EFFECT OF HAEMIN SUPPLEMENTATION ON PORPHYRIN ACCUMULATION AND CATALASE SYNTHESIS DURING ANAEROBIC GROWTH OF STAPHYLOCOCCUS.
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Biomedical subjects
Publications and source records attributed to R H DEIBEL.
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The addition of feces to selenite broth significantly enhanced the ability of this medium to select for salmonellae in an environment initially containing overwhelming numbers of coliform bacteria. Either heat-sterilized or Seitz-filtered feces produced this effect. In most experiments, the selectivity of selenite broth was unaffected by unsterile feces. Human blood and plasma markedly reduced selenite efficiency. In a base medium supporting both coliform and Shigella growth, heat-sterilized feces imposed a measure of selectivity for Shigella.
THE RECOVERY OF SALMONELLAE FROM EGG PRODUCTS WAS STUDIED, BY USE OF THREE DIFFERENT ENRICHMENT PROCEDURES: (i) selenite broth, (ii) selenite broth containing 10% sterile feces, and (iii) the lactose pre-enrichment procedure. Brilliant Green Agar was used throughout as the recovery medium. Although the lactose pre-enrichment methodology promoted Salmonella recovery from samples containing small numbers of dormant organisms, the efficiency of this enrichment method is adversely affected by unfavorable coliform-Salmonella ratios. Under such conditions, early subculture of lactose broth into selenite broth is indicated. Selenite broth containing 10% sterile feces was more efficient than the lactose pre-enrichment methodology in promoting the growth of "dormant" salmonellae. Albumen adversely affected recovery of salmonellae from selenite broth, whereas whole egg and egg yolk enhanced Salmonella recovery from this medium. The selenite-feces medium presents a solution to the major problems encountered in the detection of salmonellae in egg products and offers an approach to a single medium in which food-borne salmonellae will manifest themselves with a minimum of laboratory manipulation.
The occurrence and incidence of gram-positive organisms which possess some biochemical and serological characteristics in common with salmonellae are described. These organisms were encountered as contaminants in various food products as well as in feces. Their growth in various Salmonella enrichment media, typical Salmonella-like growth on differential selective media, biochemical similarities to Salmonella, and agglutination in Salmonella polyvalent O antiserum serve as a source of possible confusion in analyses for Salmonella. Although a definitive identification was not performed, preliminary results indicate that these bacteria closely resemble members of the genus Brevibacterium. No sanitary significance could be associated with these bacteria, because they were never found to coexist with Salmonella. The occurrence of these bacteria illustrates the pitfalls of a cursory examination for Salmonella. On the premise that ultimate identification of Salmonella rests upon serological procedures, a rapid and definitive identification of suspect Salmonella isolates is proposed, based upon reactions with polyvalent group O, group O, and Spicer-Edwards H antisera.
Jacobs, N. J. (American Meat Institute Foundation, Chicago, Ill.), R. E. Heady, J. M. Jacobs, K. Chan, and R. H. Deibel. Effect of hemin and oxygen tension on growth and nitrate reduction by bacteria. J. Bacteriol. 87:1406-1411. 1964.-The effect of hemin supplementation of growth media on the ability of several bacteria to reduce nitrate was studied. Added hemin had no detectable effect on the ability of these organisms to reduce nitrate when grown in stationary cultures exposed to air. However, under anaerobic conditions, six strains of facultatively anaerobic staphylococci required hemin for nitrate reduction and growth stimulation in complex, nitrate-containing media. In a nutritionally defined medium, one strain of Staphylococcus required both hemin and nitrate for anaerobic growth. Anaerobic growth and nitrite production of the aerobe Bacillus subtilis was stimulated by addition of hemin. However, the anaerobic growth response was markedly de-decreased as compared with that obtained under static atmospheric conditions. Hemin had no detectable effect on anaerobic nitrate reduction or growth of the obligate aerobe Pseudomonas denitrificans, or of the facultative anaerobes Escherichia coli, B. polymyxa, and Corynebacterium diphtheriae.
Deibel, R. H. (American Meat Institute Foundation, Chicago, Ill.), and C. F. Niven, Jr. Pyruvate fermentation by Streptococcus faecalis. J. Bacteriol. 88:4-10. 1964.-Streptococcus faecalis, as opposed to S. faecium, utilizes pyruvate as an energy source for growth. The fermentation is adaptive, as demonstrated by growth experiments in a casein-hydrolysate medium and the fermentation of pyruvate by cell suspensions. The principal products of pyruvate catabolism were acetoin, CO(2), and lactic, acetic, and formic acids, although carbon recoveries were low due to the formation of slime. End-product analyses suggested that both the phosphoroclastic and dismutation systems were active in pyruvate breakdown. Studies with cell-free extracts indicated a thiamine diphosphate requirement for active pyruvate catabolism. The involvement of lipoic acid in the phosphoroclastic system was investigated, and, although inconclusive results were obtained, no association of this cofactor with phosphoroclastic activity could be made.
Jones, Dorothy (American Meat Institute Foundation, Chicago, Ill.), R. H. Deibel, and C. F. Niven, Jr. Catalase activity of two Streptococcus faecalis strains and its enhancement by aerobiosis and added cations. J. Bacteriol. 88:602-610. 1964.-The nature of catalase activity noted in two unusual Streptococcus faecalis strains was determined. Enzyme activity was lost slowly when cultures were maintained by daily transfer in test tubes of broth media. Loss of activity could be prevented by aerobic culture. Supplementation of the growth medium with ferric, manganese, and zinc ions, as well as aerobiosis, enhanced catalase activity. However, addition of these cations to cell suspensions or to cell-free extracts did not increase catalase activity. Although oxygen was observed to be one of the reaction end products, the catalase activity was not inhibited by cyanide or azide, and the iron-porphyrin coenzyme of classical catalase was not detected. The enzyme was purified 185-fold by precipitation with ammonium sulfate, followed by chromotography on a diethylaminoethyl cellulose column.
Deibel, R. H. (St. James Hospital, Chicago Heights, Ill.), J. H. Silliker, and P. T. Fagan. Some characteristics of an oleate-requiring, hemolytic Pediococcus. J. Bacteriol. 88:1078-1083. 1964.-A gram-positive, catalase-negative, tetradforming coccus, isolated from a bronchial aspirate, is described. The organism was classified as a member of the genus Pediococcus on the basis of morphological and physiological characteristics. It differed from previously described members of this genus in that it evidenced a requirement for oleic acid, even in complex culture media. This isolate produced hemolysis on blood-agar formulated with certain specimens of human blood and not with others. It was demonstrated that hemolysis required oleate supplementation and that the level of this fatty acid required for hemolysis is greater than that necessary for growth of the organism. A cell-free hemolysin could not be demonstrated in the culture supernatant fluid of oleate-grown cultures. The organism was isolated from a patient suffering from tuberculosis; however, attempts to isolate similar organisms from other tuberculous patients as well as from routine throat and sputum specimens were negative. No pathogenicity was manifested when laboratory animals were inoculated with the hemolytic Pediococcus.
Deibel, R. H. (American Meat Institute Foundation, Chicago, Ill.), and M. J. Kvetkas. Fumarate reduction and its role in the diversion of glucose fermentation by Streptococcus faecalis. J. Bacteriol. 88:858-864. 1964.-Fumarate diverts the normal fermentation of glucose by Streptococcus faecalis FB82, as shown by the production of increased amounts of CO(2), formate, acetate, and acetoin, and decreased formation of lactate and ethanol. Experiments with d-glucose-1-C(14), in which low levels of labeled CO(2) were recovered, indicated that C-1 cleavage of the glucose molecule was not involved. The presence of fumarate afforded consistently larger cell crops in growth studies with glucose and other energy sources. On a molar growth-yield basis, anaerobically grown, glucose-fumarate cultures were equivalent to aerobically grown, glucose cultures. The reduction of fumarate by cell suspensions indicated that glucose, gluconate, and, to a lesser extent, glycerol and mannitol could serve as hydrogen donors. Several common metabolic inhibitors had no effect upon the fumarate reductase system in cell suspensions, although some sensitivity to acidic pH was noted. Significant levels of succinate oxidation activity were not detected. Fumarate reductase activity was demonstrated in all five S. faecalis strains tested. Distribution of this ability in S. faecium strains was variable, ranging from activity comparable with that of S. faecalis to total inactivity. The observations support the conclusion that fumarate functions as an alternate hydrogen acceptor, thus allowing pyruvate to participate in the energy-yielding phosphoroclastic and dismutation pathways.
Jones, Dorothy (American Meat Institute Foundation, Chicago, Ill.), R. H. Deibel, and C. F. Niven, Jr. Identity of Staphylococcus epidermidis. J. Bacteriol. 85:62-67. 1963.-The species Staphylococcus epidermidis is defined more precisely, and compared with other staphylococci. The physiological characteristics considered to be of primary import are: (i) ability to grow anaerobically in a defined complex medium with glucose or pyruvate as the energy source; (ii) inability to produce coagulase; (iii) ability to ferment serine as an energy source; (iv) the requirement for biotin and uracil (under anaerobic conditions) in a semisynthetic medium; and (v) inability to reduce nitrate beyond nitrite.
Jacobs, N. J. (American Meat Institute Foundation, Chicago, Ill.), J. Johantges, and R. H. Deibel. Effect of anaerobic growth on nitrate reduction by Staphylococcus epidermidis. J. Bacteriol. 85:782-787. 1963.-In anaerobic cultures, a strain of Staphylococcus epidermidis failed to reduce nitrate, although nitrate was reduced in stationary cultures grown in the presence of air. Resting suspensions of cells grown in air reduced nitrate. Although suspensions of cells grown anaerobically did not reduce nitrate, the nitrate reductase enzyme was shown to be present. When the artificial electron carrier, benzyl viologen, was added to suspensions of cells grown anaerobically, nitrate was reduced, indicating that an electron carrier required for nitrate reduction was missing. The addition of hemin to the growth medium enabled anaerobic cultures to reduce nitrate.
Deibel, R. H. (American Meat Institute Foundation, Chicago, Ill.), and J. H. Silliker. Food-poisoning potential of the enterococci. J. Bacteriol. 85:827-832. 1963.-A total of 23 enterococcus strains were fed to two and sometimes three human volunteers in an effort to elicit food-poisoning symptoms. Each culture was consumed after it was grown in whole sterile milk or on the surface of commercially sterile ham slices. Six strains of Streptococcus faecalis var. liquefaciens were consumed after complete liquefaction of gelatin. In addition, strains of S. faecalis were consumed after having been grown in media which altered the energy metabolism (arginine, gluconate, malate, and pyruvate). In no instance were any of the above conditions of growth conducive to the production of food-poisoning symptoms in the volunteers. Moreover, no evidence was found to indicate that either the age of the culture or the disruption of the cell was a factor in the production or release of a toxic principle. It would appear that until the environmental conditions (if any) for food poisoning are defined the evidence obtained suggests that the association of enterococci and food poisoning is questionable.
Goldman, Manuel (American Meat Institute Foundation, Chicago, Ill.), R. H. Deibel, and C. F. Niven, Jr. Interrelationship between temperature and sodium chloride on growth of lactic acid bacteria isolated from meat-curing brines. J. Bacteriol. 85:1017-1021. 1963.-An elevation of the temperature limit for growth of some Pediococcus homari (Gaffkya homari) and motile Lactobacillus strains could be effected by the addition of sodium chloride to the growth medium. At the optimal temperature for growth, sodium chloride was stimulatory, and as the temperature of incubation was increased a mandatory requirement for sodium chloride was manifested. At the optimal temperature for growth (30 C), the highest sodium chloride concentrations were tolerated; as the temperature was increased, this tolerance decreased, although the optimal sodium chloride concentration increased. No other substances were found that would replace the sodium chloride requirement at higher temperatures of incubation.
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Deibel, R. H. (American Meat Institute Foundation, Chicago, Ill.). Hydrolysis of proteins and nucleic acids by Lancefield group A and other streptococci. J. Bacteriol. 86:1270-1274. 1963.-Classically, group A streptococci have been considered to be nonproteolytic in spite of the observation that a proteinase is produced in broth cultures. When tested by the plate method, it was observed that 78% of 47 strains were proteolytic when the cultures were incubated anaerobically. Some strains hydrolyzed gelatin aerobically; however, the hydrolysis of casein and pepsin required anaerobic conditions of growth. Other proteins hydrolyzed included wheat gluten and a beef-muscle preparation. The hydrolysis of bovine serum albumin, lactalbumin, sheep plasma, and lysozyme was not detected in plate cultures. The hydrolysis of deoxyribonucleic acid and ribonucleic acid was characteristic of the majority of the strains. Unlike the proteolytic activity, the nucleolytic activity was independent of oxygen tension. Other streptococci were screened for hydrolytic activity; among these strains, only the single strain of Streptococcus sanguis tested possessed proteolytic activity. Deoxyribonuclease activity was observed in "animal" but not "human" group C strains, and in some strains of groups G and L.