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Biomedical subjects

M S Bergdoll

Publications and source records attributed to M S Bergdoll.

At least 19 recordsLinked to original sources

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

Production of staphylococcal enterotoxins C1 and C2 and thermonuclease throughout the growth cycle.

Synthesis of enterotoxins C1 and C2 and thermonuclease throughout the growth cycle was investigated with Staphylococcus aureus type strains FRI137 and FRI361 and S. aureus isolates M5 (C1) and L2 (C2) of animal origin. Both enterotoxins were produced during the exponential growth phase or at the beginning of the stationary phase. The minimal incubation time (7 to 12 h) and the lowest population (10(7) to 2 x 10(9) CFU/ml) associated with detectable enterotoxin (1 to 6.5 ng/ml) were related to the total amount of toxin produced after 24 h. Thermonuclease was detected in all samples whenever enterotoxins were detected. Furthermore, strain FRI137 produced thermonuclease earlier and at lower cell populations than it did enterotoxin C1. Patterns of enterotoxin and thermonuclease synthesis did not correlate. The concentration of toxins increased throughout the growth cycle, while the concentration of thermonuclease remained constant during the last hours of the growth cycle.

Animals

Effect of environmental conditions on production of toxic shock syndrome toxin 1 by Staphylococcus aureus.

The kinetics of toxic shock syndrome toxin 1 (TSST-1) production by Staphylococcus aureus was studied in a fermentor in which aeration rate, atmospheric composition, pH, and temperature were controlled. The toxin was synthesized at a maximal rate during the exponential phase. High bacterial populations were not necessarily accompanied by high TSST-1 yields. Aerobiosis increased TSST-1 production, but excessive aeration had an adverse effect. Addition of CO2 enhanced TSST-1 yield by increasing toxin production rate and efficiency. Cultures with no pH control made more TSST-1 than those maintained at pH 5.5 to 7.5. Maximum TSST-1 yields were obtained when cultures were supplied with air (20 cm3/min) and CO2 (5 cm3/min) via a sintered glass sparger.

Bacterial Toxins

Effect of chemical modification of histidine and tyrosine residues in toxic shock syndrome toxin 1 on the serologic and mitogenic activities of the toxin.

Modification of three or four of the five histidine residues in the toxic shock syndrome toxin 1 (TSST-1) with diethylpyrocarbonate did not inhibit the precipitin reaction of the modified TSST-1 with polyvalent antisera to the toxin. Monoclonal antibody 7T did not react with the modified TSST-1, but monoclonal antibody 8T did react with the toxin. Up to 50% of the mitogenic reaction of TSST-1 was inhibited by the histidine modification. Modification of one or two of the nine tyrosine residues in TSST-1 did not inhibit the precipitin reaction with polyclonal antisera to the toxin but did inhibit 85% of the mitogenic reaction.

Animals

Production of a toxic shock syndrome toxin variant by Staphylococcus aureus strains associated with sheep, goats, and cows.

A toxic shock syndrome toxin (TSST) variant with an isoelectric point (pI) of 8.6 produced by an ovine-associated Staphylococcus aureus strain was described previously. Analysis of additional strains associated with sheep, goats, cows, and humans by isoelectric focusing with immunoblotting using monoclonal antibodies revealed that all 18 strains associated with sheep and all 12 strains associated with goats produced the TSST variant. Only 1 of 10 bovine-associated strains and no human-associated strains produced the variant, whereas the others produced TSST-1 (pI between 7.0 and 7.2). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis with immunoblotting indicated that both TSST-1 and the TSST variant had a molecular size of 24 kilodaltons.

Animals

Estimation of human dose of staphylococcal enterotoxin A from a large outbreak of staphylococcal food poisoning involving chocolate milk.

An outbreak of gastroenteritis in a school district in the United States was determined to be staphylococcal food poisoning due to 2% chocolate milk containing staphylococcal enterotoxin A (SEA). Twelve one-half pint (approx 0.28 l) cartons of the 2% chocolate milk from this outbreak were analyzed for the quantity of SEA present in the milk. The amount of SEA in the cartons varied from 94 to 184 ng with the average being 144 ng (mean = 139 +/- 45). The attack rate for vomiting among those who consumed more than one carton was greater (38.3%) than among those who consumed only one carton (31.5%) with the highest attack rate among those who consumed three or more cartons (44.4%).

Animals

Molecular topography of toxic shock syndrome toxin 1 as revealed by spectroscopic studies.

Molecular characterization of toxic shock syndrome toxin 1 has been carried out and compared with a group of functionally related staphylococcal enterotoxins. The secondary structure analysis of the far-UV circular dichroic spectrum of toxic shock syndrome toxin 1 revealed 6.25% alpha-helix, 51.25% beta-pleated sheets, 9.0% beta-turns, and 33.5% random coils. The pattern, in general, was similar to the staphylococcal enterotoxins. Four antigenic sites have been predicted for toxic shock syndrome toxin 1 by using the secondary structure information in combination with the hydrophilicity calculation. The location of the antigenic sites, in general, agrees with the experimental results. Topographical analysis of the tyrosine residues as determined by second-derivative UV spectroscopy [Ragone, R., Colonna, G., Balestrieri, C., Servillo, L., & Irace, G. (1984) Biochemistry 23, 1871-1875] showed that six of nine tyrosine residues are exposed to aqueous solvent. Tryptophan fluorescence quenching studies with an anionic surface quencher, I-, and a neutral quencher, acrylamide, revealed that almost all of the tryptophan residues are buried in the protein matrix as their accessibility to the surface quencher is very low (17%). Since there are only three tryptophan residues in the amino acid sequence of the toxic shock syndrome toxin 1 and there is a tyrosine residue (Tyr-15, Tyr-115, and Tyr-153) next to each of the tryptophan residues (Trp-14, Trp-116, and Trp-154), it appears the tyrosine residues not exposed to the aqueous solvent are those next to the tryptophan residues. Functional implications of the topography of the tryptophan and tyrosine residues are assessed.

Bacterial Toxins

Structural analysis of staphylococcal enterotoxins B and C1 using circular dichroism and fluorescence spectroscopy.

Secondary and tertiary structural parameters of two functionally and serologically related proteins, staphylococcal enterotoxins B and C1, have been determined by using circular dichroism and fluorescence spectroscopy. The secondary structures derived from the respective far-UV circular dichroic spectra were 9.5% alpha-helix, 55.0% beta-pleated sheets, 16.5% beta-turns, and 19.0% random coils for enterotoxin B and 15.0% alpha-helix, 38.0% beta-pleated sheets, 25.5% beta-turns, and 21.5% random coils for staphylococcal enterotoxin C1. The values matched well with the secondary structures derived from the amino acid sequences (Chou and Fasman method). Seven antigenic sites have been predicted for both staphylococcal enterotoxins B and C1 by using the hydrophilicity and the secondary structure information. Three of these antigenic sites appear similar. Fluorescence quantum yield of the single tryptophan residue (Trp-197) of both the enterotoxins showed the tryptophan residue in staphylococcal enterotoxin B to be approximately 46% more fluorescent than in staphylococcal enterotoxin C1. Tryptophan fluorescence quenching by the surface quencher I- and the neutral quencher acrylamide revealed that the single tryptophan residue in each of the enterotoxins is buried in the protein matrix and is not accessible to the surface quencher I-. The tryptophan residue in staphylococcal enterotoxin C1 is 14% less accessible to acrylamide than in staphylococcal enterotoxin B. The data, in general, reflect several similarities and significant differences between the two related enterotoxins.

Amino Acid Sequence

Enterotoxigenicity of Staphylococcus intermedius of canine origin.

Seventy-three staphylococcal strains isolated from pyrodermatitis in dogs were classified as Staphylococcus intermedius (52 strains) or Staphylococcus aureus (21 strains) on the basis of acetoin formation, anaerobic mannitol fermentation, aerobic maltose fermentation, pigmentation, coagulation of human plasma, and reaction on crystal violet agar. Enterotoxin was produced by 13 of the 52 S. intermedius strains and 6 of the S. aureus strains. The highest percentage of enterotoxigenic strains produced enterotoxins C (6 strains), D (7 strains), and E (6 strains). Four strains produced the toxic shock syndrome toxin-1. There was little difference in the antibiotic susceptibility between the enterotoxigenic and non-enterotoxigenic strains.

Animals

Characterization of staphylococci from patients with toxic shock syndrome.

Fifty staphylococcal strains that produced toxic shock syndrome (TSS) toxin 1 and that were isolated from patients with TSS were characterized. One strain had more properties that were characteristic of Staphylococcus hyicus than of Staphylococcus aureus. Forty-four strains had the same properties or differed in only one property. Thirty-five of the 50 strains produced either enterotoxin A or C or both in addition to TSS toxin 1.

Humans

Effects of blood and different media on the production of toxic shock syndrome toxin 1 by Staphylococcus aureus in the tampon sac method.

The use of three different agar concentrations in the tampon sac method resulted in slightly higher fluid uptake by the tampons when a 0.5% agar concentration was used. However, there was essentially no difference in the total amount of toxin produced. The largest amount of toxic shock syndrome toxin 1 was produced with brain heart infusion agar, followed closely by 3% NZ-amine NAK-1% yeast extract medium. The addition of plasma and serum to the inoculum resulted in increases (62 and 73%, respectively) in toxin production. The addition of whole blood to the inoculum had a variable effect on toxin production, with an increase in the amount of toxin produced with some tampons and not with others. Over fivefold differences in the amount of toxin produced were obtained when duplicate experiments were done on successive days, whereas the differences were less than twofold for experiments done on the same day. This was related to the effect of small changes in the parameters on toxic shock syndrome toxin 1 production.

Bacterial Toxins

Chromatofocusing in the purification of staphylococcal enterotoxin D.

A chromatofocusing procedure for the purification of staphylococcal enterotoxin D was developed. The purification included the removal of the toxic protein from culture supernatant fluids of Staphylococcus aureus 1151m by batch adsorption with CG-50 resin, chromatofocusing on Polybuffer Exchanger 94, and gel permeation chromatography on Sephacryl S-200. The purity of the staphylococcal enterotoxin D obtained was approximately 98%.

Chromatography, Gel

Prevalence and characterization of Staphylococcus aureus in young goats.

Thirty-six Staphylococcus aureus isolates recovered from 35 of 204 young goats at slaughter were characterized. All isolates were susceptible to cephalothin, clindamycin, chloramphenicol, gentamicin, kanamycin, and amikacin. All but 2 were susceptible to erythromycin and tetracycline, and 19 and 20 were susceptible to penicillin and ampicillin, respectively. Thirteen isolates were classified as biotype A, 9 isolates were classified as biotype B, 8 isolates were classified as biotype C, and 6 isolates were classified as intermediate between B and C or were not biotypable. Six biotype A isolates were enterotoxigenic; 4 produced enterotoxin B, 1 produced enterotoxin C, and 1 produced enterotoxin D. Two biotype B strains produced enterotoxin B, and all 8 biotype C isolates produced enterotoxin C and the toxic shock syndrome toxin-1.

Animals

Complete amino acid sequence of staphylococcal enterotoxin A.

The amino acid sequence of staphylococcal enterotoxin A is presented. Staphylococcal enterotoxin A is a single-chain polypeptide which consists of 233 amino acid residues with a molecular weight of 27,078 and has the amino acid composition Cys2, Asp17, Asn19, Thr16, Ser13, Glu15, Gln12, Pro4, Gly15, Ala7, Val13, Met2, Ile10, Leu23, Tyr18, Phe8, His6, Lys24, Arg7, Trp2, with serine as both amino- and carboxyl-terminal amino acids. Automated sequence analysis of intact enterotoxin A, as well as characterization of the peptides obtained from cyanogen bromide treatment and trypsin and chymotrypsin digestion, led to the elucidation of the complete primary structure of this protein. Less structural homology is observed among staphylococcal enterotoxins A, B (Huang, I-Y., and Bergdoll, M. S. (1970) J. Biol. Chem. 245, 3518-3525), and C1 (Schmidt, J. J., and Spero, L. (1983) J. Biol. Chem. 258, 6300-6306) than that seen between enterotoxins B and C1.

Amino Acid Sequence