Biofilm formation and exopolysaccharide (EPS) production by Cronobacter sakazakii depending on environmental conditions

Jung, Jinho; Choi, Na-Young; Lee, Sun-Young

doi:10.1016/j.fm.2012.11.008

Cited by 81 publications

(36 citation statements)

References 26 publications

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“…Comparison of the amount of proteins and saccharides after 5 and 7 days of growth on cellulose support showed that: (1) proteins were more abundant than carbohydrates in all media except YEM, where polysaccharides predominated; (2) the absolute amount of proteins and carbohydrates increased from day 5 to day 7; and (3) the relative percentages of the two compounds did not change from day 5 to day 7. These experiments not only showed a medium-dependent biosynthetic behaviour for B. cenocepacia BTS2, as similarly reported for other bacterial species (Kives et al, 2006;Chen et al, 2013;Jung et al, 2013;Moryl et al, 2014), but also evidenced differences related to the support used. Since the biomass produced on YEM was different from those developed on all other media, some experiments were conducted using only YEM and MH media.…”

Section: Biofilm Formed On Cellulose Membranessupporting

confidence: 86%

Biofilms produced by Burkholderia cenocepacia: influence of media and solid supports on composition of matrix exopolysaccharides

et al. 2016

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Section: Biofilm Formed On Cellulose Membranessupporting

confidence: 86%

Biofilms produced by Burkholderia cenocepacia: influence of media and solid supports on composition of matrix exopolysaccharides

et al. 2016

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“…In general, LPS and various other polysaccharides, such as CPS and EPS, have been identified as major virulence factors. They contribute to capsule and biofilm formation of Gram-negative pathogens (48,49) and are essential for the persistence of C. sakazakii in harsh environments (30). The highly varied polysaccharide structures are strong immunogens showing a high degree of antigenic variation and are often targets for the production of antibodies.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies

Scharinger

Dietrich

Kleinsteuber

et al. 2016

Appl Environ Microbiol

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Cronobacter sakazakii is a foodborne pathogen associated with rare but often lethal infections in neonates. Powdered infant formula (PIF) represents the most frequent source of infection. Out of the identified serotypes (O1 to O7), O1, O2, and O3 are often isolated from clinical and PIF samples. Serotype-specific monoclonal antibodies (MAbs) suitable for application in enzyme immunoassays (EIAs) for the rapid detection of C. sakazakii have not yet been developed. In this study, we created specific MAbs with the ability to bind to C. sakazakii of serotypes O1, O2, and O3. Characterization by indirect EIAs, immunofluorescence, motility assays, and immunoblotting identified lipopolysaccharide (LPS) and exopolysaccharide (EPS) as the antigenic determinants of the MAbs. The established sandwich EIAs were highly sensitive and were able to detect between 2 ؋ 10 3 and 9 ؋ 10 6 CFU/ml. Inclusivity tests confirmed that 93% of serotype O1 strains, 100% of O2 strains, and 87% of O3 strains were detected at low cell counts. No cross-reactivity with >100 strains of Cronobacter spp. and other Enterobacteriaceae was observed, except for that with C. sakazakii serotype O3 and Cronobacter muytjensii serotype O1. Moreover, the sandwich EIAs detected C. sakazakii in PIF samples artificially contaminated with 1 to 10 bacterial cells per 10 g of sample after 15 h of preenrichment. The use of these serotype-specific MAbs not only allows the reliable detection of C. sakazakii strains but also enables simultaneous serotyping in a simple sandwich EIA method. Cronobacter spp. are Gram-negative opportunistic foodborne pathogens of the family Enterobacteriaceae that cause rare but severe infections in patients of all age groups. In adults, Cronobacter spp. are often associated with nosocomial infections, including pneumonia, septicemia, wound infections, and osteomyelitis, while causing invasive disease in young infants and neonates (1-4). Among the seven identified Cronobacter species, C. sakazakii, C. malonaticus, C. muytjensii, C. turicensis, C. dublinensis, C. condimenti, and C. universalis (5-8), C. sakazakii plays a prominent role due to it causing life-threatening infections in neonates (9-11). Clinically manifested infections present as necrotizing enterocolitis, sepsis, and meningitis, with a mortality rate as high as 80% (1, 12, 13). Although C. sakazakii has been isolated from a variety of different plant-and animal-based food products (14, 15), the presence in powdered infant formula (PIF) seems crucial in the infection of neonates (9, 12, 16). According to an established O-antigen serotyping scheme based on rabbit antisera and a PCRbased serotyping method (17-21), seven serotypes (O1 to O7) have been identified for C. sakazakii. Serotypes O1 and O2 seem to be most prevalent in PIF samples and in clinical cases, whereas serotype O3 has been isolated quite frequently from PIF but not as often from clinical cases (19,(22)(23)(24).Today, the contamination of PIF by C. sakazakii is being detected using conventional microbiological m...

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“…Some bacteria exist as sessile forms only in rich environments, e.g. EPS production by Cronobacter sakazakii is enhanced after biofilm cultivation in TSB (Jung et al, 2013). Other species better form biofilms in adverse conditions.…”

Section: Clsm Analysismentioning

confidence: 99%

Effect of nutrient and stress factors on polysaccharides synthesis in Proteus mirabilis biofilm.

et al. 2014

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The extracellular matrix in biofilm consists of water, proteins, polysaccharides, nucleic acids and phospholipids. Synthesis of these components is influenced by many factors, e.g. environment conditions or carbon source. The aim of the study was to analyse polysaccharides levels in Proteus mirabilis biofilms after exposure to stress and nutritional conditions. Biofilms of 22 P. mirabilis strains were cultivated for 24, 48, 72 hours, 1 and 2 weeks in tryptone soya broth or in modified media containing an additional amount of nutrients (glucose, albumin) or stress factors (cefotaxime, pH 4, nutrient depletion). Proteins and total polysaccharides levels were studied by Lowry and the phenol-sulphuric acid methods, respectively. Glycoproteins levels were calculated by ELLA with the use of selected lectins (WGA and HPA). For CLSM analysis dual fluorescent staining was applied with SYTO 13 and WGA-TRITC. In optimal conditions the levels of polysaccharides were from 0 to 442 μg/mg of protein and differed depending on the strains and cultivation time. The agents used in this study had a significant impact on the polysaccharides synthesis in the P. mirabilis biofilm. Among all studied components (depending on tested methods), glucose and cefotaxime stimulated the greatest production of polysaccharides by P. mirabilis strains (more than a twofold increase). For most tested strains the highest amounts of sugars were detected after one week of incubation. CLSM analysis confirmed the overproduction of N-acetyloglucosamine in biofilms after cultivation in nutrient and stress conditions, with the level 111-1134%, which varied depending on the P. mirabilis strain and the test factor. *Presented at the 3-rd Workshop on Microbiology "MIKROBIOT 2013" in Łódź, Poland. Abbreviations: ABTS 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), CAUTI catheter associated urinary tract infection, CLSM confocal laser scanning microscope, CTX cefotaxime, DMSO dimethyl sulfoxide, ECM extracellular matrix, ELLA enzyme linked lectinosorbent assay, EPS extracellular polysaccharides, GalNAc N-acetylgalactosamine, GlcNAc N-acetylglucosamine, HPA Helix pomatia agglutinin, PBS phosphate buffered saline, TRITC tetramethylrhodamine-5-(and-6)-isothiocyanate, TSB tryptone soya broth, UTI urinary tract infection, WGA wheat germ agglutinin

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Biofilm formation and exopolysaccharide (EPS) production by Cronobacter sakazakii depending on environmental conditions

Cited by 81 publications

References 26 publications

Biofilms produced by Burkholderia cenocepacia: influence of media and solid supports on composition of matrix exopolysaccharides

Biofilms produced by Burkholderia cenocepacia: influence of media and solid supports on composition of matrix exopolysaccharides

Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies

Effect of nutrient and stress factors on polysaccharides synthesis in Proteus mirabilis biofilm.

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