Characterization and purification of porcine small intestinal mucus receptor for Escherichia coli K88ac fimbrial adhesin

Jin, L

doi:10.1016/s0928-8244(99)00155-8

Cited by 6 publications

(10 citation statements)

References 20 publications

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“…Pig mucin from the ileum of a healthy 14‐day‐old piglet was prepared according to the methods described by Jin et al . () and Li et al . (), adjusted to a concentration of 1 mg ml −1 and stored at −20°C prior to use.…”

Section: Methodsmentioning

confidence: 88%

In vitroevaluation of the mucin-adhesion ability and probiotic potential ofLactobacillus mucosaeLM1

2014

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Aims: In this report, we characterized the probiotic potential of Lactobacillus mucosae LM1, focusing on its in vitro mucin-adhesion abilities. Methods and Results: Screening assays were used to evaluate LM1. Previous studies on Lact. mucosae species have been performed, but few have examined the ability of this species to adhere to and colonize the intestinal mucosa. Thus, adhesion, aggregation and pathogen inhibition assays of LM1 along with microbial adhesion to solvents (MATS) assay were carried out in comparison with another putative probiotic, Lactobacillus johnsonii PF01, and the commercial strain, Lactobacillus rhamnosus GG. Based on MATS assay, the cell surfaces of the lactobacilli strains were found to be hydrophobic and highly electron-donating, but the average hydropathy (GRAVY) index of predicted surface-exposed proteins in the LM1 genome indicated that most were hydrophilic. LM1 showed the highest adhesion, aggregation and hydrophobicity among the strains tested and significantly inhibited the adhesion of Escherichia coli K88 and Salmonella enterica serovar Typhimurium KCCM 40253. Correlations among adhesion, aggregation and hydrophobicity, as well as between coaggregation and displacement of E. coli, were observed. Conclusions: Increased adhesion may not always correlate with increased pathogen inhibition due to various strain-specific mechanisms. Nevertheless, LM1 has promising probiotic properties that can be explored further using a genomics approach. Significance and Impact of the Study: Our data on adhesion of LM1 strain showed a significant correlation between adhesion, hydrophobicity of cell surface and autoaggregation. This study gives basic knowledge for the elucidation of the adhesion mechanism of Lactobacillus sp. and prediction of its adherence in specific host models.

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Section: Methodsmentioning

confidence: 88%

In vitroevaluation of the mucin-adhesion ability and probiotic potential ofLactobacillus mucosaeLM1

2014

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“…The results of the present study support the idea that E. faecium 18C23 cells or the substances released in its culture might occupy the binding sites, although these binding sites are not necessarily the same epitope as that for E. coli K88. The mucus receptor for E. coli K88 is involved with glycoprotein and had been characterized as an 80-kDa protein in our previous study (17). Apparently the nature of the mucus receptor for E. faecium 18C23 seems to be a glycoprotein, at least in part based on the results that (i) treatment with trypsin reduced the adhesion of this organism, indicating the protein nature of the receptor, and (ii) treatment with metaperiodate decreased the adhesion of E. faecium 18C23 to the mucus receptor, also indicating the carbohydrate is involved with the attachment to the receptor.…”

Section: Discussionmentioning

confidence: 99%

“…It has been established that receptors for the K88 fimbriae are detectable in the mucus overlying the epithelial cells of the piglet small intestine (17). The presence of receptors in mucus may play a role in the pathogenesis of K88-bearing strains (7).…”

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confidence: 99%

A Strain of Enterococcus faecium (18C23) Inhibits Adhesion of Enterotoxigenic Escherichia coli K88 to Porcine Small Intestine Mucus

Jin

Marquardt

Zhao

2000

Appl Environ Microbiol

105

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Few studies, if any, have addressed the adhesion of enterococci to the intestinal mucosa and their interference with the adhesion of pathogens, although more than 60% of probiotic preparations in the market contain strains of enterococci. The objective of this study was to investigate if Enterococcus faecium 18C23 has the ability to inhibit the adhesion of Escherichia coli K88ac and K88MB to the small intestine mucus of piglets. Approximately 9% of E. faecium 18C23 organisms adhered to the small intestine mucus, and the adhesion was found to be specific. Living E. faecium 18C23 culture efficiently inhibited the adhesion of E. coli K88ac and K88MB to the piglet intestine mucus. Inhibition of the adhesion of E. coli K88ac to the small intestine mucus was found to be dose dependent. Inhibition of >90% was observed when 10 9 CFU or more of living E. faecium 18C23 culture per ml was added simultaneously with E. coli to immobilized mucus. The substances from both the 18C23 cells and the spent culture supernatant contributed to the inhibition of adhesion of E. coli K88 to the small intestine mucus receptors. The inhibiting effect was not solely a pH effect since considerable inhibitory action was demonstrated after neutralizing the mixture or spent culture supernatant to pH 7.0. Part of the inhibition of adhesion of E. coli K88ac by E. faecium 18C23 or its supernatant might occur through steric hindrance.Enterotoxigenic Escherichia coli (ETEC) strains are frequent causes of piglet diarrhea during the preweaning and immediate postweaning periods. Among the different ETEC strains (K88-, K99-, or 987P-expressing strains), those expressing K88 fimbrial antigen are the most prevalent (1, 13). These fimbriae mediate the adhesion of E. coli K88-expressing strains to the intestinal epithelial mucosa and also to the mucus layer lining the small intestine (7), and thereafter the organism elaborates one or two enterotoxins, heat-stable toxin and heatlabile toxin, which induce massive fluid and electrolyte secretion into the gut lumen (1, 13). Among ETEC variants expressing the ab, ac, or ad K88 fimbriae, those possessing the K88ac fimbrial antigen are the most common variant found in pathogenic E. coli isolates in the United States (26). Antibiotics are routinely used in an attempt to control pathogens, but the organisms are becoming resistant to the more commonly used treatments, making antibiotic therapy unreliable (10). Furthermore, the use of antimicrobial growth promoters may cause the development of resistance in a number of important pathogenic bacterial species (28). Recently, the European Union decided to ban the use of four widely used antibiotics, i.e., tylosin, virginiamycin, spiramycin, and zinc bacitracin, as growth promoters from July 1999. As a consequence, there is an urgent need to seek an alternative to antibiotics for the purpose of enhancing the health status and production performance of domestic animals.Probiotics have been used to reduce the colonization of the intestines of animals by pathogens. This, in ...

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“…Taking advantage of modern molecular techniques, one could design useful biovaccine armed with other bacterial antigen, for instance fimbriae, since the very first step in bacterial attachment to the host epithelium is usually mediated by fimbriae (13)(14)(15). Therefore, in the present study, known fimbrial genes of E. coli K99 are sub-cloned, cloned into L. acidophilus and subsequently produced targeted bacterial fimbriae which can compete with E. coli for the same receptor.…”

Section: Discussionmentioning

confidence: 99%

Lactobacillus acidophilus Expressing Recombinant K99 Adhesive Fimbriae Has an Inhibitory Effect on Adhesion of Enterotoxigenic Escherichia coli

Chu

Kang

et al. 2005

Microbiology and Immunology

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The most common enteric colibacillosis in neonatal and newborns is caused by enterotoxigenic Escherichia coli(ETEC). Colonization of ETEC in the small intestine is associated with adhesions using fimbriae, which is known as a specific adhesion factor and provides highly specific means for anchoring and prerequisite for an infectious agent. In the present study we have engineered Lactobacillus acidophilus to produce recombinant K99 fimbriae, which is used for the colonization to the intestine of pigs. The expression of K99 fimbrial protein was confirmed using SDS-PAGE, immunoblot and agglutination analyses. To evaluate a function of the K99 fimbrial protein, inhibition and competition tests were performed on pre-screened intestinal brush border from pigs. The tests showed that recombinant L. acidophilus, not control L. acidophilus, had a significant inhibitory effect to and competition against K99+ E. coli in a dose dependent manner. In conclusion, we demonstrated that recombinant K99 fimbriae producing L. acidophilus was able to prevent E. coli binding to intestinal brush border.

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Characterization and purification of porcine small intestinal mucus receptor for Escherichia coli K88ac fimbrial adhesin

Cited by 6 publications

References 20 publications

In vitroevaluation of the mucin-adhesion ability and probiotic potential ofLactobacillus mucosaeLM1

In vitroevaluation of the mucin-adhesion ability and probiotic potential ofLactobacillus mucosaeLM1

A Strain of Enterococcus faecium (18C23) Inhibits Adhesion of Enterotoxigenic Escherichia coli K88 to Porcine Small Intestine Mucus

Lactobacillus acidophilus Expressing Recombinant K99 Adhesive Fimbriae Has an Inhibitory Effect on Adhesion of Enterotoxigenic Escherichia coli

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