An<i>In vitro</i>Study of Ileal Colonisation Resistance to<i>Escherichia coli</i>Strain Bd 1107/75 08 (K88) in Relation to Indigenous Squamous Gastric Colonisation in Piglets of Varying Ages

Blomberg, Lena; Conway, Patricia L.

doi:10.3109/08910608909140232

Cited by 28 publications

(13 citation statements)

References 15 publications

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“…Blomberg et al [14] later found that piglet ileal mucus contains protein and glycolipid receptors speci¢c for E. coli K88 ¢mbriae. The amount of K88ab receptor in the mucus was shown to be age related [10,12,13], as is susceptibility to E. coli K88 induced diarrhoea, and it was therefore proposed that the presence of receptors in mucus may play a role in the pathogenesis of K88-bearing strains [11,15]. While there are some published reports that eggyolk antibodies inhibited the adhesion of E. coli to intestinal cells of piglets in vitro [8,16], few studies have been directed to the ability of antibodies to interfere with the adhesion of the pathogen to host epithelial mucosa.…”

Section: Introductionmentioning

confidence: 96%

In vitro inhibition of adhesion of enterotoxigenic Escherichia coli K88 to piglet intestinal mucus by egg-yolk antibodies

Jin

1998

FEMS Immunology and Medical Microbiology

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The objective of the study was to determine if the adhesion of E. coli K88 to piglet intestinal mucus could be inhibited in vitro by spray-dried egg-yolk anti-K88 antibodies. Binding of E. coli was monitored using a radioactive assay. Four 14+/-2-day-old healthy piglets were used for the preparation of mucus from the small intestine. Competition and displacement phenomena were investigated by incubating (a) egg-yolk antibodies and E. coli together prior to adding to the mucus and (b) E. coli and mucus, followed by egg-yolk antibodies. The results demonstrated that egg-yolk antibodies inhibited the adhesion of 3H-labeled local strain of hemolytic E. coli K88+ (E. coli K88+ MB) to piglet small intestinal mucus by 84.6-97.0% when the egg-yolk antibodies were diluted 10, 20, 40 or 100 times. The adhesion inhibiting effects of egg-yolk antibodies declined dramatically when the antibody dilution was more than 250-fold. A similar adhesion inhibiting effect was observed when egg-yolk antibodies were incubated with E. coli K88+ MB for 15, 30 and 60 min prior to the adhesion test. Egg-yolk antibodies when diluted 50- and 100-fold had a very strong inhibiting ability against E. coli K88+ MB at a concentration of 10(9) colony forming units (cfu) ml(-1) (adhesion was < 6%). However, dilution of 100 times for egg yolk antibodies was insufficient to inhibit the adhesion of E. coli K88+ MB to intestinal mucus when the concentration of E. coli K88+ MB was 10(10) cfu ml(-1). The displacement test indicated that there was no significant reduction in the adhesion of E. coli K88+ MB to the small intestinal mucus when egg-yolk antibodies were added after adhesion of the organism to the mucus. These studies demonstrate that anti-K88+ MB fimbriae antibodies from chicken egg-yolk when added to E. coli K88+ MB prevented their binding to receptors in the mucus isolated from the intestine of piglets.

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Section: Introductionmentioning

confidence: 96%

In vitro inhibition of adhesion of enterotoxigenic Escherichia coli K88 to piglet intestinal mucus by egg-yolk antibodies

Jin

1998

FEMS Immunology and Medical Microbiology

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“…It is also well documented that genetics play a significant role in the susceptibility of piglets to infectious diarrhea. Piglets are resistant to infection if they are genetically defective in their ability to express K88-specific brush border receptors (9,18); furthermore, the presence of K88-specific receptors in porcine ileal mucus is age dependent (4,5). The receptors of porcine small intestine mucus that bind to K88ab fimbriae have been isolated and identified by using gel filtration chromatography and 3 Hlabeled E. coli (15), while the K88ac fimbrial receptors of porcine intestinal brush border have been identified using by 35 SO 4 -labeled E. coli (3,6).…”

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confidence: 99%

Isolation, Affinity Purification, and Identification of Piglet Small Intestine Mucosa Receptor for Enterotoxigenic Escherichia coli K88ac+ Fimbriae

2000

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An affinity chromatography technique was utilized to isolate and purify the receptors of Escherichia coli K88ac؉ fimbriae from the mucus of the small intestines of newborn piglets. Purified K88ac؉ fimbriae were covalently immobilized onto a beaded agarose matrix (Sepharose 4B). The immobilized fimbriae were used for the affinity purification of the K88ac؉ receptors. Only two major proteins were tightly and specifically bound to the immobilized fimbriae after the column containing bound receptor was washed exhaustively with a buffer containing a high concentration of salt and a detergent. The receptors were eluted as a single component at a low pH. The isolated proteins were then subjected to enzyme-linked immunosorbent assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot (immunoblot) analyses. The two proteins were of high purity, were responsible for nearly all of the fimbrial binding capacity of the crude mucus, and had molecular masses of 26 and 41 kDa. The method for isolation of E. coli binding proteins is simple and yields purified intestinal receptors in a single chromatographic run. The intestinal mucus of different piglets has different proportions of the two receptor proteins.Enterotoxigenic Escherichia coli (ETEC) strains bearing K88 fimbriae can specifically adhere to receptors in the mucus of the small intestines of piglets, causing diarrhea (14, 15). A layer of mucus covers the epithelial cells in the mammalian small intestine, is secreted by specialized goblet cells, and contains receptors that recognize specific adhesion proteins (2, 5, 15, 16). K88 fimbriae are filamentous surface appendages that enable ETEC to bind to the receptor in the mucus layer of the small intestine. This binding prevents the removal of the bacteria by intestinal peristalsis and is a prerequisite for virulence (6,10,18,19,20). Neonatal piglets are extremely sensitive to infection by ETEC but can be protected by specific anti-K88 egg yolk antibodies (11,14,17). Piglets are commonly infected by E. coli strains expressing several serological types of the K88 fimbrial antigens. The K88 fimbrial antigens are classified into three sets designated K88ab, K88ac, and K88ad. Each variant shares a common antigen (a) and expresses one of three variant-specific antigens (b, c, or d, respectively) (6, 7, 9, 12). It is also well documented that genetics play a significant role in the susceptibility of piglets to infectious diarrhea. Piglets are resistant to infection if they are genetically defective in their ability to express K88-specific brush border receptors (9, 18); furthermore, the presence of K88-specific receptors in porcine ileal mucus is age dependent (4, 5). The receptors of porcine small intestine mucus that bind to K88ab fimbriae have been isolated and identified by using gel filtration chromatography and 3 Hlabeled E. coli (15), while the K88ac fimbrial receptors of porcine intestinal brush border have been identified using by 35 SO 4 -labeled E. coli (3, 6). However, little information is av...

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“…It is possible that at least some of the mucus receptor glycoconjugates seen in high numbers in preweaned pigs (Conway et al 1990) are derived from maternal milk or are reIeased from sloughed epithelial cells. Blomberg & Conway (1989) observed that K88 ETEC adhered more poorly to ileal mucus from piglets in which the gastric tissue was densely colonized by lactobacilli compared with mucus from piglets with sparsely colonized gastric tissue. Recent in vitro investigations have shown that Lactobacillus fermenturn releases into the culture supernatant one or more proteinaceous components which affect mucus so that adhesion of cells bearing K88ab and K88ac is reduced (Blomberg et al 1993).…”

Section: Specific Receptors For Several Etec Fimbrial Adhesins Have Bmentioning

confidence: 87%

Nutritional Influences on Interactions Between Bacteria and the Small Intestinal Mucosa

Kelly¹,

Begbie²,

King³

1994

Nutr. Res. Rev.

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AnIn vitroStudy of Ileal Colonisation Resistance toEscherichia coliStrain Bd 1107/75 08 (K88) in Relation to Indigenous Squamous Gastric Colonisation in Piglets of Varying Ages

Cited by 28 publications

References 15 publications

In vitro inhibition of adhesion of enterotoxigenic Escherichia coli K88 to piglet intestinal mucus by egg-yolk antibodies

In vitro inhibition of adhesion of enterotoxigenic Escherichia coli K88 to piglet intestinal mucus by egg-yolk antibodies

Isolation, Affinity Purification, and Identification of Piglet Small Intestine Mucosa Receptor for Enterotoxigenic Escherichia coli K88ac+ Fimbriae

Nutritional Influences on Interactions Between Bacteria and the Small Intestinal Mucosa

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