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At least 19 recordsLinked to original sources

PubMed1983

Evidence that a non-O1 Vibrio cholerae produces enterotoxin that is similar but not identical to cholera enterotoxin.

Cholera-like enterotoxin produced by a non-O1 strain of Vibrio cholerae, S7 (S7 enterotoxin), isolated from human diarrheal stool, was purified, and its physicochemical, biological, and immunological properties were compared with those of cholera enterotoxin from V. cholerae O1 569B (CT) and an enterotoxin produced by another non-O1 V. cholerae (E8498 enterotoxin) reported previously (Yamamoto et al., Infect. Immun. 39:1128-1135, 1983). The purified S7 enterotoxin had physicochemical properties different from those of CT and E8498 enterotoxin. S7 enterotoxin had greater relative mobility in conventional polyacrylamide gel disc electrophoresis and a lower isoelectric point, and its B subunit was smaller than those of CT and E8498 enterotoxin. The results of sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis suggested that the size of the aggregate of the B subunits of S7 enterotoxin is larger than that of CT and E8498 enterotoxin. The biological and immunological properties of S7 enterotoxin were indistinguishable from those of CT and E8498 enterotoxin. These results indicate that non-O1 vibrios may produce more than one kind of cholera-like enterotoxin: one which is identical to CT (E8498 enterotoxin type) and another which is not identical to CT (S7 enterotoxin type).

Animals↗

PubMed2002

Survival and growth of enterotoxin-positive and enterotoxin-negative Clostridium perfringens in laboratory media.

The survival and growth characteristics of enterotoxin-positive and enterotoxin-negative Clostridium perfringens were compared. Spores of C. perfringens were heated and incubated in laboratory media to simulate the cooking and inadequate storage of cooked food. In our experiment, enterotoxin-positive and enterotoxin-negative spores were heated and incubated individually and it was found that spores of enterotoxin-positive strains were more heat-resistant than spores of the enterotoxin-negative strains. In another experiment, cocktails of enterotoxin-positive and enterotoxin-negative spores were heated and incubated. At the time of inoculation, the ratio of enterotoxin-positive spores to enterotoxin-negative spores was about 1:100. In the case of high-temperature heat treatment, the number of enterotoxin-negative spores decreased to below that of enterotoxin-positive spores after heating, and enterotoxin-positive strains grew to high levels during incubation. In the case of low-temperature heat treatment, the number of enterotoxin-negative spores remained almost unchanged after heating and enterotoxin-positive strains did not grow to high CFU levels during incubation.

Clostridium perfringens↗

PubMed2002

Role of various enterotoxins in Aeromonas hydrophila-induced gastroenteritis: generation of enterotoxin gene-deficient mutants and evaluation of their enterotoxic activity.

Three enterotoxins from the Aeromonas hydrophila diarrheal isolate SSU have been molecularly characterized in our laboratory. One of these enterotoxins is cytotoxic in nature, whereas the other two are cytotonic enterotoxins, one of them heat labile and the other heat stable. Earlier, by developing an isogenic mutant, we demonstrated the role of a cytotoxic enterotoxin in causing systemic infection in mice. In the present study, we evaluated the role of these three enterotoxins in evoking diarrhea in a murine model by developing various combinations of enterotoxin gene-deficient mutants by marker-exchange mutagenesis. A total of six isogenic mutants were prepared in a cytotoxic enterotoxin gene (act)-positive or -negative background strain of A. hydrophila. We developed two single knockouts with truncation in either the heat-labile (alt) or the heat-stable (ast) cytotonic enterotoxin gene; three double knockouts with truncations of genes encoding (i) alt and ast, (ii) act and alt, and (iii) act and ast genes; and a triple-knockout mutant with truncation in all three genes, act, alt, and ast. The identity of these isogenic mutants developed by double-crossover homologous recombination was confirmed by Southern blot analysis. Northern and Western blot analyses revealed that the expression of different enterotoxin genes in the mutants was correspondingly abrogated. We tested the biological activity of these mutants in a diet-restricted and antibiotic-treated mouse model with a ligated ileal loop assay. Our data indicated that all of these mutants had significantly reduced capacity to evoke fluid secretion compared to that of wild-type A. hydrophila; the triple-knockout mutant failed to induce any detectable level of fluid secretion. The biological activity of selected A. hydrophila mutants was restored after complementation. Taken together, we have established a role for three enterotoxins in A. hydrophila-induced gastroenteritis in a mouse model with the greatest contribution from the cytotoxic enterotoxin Act, followed by the Alt and Ast cytotonic enterotoxins.

Aeromonas hydrophila↗

PubMed1991

Production of staphylococcal enterotoxin D in foods by low-enterotoxin-producing staphylococci.

The goal of this investigation was to determine whether staphylococcal strains producing enterotoxins at nanogram levels per milliliter in laboratory medium, not detectable by gel diffusion methods, could produce sufficient enterotoxin in foods to result in food poisoning. Three low-enterotoxin D (SED)-producing strains were selected for this research because this enterotoxin is produced in smaller amounts than the other enterotoxins. The foods used were cream pie and cooked ham, divided into two portions, sterile and non-sterile. Each portion was inoculated with known concentrations of the staphylococcal strains under study and incubated for 48 h at 25, 30, and 37 degrees C. Samples were taken after 24 and 48 h. Enterotoxin was detectable in both sterilized and unsterilized cream and ham after 24 h at 37 degrees C with an inoculum of 10(3)/g. Some strains produced detectable amounts of enterotoxin in the sterilized foods after 24 h at 30 degrees C and some produced detectable amounts of enterotoxin in the sterilized foods after 24 h at 25 degrees C with inocula of 10(4)/g. It can be concluded that staphylococcal strains producing enterotoxin at ng/ml levels in laboratory medium, not detectable by gel diffusion methods, can produce sufficient enterotoxin (ng/g) in foods to cause food poisoning.

Animals↗

PubMed1995

Immunogenicity and antigenic relationship of Salmonella enterotoxin with other enterotoxins.

The immunogenicity and antigenic relationship of Salmonella enterotoxin with other enterotoxins were studied. The purified enterotoxin of Salmonella typhimurium strains was immunogenic in rabbits and antiserum produced against it completely neutralized the enterotoxic activity of purified as well as crude enterotoxins. The Salmonella enterotoxin did not cross-react with cholera toxin, heat-labile enterotoxin of Escherichia coli or Shiga toxin. On the other hand, enterotoxins produced by different serotypes of Salmonella were antigenically related. Heat, pH and proteolytic enzymes had not only adversely affected the enterotoxigenicity but also the antigenicity of the Salmonella enterotoxin. However, formalin was found to completely destroy the enterotoxigenicity of enterotoxin without affecting the antigenicity. Formalin-treated enterotoxin may thus be tried as a vaccine to prevent the diarrhoeal syndrome induced by enterotoxigenic salmonellae.

Animals↗

PubMed1983

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin.

Cholera-like enterotoxin was isolated and purified from the culture supernatant of a non-O1 strain of Vibrio cholerae, E8498, isolated from the environment. Enterotoxin was purified by aluminum hydroxide absorption and elution and successive gel filtrations on Sephadex G-100, Bio-Gel A-5m, and Sephadex G-75. Purified enterotoxin gave a single stained band on polyacrylamide gel disc electrophoresis, and the mobility was the same as that of cholera enterotoxin. The specific biological activity of the purified enterotoxin was almost the same as that of cholera enterotoxin in the Chinese hamster ovary cell assay, fluid accumulation in mouse ligated intestine, increase in vascular permeability in rabbit skin, and passive immune hemolysis. Sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis showed that the purified enterotoxin consisted of subunits A and B, identical to those of cholera enterotoxin, and Ouchterlony double gel diffusion tests indicated that the two toxins were immunologically identical. Enterotoxins prepared from several non-O1 strains isolated from human patients were also immunologically identical to cholera enterotoxin.

Biological Assay↗

PubMed1967

Identification of a fourth staphylococcal enterotoxin, enterotoxin D.

A fourth staphylococcal enterotoxin was identified serologically with antiserum to the very crude enterotoxic products of growth of a strain which also produces enterotoxin C, and then with antiserum to the considerably purified enterotoxic antigen of a strain which produces only the new enterotoxin. The identification of this antigen as enterotoxin D was based on the following observations. It was produced by strains which do not produce enterotoxins A, B, or C; it was absent in the growth products of nonenterotoxigenic strains; when appreciably purified, it was associated with emetic activity in the cat, and its biological activity was neutralized only by antisera containing its specific antibody and not by antibodies to enterotoxins A, B, and C. Staphylococcal strain 494 (ATCC 23235) was selected as the prototype strain. The production of this enterotoxin alone and together with enterotoxin A by strains of food-poisoning origin indicates that its role in food poisoning is second in frequency only to that of enterotoxin A. The incidence of production of enterotoxins A, B, C, and D, and of unidentified cat emetic substances by strains from several source categories, is presented.

Animals↗

PubMed1998

The enterotoxin D plasmid of Staphylococcus aureus encodes a second enterotoxin determinant (sej).

Staphylococcus aureus enterotoxin D is one of the serotypes most commonly associated with food poisoning. Further characterization of the enterotoxin D-encoding plasmid revealed the presence of an open reading frame which encodes a previously unidentified enterotoxin, designated staphylococcal enterotoxin J (SEJ). SEJ is a protein of 269 amino acid residues which has substantial sequence similarity to the staphylococcal A, E, D family of enterotoxins. The enterotoxin D and J open reading frames are transcribed in opposite directions and are separated by an 895 nucleotide intergenic region which contains a perfect inverted repeat, with each arm of the repeat having a length of 21 nucleotides. Chloramphenicol acetyl transferase (cat) transcriptional fusions were used to quantify expression from the enterotoxin gene promoters. Both enterotoxin genes are expressed in S. aureus. However, only sed is regulated by the agr virulence gene signal transduction pathway. Western blot analyses utilizing anti-enterotoxin antisera have confirmed the results obtained with the cat reporter system. PCR amplification studies suggest that the sej determinant may be present on all sed-encoding plasmids.

Amino Acid Sequence↗

PubMed1990

Nucleotide sequence of the staphylococcal enterotoxin C3 gene: sequence comparison of all three type C staphylococcal enterotoxins.

The structural gene entC3, which encodes staphylococcal enterotoxin C3 was cloned from the genome of Staphylococcus aureus FRI-913 and sequenced. The primary amino acid sequence of the toxin was deduced from the nucleotide sequence data. entC3 contains 801 bp and encodes a precursor protein of 266 amino acids. Glutamic acid was found to be the N-terminus of mature enterotoxin C3. Thus, the first 27 residues of the toxin precursor comprise the signal peptide, and the mature toxin contains 239 amino acids with a molecular weight of 27,563 daltons. Enterotoxin C3 differs from enterotoxin C2 by four amino acids and from enterotoxin C1 by nine residues. The 167 C-terminal residues of the three toxins are identical, except for one conservative amino acid substitution in enterotoxin C3. The degree of immunological relatedness among the three Type C enterotoxins is proportional to their molecular relatedness. This study also provides evidence that the N-termini of Type C enterotoxins determine subtype-specific antigenic epitopes, while more conserved C-terminal regions determine biological properties and cross-reactive antigenic epitopes shared with other pyrogenic toxins.

Amino Acid Sequence↗

PubMed1986

Specificity and cross-reactivity of staphylococcal enterotoxin A monoclonal antibodies with enterotoxins B, C1, D, and E.

The cross-reactivity of monoclonal antibodies produced against staphylococcal enterotoxin A with purified and crude enterotoxins B, C1, D, and E and the specificity of such reactions were evaluated by the indirect enzyme-linked immunosorbent assay and immunoblotting of Western blots (from sodium dodecyl sulfate-polyacrylamide gel electrophoresis) followed by autoradiography. Purified and crude enterotoxins B were also tested with polyclonal antibodies. Specificity of reactivity was demonstrated by immunoblotting of crude enterotoxin A, crude enterotoxin A treated with trypsin, crude enterotoxin E, and also with crude A, B, C1, and D that were pretreated with Sepharose-4B-linked normal rabbit immunoglobulin G to remove protein A. A band corresponding to each staphylococcal enterotoxin was seen with monoclonal antibodies under all conditions tested and also with crude and purified enterotoxin B with two different (rabbit and goat) polyclonal antisera.

Antibodies, Bacterial↗

PubMed1965

Identification of a new enterotoxin as enterotoxin C.

Bergdoll, Merlin S. (University of Chicago, Chicago, Ill.), Concordia R. Borja, and Remedios M. Avena. Identification of a new enterotoxin as enterotoxin C. J. Bacteriol. 90:1481-1485. 1965.-Identification of a new enterotoxin was accomplished by purification of the enterotoxins produced by staphylococcal strains 137 and 361 and by the preparation of specific antitoxin to the enterotoxin. Toxicity of the preparations was determined in rhesus monkeys, and specificity of the enterotoxin-antitoxin reaction was determined in gel-diffusion plates. The enterotoxin has been designated enterotoxin C, and staphylococcal strain 137 (ATCC 19095) was selected as the prototype strain.

Animals↗

PubMed1983

Serum antibodies to enterotoxins produced by Staphylococcus aureus with special reference to enterotoxin F and toxic shock syndrome.

The presence of antibodies to staphylococcal enterotoxins (enterotoxins A through F) in sera of healthy subjects (n = 567) and in sera of patients with toxic shock syndrome (n = 20) was determined. Furthermore, production of enterotoxins by Staphylococcus aureus isolated from humans was investigated. In 46, 86, 78, 41, 20, and 91% of the sera of healthy subjects, antibodies were found against enterotoxins A, B, C, D, E, and F, respectively. The high percentage of sera with antibodies against enterotoxin F correlated with the relatively high frequency of enterotoxin F-producing S. aureus isolated from humans (one-third of the isolates produced enterotoxin F). In patients with toxic shock syndrome, antibodies against enterotoxin F were not present or were present only at very low levels. An increase of antibodies after onset of the disease was observed in two of eight patients investigated. From the results, it can be concluded that only those humans who show low levels of antibodies are susceptible to toxic shock syndrome.

Adolescent↗

PubMed1982

The stability of enterotoxin production in Yersinia enterocolitica and the methanol solubility of heat-stable enterotoxin.

Twenty five strains of Yersinia enterocolitica serogroup O3, were isolated from human enteritis and studied for heat-stable enterotoxin production. Enterotoxin production was found even in the crude supernatant fluid of cultures that had been stored in stock agar for a year. According to the suckling mice and rabbit gut loop tests, after 1 to 5 years storage the filtrate showed heat-stable enterotoxin activity only in a purified and concentrated form. Following more than 5 years storage positive results could be obtained only in rabbit gut loop test. After 9 years the freeze dried strains still showed a full capacity of heat-stable enterotoxin production. Studies with concentrated substances showed that even after more than 9 years, there was no spontaneous loss of heat-stable enterotoxin production, only quantitative changes occurred. The methanol solubility of the heat-stable enterotoxin of Y. enterocolitica is--as distinct from the heat-stable enterotoxin of Escherichia coli--homogeneous and only the methanol soluble fractions showed any activity. The activity of methanol soluble enterotoxin from several years old subcultures could be demonstrated in an isolated rabbit gut loop model even when it failed to show any activity in suckling mice.

Animals↗

PubMed1975

The application of QAE-Sephadex for the purification of two staphylococcal enterotoxins. I. Purification of enterotoxin C2.

A new method developed for purification of enterotoxin C2 from Staphylococcus aureus strain 361 consisted of four steps: batchwise adsorption from culture supernatant on QAE-Sephadex; gel filtration on Sephadex G-100; chromatography on QAE-Sephadex using a buffer of constant pH and molarity; and gel filtration using a volatile buffer of constant pH and molarity; and gel filtration using a volatile buffer as the eluting solvent. The purified enterotoxin appeared homogeneous by gel immunodiffusion, gel chromatography and in the analytical ultracentrifuge, although an apparent heterogeneity was noted on QAE-Sephadex chromatography and polyacrylamide disc electrophoresis at pH 4.5. The emetic dose, ED50, by intravenous route in cynomolgus monkeys was 0.04 mug/kg of animal weight. Upon treatment with sodium dodecylsulfate, beta-mercaptoethanol and urea, enterotoxin C2 separated into 3 bands in sodium dodecylsulfate-electrophoresis. One band mol. wt 29000, and two bands of lower molecular weight were so close that they moved as a single zone. After elution from gels, the zone of lower molecular weight were so close that they moved as a single zone. After elution from gels, the zone of lower molecular weight oligopeptides emerged as a single peak at the same position as untreated enterotoxin C2 during gel filtration with buffer lacking thiol and denaturant, and gave a reaction of complete identify to enterotoxin C2 in Ouchterlony immunodiffusion. The results suggest that enterotoxin C2 is a mixture composed of intact polypeptide chains, mol. wt 29000, and two fragments cleaved in the disulfide region of molecular weight of approx. 15400 and 12800 linked by the single disulfide bond in the toxin molecule. Amino acid analysis indicates that enterotoxin C2 consists of 255 amino acid residues.

Amino Acids↗

PubMed2001

Detection of genes encoding for enterotoxins and determination of the production of enterotoxins by HBL blood plates and immunoassays of psychrotrophic strains of Bacillus cereus isolated from pasteurised milk.

The presence of genes for the production of the three components of the HBL enterotoxin complex and enterotoxin-T in Bacillus cereus was evaluated by PCR tests for strains isolated from milk. In addition enterotoxin production of B. cereus was evaluated by means of the HBL blood agar plate and two commercially available toxin tests. All three genes for the HBL enterotoxin complex were detected in 55% of the 86 strains tested, the enterotoxin-T gene was detected in 62% of the strains. A few strains showed a weak reaction in the PCR tests for the L1 or L2 components of the HBL enterotoxin complex. Many strains that were found to contain the genes for the HBL complex gave negative or doubtful results in the HBL blood agar plate test. All strains that contain the L2 part of the HBL complex showed a titer of at least 8 in the Oxoid RPLA test. Two strains that did not contain the L2 part of the HBL enterotoxin complex gave high titers (= 64) in the RPLA test.

Animals↗

PubMed1978

Staphylococcal enterotoxin and thermonuclease production during induced bovine mastitis and the clinical reaction of enterotoxin in udders.

Enterotoxin A- and C-producing strains of Staphylococcus aureus and partially and extensively purified enterotoxin A were inoculated into the udder quarters of cows. In the course of experimentally induced mastitis caused by the inoculated S. aureus strain, enterotoxin C but not A was detected in the infected udder. Enterotoxin C was observed in mastitic milk samples at very low S. aureus population levels (10(2) to 10(3) colony-forming units per ml). The results suggest that either the synthesis of enterotoxin C is stimulated in vitro or that growth of S. aureus cells in udders was, in fact, higher than the colony-forming unit values indicated. Thermonuclease was shown to be excreted into mastitic milk at a slower rate than was enterotoxin. An inoculation of 1 microgram of enterotoxin A in autogenic milk returned to the udder caused clinical reactions (swelling, palpation sensitivity, and increase in the level of somatic cells) within 6 h.

Animals↗

PubMed1989

Porcine Clostridium perfringens type A spores, enterotoxin and antibody to enterotoxin.

Forty-two Clostridium perfringens type A strains isolated from cases of diarrhoea in pigs were tested for their ability to sporulate and produce enterotoxin in three different sporulation media. Enterotoxin was produced by 11 of the 42 C perfringens type A isolates (26.2 per cent). Thirteen isolates (30.9 per cent) produced spores at a frequency of 10 per cent or more. Spore production was recorded in 24 (57.1 per cent) of the isolates. The titres of enterotoxin produced by the isolates ranged from 1:2 to 1:64. The enterotoxin produced was compared with that produced by a reference strain and found to be identical. Ninety-eight of 106 sow sera from four different farms were found to possess antibodies to C perfringens type A enterotoxin with titres ranging from 1:2 to 1:64. Spores of C perfringens type A were detected in pig faeces and intestinal contents in 20 of 23 cases of enteritis at levels of up to 5 x 10(6) cells/g of faeces. Smaller numbers of spores, up to 2 x 10(4)/g were present in five of 10 samples from non-diarrhoeic pigs. Enterotoxin was demonstrated by Vero cell assay in five of the 23 samples from diarrhoeic pigs but in none of the 10 samples from non-diarrhoeic animals. It was clear from these studies that C perfringens type A strains in pigs could sporulate and produce enterotoxin in vitro and in vivo and that enteritis might be associated with sporulating organisms in vivo.

Animals↗

PubMed2006

Staphylococcus aureus isolates from Irish domestic refrigerators possess novel enterotoxin and enterotoxin-like genes and are clonal in nature.

A previous study carried out by the National Food Centre in Dublin on bacterial contamination of Irish domestic refrigeration systems revealed that 41% were contaminated with Staphylococcus aureus. One hundred fifty-seven S. aureus isolates were screened by multiplex PCR analysis for the presence of 15 staphylococcal enterotoxin and enterotoxin-like genes (sea-see, seg-sei, selj-selo, and selq) and the toxic shock toxin superantigen tst gene. Of the refrigerator isolates, 64.3% possessed more than one staphylococcal enterotoxin or staphylococcal enterotoxin-like gene. All bar one of the 101 staphylococcal enterotoxin or staphylococcal enterotoxin-like gene-positive strains possessed the egc locus bearing the seg, sei, selm, seln, and selo genes. Twelve random amplified polymorphic DNA (RAPD) types accounted for 119 (75.8%) of the strains, two of these types accounting for 25 (RAPD type 1, 15.9%) and 52 (RAPD type 5, 33.1%), respectively. All of the RAPD type 5 isolates possessed the egc gene cluster only. The RAPD type 5 amplicon profile was identical to that of S. aureus isolates associated with osteomyelitis in broiler chickens in Northern Ireland that also possessed the egc locus only. However, the RAPD type 5 domestic refrigerator and chicken isolates differed in penicillin G sensitivity, production of Protein A and staphylokinase, and crystal violet agar growth type. These findings highlight that the average Irish household refrigerator harbors potential enterotoxin-producing S. aureus that may or may not be of animal origin and, accordingly, is a potential reservoir for staphylococcal food poisoning.

Animals↗