Screening of pig intestines for K88 non-adhesive phenotype by enzyme immunoassay

Chandler, David; Chandler, Howard M.; Luke, R. K. J.; Tzipori, Saul; Craven, John A.

doi:10.1016/0378-1135(86)90015-5

Cited by 9 publications

(3 citation statements)

References 14 publications

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“…Intestines were processed within two hours of death and the K88 phenotype of animals was determined by enzyme immunoassay (EIA) as previously described 810 13 Briefly, K88 + ETEC were immobilised to wells of a microtitre plate and incubated with piglet intestinal mucosa samples. Mucosal material bound to the bacteria was detected with antibody (rabbit IgG) raised against porcine intestine followed by urease conjugated goat antirabbit IgG (Sigma) and urea substrate.…”

Section: Methodsmentioning

confidence: 99%

Bromelain protects piglets from diarrhoea caused by oral challenge with K88 positive enterotoxigenicEscherichia coli

Chandler

Mynott

1998

Gut

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Section: Methodsmentioning

confidence: 99%

Bromelain protects piglets from diarrhoea caused by oral challenge with K88 positive enterotoxigenicEscherichia coli

Chandler

Mynott

1998

Gut

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“…This was recently investigated in experiments observing the instability of K88 receptor within pig intestines (3a). The variability of receptor activity in intestinal contents was confirmed by EIA as described previously (3), and instability of receptor activity could be controlled by addition of trypsin inhibitor to sample collection buffers. This technique was also used to demonstrate an effect of an exogenous enzyme (Detach) on a K88 intestinal glycoprotein receptor, thereby confirming the disruptive influence of protease on the binding of K88 adhesin to intestinal tissue.…”

Section: Discussionmentioning

confidence: 88%

Efficacy of enteric-coated protease in preventing attachment of enterotoxigenic Escherichia coli and diarrheal disease in the RITARD model

1991

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In this study, we report on a novel approach based on modification of the intestinal surface to prevent diarrhea caused by enterotoxigenic Escherichia coli (ETEC). The removable intestinal tie adult rabbit diarrhea (RITARD) model was used to test the efficacy of an enteric-coated protease preparation (Detach; Enzacor Technology Pty. Ltd.) in the prevention of bacterial attachment and diarrheal disease caused by colonization factor antigen I-positive (CFA/I+) E. coli H10407. Protease was administered orally to rabbits 18 h prior to challenge with 10" bacteria. Four groups of rabbits were inoculated with different ETEC strains which produced different combinations of adhesin and enterotoxin or with sterile phosphate-buffered saline. Occurrence of diarrhea during the subsequent 24-h incubation period was recorded. Oral administration of protease was successful in reducing diarrhea and diarrhea-induced death in six of seven (86%) rabbits infected with CFA/I+, heat-stable and heat-labile toxin-positive E. coli (H10407). Seven of eight (87%) rabbits not protected by protease treatment died or developed severe diarrhea. Quantitative analysis of bacterial cultures obtained from the small intestine of rabbits showed a significant (P < 0.001) 2,000-fold reduction in CFU per centimeter of intestine following treatment with protease. The efficacy of protease treatment was 99.5%, with very wide confidence limits (>0 to 99.9%). The data indicate that the use of protease to prevent ETEC diarrheal disease has considerable potential.

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“…Ofek et al 1331 used EIA to quantitate binding of S. pyogenes and E. coli 987P fimbriae to human oral epithelial cells and to pig intestinal epithelial cells. Chandler et al [36] reported on the use of EIA for identifying K88-ETEC phenotypes in pigs. Tests based on this EIA were subsequently used to study the distribution and stability of the K88 receptor [31].…”

Section: Discussionmentioning

confidence: 99%

Detection of attachment of enterotoxigenicEscherichia coli(ETEC) to human small intestinal cells by enzyme immunoassay

Mynott

Luke

Chandler

1995

FEMS Immunology &Amp; Medical Microbiology

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Simple immunoassays were developed to study the binding between enterocytes of the small intestine and other cell types, and enterotoxigenic Escherichia coli (ETEC). CFA/I or CFA/II pilus protein or CFA-positive E. coli bacteria were immobilised in wells of microtitre plates and incubated with vesicles or crude mucus prepared from human brush border enterocytes. Binding of the cell preparations was detected by adding specific rabbit anti-brush border IgG followed by urease-labelled goat anti-rabbit IgG and urea substrate. The binding of purified CFA/I to human or rabbit small intestine, human oral epithelial cells or Caco-2 cells was detected with specific anti-CFA/I IgG. Both human brush border and mucus-derived preparations were able to attach to ETEC. The binding was CFA-specific and strong enough to withstand several washings. In contrast, CFA/I did not bind to small intestinal cells of non-human small intestinal origin, indicating that there may be important differences in affinity between receptors present on human small intestinal cells and cells of non-human small intestinal origin. Antibodies directed against human small intestinal and non-small intestinal cells did not cross-react with either preparation, indicating that receptors between these different cell sources are different. The EIA proved useful during the identification of a newly-recognised 15 kDa bacterial surface component of ETEC strain H10407P, which may function as a putative attachment factor. The EIAs developed in this study were easy to perform and multiple tests could be performed on small samples, including biopsy samples obtained during endoscopy.

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Screening of pig intestines for K88 non-adhesive phenotype by enzyme immunoassay

Cited by 9 publications

References 14 publications

Bromelain protects piglets from diarrhoea caused by oral challenge with K88 positive enterotoxigenicEscherichia coli

Bromelain protects piglets from diarrhoea caused by oral challenge with K88 positive enterotoxigenicEscherichia coli

Efficacy of enteric-coated protease in preventing attachment of enterotoxigenic Escherichia coli and diarrheal disease in the RITARD model

Detection of attachment of enterotoxigenicEscherichia coli(ETEC) to human small intestinal cells by enzyme immunoassay

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