PubMed Health⌕ Search

Biomedical subjects

E S Wynne

Publications and source records attributed to E S Wynne.

13 recordsLinked to original sources

Persistence of an Escherichia coli-Shigella flexneri hybrid in the intestinal tract of Macaca mulatta.

A live oral vaccine prepared from an Escherichia coli-Shigella flexneri 4b hybrid was administered by gavage to Macaca mulatta. In the three main studies involving 160 monkeys, three doses were used, generally consisting of 5 x 10(9) to 60 x 10(9) cells per dose, with an interval between doses of 6, 7, or 14 days. Rectal swab cultures at the time of the last vaccine dose, and 14 to 58 days later, revealed that the hybrid persisted in the intestinal tract for at least 7 days in 16%, and for at least 21 days in 8%, of the monkeys. Our findings are comparable to those of Formal et al. for shedding of an E. coli-S. flexneri 2a hybrid. Protection studies with the E. coli-S. flexneri 4b hybrid are indicated.

Administration, Oral↗

Sporicidal action of auto-oxidized ascorbic acid for Clostridium.

Neutralized ascorbic acid (AA), buffered or unbuffered and autoclaved or filter-sterilized, was sporicidal for Clostridium. A 0.2% concentration of AA was generally employed, and spore counts were made in a soft-agar modification of Wynne's medium in Prickett tubes. Spores of Clostridium botulinum 115B were less susceptible than those of C. sporogenes PA 3679, whereas C. bifermentans spores were by far the most sensitive. At 75 C, spores of PA 3679 were killed at a rate of about 9% at 0 min (warm-up) to 99+% at 100 min. The lower the temperature, the longer the time needed for a given lethality. The percentage of killing increased with increasing concentrations of AA, and the rate of killing was lower at a higher concentration of spores. At least two mechanisms were operative: a major mechanism involving a product(s) of AA auto-oxidation, and a minor mechanism involving copper-ascorbate toxicity. AA reduced in natural gas was not sporicidal after 18.5 hr at 25 C, whereas 92% of the spores were killed by oxidized AA. Although H(2)O(2) per se was sporicidal, catalase did not reverse lethality of fresh or oxidized AA. Dehydroascorbate was as sporicidal as any AA preparation. Added copper (0.00001%) increased the rate of lethality of freshly prepared AA from 66 to 83% but was not effective with thoroughly oxidized AA. Ethylenediaminetetraacetic acid, NH(4) (+), and phosphate partially reversed AA toxicity, deionized water had no effect, and complex media, as well as thioglycolate, eliminated AA lethality. Since the percentage of killing was affected by spore concentration, AA did not seem to stimulate "lethal germination."

Ascorbic Acid↗

Agar concentration in counting Clostridium colonies.

Decreasing the agar concentration of a counting medium from the usual 1.5% resulted in larger colonies with less interference from gas in Clostridium botulinum 115B and C. sporogenes PA 3679. Optimal agar concentration was 0.65% for C. botulinum with 24-hr incubation and 0.50% for C. sporogenes with 48-hr incubation. Lower concentrations yielded growth too diffuse for counting. Motility was considered the explanation for increased colony size in softer agar. The greater the degree of motility, the greater would be the diffusibility expected, and thus the higher the agar concentration required to insure discrete colonies. For quantitating motility, evaluations were made by use of microscopic examination of liquid cultures and rate of diffusion in a semisolid medium. With both criteria, the degree of motility of C. botulinum 115B clearly exceeded that of C. sporogenes PA 3679. Small-colony variants of C. botulinum in 0.65% agar yielded only small colonies on subculture, with a corresponding decrease in degree of motility of the cells by both criteria. Colony size of the nonmotile C. perfringens ATCC 3624 was unaffected by lowered agar concentrations.

Agar↗