Measurement of bacterial adhesion—in vitro evaluation of different methods

Vesterlund, Satu; Paltta, J.; Karp, Matti; Ouwehand, Arthur C.

doi:10.1016/j.mimet.2004.09.013

Cited by 141 publications

(114 citation statements)

References 25 publications

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“…The crystal violet method was used to determine the adhesion ability of the four selected strains and the five indicator strains separately (Vesterlund et al 2005). The test cultures (100 µL) were added into micro titer polystyrene plate wells previously coated with 150 µL of fish intestinal mucus.…”

Section: In Vitro Adhesion Assaymentioning

confidence: 99%

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In vitro evaluation of adhesion and aggregation abilities of four potential probiotic strains isolated from guppy (Poecilia reticulata)

Balakrishna¹

2013

Braz. arch. biol. technol.

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The aim of this work was to study the probiotic-related characteristics of four strains of bacteria isolated from the normal flora of the guppy, Poecilia reticulata. In vitro results showed that the strains, namely, MBTU_PB1, MBTU_PB2, MBTU_PB3 and MBTU_PB4 had higher adhesion abilities than the tested indicator strains. However, an association between the cell-surface hydrophobicity and the ability to adhere to the intestinal mucus was not observed for these strains. Further, the selected strains were strongly autoaggregating (autoaggregation percentage ≥ 80) and also showed strain-specific coaggregation abilities with the tested indicator strains.

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Section: In Vitro Adhesion Assaymentioning

confidence: 99%

“…Stained mucus without the cells was used as negative control. Results were expressed by subtracting the absorbance value of this negative control from the absorbance value recorded for all the samples according to Vesterlund et al (2005). Each experiment was performed in triplicate.…”

Section: In Vitro Adhesion Assaymentioning

confidence: 99%

In vitro evaluation of adhesion and aggregation abilities of four potential probiotic strains isolated from guppy (Poecilia reticulata)

Balakrishna¹

2013

Braz. arch. biol. technol.

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“…For use as controls in mucus-binding experiments, recombinant SpaC pilin (positive control) and ovalbumin (Sigma) (background control) were radiolabeled using the aforementioned procedure as described previously (21,41). The L. rhamnosus GG and LC705 strains were metabolically radiolabeled with tritiated thymidine using the procedure described previously (40). Briefly, L. rhamnosus cells were grown overnight in MRS liquid medium containing 10 l/ml [5Ј-the resected human intestinal tissue in this study.…”

Section: Bacterialmentioning

confidence: 99%

“…In addition, participating patients were fully informed beforehand and had provided their written consent. Noncancerous tissue segments containing intact mucosa were selected, and the mucus gel layer was recovered as described previously (40).…”

Section: Bacterialmentioning

confidence: 99%

“…As an additional control, radiolabeled cells of the L. rhamnosus LC705 strain were treated with or without anti-MBF serum (diluted 1:100). Aliquots (100 l) of the antiserum-cell suspension were then added to intestinal mucus-coated microtiter plate wells (40) and incubated at 37°C for 1 h. After the wells were rinsed twice with PBS to remove loosely mucus-adhering cells, cells still remaining attached to bound mucus were incubated in lysis solution (1% SDS-0.1 N NaOH) at 60°C for 1 h. The amount of radioactivity in the lysed cell suspensions was then determined by liquid scintillation counting. Adhesion to mucus was expressed as a percentage calculated from the ratio between the amounts of radioactivity measured for the lysed cell suspension and the cell suspension added originally to the wells.…”

Section: Bacterialmentioning

confidence: 99%

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Functional Characterization of a Mucus-Specific LPXTG Surface Adhesin from Probiotic Lactobacillus rhamnosus GG

Ossowski

Satokari

Reunanen

et al. 2011

Appl Environ Microbiol

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In spite of the wealth of clinical evidence supporting the health benefits of Lactobacillus rhamnosus GG in humans, there is still a lack of understanding of the molecular mechanisms behind its probiosis. Current knowledge suggests that the health-promoting effects of this probiotic strain might be partly dependent on its persistence in the intestine and adhesion to mucosal surfaces. Moreover, L. rhamnosus GG contains mucusbinding pili that might also explain the occupation of its ecological niche as a comparatively less stringent allochthonous intestine-dwelling bacterium. To uncover additional surface proteins involved in mucosal adhesion, we investigated the adherence properties of the only predicted protein (LGG_02337) in L. rhamnosus GG that exhibits homology with a known mucus-binding domain. We cloned a recombinant form of the gene for this putative mucus adhesin and established that the purified protein readily adheres to human intestinal mucus. We also showed that this mucus adhesin is visibly distributed throughout the cell surface and participates in the adhesive interaction between L. rhamnosus GG and mucus, although less prominently than the mucus-binding pili in this strain. Based on primary structural comparisons, we concluded that the current annotation of the LGG_02337 protein likely does not accurately reflect its predicted properties, and we propose that this mucus-specific adhesin be called the mucus-binding factor (MBF). Finally, we interpret our results to mean that L. rhamnosus GG MBF, as an active mucus-specific surface adhesin with a presumed ancillary involvement in pilus-mediated mucosal adhesion, plays a part in the adherent mechanisms during intestinal colonization by this probiotic.The commensal Gram-positive lactobacilli are one of the first groups of bacteria to inhabit the human gastrointestinal (GI) tract (16) and include some strains (autochthonous) that colonize the intestine stably throughout the lifetime of the host (31). In addition, there are those strains, called allochthonous, that persist only briefly in the intestine, many of these being probiotics (1, 9, 13) and understood to stimulate health-benefiting immune responses in host intestinal cells (for a review, see reference 14) or cause competitive displacement of invading pathogens (for a review, see reference 35). Thus far, the precise molecular mechanisms that differentiate the colonization ability between autochthonous and allochthonous intestinal lactobacilli remain undefined (42), although they are likely to be partly dependent on a diverse range of cell surface adhesion molecule-mediated interactions with the host intestinal mucosa. With that being said, there are a growing number of reports in the literature that indicate that lactobacillar adherence to the intestinal mucosal layer is mediated by surface proteins with a mucus-binding capacity (15,21,22,25,30,32,33,41). Moreover, homology-driven genome mining in several Lactobacillus spp. (3,7,8,11,33) has identified the presence of various-sized putative mucus ...

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