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Showing papers in "Applied and Environmental Microbiology in 1966"


Journal ArticleDOI
TL;DR: Serum specimens are tested directly rather than diluting them to a narrow range of antibiotic concentrations, possible because of a procedure for calculations that recognizes the curvilinear relationship between zone sizes and antibiotic concentrations.
Abstract: Large glass plates are used for this modified agar-well diffusion assay method, allowing up to 81 replications on a single plate With a specially designed agar punch, it is possible to prepare the small agar wells very quickly The saving in serum resulting from fewer replications of standards with the large plates, and the small volume of the agar wells, makes it economically feasible to use pooled human serum for the standard antibiotic solutions Methods are described for preparing the standard solutions, and for providing controls for the deterioration of standards and unknowns Procedures for preparing and maintaining the commonly used assay organisms are presented Serum specimens are tested directly rather than diluting them to a narrow range of antibiotic concentrations This is possible because of a procedure for calculations that recognizes the curvilinear relationship between zone sizes and antibiotic concentrations Adaptation of this method to a number of the commonly used antibiotics is described With this method, it has been possible to test large numbers of clinical specimens in a minimal time, and with accuracy consistently better than 10% Images

997 citations


Journal ArticleDOI
TL;DR: The results show that medium 10 is suitable for enumeration and isolation of many predominant rumen bacteria and Regardless of differences in the predominant flora associated with diet, medium 10 and the RFM supported growth of similar bacterial populations.
Abstract: Colony counts which approximated those in a habitat-simulating, rumen fluid-agar medium (RFM) were obtained in medium 10, a medium identical to the RFM except for the replacement of rumen fluid with 1.5 x 10(-6)m hemin, 0.2% Trypticase, 0.05% yeast extract, and a 6.6 x 10(-2)m volatile fatty acid mixture qualitatively and quantitatively similar to that in rumen fluid. Single deletion of Trypticase, yeast extract, or the volatile fatty acid mixture from medium 10 significantly reduced colony counts. Colony counts were also reduced when medium 10 was modified to contain higher concentrations of Trypticase or volatile fatty acids. Significant differences were found between colony counts obtained from diluted rumen contents of animals fed a cracked corn-urea diet, and the colony counts obtained from animals fed either a cracked corn-soyean oil meal or an alfalfa hay-grain diet. Qualitative differences were found between the predominant bacterial strains isolated from rumen contents of animals fed cracked corn diets and strains isolated from animals fed alfalfa hay-grain. Regardless of differences in the predominant flora associated with diet, medium 10 and the RFM supported growth of similar bacterial populations. The results show that medium 10 is suitable for enumeration and isolation of many predominant rumen bacteria.

669 citations


Journal ArticleDOI
TL;DR: A method has been developed for the production of aflatoxin by growing Aspergillus flavus strain NRRL 2999 on the solid substrate rice using chloroform-hexane mixtures.
Abstract: A method has been developed for the production of aflatoxin by growing Aspergillus flavus strain NRRL 2999 on the solid substrate rice. Optimal yields, more than 1 mg of aflatoxin B(1) per g of starting material, were obtained in 5 days at 28 C. A crude product containing aflatoxins was isolated by chloroform extraction and precipitation with hexane from concentrated solutions. The crude product consisted of 50% aflatoxin in the following ratio: B(1)-B(2)-G(1)-G(2), 100:0.15:0.22:0.02. Aflatoxin B(1) was separated from almost all the impurities and from the other aflatoxins by chromatography on silica gel with 1% ethyl alcohol in chloroform. Analytically pure aflatoxin B(1) was recrystallized from chloroform-hexane mixtures.

605 citations


Journal ArticleDOI
TL;DR: The maximal amount of toxin was produced by ATCC culture 15548 in 1-liter flasks containing 100 ml of medium incubated as stationary cultures for 6 days at 25 C.
Abstract: Isolates of Aspergillus flavus produced 0.2 to 63 mg of aflatoxins B1 and G1 per 100 ml in a nutrient solution consisting of 20% sucrose and 2% yeast extract. Various factors influencing the fermentation were studied. The maximal amount of toxin was produced by ATCC culture 15548 in 1-liter flasks containing 100 ml of medium incubated as stationary cultures for 6 days at 25 C.

313 citations


Journal ArticleDOI
TL;DR: The turnover rate of the extracellular pool was rapid, and it was concluded that most of the acetic acid must be metabolized to methane by a specialized microflora not involved in the formation of acetic Acid.
Abstract: The quantitative contribution of acetic acid to methane production was studied by use of C(14)-labeled acetic acid. Samples of domestic sewage sludge were incubated anaerobically in Warburg vessels. The rate of methane production in the vessels was 0.033 mumoles per ml per min. C(14)-labeled acetic acid was added, and the turnover rate was calculated. The pool size of acetic acid in the sludge was 4.7 mumoles/ml. The turnover rate was 0.0052 min(-1), giving a rate of formation of acetic acid of 0.024 mumoles per ml per min. Under these conditions, acetic acid would account for approximately 73% of the methane produced by the sludge. Acetic acid was found to exist primarily in an extracellular pool. The turnover rate of the extracellular pool was rapid, and it was concluded that most of the acetic acid must be metabolized to methane by a specialized microflora not involved in the formation of acetic acid.

270 citations


Journal ArticleDOI
TL;DR: Duckling assays showed that detoxification of aflatoxin solutions by B-184 was complete, with no new toxic products being formed.
Abstract: Yeasts, molds, bacteria, actinomycetes, algae, and fungal spores were screened for their ability to degrade aflatoxin. Some molds and mold spores partially transformed aflatoxin B1 to new fluorescing compounds. Only one of the bacteria, Flavobacterium (aurantiacum?) NRRL B-184, removed aflatoxin from solution. Both growing and resting cells of B-184 took up toxin irreversibly. Toxin-contaminated milk, oil, peanut butter, peanuts, and corn were completely detoxified, and contaminated soybean was partially detoxified by addition of B-184. Duckling assays showed that detoxification of aflatoxin solutions by B-184 was complete, with no new toxic products being formed.

266 citations


Journal ArticleDOI
TL;DR: Evidence is presented for the isolation and identification of bacteria able to synthesize an unusual antibiotic containing five bromine atoms per molecule, given the name Pseudomonas bromoutilis because of its distinctive capability.
Abstract: Evidence is presented for the isolation and identification of bacteria able to synthesize an unusual antibiotic containing five bromine atoms per molecule The identification and taxonomic position of these bacteria was made by use of a computer in conjunction with traditional methods These microorganisms and closely related strains have been isolated on various occasions from tropical water in the vicinity of Puerto Rico One bacterium, a pseudomonad, has been given the name Pseudomonas bromoutilis because of its distinctive capability The antibiotic has been extracted, purified, and obtained in crystal form, and its structure has been determined Although clinical tests of its properties were not encouraging, it may be of significant value and interest from an ecological standpoint Images

239 citations


Journal ArticleDOI
TL;DR: This method is based on the absorbance of the fluorescent antibody at those wavelengths primarily associated with the fluorescein and gamma-globulin fractions, and permits these materials to be determined by a single nondestructive analytical procedure.
Abstract: A method is presented for the rapid determination of the fluorescein content, protein content, and fluorescein-to-protein ratio for immune globulin conjugates with fluorescein isothiocyanate as the fluor. This method is based on the absorbance of the fluorescent antibody at those wavelengths primarily associated with the fluorescein and γ-globulin fractions, and permits these materials to be determined by a single nondestructive analytical procedure. A small sample of the fluorescent antibody, in many cases 0.1 ml or less, is adequate for the above determinations. A nomograph is presented which allows simultaneous determination of the materials from the observed absorbance at the two wavelengths. The method is sufficiently accurate for most applications of the fluorescent-antibody techniques. Although this procedure has been developed primarily for fluorescent-antibody conjugates prepared from rabbit γ-globulin, it can be used directly for antibodies prepared from other animals provided the γ-globulin is relatively free from albumin.

138 citations


Journal ArticleDOI
TL;DR: In comparison with conventional assay procedures, the clearing method offered several advantages: (i) results were at least as well correlated with the capacity to utilize native cellulose as a substrate; (ii) the method measured activity of growing cultures rather than culture filtrates, thus involving less risk of losses due to product inhibition, binding, or denaturation of enzymes.
Abstract: A simple method is described for determining the relative cellulolytic activity of fungi Opaque columns of an agar medium containing a partially crystalline cellulose preparation were inoculated with the fungi Depth of the clear zone that developed beneath the growing cultures provided a visual measure of cellulolytic activity on a continuous, cumulative basis Depth of clearing (DC) was determined for 25 species of fungi differing widely in cellulolytic activity, and compared by correlation analysis with results of three other methods for measuring cellulolytic activity Relatively high coefficients of correlation (greater than 06) were obtained between DC and weight loss of cotton sliver, loss in tensile strength of cotton duck, and carboxymethyl cellulase activity in culture filtrates In comparison with conventional assay procedures, the clearing method offered several advantages: (i) results were at least as well correlated with the capacity to utilize native cellulose as a substrate; (ii) the method measured activity of growing cultures rather than culture filtrates, thus involving less risk of losses due to product inhibition, binding, or denaturation of enzymes; (iii) repeated measurements were made on the same experimental set up, so that errors due to arbitrarily selected times of harvest were avoided conveniently; and (iv) the method required less working time and very simple equipment, making it convenient for large-scale screening tests Images

110 citations



Journal ArticleDOI
TL;DR: Replacement of the bicarbonate-phosphate buffer used to maintain fermentor pH at 6.7 with phosphate alone did not greatly alter the fermentation products produced from a hay-concentrate substrate.
Abstract: The effect of pH on rumen fermentation and microbial population was studied in a continuously cultured rumen ecosystem. A marked decrease in the production of volatile fatty acids and methane from alfalfa hay occurred when the cultures were maintained at pH values below 6.0. The decrease in acetate and methane production was greater than that of propionate production. The culture maintained at pH 6.7 contained the types of bacteria often found in high concentration in the rumen, whereas the culture maintained at pH 5.0 had a high percentage of bacteria which could not be identified with the major rumen bacteria found in rumens of animals fed alfalfa hay. Replacement of the bicarbonate-phosphate buffer used to maintain fermentor pH at 6.7 with phosphate alone did not greatly alter the fermentation products produced from a hay-concentrate substrate.

Journal ArticleDOI
TL;DR: In this article, a study was made of the occurrence, distribution, and persistence of coliform, fecal coliforms, and fecal streptococci in the intestinal tract of freshwater fish.
Abstract: A study was made of the occurrence, distribution, and persistence of coliforms, fecal coliforms, and fecal streptococci in the intestinal tract of freshwater fish. A total of 132 fish representing 14 different species were used in various phases of these experiments. Examination of the intestinal contents of 78 fish from moderately polluted sections of the Little Miami River indicated that fecal coliform densities were lowest in bluegills (less than 20 per gram) and highest in catfish (1,090,000 per gram). Levels of fecal streptococci for these two species were 220 and 240,000 per gram, respectively. The occurrence of fecal coliforms in fish caught in this stream reflected the warm-blooded-animal-pollution level of the water. All fish used in this phase of the study were caught during July, August, and September when the water temperatures were between 13 and 18 C. The fate of fecal coliforms and Streptococcus faecalis in the fish intestine indicated that these organisms can probably survive and multiply when fish and water temperatures are 20 C or higher, but only when the organisms are retained in the gut for periods beyond 24 hr. Based on the biochemical reactions for 3,877 coliform strains isolated from 132 freshwater fish of 14 different species, 91.4% of all strains were composed of five IMViC types. In a similar study of the biochemical reactions of 850 streptococci isolated from the intestinal tract of 55 freshwater fish, the predominant strains included S. faecalis and various closely associated biotypes. No consistently recurring pattern for either coliforms or streptococci could be developed to identify species of fish investigated. The composition of the intestinal flora is, however, related in varying degree to the level of contamination of water and food in the environment.

Journal ArticleDOI
TL;DR: The direct plate count and MPN method were found to be in good correlation for T2 bacteriophage and bulk T bacteriophile in samples obtained from a sewage treatment plant and from contaminated seawater.
Abstract: The estimation of low numbers of the Escherichia coli bacteriophage was made possible by use of the most probable number (MPN) method. This method is similar to the technique used for counting coliform bacteria. The statistical results were computed by referring to tables. The method makes it possible to record values as low as two particles per 100 ml of sample. The direct plate count and MPN method were found to be in good correlation for T2 bacteriophage and bulk T bacteriophage in samples obtained from a sewage treatment plant and from contaminated seawater.

Journal ArticleDOI
TL;DR: An anaerobic system is described which allows the microbiologist or hospital technician ease of operation not previously possible with other systems, and incorporates unique safety features which eliminate the possibility of a laboratory explosion.
Abstract: An anaerobic system is described which allows the microbiologist or hospital technician ease of operation not previously possible with other systems. Gas cylinders, vacuum pumps, valves, and gauges have been eliminated. A new anaerobic lid was developed that is fitted only with a snap-in rubber gasket, a double stainless-steel gauze flash arrestor, and a catalyst holder. The holder contains a palladium pellet catalyst which is active at room temperature and requires no heat for activation. This system was made specifically for use with a disposable hydrogen-carbon dioxide generator and a disposable methylene blue anaerobic indicator. In addition to ease of operation, this jar incorporates unique safety features which eliminate the possibility of a laboratory explosion. An oxygen-free atmosphere composed primarily of nitrogen and carbon dioxide was quickly achieved within the jar to insure maximal growth.

Journal ArticleDOI
TL;DR: With a modified Moore swab for sampling bacteria, Salmonella organisms were recovered consistently from surface waters when the enrichment broths of SBG-sulfa and tetrathionate and Brilliant Green Agar plates were incubated at 41.5 C.
Abstract: With a modified Moore swab for sampling bacteria, Salmonella organisms were recovered consistently from surface waters when the enrichment broths of SBG-sulfa and tetrathionate and Brilliant Green Agar plates were incubated at 41.5 C. When an equal portion of the same swab sample was incubated at 37.0 C, no salmonellae were detected. By use of the elevated-temperature technique, salmonellae were recovered from stream sites having relatively low total coliform densities of 2,200 per 100 ml and fecal coliform densities of 220 per 100 ml. These pathogens were isolated in water sampled as far as 73 river miles and 4 days time of travel downstream from the source of pollution under an ice cover that averaged 2.5 ft (76.2 cm) in thickness.

Journal ArticleDOI
TL;DR: The sodium chloride inhibition of spore outgrowth of four strains of type E Clostridium bolulinum was determined in a Trypticase-peptone-glucose (TPG) medium and the minimal pH permitting outgrowthof type E spore inocula was affected by the concentration of reducing compound present in the system.
Abstract: The sodium chloride inhibition of spore outgrowth of four strains of type E Clostridium bolulinum was determined in a Trypticase-peptone-glucose (TPG) medium. At 16, 21, and 30 C, spores of three strains required 5.0% and one strain 4.5% salt for complete inhibition during 1 year of incubation. At 8 and 10 C, spores of the four strains required 4.5% salt for definite inhibition. Salt concentrations slightly lower than those providing inhibition tended to extend spore outgrowth time at low temperatures. The minimal pH permitting outgrowth of type E spore inocula was affected by the concentration of reducing compound present in the system. When either 0.02% sodium thioglycolate or 0.05% L-cysteine hydrochloride was used, outgrowth at 30 and 8 C occurred at much lower pH levels than when 0.2% thioglycolate was added. At 30 C, spores of one strain showed outgrowth in TPG medium as low as pH 5.21 with an inoculum of 2 million spores per replicate tube. At a 10-fold higher inoculum, the same strain showed outgrowth at pH 5.03 in one of five replicate tubes. At 8 C, spore outgrowth of the four strains occurred at pH 5.9, but not at pH 5.7, in TPG medium containing L-cysteine hydrochloride.

PatentDOI
TL;DR: The heat-shock treatment not only proved effective in ridding the fruit of naturally occurring, interfering, and competitive microbial groups prior to brining and inoculation, but also made the olives highly fermentable with respect to growth and acid production by the introduced culture, particularly L. plantarum.
Abstract: The method previously developed by us for the pure-culture fermentation of brined cucumbers and other vegetables has been applied successfully to Manzanillo variety olives. Field-run grade fruit was processed first by conventional procedures to remove most of the bitterness. Then the relative abilities of Lactobacillus plantarum, L. brevis, Pediococcus cerevisiae, and Leuconostoc mesenteroides to become established and produce acid in both heat-shocked (74 C for 3 min) and unheated olives, brined at 4.7 to 5.9% NaCl (w/v basis), were evaluated. The heat-shock treatment not only proved effective in ridding the fruit of naturally occurring, interfering, and competitive microbial groups prior to brining and inoculation, but also made the olives highly fermentable with respect to growth and acid production by the introduced culture, particularly L. plantarum. Of the four species used as inocula, L. plantarum was by far the most vigorous in fermentation ability. It consistently produced the highest levels of brine acidity (1.0 to 1.2% calculated as lactic acid) and the lowest pH values (3.8 to 3.9) during the fermentation of heat-shocked olives. Also, L. plantarum completely dominated fermentations when used in two-species (with P. cerevisiae) and three-species (with P. cerevisiae and L. brevis) combinations as inocula. In contrast, when L. plantarum was inoculated into the brines of unheated olives it failed to become properly established; the same was true for the other species tested, but even to a more pronounced degree. L. brevis was the only species used that failed to develop in brines of both heat-shocked and unheated olives. Modification of the curing brine by the addition of lactic acid at the outset, either with or without dextrose, led to a much earlier onset of fermentation with accompanying acid development, as compared to treatments with dextrose alone or nonadditive controls. Reasons for the marked improvement of the fermentability of Manzanillo olives receiving the prebrining heat-shock treatment are discussed.

Journal ArticleDOI
TL;DR: Evidence was adduced to show that the natural fermentation of coffee was the result of activity of microflora from the cherry surface itself rather than that of flora of air or water.
Abstract: Pectinolytic yeasts, Saccharomyces marxianus, S. bayanus, S. cerevisiae var. ellipsoideus, and Schizosaccharomyces sp., predominated in the natural fermentation of coffee cherries of Coffea robusta variety grown in Chikmagalur district of Mysore State, India. These yeast species were found on the cherry surfaces, and evidence was adduced to show that the natural fermentation of coffee was the result of activity of microflora from the cherry surface itself rather than that of flora of air or water. Incorporation of pure cultures of Saccharomyces species was shown to aid the process when a mixture of all three species was used. An enzyme preparation from the Saccharomyces species was observed to hasten the mucilage-layer degradation.

Journal ArticleDOI
TL;DR: Twelve species of the genus Bacillus, a clostridium, and a streptomycete were inhibited when 30 mug/ml of crude aflatoxin was incorporated into the growth substrate.
Abstract: Among the 329 microorganisms tested for aflatoxin sensitivity were 30 genera of bacteria, 34 genera of fungi, 4 genera of algae, and 1 protozoan. Twelve species of the genus Bacillus, a clostridium, and a streptomycete were inhibited when 30 μg/ml of crude aflatoxin (36% pure) was incorporated into the growth substrate. A strain of Bacillus brevis and two of B. megaterium were most sensitive to aflatoxin, being inhibited at 10 and 15 μg/ml, respectively.

Journal ArticleDOI
TL;DR: Agreement between the rate of bacterial growth (DAP) and total microbial growth (fermentation capacity) suggests that the protozoa also grow at about this rate.
Abstract: Diaminopimelic acid (DAP) was measured in rumen samples for which growth rate had previously been estimated from changes in fermentation capacity. The growth measurements by the two methods agreed within the experimental error. Agreement between the rate of bacterial growth (DAP) and total microbial growth (fermentation capacity) suggests that the protozoa also grow at about this rate.

Journal ArticleDOI
TL;DR: A method for production of enterotoxin A in multiple liter lots is described, and production of 4 to 6 mug of enterOToxin A per ml occurred.
Abstract: A method for production of enterotoxin A in multiple liter lots is described. The medium contained 4% N-Z Amine NAK supplemented with 0.001% niacin and 0.00005% thiamine, and was adjusted to pH 6. The inoculated medium in lots of 400 to 600 ml, in 2-liter Erlenmeyer flasks, was incubated at 37 C for 24 hr on a gyrotory shaker at 280 rev/min. Production of 4 to 6 μg of enterotoxin A per ml occurred.

Journal ArticleDOI
Shuh-Wei Hwang1
TL;DR: The results indicated that liquid nitrogen storage of frozen specimens may be used as an alternative to lyophilization for long-term preservation of stock cultures of fungi.
Abstract: A preservation technique was tested on 162 strains of culturally fastidious fungi sensitive to lyophilization, representing five classes. The results indicated that liquid nitrogen storage of frozen specimens may be used as an alternative to lyophilization for long-term preservation of stock cultures of fungi. The fungus was frozen in 10% (v/v) glycerol-water menstruum in heat-sealed ampoules. The cooling from ambient temperatures to -35 C was controlled at a rate of approximately 1 C per minute. Further cooling to the storage temperature of -165 to -196 C was uncontrolled and took place at an accelerated rate. Frozen ampoules were thawed in a water bath at 38 to 40 C. Viable and unmutated cultures were developed from reactivated specimens after storage for as long as 5 years.

Journal ArticleDOI
TL;DR: The length of time that poliovirus could be recovered from wool gabardine and blanket, and from cotton sheeting, terry cloth, and knit jersey fabrics was determined under conditions of controlled temperature and humidity.
Abstract: The persistence of vaccinia virus on wool (blanket and gabardine) and cotton (sheeting, terry cloth, and knit jersey) fabrics was studied. The fabrics were exposed to the virus by three methods: direct contact, aerosol, and virus-containing dust having a high content of textile fibers. Fabrics exposed to virus by each method were held in 35 and 78% relative humidities at 25 C. Virus was recovered for up to 14 weeks from wool fabrics exposed to virus and held in the low humidity. In contrast, virus persisted for shorter periods of time on the cotton fabrics. No virus was detected on terry cloth as early as 3 days after exposure to virus. The virus appeared to be less stable in the high humidity, and the method of exposure of the fabrics to virus apparently had an effect upon the persistence of the agent. On all fabrics, viral persistence was of sufficient duration to be of epidemiological significance.

Journal ArticleDOI
TL;DR: Experiments showed that essentially all the sulfur in the cephalosporins was derived from methionine, consistent with the hypothesis that a cystathionine-mediated pathway is operative in the transfer of sulfur between methionines and cysteine.
Abstract: Methionine has an almost unique stimulatory effect on biosynthesis of cephalosporins (by Cephalosporium acremonium). No other sulfur-containing compound tested, except dl-methionine-dl-sulfoxide, replaced methionine. dl-Methionine stimulated the synthesis of cephalosporins when added after the growth phase. The utilization of inorganic sulfate was repressed by methionine. Experiments with l-methionine-S(35) showed that essentially all the sulfur in the cephalosporins was derived from methionine. Sulfur-labeled compounds found in the soluble pool from cells grown with methionine-S(35) were methionine, homocysteine, taurine, cystathionine, cysteic acid, glutathionine, and cysteine. dl-Serine-3-C(14) was incorporated into the antibiotics, and its utilization was stimulated by methionine. l-Cysteine had a sparing effect on the incorporation of methionine-S(35) and serine-C(14) into the antibiotics. The data are consistent with the hypothesis that a cystathionine-mediated pathway is operative in the transfer of sulfur between methionine and cysteine.

Journal ArticleDOI
TL;DR: On all fabrics, viral persistence was of sufficient duration to be of epidemiological significance and the method of exposure of the fabrics to virus apparently had an effect upon the persistence of the agent.
Abstract: The persistence of vaccinia virus on wool (blanket and gabardine) and cotton (sheeting, terry cloth, and knit jersey) fabrics was studied. The fabrics were exposed to the virus by three methods: direct contact, aerosol, and virus-containing dust having a high content of textile fibers. Fabrics exposed to virus by each method were held in 35 and 78% relative humidities at 25 C. Virus was recovered for up to 14 weeks from wool fabrics exposed to virus and held in the low humidity. In contrast, virus persisted for shorter periods of time on the cotton fabrics. No virus was detected on terry cloth as early as 3 days after exposure to virus. The virus appeared to be less stable in the high humidity, and the method of exposure of the fabrics to virus apparently had an effect upon the persistence of the agent. On all fabrics, viral persistence was of sufficient duration to be of epidemiological significance.

Journal ArticleDOI
TL;DR: Cultures of lactic acid bacteria, mostly from foods, were tested for their effect on the growth of Staphylococcus aureus in Trypticase Soy Broth, and the more effector bacteria there were in the inoculum, the greater was the overall inhibition (or stimulation) of S. aUREus.
Abstract: Cultures of lactic acid bacteria, mostly from foods, were tested for their effect on the growth of Staphylococcus aureus in Trypticase Soy Broth (BBL). Some of the effectors, e.g., Streptococcus faecalis, S. faecium, Lactobacillus lactis, L. brevis, and Leuconostoc mesenteroides, stimulated growth of S. aureus during early hours of growth, especially at higher temperatures of incubation, but most cultures were inhibitory, and some (S. faecium and L. mesenteroides) were even killing by the time of attainment of the maximal phase of growth of the Staphylococcus. Low-temperature meat lactobacilli and Leuconostoc dextranicum inhibited S. aureus at 10, 15, 20, and 25 C throughout its growth. Streptococcus faecalis var. liquefaciens inhibited at these temperatures and at 30 and 37 C, as well. When the ratio of effectors to staphylococci in the inoculum was 100:1, the three enterococci, the meat Lactobacillus, and L. dextranicum prevented the attainment of 5 × 106 staphylococci per milliliter at 15 C, and all but the meat Lactobacillus did so at 22 C. A ratio of 1:1 accomplished similar results at 15 C, except that S. aureus was only delayed for 12 hr by S. faecalis. A ratio of 1:100 usually was ineffective. In general, the more effector bacteria there were in the inoculum, the greater was the overall inhibition (or stimulation) of S. aureus. Inhibition was most effective at 10 or 15 C, less so at 20 or 25 C, and least at 30 or 37 C, whereas stimulation during early growth was greater at the higher temperatures. Results with different strains of the effectors and with two strains of S. aureus were similar, for the most part.

Journal ArticleDOI
TL;DR: Cultures previously reported not to degrade aflatoxin could be induced to do so under these conditions and the percentage and rate of toxin degradation were independent of toxin concentration, and appeared to be nonenzymatic and nonspecific.
Abstract: Yields of from 200 to 300 mg per liter of aflatoxins B1 and G1 were produced by two strains of Aspergillus flavus in 20-liter fermentors under proper conditions of inoculum (well-dispersed growth) and aeration (0.5 volume per volume per min of air, 300 rev/min, 30 psi back pressure, baffles). Peak yields were usually attained in 72 hr, after which the aflatoxin concentration declined rapidly. Degradation of aflatoxin depended primarily on mycelial lysis and high-aeration conditions. Cultures previously reported not to degrade aflatoxin could be induced to do so under these conditions. The percentage and rate of toxin degradation were independent of toxin concentration, and appeared to be nonenzymatic and nonspecific. Degradation simulating that occurring in the fermentor was achieved by reacting aflatoxin with peroxidized methyl esters of vegetable oil; initial degradation was rapid and appeared to involve a complex series of reactions.

Journal ArticleDOI
TL;DR: Ether extracts of cultures of 29 strains representing 6 species of Bacillus, and of individual strains of Escherichia coli, Aerobacter aerogenes, and Pseudomonas aeruginosa were examined in a gas chromatograph by use of flame ionization and electron capture detectors, and qualitative and quantitative differences between strains were noted.
Abstract: Ether extracts of cultures of 29 strains representing 6 species of Bacillus , and of individual strains of Escherichia coli, Aerobacter aerogenes , and Pseudomonas aeruginosa were examined in a gas chromatograph by use of flame ionization and electron capture detectors. Among the products detected were compounds with the chromatographic characteristics of acetic, propionic, and butyric acids, ethyl alcohol, diacetyl, acetoin, and 2,3-butanediol. The differences in peak areas of the various products formed by the bacteria were determined statistically for the chromatograms obtained with the two detectors, and the peaks were arranged in order of decreasing areas to yield a signature for each bacterial strain. Different signatures were obtained for the various genera and species and for strains of the same species. B. licheniformis, B. subtilis , and A. aerogenes formed significant quantities of a number of volatile compounds, and qualitative and quantitative differences between strains were noted. The electron capture detector was particularly sensitive to diacetyl and acetoin as well as to unknown compounds. By use of this detector, the presence of 5 pg of diacetyl and 20 pg of acetoin could be demonstrated. The quantity of acetoin detected in B. subtilis and B. licheniformis cultures was present in as little as 6.3 × 10 -3 μliters of medium. Images

Journal ArticleDOI
TL;DR: A total of 164 fatty nitrogen compounds, consisting of quaternary ammonium compounds, alkylamines, N-alkyl-1, 3-propylene diamines, substituted amino hydroxystearonitriles, and nitrogen-containing surfactants, were screened and a number of compounds were very active against gram-negative bacteria.
Abstract: A total of 164 fatty nitrogen compounds, consisting of quaternary ammonium compounds, alkylamines, N-alkyl-1, 3-propylene diamines, substituted amino hydroxystearonitriles, substituted amino hydroxystearyl amines, and nitrogen-containing surfactants, were screened for bacteriostatic, fungistatic, and algistatic activity. The most active compounds were dodecylamine and dodecylamine acetate. A number of compounds were very active against gram-negative bacteria. Most of the surfactants were virtually nontoxic to all of the test organisms.

Journal ArticleDOI
TL;DR: A bacteriocin-like substance, active against strains of Clostridium botulinum type E, is produced by certain nontoxic organisms whose biochemical properties and morphological characteristics are similar to type E.
Abstract: A bacteriocin-like substance, active against strains of Clostridium botulinum type E, is produced by certain nontoxic organisms whose biochemical properties and morphological characteristics are similar to type E. The substance, for which the name “boticin E” is proposed, is bacteriolytic for vegetative cells and bacteriostatic for spores of type E. Its spectrum of activity is somewhat strain-specific. Of the clostridial species tested, only C. botulinum type E and, to a lesser extent, C. perfringens and C. acetobutylicum, but not C. botulinum types A, B, or F, are sensitive. Irreversibly resistant variants originating from both vegetative cells and spores of certain strains were obtained. The active substance is heat-stable and dialyzable, and is not inactivated by chloroform but is digested by trypsin. Ethyl alcohol and acetone precipitates are fully active, whereas trichloroacetic acid precipitates are only partially active. Other nontoxic organisms producing similar antagonistic substances are discussed.