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[Expression of Staphylococcus aureus enterotoxin A gene in heterologous systems].

The genomic library of Staphylococcus aureus genes on the plasmid vector pSL5 has been constructed. The library contains a 2.5 kb HindIII DNA fragment including the gene for enterotoxin A. The entA gene on the high copy number plasmids in the Escherichia coli cells deficient in proteolysis determines the synthesis of enterotoxin A in the amounts comparable to the ones in the parent strain Staphylococcus aureus FRI 722(H).

Bacillus subtilis↗

The effects of Clostridium perfringens enterotoxin on rat and rabbit ileum: an electron microscopic study.

Intestinal epithelial damage caused by Clostridium perfringens enterotoxin in rats and rabbits was identified by light microscopy and compared at the surface (scanning electron microscopy), and the ultrastructural (transmission electron microscopy) levels. Under the light microscope damage to the epithelial layer of villus tips was clearly evident in cross-sections. Whole tissue viewed under the scanning electron microscope showed comparable tip localization of morpholigic damage in the form of collapsed tips and a dense covering of rounded blebs on the tips. Ulstructuctural observations included partial and sometimes complete disappearance of microvilli structures, budding of the terminal web region into the lumen, and even complete destruction of epithelial cells. These data suggest that C. perfringens enterotoxin attacks the epithelial cells with a preference for cells at the villus tips and causes damage at least in part by altering the cells' apical membranes. This then leads to cellular sloughing, death, and lysing.

Animals↗

Effect of cholera enterotoxin on pacemaker rate and cyclic adenosine 3':5'-monophosphate in isolated rabbit sinoatrial node.

The positive chronotropic effect of cholera enterotoxin on isolated rabbit sinoatrial (S-A) node was investigated. This toxin produced a time and dose-related increase in pacemaker rate and cyclic adenosine 3':5'-monophosphate (cyclic AMP) levels and shifted to the left the dose-response curves of norepinephrine on the pacemaker rate and cyclic AMP content in the S-A node. Thus, the sensitivity of the S-A node to norepinephrine was increased by the toxin. These effects of the toxin were not blocked by propranolol. There was a linear relation between the increases in pacemaker rate and cyclic AMP content after application of both the toxin and norepinephrine. However, the slope of the regression line relating pacemaker rate to cyclic AMP level differed with these two agents. These results suggest that cholera enterotoxin increases pacemaker rate with activation of the adenylate cyclase system in the S-A node, but the mechanism differs from that of norepinephrine.

Adenylyl Cyclases↗

Testing strains of Clostridium perfringens type A isolated from diarrhoeic piglets for the presence of the enterotoxin gene.

A diarrhoeic syndrome in piglets has been linked to Clostridium perfringens type A because this organism has been isolated in large numbers from all cases. The strains isolated from these cases and strains isolated from healthy piglets were screened for the enterotoxin gene of C perfringens by DNA-hybridisation. Using two different synthetic DNA-probes, none of the strains isolated from diseased pigs was positive in this reaction, indicating that the enterotoxin of C perfringens is not involved in the syndrome.

Animals↗

[Enzyme immunoassay (ELISA) for detecting S. aureus enterotoxins B and C in milk].

Staphylococcus aureus enterotoxins B and C, were obtained from strains 243 and 361 respectively. An immunoenzymatic technique was standardized both quantitative and qualitatively. We were able to detect enterotoxin to nanogram level on extracts from artificially contaminated milk. This ELISA immunoenzymatic technique implemented by us under the operative conditions of our working environment, resulted to be easy to handle, consumes only a short time, has a low cost and great diagnostic sensitivity.

Animals↗

Studies on Streptococcus faecalis enterotoxin.

Streptococcus faecalis has been reported to cause food poisoning. Six strains of S. faecalis were tested for Sherman's criteria. These strains were non-hemolytic, DNase+ and Ent+. The enterotoxin was purified on Sephadex G-200 column and maximum activity was observed at 37 C and pH 7.0. Enterotoxin treated with trypsin and papain elicited very poor response to fluid accumulation. The sensitivity of all the strains against different antibiotics was determined. Strain 53 M was treated with acridine orange and ethidium bromide and a total of 44 Amps Strr and 3 Amps Strs mutants were tested for toxin production. Out of these only 4 were toxin negative, amongst which 3 were also DNase negative and 1 showed partial DNase activity.

Acridine Orange↗

Incidence of several pathogens and staphylococcal enterotoxins in Kariesh cheese.

One hundred samples of Kariesh cheese were randomly collected from Fayoum markets and examined for the incidence of enterococci, coliform organisms, staphylococci, Salmonella and detection of staphylococcal enterotoxins A, B, C, and D. The average total count was 16.4 x 10(7) c.f.u./gm. Coliform bacteria were present in the all the examined, samples with an average of 18.9 x 10(3) c.f.u./gm. However, all samples were free from salmonella. Faecal streptococci and staphylococci were also found in average numbers of (16 x 10(5) c.f.u./gm and 92 x 10(3) c.f.u./gm). Enterotoxins A, B, C and D were not detected in any of the samples tested.

Cheese↗

An evaluation of the pooling culture method for the detection of Escherichia coli enterotoxins.

The efficiency of the pooling method for the detection of enterotoxigenic Escherichia coli (ETEC) strains associated with children acute diarrhea, was evaluated. This study involved the analysis of 6989 E. coli strains corresponding to 1485 cases, coming from 7 hospitals at different geographic locations. Three to five strains from each case were inoculated in pool in Casamino-acids-yeast extract-salts medium plus lincomycin (30 micrograms/ml) and incubated at 37 degrees C during 18 hours with shaking. Polymyxin sulphate B (2200 U/ml) was added to the cultures, and incubation with shaking continued for additional 30 minutes. The culture was then centrifuged and the supernatant tested by a) the enzyme-linked immunosorbent assay to detect heat-labile enterotoxin (LT) and b) the suckling mouse assay for the detection of heat-stable enterotoxin (ST). Strains from a pool were individually studied in all positive cases as shown by the pooling methodology. Also, in 1 out of every 15 negative cases by the pool method, their component strains were individually analyzed, to confirm that there were no false negatives. Fifty seven LT-ETEC, 61 ST-ETEC and 15 LT-ST cases were detected. From the 89 negatives selected by the pool method, 2 cases were positive when the component strains were individually tested (p1 = 2.25%, 95% confidence interval from 0.27 to 7.88%). In both cases one of the non ETEC strains inhibited the ETEC strain. In other 5 negative cases (p2 = 5.62%, 95% confidence interval from 1.85% to 12.63%), ETEC strains were detected when analyzed individually, and gave a positive result when the pooling method was repeated.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriological Techniques↗

Binding of Vibrio cholera toxin and the heat-labile enterotoxin of Escherichia coli to GM1, derivatives of GM1, and nonlipid oligosaccharide polyvalent ligands.

Vibrio cholera toxin and the heat-labile enterotoxin of Escherichia coli have been shown to differ somewhat in their ligand specificity and in the antigenicity of their binding sites. Therefore, the components of the oligosaccharide portion of GM1 bound by cholera toxin and the heat-labile enterotoxin of E. coli were identified by determining the concentration of GM1, derivatives of GM1, oligosaccharide isolated from GM1, or clustered oligosaccharide needed to inhibit toxin binding to GM1-coated plastic wells. The KIs for GM1, the C(7) sialosyl alcohol [corrected] of GM1, and ethanolamine-sialosyl-GM1 were similar (approximately 30-50 nM) for both toxins. N-Deacetylation of GM1 resulted in a small increase in KI; formation of the sialosyl methyl ester increased the KI 2-5 fold; loss of the terminal galactosyl residue (GM2) increased the KI by 10-15-fold; and removal of the sialosyl moiety (asialo-GM1) resulted in loss of inhibition of both toxins. Oligosaccharide isolated from GM1 had a KI for both toxins that was approximately 100-fold greater than that obtained for GM1 and approximately 1000-fold greater than that for a clustered oligosaccharide derivative having an average of 8 oligosaccharide residues (isolated from GM1) per molecule of poly-L-lysine. These results indicate that both toxins are functionally quite similar in their recognition of GM1 as a ligand in that each requires the free carboxyl group of sialic acid for optimum binding, does not need carbons 8 and 9 of the sialosyl moiety nor the acetyl groups associated with the sialic acid and galactosamine residues, and can have its binding to GM1 blocked by a nonlipid compound, i.e. oligo-GM1-poly-L-lysine.

Bacterial Toxins↗

[Detection of heat-labile enterotoxin produced by E. coli with reversed passive latex agglutination].

From June 1987 to March 1988, 103 E. coli strains were collected from non-foodborne samples, foodborne samples, FDA (USA) etc. and analyzed for heat-labile E. coli enterotoxin (LT) by reversed passive latex agglutination (RPLA). The results showed that 84 strains were LT-positive, 19 strains were LT-negative. Forty six strains of enteropathogenic E. coli isolated from foodborne samples showed that 34 strains were LT-positive, 12 strains were negative. Four enterotoxigenic E. coli strains from FDA (USA) used as positive controls, can produce heat-labile E. coli enterotoxin and give positive results.

Bacterial Toxins↗

[Escherichia coli enterotoxins--their significance for food hygiene and recent findings on the structure and mode of action (review)].

Enterotoxin-forming Escherichia (E.) coli germs may be a cause of food poisoning. Any assessment of E. coli in various types of foodstuff together with an appraisal of its relevance to food hygiene will be of inadequate reliability unless virulence properties of germs are investigated, as well. No comprehensive evaluation can be made of quantitative occurrence of food poisoning caused by enterotoxigenic E. coli strains. Literature data are now being compiled on the occurrence of such strains in food items and on the incidence of intoxications. Reference is made in great detail to recent findings on structure and mode of action of heat-labile and heat-stable enterotoxins of E. coli.

Enterotoxins↗

Neutralisation of the new cholera toxin by antiserum against crude enterotoxin of cholera toxin gene-positive Vibrio cholerae 01 in rabbit ileal loop model.

The enterotoxicity of the new cholera toxin (NCT) prepared from cholera toxin gene-negative (CT-) V. cholerae 01 strains isolated from human diarrhoeal and environmental sources was assayed in rabbit ileal loops and the toxin unit was calculated to be 24 micrograms of protein. The enterotoxicity of the NCT preparations were completely neutralised by the antiserum raised against the enterotoxin preparation from the CT+ V. cholerae 01 strain 569B at 1 in 16 dilution in ileal loops. The antiserum contained 1 unit of antitoxin in 85 x 10(-4) ml amount. The data indicate that the antiserum prepared against the enterotoxin of CT+ strain contains antibody against the NCT and can neutralise the toxin in vivo. The observations also suggest that CT+ strains liberate the NCT simultaneously with CT and the latter gets eliminated during the process of purification.

Animals↗

Incidence of enterotoxin producing Staphylococcus aureus among pyogenic skin infections.

Out of the total 68 S. aureus strains isolated and studied from pyoderma patients, 33 (48.5%) strains produced enterotoxin. Isolates from IED, impetigo and folliculitis exhibited high degree of enterotoxigenicity. SE-C and its combinations with other enterotoxins was common. 60.6% of the SE producers were found phage nontypable. Typable enterotoxigenic strains were associated with III, IV and mixed phage groups. S. aureus var. hominis and S. aureus var. bovis are the prevalent subspecies types and potent SE producers among pyogenic skin infections.

Bacteriophage Typing↗

Fimbriae and enterotoxins associated with Escherichia coli serogroups isolated from pigs with colibacillosis.

A comprehensive study of 223 Escherichia coli isolates from pigs with colibacillosis included determination of O serogroups, detection of heat-labile enterotoxin, heat-stable enterotoxin (STa and STb), and identification of K88, K99, 987-P, F-41, and type 1 fimbriae. The incidence of the various E coli types among isolates of pigs of different ages was also determined. Escherichia coli bearing K88 fimbriae accounted for 48% of all isolates studied, were most often of serogroup O157, O149, or O8, and usually produced labile toxin alone or in combination with STa or STb. These E coli were commonly isolated from pigs in each age group studied (0 to 5 days, 6 to 10 days, 11 to 24 days, and greater than 24 days). Escherichia coli bearing 987-P accounted for 30% of the isolates, were most often of serogroup O141 or O20, and usually produced STa. Escherichia coli bearing K99 accounted for 13% of the isolates, usually were of serogroup O101 or O8, and almost always produced STa. Escherichia coli bearing 987-P or K99 were most often isolated from pigs less than 6 days of age. Fimbriae F-41, when identified, were usually on E coli of serotype O101:K99. Although infrequently found, type 1 fimbriae were on E coli of most of the serogroups identified in this study.

Animals↗

[Comparative study of the action of Vibrio cholerae enterotoxin, thermolabile Escherichia coli toxin and a Salmonella typhimurium filtrate on the small intestine of laboratory animals].

The results of the comparative study of the effects produced by V. cholerae enterotoxin E. coli thermolabile toxin (TL-toxin) and S. typhimurum 415 filtrate on the loops of the small intestine of rabbits and C57BL/6 mice are presented. The results obtained in mice were considerably more uniform than those obtained in rabbits. For the first time the dose-effect curve for V. cholerae enterotoxin and TL-toxin, obtained on both animal species, was found to be bell-shaped. The concentration relationship for dibutyryl-cAMP proved to be similar in character. The authors suggest that these effects are based on the specific relationship between the cellular processes, controlling the transport of electrolytes and water, and the level of cAMP.

Animals↗

In vitro effects of a mixture of Escherichia coli heat-stable enterotoxins on chloride flux in everted jejunal sacs of male pigs.

In vitro effects of a mixture of Escherichia coli heat-stable enterotoxins (STa and STb) on isolated jejunum of 3-week-old male pigs were studied, using everted intestinal sac techniques. Heat-stable enterotoxins increased chloride secretion and chloride absorption in everted intestinal sacs. The increase of secretory flux was greater than that for absorptive flux. Vasoactive intestinal peptide (6 x 10(-9) M) increased chloride secretion, but had no effect on chloride absorption. Neither vasoactive intestinal peptide nor pilocarpine (10(-5) M) had additive effect to ST. Secretory effects of ST were not blocked by atropine (2 x 10(-5) M), clonidine (10(-6) M), or morphine (4.2 x 10(-6) M).

Animals↗

[Non-specific antitumoral activity of staphylococcal enterotoxins].

St. aureus enterotoxins A and B possess an antitumour effect. After intraperitoneal inoculation they decrease the size and in some cases prevent the development of the human hypernephroma in the cheek pouch of golden hamsters. The effect of enterotoxins may possibly consist in inducing the production of endogenous immune interferon which activates the host immune system and enhances the rejection of heterologous tumour cells.

Animals↗

[Use of filter-adsorbed enterotoxins for isolating human immune interferon].

The use of A and B enterotoxins of St. aureus adsorbed on Millipore nitrocellulose filters for induction of donor blood leukocytes resulted in production of immune interferon free of the inducer. The interferon produced by this method is acid-labile and inactivated by antiserum to human immune interferon but not to human leukocyte interferon. The advantage of this type of induction consists in the possibility of multiple use of the filters with adsorbed enterotoxins.

Adsorption↗