PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “ENTEROTOXINS”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,243 records · Page 69Linked to original sources

Rapid solid-phase radioimmunoassay for staphylococcal enterotoxin A.

A rapid solid-phase radioimmunoassay for staphylococcal enterotoxin A is described. The assay procedure requires 3 to 4 h for completion by using a competitive inhibition system in which the antibody is attached to bromacetyl cellulose particles. It is accurate to a level of 0.01 mug of enterotoxin A/ml in a variety of media such as ham, milk products, crab meat, custard, etc. No significant interference was found with any media or food product tested.

Antibodies, Bacterial↗

Staphylococcal enterotoxin B and nuclease production under controlled dissolved oxygen conditions.

Enterotoxin B, nuclease, and total exoprotein production by Staphylococcus aureus strain S-6 was studied in a 0.5-liter fermentor system. While these extracellular products were elaborated over a wide range of aeration rates, maximal production occurred within the very narrow range of 125 to 150 cm(3) of air per min. The levels attained at the optimal aeration rate were not increased by maintaining a constant pH, although yield of enterotoxin:cell mass was highest at a constant pH of 7.0. During the growth cycle of the cultures, when aeration rate alone or aeration rate and pH were held constant, the dissolved oxygen (DO) levels, initially set at 100% of saturation, decreased to 5 to 10% 4 to 5 h after inoculation. The oxygen demand of the culture then maintained this level for an additional 4 to 6 h. This interval of low DO was characterized by maximal growth and exoprotein production. When the DO was controlled at a constant value throughout growth (by increasing or decreasing the airflow rate as appropriate), the culture demonstrated different optima for maximal growth and exoprotein production. A constant DO of 100% stimulated growth to extremely high densities, but the accumulation of toxin and nuclease was not observed. On the other hand, maintaining constant DO levels at 50 or 10% raised exoprotein levels higher than those achieved in a culture grown at the optimal aeration rate. Compared to the optimal aeration rate culture, the 10% DO culture yielded 20% more nuclease, 25% more toxin, and 40 to 50% more total exoprotein. These results indicate that it is the DO and not the aeration rate, per se, that is influential in controlling growth, toxin, nuclease, and total exoprotein production.

Bacterial Proteins↗

Response of ligated intestinal loops in chickens to the enterotoxin of Clostridium perfringens.

Approximately 45-cm length of jejunoileum of 7-week-old chickens was found to be responsive and suitable for testing the enterotoxin of Clostridium perfringens by the ligated intestinal loop technique. Injections of 20 to 30 mug of enterotoxin per loop caused positive response of fluid accumulation. Chickens were found to be more convenient and economical for this purpose than other laboratory and domestic animals.

Animals↗

Production of antisera against the enterotoxin of Bacteroides fragilis and their use in a cytotoxicity neutralization assay of HT-29 cells.

To study the enterotoxin of Bacteroides fragilis, the colon carcinoma cell line HT-29 was used in a standard cytotoxicity assay. We produced high-titer neutralizing antisera in rabbits and goats against both crude and purified toxin and developed a cytotoxicity neutralization assay for use in confirming enterotoxin activity in culture filtrates and stools. The neutralization titers of the antisera on the colon carcinoma cell line HT-29 ranged from 1,600 to 2,400. In an antibody screening enzyme-linked immunosorbent assay, titers ranged from 10(4) to 10(5). The antisera produced against the highly purified toxin also neutralized the enterotoxic activity of the toxin and were monospecific by immunoelectrophoresis.

Animals↗

Assay, characterization, and localization of an enterotoxin produced by Salmonella.

An enterotoxic factor isolated from cultures of Salmonella yielded reproducible results in the suckling mouse model in contrast to other animal models. The enterotoxin appears to possess properties similar to both the heat-stable and heat-labile enterotoxins of Escherichia coli. Preliminary results indicate that the toxin is a protein, is located in the cell wall or outer-membrane fraction, and is difficult to separate from other cell wall constituents.

Amylases↗

Inhibition of the steroidogenic effects of cholera and heat-labile Escherichia coli enterotoxins by GM1 ganglioside: evidence for a similar receptor site for the two toxins.

The effects of three different ganglioside preparations on cholera enterotoxin (CT) and heat-labile Escherichia coli enterotoxin (ECT)-induced steroidogenesis in Y1 and OS3 adrenal tumor cells in tissue culture were examined. Only with GM1 ganglioside was any inhibition of the toxins' effects noted. Concentrations of the crude ECT preparation that gave similar morphogenic and steroidogenic effects as CT were inhibited by the same amount or less of GM1 as that required to inhibit the effects of CT. The results of competition experiments also demonstrated that previous incubation of GM1 with one toxin could inhibit the ganglioside's ability to inactivate the other toxin. These findings indicate that at least for Y1 and OS3 adrenal tumor cells, GM1 may resemble or be the receptor for both CT and ECT.

Adrenal Gland Neoplasms↗

Intrinsic and chemically produced microheterogeneity of Staphylococcus aureus enterotoxin type C.

Staphylococcus aureus enterotoxins C1 (SEC1) and C2 (SEC2) produced from 50-liter quantities of crude culture supernatants were purified chromatographically in a neutral or acid milieu. Microheterogenity of SEC1 was markedly increased by treatment of the purified toxin with alkali, and new more acidic charged species appeared. SEC2 was more heterogenous than any of the other S. aureus enterotoxins and was affected only slightly by treatment with alkali. Prolonged incubation of the organism during production of the SEC2 produced changes in charged species that may be related to a bacterial deamidase, since similar changes were not seen with alkaline treatment of the purified toxin. Although SEC1 and SEC2 showed complete identity immunologically, they are separate, distinct toxins, and alkali treatment of SEC1 did not produce SEC2.

Alkalies↗

Purification and properties of Klebsiella pneumoniae heat-stable enterotoxin.

The enterotoxic material in cell-free growth preparations of Klebsiella pneumoniae serotype 5 was purified by sequential ultrafiltration and gel filtration (GF) procedures and the fractions were assayed for enterotoxic activity by determining their ability to induce in vivo net water secretion in the rat jejunum. Whole-cell lysates were inactive. Anaerobic broth culture conditions yielded a 10-fold increase in toxin production over aerobic conditions. Enterotoxic activity was absent in the UM-10 retentate of the broth filtrate but present in both the retentate and filtrate of the UM-2 membrane. GF of the two UM-2 ultrafiltration fractions through a Sephadex G-25 column yielded an active eluate, whose potency was increased by 10- or 200-fold, in or adjacent to the void volume. When subsequently passed through a G-50 column, these pools eluted at a Kav of between 0.4 and 0.6 and were further increased in potency by two- or fivefold. A second equally potent fraction was also recovered in the void volume of the G-50 eluate of the UM-2 filtrate; this may represent a polymer. Progressive purification by GF was associated with an increased protein and decreased carbohydrate content of the most active fractions. The most active G-50 eluate of the UM-2 retentate had a minimal effective enterotoxic dose of 5 mug/ml and that of the filtrate was less than 0.1 mug/ml. Heating the active GF eluates to 100 C for 30 min did not abolish enterotoxic activity and lowering the pH to 1 or incubation with either Pronase or trypsin had no effect on activity. These observations indicate that K. pneumoniae heat-stable enterotoxin is probably a single toxin with an apparent molecular weight in the range of 5,000. The elution characteristics during GF as well as the chemical composition of the most purified enterotoxin fractions indicate that the toxin is not associated with endotoxin.

Animals↗

Partial purification and properties of Enterobacter cloacae heat-stable enterotoxin.

Cell free preparations of the whole-cell lysate and ultrafiltration (UF) fractions of broth cultures of a strain of Enterobacter cloacae, isolated from a Puerto Rican with tropical sprue, were assayed for their ability to induce in vivo net water secretion in the rat jejunum. The whole-cell lysate and UM-10 retentate of broth cultures were inactive. The UM-2 retentate and filtrate were active at a concentration of 100 mug/ml or more; the toxigenic activity was entirely retained, and increased to 1 mug/ml, by a UM-05 membrane; washing this retentate yielded a fraction with an activity of 10 ng/ml. Stationary aerobic culture conditions yielded the most active UF fractions when ammonium sulfate was used as the precipitating agent, whereas anaerobic culture conditions produced the most active fractions in broth cultures precipitated by acetone. Passage of the active acetone-precipitated UF fractions through a Sephadex G-25 column yielded eluate pools with enhanced toxigenic activity in, or adjacent to, the void volume, but maximum activity of the ammonium sulfate-precipitated UM-05 retentate eluated at a Kav of 0.38 to 0.52. Neither of the most active gel filtration elution fractions of the UM-05 retentates contained detectable carbohydrate, suggesting that the toxin is not associated with endotoxin. Toxigenic activity was unaltered by exposure to a temperature of 100C for 30 min, lowering the pH to 1, or incubation with either Pronase or trypsin. These observations indicate that the strain of E. cloacae under study elaborates a heat-stable enterotoxin htat has approximately the same molecular weight and shares many of the characteristics of the heat-stable enterotoxin produced by some strains of Escherichia coli and Klebsiella pneumoniae.

Aerobiosis↗

Partial purification and characterization of a heat-labile enterotoxin of Escherichia coli.

A partially purified enterotoxin was obtained from the growth medium of Escherichia coli strain 711 (P307), a derivative of E. coli K-12, by ultrafiltration, precipitation with ammonium sulfate, molecular sieving, and anion exchange column chromatography. The active moiety, which is heat-labile, behaved like a protein particle of 180,000 to 200,000 daltons during molecular sieving and ultracentrifugation. During polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE), it dissociated into two subunits with apparent molecular weights of 68,000 to 70,000 and 14,000 to 15,000. SDS-PAGE after heating in SDS changed the larger subunit to an apparent molecular weight of about 40,000; the smaller subunit did not change. The intact particle induced rounding of the cells in Y-1 mouse adrenal tumor cells used for assay. The detergent-dissociated molecules were not active. Proteolysis of the purified toxin by tolylsulfonyl phenylalanyl chloromethyl ketone-trypsin appeared to enhance its activity. The addition of serum to the assay medium resulted in partial depression of the activity. Activity was also abolished by preincubation of the toxin with either a rabbit antiserum to it or solutions containing GM1 ganglioside. The length of time needed to evoke a response in the assay system by fractions from different stages in the purification of the enterotoxin was a useful parameter in the evaluation of specific activity.

Bacterial Proteins↗

Suckling mouse model for detection of heat-stable Escherichia coli enterotoxin: characteristics of the model.

Although the suckling mouse assay is widely used for the detection of heat-stable Escherichia coli enterotoxin (ST), few data have been published concerning the reproducibility, optimal growth, and test conditions of this assay. Four strains of toxigenic E. coli known to elaborate both heat-labile enterotoxin and ST or ST alone were used to study these parameters. ST activity after heat treatment and the effect of purified choleragen were also examined. ST production was optimal in Casamino Acids-yeast extract media, but both Trypticase soy and brain heart infusion broths resulted in several false negative reactions. Growing cultures in roller tubes was the most reliable method of ST production. Shaking-flask cultures and stationary-grown cultures resulted in suboptimal ST production in several strains. Optimal mouse incubation time was 3 h, and fluid secretion did not rise thereafter. Adequate toxin production occurred after 16 to 24 h of incubation. The coefficient of variation of various toxins tested on many occasions varied between 10.5 and 15.7%. Toxin activity was stable for 6 months when frozen at - 20 C. There was no decrease in ST activity when heated at 65 C for 15 min, but a small decrease was observed in two of four strains after heating at 100 C for 30 min. Choleragen, tested at various doses and at multiple times, gave uniformly negative results. These studies indicate that when done under the proper conditions, the suckling mouse assay is a simple, rapid, and reproducible assay for E. coli ST.

Animals↗

Assay of Escherichia coli enterotoxins by in vivo perfusion in the rat jejunum.

Assay of Escherichia coli enterotoxins by in vitro perfusion in rats was evaluated by examining the effects of variously prepared fractions of heat-labile (LT) and heat-stable (ST) toxins on water transport in this system. The assay was found to respond equally well, in a dose-related manner, to both LT and ST; it was sufficiently sensitive to detect the toxigenic effect of concentrations as small as 1 ng/ml. With the assay, it was found that LT is produced in cultures grown under aerobic, but not anaerobic, conditions; in contrast, ST is elaborated in stationary aerobic and anaerobic broth cultures but not in those grown under agitated aerobic conditions. Both toxins can be precipitated by either ammonium sulfate or acetone. The two toxin forms were completely separated from each other by sequential ultrafiltration. LT alone (thermolabile after exposure to 100 degrees C for 30 min) was retained by a PM-30 membrane, and ST alone was present in UM-2 retentates; ST was retained more effectively by a UM-05 membrane, with a 1,000-fold increase in activity over that of the UM-2 retentate. Washed ultrafiltration retentates containing either LT or ST derived from the proper culture conditions all induced water secretion at concentrations of 100 ng or less per ml. These results indicate that in vivo perfusion in rats is a sensitive, duplicable assay for both the LT and ST forms of E. coli enterotoxin.

Animals↗

Diarrhea caused by Escherichia coli that produce only heat-stable enterotoxin.

To determine the role of Escherichia coli heat-stable enterotoxin (ST) as a virulence factor in human diarrhea, a strain that elaborates only ST (E. coli 214-4) was fed to free-living volunteers in doses of 10(6), 10(8), and 10(10) organisms. Short-lived (1 day) mild illness consisting of abdominal cramps with vomiting or diarrhea occurred in three of five individuals fed 10(8). Typical travelers' diarrhea (loose stools, abdominal cramps, and low-grade fever for 2 to 3 days) was seen in four of five volunteers given 10(10); two had brief cholera-like purging of rice-water stools. Despite fever, there was no evidence of mucosal invasion. E. coli 214-4 became the predominant coliform in stools; coproculture isolates were uniformly negative for heat-labile enterotoxin (LT), whereas most produced ST. Ten of 13 individuals developed rises in antibody to somatic E. coli antigen, and none had rises in LT antitoxin. E. coli that elaborate only ST can cause diarrheal disease in adults.

Adolescent↗

Preparative polyacrylamide gel electrophoresis purification of Clostridium perfringens enterotoxin.

Preparative polyacrylamide gel electrophoresis has been used to purify the enterotoxin of Clostridium perfringens from Sephadex G-100 extracts. Purified toxin of high specific activity was eluted in 1 to 3 h, depending upon the length of the acrylamide gel used. Recovery of biological activity with this technique ranged from 80 to 90%. The purity and physical characteristics of the toxin were similar to those previously reported for the protein purified by other methods. Use of preparative electrophoresis will enable the production of larger amounts of high-specific-activity toxin in a shorter time than other currently available procedures. This method was also used to isolate a form of enterotoxin that has a mobility, relative to bromophenol blue tracking dye, of 0.87 to 0.90 in 7% acrylamide gels.

Clostridium perfringens↗

Microtiter solid-phase radioimmunoassay for detection of Escherichia coli heat-labile enterotoxin.

The development of a microtiter solid-phase radioimmunoassay for the detection of Vibrio cholerae enterotoxin and heat-labile Escherichia coli enterotoxin is described. The test is based on the immunological similarity between V. cholerae toxin and E. coli heat-labile toxin. The assay is easy to perform, quantitative, and at least as sensitive and specific as the Y-1 adrenal cell system.

Adrenal Glands↗

Detection of heat-labile enterotoxin-like activity in stools of patients with cholera and Escherichia coli diarrhea.

The Y1 adrenal cell tissue culture assay was used to detect heat-labile enterotoxin-like activity in the stools of 14 of 74 patients with diarrhea. A positive effect of the stool on the adrenal cells was heat-labile and neutralized by cholera antitoxin. Enterotoxin-like activity was detected in the stools of 10 of 30 patients with cholera and in those of 2 of 4 from whom heat-labile Escherichia coli were isolated. None of the stools from nine individuals with Vibrio parahaemolyticus, Salmonella, or Shigella infections were positive. Two of 31 individuals from whom no pathogens were isolated had detectable toxin-like activity in their stools. The Y1 adrenal cell assay provides a rapid method of diagnosing heat-labile enterotoxigenic diarrhea and could be an adjunct in epidemiological studies of gastroenteritis.

Bacterial Toxins↗