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M D Pierson

Publications and source records attributed to M D Pierson.

33 records · Page 2Linked to original sources

Mechanisms of sorbate inhibition of Bacillus cereus T and Clostridium botulinum 62A spore germination.

The mechanism by which potassium sorbate inhibits Bacillus cereus T and Clostridium botulinum 62A spore germination was investigated. Spores of B. cereus T were germinated at 35 degrees C in 0.08 M sodium-potassium phosphate buffers (pH 5.7 and 6.7) containing various germinants (L-alanine, L-alpha-NH2-n-butyric acid, and inosine) and potassium sorbate. Spores of C. botulinum 62A were germinated in the same buffers but with 10 mM L-lactic acid, 20 mM sodium bicarbonate, L-alanine or L-cysteine, and potassium sorbate. Spore germination was monitored by optical density measurements at 600 nm and phase-contrast microscopy. Inhibition of B. cereus T spore germination was observed when 3,900 micrograms of potassium sorbate per ml was added at various time intervals during the first 2 min of spore exposure to the pH 5.7 germination medium. C. botulinum 62A spore germination was inhibited when 5,200 micrograms of potassium sorbate per ml was added during the first 30 min of spore exposure to the pH 5.7 medium. Potassium sorbate inhibition of germination was reversible for both B. cereus T and C. botulinum 62A spores. Potassium sorbate inhibition of B. cereus T spore germination induced by L-alanine and L-alpha-NH2-n-butyric acid was shown to be competitive in nature. Potassium sorbate was also a competitive inhibitor of L-alanine- and L-cysteine-induced germination of C. botulinum 62A spores.

Alanine↗

Virulence prediction of Yersinia enterocolitica by pyrolysis gas-liquid chromatography.

Pyrolysis gas-liquid chromatography (PGLC) was used to differentiate between HeLa cell-invasive and noninvasive strains of Yersinia enterocolitica and between Sereny-positive and -negative strains. A temperature-programmed gas-liquid chromatograph, equipped with a high-resolution Carbowax 20M coated capillary column, separated the volatiles from pyrolyzed whole cells preparations and cell wall fractions. The resulting pyrolysis elution patterns (pyrograms) were divided into 313 30-s time interval areas. The time interval areas were normalized in relation to the entire pyrogram area and were evaluated by stepwise linear discriminant analysis. The results of the PGLC-statistical analyses showed good correlation in prediction of the HeLa cell invasivity test. The technique of PGLC coupled with statistical analyses is objective, in contrast to traditional methods of determining pathogenicity of Y. enterocolitica.

Bacteriological Techniques↗

Effect of reducing agents on oxidation-reduction potential and the outgrowth of Clostridium botulinum type E spores.

Oxidation-reduction potential (Eh) levels were measured and standardized to pH (Eh7) for Trypticase soy broth containing various concentrations of reducing agents. Prereduced Trypticase soy broth with no added reducing agents exhibited a potential of -141 mV. Ascorbic acid at 0.2 to 0.005% and sodium thioglycolate at concentrations below 0.05% produced an Eh7 higher than the prereduced Trypticase soy broth containing no added reducing agents. The addition of cysteine hydrochloride,2-mercaptoethanol, and sodium formaldehyde sulfoxylate to prereduced Trypticase soy broth resulted in a reduction of Eh7 compared to the system without added reducing agents. The order of relative reducing intensity (from highest to lowest) for the reducing agents when comparing molar concentration was: sodium formaldehyde sulfoxylate,2-mercaptoethanol, cysteine hydrochloride, sodium thioglycolate, and ascorbic acid. Optimal growth of the test organism occurred at low Eh7 and low concentration of the reducing agents. A direct correlation existed between growth of the test organism and -Eh7 x -log concentration of the reducing agent.

Anaerobiosis↗

Effect of sample transport systems on survival of bacteria in ground beef.

The effects of two transport systems and cryoprotective agents on the survival of bacteria in ground beef samples were evaluated. Survival of Clostridium perfringens in ground beef samples after simulated transport (72 h) was higher (about 99%) in Dry Ice than in Trans Temp shipping units (-3 degrees C). There were no significant differences between the two transport systems in survival of coliforms, Escherichia coli, Staphylococcus aureus, or aerobic bacteria. Mixing ground beef samples at a ratio of 1:1 (wt/vol) with 10, 20, or 30% buffered solutions of dimethyl sulfoxide or glycerol before freezing improved the survival of C. perfringens and coliforms in both transport systems. Recovery of E. coli was significantly higher with the addition of 10% dimethyl sulfoxide before Dry Ice transport. Addition of 10% dimethyl sulfoxide resulted in a 100% recovery of both S. aureus and aerobic bacteria from ground beef after simulated transport in Trans Temp shipping units. The use of cryoprotective agents can improve the survival of bacteria during transport of ground beef samples.

Animals↗

Differentiation of selected Enterobacteriaceae by pyrolysis-gas-liquid chromatography.

Pyrolysis-gas-liquid chromatography was used to differentiate selected species of Enterobacteriaceae. Individual cultures of Salmonella typhi, Hafnia alvei, and Proteus vulgaris, and 12 strains of Yersinia enterocolitica were grown in nutrient broth. After harvest and lyophilization, the bacterial samples were pyrolyzed at 900 degrees C, and their volatile fractions were separated on a 50-m capillary column coated with Carbowax 20M. The resulting pyrolysis elution patterns (pyrograms) of the four species were monitored on an integrating console, which was coupled with the chromatographic detector. The pyrograms were divided into 312 30-s time interval areas, and each interval area was normalized in relation to the area of the entire curve. The normalized areas were evaluated by stepwise linear discriminant analysis, and the discriminating component coordinates were used to generate a plot of the canonical variables. Distinct clustering patterns allowed discrimination among the four genera of Enterobacteriaceae studied. The tight clustering of the 12 Y. enterocolitica strains suggests the advantage of pyrolysis-gas-liquid chromatography over traditional approaches for species identification.

Bacteriological Techniques↗

Sublethal heat stress of Vibrio parahaemolyticus.

When Vibrio parahaemolyticsu ATCC 17802 was heated at 41 degrees C for 30 min in 100 mM phosphate-3% NaCl buffer (pH 7.0), the plate counts obtained when using Trypticase soy agar containing 0.25% added NaCl (0.25 TSAS) were nearly 99.9% higher than plate counts using Trypticase soy agar containing 5.5% added NaCl (5.5 TSAS). A similar result was obtained when cells of V. parahaemolyticus were grown in a glucose salts medium (GSM) and heated at 45 degrees C. The injured cells recovered salt tolerance within 3 h when placed in either 2.5 TSBS or GSM at 30 degrees C. The addition of chloramphenicol, actinomycin D, or nalidixic acid to 2.5 TSBS during recovery of cells grown in 2.5 TSBS indicated that recovery was dependent upon protein, ribonucleic acid (RNA, and deoxyribonucleic acid (DNA) synthesis. Penicillin did not inhibit the recovery process. Heat-injured, GSM-grown cells required RNA synthesis but not DNA synthesis during recovery in GSM. Chemical analyses showed that total cellular RNA decreased and total cellular DNA remained constant during heat injury. The addition of [6-3H]uracil, L-[U-14C]leucine, and [methyl-3H]thymidine to the recovery media confirmed the results of the antibiotic experiments.

Bacterial Proteins↗

"Phoenix phenomenon" in the growth of Clostridium perfringens.

The "Phoenix phenomenon" was observed with Clostridium perfringens Hobbs' serological type 9 (HT9) in a cooked-meat medium at 81.7 degrees C by a decrease in plate count (phase I), followed by an increase in count to the intiial level (phase II) and a continued increase above the initial count (phase III). The effects of sporulation, age of inoculum, assay medium, anaerobiosis, diluent, and growth inhibitor were studied. This phenomenon was reproduced in experiments with sporulation-negative mutants derived from HT9 inocula of various cell ages, and different assay media (sulfite-iron agar, tryptose-soytone-yeast extract agar, prereduced peptone-yeast extract agar, prereduced veal agar, and veal agar). When strict anaerobic conditions were employed, it was necessary to increase the heating temperature to 52.3 degrees C to observe the phenomenon. The phenomenon was eliminated at 52.3 degrees C when a combination of strict anaerobic conditions, prereduced media, and prereduced veal diluent was employed. The addition of nalidixic acid at the minimum point of the growth curve (end of phase I) had no effect on the appearance of phase II; however, phase III was completely inhibited. This indicated that phases I and II were an injury-recovery process.

Aerobiosis↗

Heat injury and recovery of vegetative cells of Clostridium botulinum type E.

Vegetative cells of Clostridium botulinum type E (Tenno) were heated at 40.5 C in a prereduced peptone-yeast extract broth (PY). During the heating period, cell numbers remained essentially constant for 3 h as indicated by roll tube counts in PY agar (PYA); however, injury and recovery from injury were observed when the cells were enumerated using PYA containing either 0.06 or 0.07% bile salts.

Anaerobiosis↗