aroE mutant compared to the parent strain. Nevertheless, mice preimmunized with aro mutant bacteria elicited good T-cell response and full protection against a subsequent challenge with the virulent wild-type strain. A total of 5 ؋ 10 6 aroA, aroB, and aroA/B mutant bacteria were sufficient to obtain a protective T-cell response, while 5 ؋ 10 8 aroE or aroA/E mutants were necessary to achieve comparable numbers of antigen-specific T cells. These numbers were well tolerated without causing any signs of disease, indicating that Listeria strains with deletions in genes of the basic branch of the aromatic amino acid pathway could be useful vaccine carriers for inducing T-cell immunity.

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“…In a variety of bacteria, such as Bacillus species, Streptococcus species, enterococci, Listeria monocytogenes, and staphylococci, Tn917 was used to identify genes related to characteristic phenotypes or metabolic pathways (15,25,36,46,49,51,55). In the era before the sequencing of complete bacterial genomes, transposon insertion sites were analyzed by cloning or inverted PCR requiring the effort of chromosomal mapping.…”

mentioning

confidence: 99%

Establishment of an Arbitrary PCR for Rapid Identification of Tn 917 Insertion Sites in Staphylococcus epidermidis : Characterization of Biofilm-Negative and Nonmucoid Mutants

Knobloch

Nedelmann

Kiel

et al. 2003

Appl Environ Microbiol

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Transposon mutagenesis with the Enterococcus faecalis transposon Tn917 is a genetic approach frequently used to identify genes related with specific phenotypes in gram-positive bacteria. We established an arbitrary PCR for the rapid and easy identification of Tn917 insertion sites in Staphylococcus epidermidis with six independent, well-characterized biofilm-negative Tn917 transposon mutants, which were clustered in the icaADBC gene locus or harbor Tn917 in the regulatory gene rsbU. For all six of these mutants, short chromosomal DNA fragments flanking both transposon ends could be amplified. All fragments were sufficient to correctly identify the Tn917 insertion sites in the published S. epidermidis genomes. By using this technique, the Tn917 insertion sites of three not-yet-characterized biofilm-negative or nonmucoid mutants were identified. In the biofilm-negative and nonmucoid mutant M12, Tn917 is inserted into a gene homologous to the regulatory gene purR of Bacillus subtilis and Staphylococcus aureus. The Tn917 insertions of the nonmucoid but biofilm-positive mutants M16 and M20 are located in genes homologous to components of the phosphoenolpyruvate-sugar phosphotransferase system (PTS) of B. subtilis, S. aureus, and Staphylococcus carnosus, indicating an influence of the PTS on the mucoid phenotype in S. epidermidis.

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Characterisation of a Listeria monocytogenes mutant deficient in D-arabitol fermentation

Cited by 10 publications

References 5 publications

Attachment of Listeria monocytogenes to Radish Tissue Is Dependent upon Temperature and Flagellar Motility

Attachment of Listeria monocytogenes to Radish Tissue Is Dependent upon Temperature and Flagellar Motility

Growth, Virulence, and Immunogenicity ofListeria monocytogenes aroMutants

Establishment of an Arbitrary PCR for Rapid Identification of Tn 917 Insertion Sites in Staphylococcus epidermidis : Characterization of Biofilm-Negative and Nonmucoid Mutants

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