Pathogenesis of Experimental Cholera in Infant Rabbits: I. Observations on the Intraintestinal Infection and Experimental Cholera Produced with Cell-Free Products

Ra, Finkelstein; Ht, Norris; Nk, Dutta

doi:10.1093/infdis/114.3.203

Cited by 101 publications

(39 citation statements)

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“…An alternative explanation may be that an increase in organized contractile activity develops that contributes to aborad fluid movement. Studies using sulfobromophalein dye placed in the stomach of infant rabbits infected with cholera demonstrated a shortened transit time (17). However, other studies using an indicator dilution technique revealed normal to decreased values during segmental perfusion (18).…”

Section: Introductionmentioning

confidence: 99%

Intestinal myoelectric activity in response to live Vibrio cholerae and cholera enterotoxin.

Mathias¹,

Carlson²,

DiMarino³

et al. 1976

J. Clin. Invest.

104

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Section: Introductionmentioning

confidence: 99%

Intestinal myoelectric activity in response to live Vibrio cholerae and cholera enterotoxin.

Mathias¹,

Carlson²,

DiMarino³

et al. 1976

J. Clin. Invest.

104

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“…Infant rabbits. were inoculated intra-intestinally (16) with three representative virulent strains of V. cholerae: CA 401 [from which the soluble HA was isolated (12)], 569B (16), and 3083 [the parent of the Texas Star-SR avirulent A-B+ mutant candidate vaccine strain (17,18)]. The infant rabbits (nine were inoculated with CA 401, three with 569B, and five with 3083) were sacrificed approximately 18 hr later and intestinal fluids ranging between 6 and 17.5 ml and containing 2.0-42 x 107 live cholera vibrios per ml, were harvested and lightly centrifuged (200 x g for 5 min) to sediment the nonbacterial solids.…”

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confidence: 99%

Vibrio cholerae hemagglutinin/lectin/protease hydrolyzes fibronectin and ovomucin: F. M. Burnet revisited

Finkelstein

Boesman-Finkelstein

Holt

1983

Proc. Natl. Acad. Sci. U.S.A.

165

136

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Cholera vibrios produce a single polymeric protein that (i) causes hemagglutination; (ii) appears to participate in their attachment to gut epithelium; (iii) may mediate their detachment from gut epithelium; and (iv) is a protease that hydrolyzes fibronectin and mucin, cleaves lactoferrin, and nicks the A subunit of the choleragen-related heat-labile enterotoxin of Escherchia coli.In 1947, F. M. Burnet reported the discovery of a mucinase, elaborated by Vibrio cholerae, which participated in the desquamation of epithelium from pieces of guinea pig intestine in vitro (1, 2). Even though Burnet meticulously avoided overstating his conclusions, the observations inspired a flurry of activity among cholera researchers, which led to recommendations that cholera mucinase be included in cholera vaccines (3-6). The concept that the cholera mucinase played a significant direct role in the pathogenesis of choleraic diarrhea was, however, short-lived. Gangarosa et al. (7) showed that the integrity of the intestinaltepithelium was unaltered during the disease in human beings; Gordon (8) found that the disease was not an "exudative enteropathy"; Phillips (9) observed. that the intestinal secretions were low in protein and isotonic; and Finkelstein et al. (10) found that the enterotoxin (choleragen) which caused the diarrhea of cholera could be separated from the cholera mucinase and that the mucinase was not diarrheagenic. Our current observations, however, indicate that Burnet's mucinase may still be involved in the pathogenesis of cholera even though it is not responsible directly for the diarrhea of cholera.Recent efforts in our laboratory have been directed toward understanding the mechanism(s) by which the cholera vibrios elude the host intestinal clearance mechanisms of mucus secretion and peristalsis (11). We have isolated (12), from broth cultures of V. cholerae, a hemagglutinin (HA), which we previously called cholera lectin (13). This protein, a polymer of 32,000 Mr subunits, which appears to participate in the attachment of cholera vibrios to intestinal epithelium, has inherent protease activity (12). Specific antibody against the purified HA inhibits its protease function and also inhibits attachment of V. cholerae to intestinal epithelium (12).The present report shows that the HA/lectin/protease is produced in both a cell-associated and a soluble form in vivo and that it hydrolyzes fibronectin, mucin, -and lactoferrin-three proteins that may participate in host defense against cholera. It also nicks the A subunit of the choleragen-related heat-labile enterotoxin (LT) of Escherichia coli (14) 18)]. The infant rabbits (nine were inoculated with CA 401, three with 569B, and five with 3083) were sacrificed approximately 18 hr later and intestinal fluids ranging between 6 and 17.5 ml and containing 2.0-42 x 107 live cholera vibrios per ml, were harvested and lightly centrifuged (200 x g for 5 min) to sediment the nonbacterial solids. The supernates were subjected to higher speed (12,000 x g for 20 min) cent...

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“…Large-scale growth of strain P307 was produced on syncase medium (Finkelstein, Norris and Dutta, 1964) modified by the substitution of glucose for sucrose and by the addition of yeast extract 0.6%(w/v).…”

Section: Methodsmentioning

confidence: 99%

Assay of the Heat-Labile Enterotoxin of Escherichia Coli in Infant Rabbits

Burgess

Melling

Mullan

et al. 1979

Journal of Medical Microbiology

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S T R A I N S o f Escherichiu coli enteropathogenic for swine have been shown to secrete enterotoxins in vitro (Smith and Halls, 19673;Gyles and Barnum, 1969). These enterotoxins cause accumulation of fluid when they are introduced into ligated intestinal loops prepared in rabbits or pigs (Smith and Halls, 1967~). The strains of E. coli involved are believed to produce two different types of enterotoxin, namely a heat-labile non-dialysable immunogenic toxin (LT) and a heat-stable dialysable toxin (ST).The use of infant rabbits for the detection of LT has been reported by Kutas, Vetesi and Semjen (1974). The present paper described the response of infant rabbits to LT in greater detail, and the evaluation of them as small-anima1 models for the detection of LT.The quantity of LT required for assessment of the infant-rabbit model necessitated a large-scale fermentation, and during the course of this work, it was reported that fermentation of strains of E. coli in the presence of lincomycin (Levner, Wiener and Rubin, 1977) or mitomycin C (Isaacson and Moon, 1975) increased LT synthesis. Hence, it was decided to investigate the effect of these agents on the yield of LT from E. coli strain P307, and these results are also reported here. The preliminary assessment of increase in yield of LT was determined in rabbit ligated-intestinal loops. MATERIALS AND METHODSOrganisms. E. coli strain P307 (08: K87: K88a, b) was obtained from Dr H. Williams Smith (Houghton Poultry Research Station, Houghton, Huntingdonshire). It produced LT and ST. Five substrains resistant to lincomycin 300 pg/ml were obtained by sequential transfer of liquid cultures into media containing increasing concentrations of lincomycin, followed by purification of resistant clones as reported by Levner et al. (1977). The Lin' phenotype was stable in the absence of the drug.Culture media. Large-scale growth of strain P307 was produced on syncase medium (Finkelstein, Norris and Dutta, 1964) modified by the substitution of glucose for sucrose and by the addition of yeast extract 0.6%(w/v).The lincomycin-resistant strains were grown on the modified casamino acids and yeast extract medium described by Evans, Evans and Gorbach (19736) in the presence and absence of linomycin 250 pg/ml.

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Pathogenesis of Experimental Cholera in Infant Rabbits: I. Observations on the Intraintestinal Infection and Experimental Cholera Produced with Cell-Free Products

Cited by 101 publications

References 0 publications

Intestinal myoelectric activity in response to live Vibrio cholerae and cholera enterotoxin.

Intestinal myoelectric activity in response to live Vibrio cholerae and cholera enterotoxin.

Vibrio cholerae hemagglutinin/lectin/protease hydrolyzes fibronectin and ovomucin: F. M. Burnet revisited

Assay of the Heat-Labile Enterotoxin of Escherichia Coli in Infant Rabbits

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