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Improved in vitro assay for determining the mucin adherence of bacteria sensitive to Triton X-100 treatment

(2015) FOLIA MICROBIOLOGICA. 60(5). p.435-442
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Abstract
Mucin-associated microbiota are in relatively close contact with the intestinal epithelium and may thus have a more pronounced effect on host health. We have previously developed a simple mucin agar assay to simulate initial mucus colonization by intestinal microbial communities. Adherence of microbiota was estimated using flow cytometry after detachment with Triton X-100. In this study, the effect of this detergent on the cultivability of both virulent and commensal strains was investigated. Mucin attachment of selected strains was evaluated using the mucin adhesion assay. Bacteria were dislodged from the mucin surface by incubation with Triton or from the whole mucin agar layer using a stomacher. Mechanical extraction resulted in 1.24 +/- 0.42, 2.69 +/- 0.44, and 1.56 +/- 0.85 log CFU/mL higher plate counts of Lactobacillus rhamnosus, Bacillus cereus, and Escherichia coli strains, respectively, than the chemical method. The sensitivity of bacteria to Triton varied among microbial species and strains. Among others, Triton inhibited the growth of Salmonella enterica LMG 10396 and Pseudomonas aeruginosa LMG 8029 on laboratory media, although these bacteria maintained their viability during this treatment. Only Gram-positive strains, Enterococcus hirae LMG 6399 and L. rhamnosus GG, were not affected by this detergent. Therefore, the mechanical method is recommended for the extraction of mucin-adhered bacteria that are sensitive to Triton, especially when followed by traditional cultivation techniques. However, this approach can also be recommended for strains that are not affected by this detergent, because it resulted in higher recovery of adhered L. rhamnosus GG compared to the chemical extraction.
Keywords
STAPHYLOCOCCUS-AUREUS, ESCHERICHIA-COLI, NONCULTURABLE STATE, METHICILLIN-RESISTANT, CHEMICAL-COMPOSITION, INTESTINAL MUCUS, CELLS, LYSIS, ADHESION, COLONIZATION

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Chicago
Tsilia, Varvara, Pieter Van den Abbeele, and Tom Van de Wiele. 2015. “Improved in Vitro Assay for Determining the Mucin Adherence of Bacteria Sensitive to Triton X-100 Treatment.” Folia Microbiologica 60 (5): 435–442.
APA
Tsilia, V., Van den Abbeele, P., & Van de Wiele, T. (2015). Improved in vitro assay for determining the mucin adherence of bacteria sensitive to Triton X-100 treatment. FOLIA MICROBIOLOGICA, 60(5), 435–442.
Vancouver
1.
Tsilia V, Van den Abbeele P, Van de Wiele T. Improved in vitro assay for determining the mucin adherence of bacteria sensitive to Triton X-100 treatment. FOLIA MICROBIOLOGICA. 2015;60(5):435–42.
MLA
Tsilia, Varvara, Pieter Van den Abbeele, and Tom Van de Wiele. “Improved in Vitro Assay for Determining the Mucin Adherence of Bacteria Sensitive to Triton X-100 Treatment.” FOLIA MICROBIOLOGICA 60.5 (2015): 435–442. Print.
@article{5870929,
  abstract     = {Mucin-associated microbiota are in relatively close contact with the intestinal epithelium and may thus have a more pronounced effect on host health. We have previously developed a simple mucin agar assay to simulate initial mucus colonization by intestinal microbial communities. Adherence of microbiota was estimated using flow cytometry after detachment with Triton X-100. In this study, the effect of this detergent on the cultivability of both virulent and commensal strains was investigated. Mucin attachment of selected strains was evaluated using the mucin adhesion assay. Bacteria were dislodged from the mucin surface by incubation with Triton or from the whole mucin agar layer using a stomacher. Mechanical extraction resulted in 1.24 +/- 0.42, 2.69 +/- 0.44, and 1.56 +/- 0.85 log CFU/mL higher plate counts of Lactobacillus rhamnosus, Bacillus cereus, and Escherichia coli strains, respectively, than the chemical method. The sensitivity of bacteria to Triton varied among microbial species and strains. Among others, Triton inhibited the growth of Salmonella enterica LMG 10396 and Pseudomonas aeruginosa LMG 8029 on laboratory media, although these bacteria maintained their viability during this treatment. Only Gram-positive strains, Enterococcus hirae LMG 6399 and L. rhamnosus GG, were not affected by this detergent. Therefore, the mechanical method is recommended for the extraction of mucin-adhered bacteria that are sensitive to Triton, especially when followed by traditional cultivation techniques. However, this approach can also be recommended for strains that are not affected by this detergent, because it resulted in higher recovery of adhered L. rhamnosus GG compared to the chemical extraction.},
  author       = {Tsilia, Varvara and Van den Abbeele, Pieter and Van de Wiele, Tom},
  issn         = {0015-5632},
  journal      = {FOLIA MICROBIOLOGICA},
  language     = {eng},
  number       = {5},
  pages        = {435--442},
  title        = {Improved in vitro assay for determining the mucin adherence of bacteria sensitive to Triton X-100 treatment},
  url          = {http://dx.doi.org/10.1007/s12223-015-0376-0},
  volume       = {60},
  year         = {2015},
}

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