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Enterotoxigenic strain of Staphylococcus aureus (ATCC 14458) was grown under various conditions with constant shaking to determine the requirements for maximum toxin production. It was evident that 3% tryptic soy broth, 3% NZ-Amine NAK + 3% casein hydrolysate, 3% NZ-Amine NAK + 1% yeast extract, and 3% NZ-Amine NAK + 1% yeast extract + 0.2% glucose are most available toxin production media. But concentration of glucose could strictly triggered the enterotoxin producing efficiency. When glucose concentration was less than 0.5%, although with higher yield, the toxin production was delayed for certain period of time. However, if glucose concentration was up to more than 0.5%, the enterotoxin production was almost inhibited. Some metabolites of glucose to elucidate the inhibitory effect have also investigated. Our results indicated that glycerol and citric acid inhibited the toxin production directly, while the inhibitory effect of lactic acid and acetic acid were due to those acidic metabolites, decreased the pH value of media, and adversely suppressed the bacterial growth.
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PMID:[Studies on staphylococcal enterotoxin B. II. Production and regulation (author's transl)]. 3 15

In cultivation of meningococcus of serological group A in fluid semisynthetic medium of simple composition prepared on the basis of purified acid casein hydrolysate with profound splitting there were obtained microbial cultures with a density of 4-5 x 10(9) microbial cells per 1 ml after 20-24 hours of cultivation with shaking. Alkalinity of the medium increased (to pH 8.0-8.2 during the stationary phase) with increase of the microbial cell concentration. A study of the accumulation of group-specific thermostable polysaccharide antigen in dynamics of meningococcus cultivation on semisynthetic medium tested showed the preparations obtained by alcoholic precipitation to be colourless and to contain much antigen (by inhibition of indirect hemagglutination), particularly at the phasees of negative growth acceleration and at the stationary phase. The suggested fluid semisynthetic medium of simple composition could be used for production of diagnostic and prophylactic meningococcus preparations belonging to the serological group A.
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PMID:[Cultivation of Neisseria meningitidis serogroup A on semisynthetic liquid nutrient media]. 5 74

Submerged cultures of Brucella abortus strain 19 were studied in shaking flasks. The influence of the sterilization methods and the medium composition on the bacterial yield and cellular dissociation were studied. The selected medium was as follows (amounts in g/l): casein pancreatic hydrolizated 30; yeast extract 10; glucose, 30; sodium phosphate dibasic anhydrous 3,3; sodium monobasic monohydrate 9. Cell concentration of 8 . 10(10) viable cell/ml was obtained after 48 hours when the medium components were separated and sterilized at 121 degrees C for 20 min in autoclave.
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PMID:[Influence of the sterilization technic and culture media components on the growth and dissociation of Brucella abortus strain 19 in submerged cultures]. 11 82

Three groups of 13 adult female rats received daily for two months 23 g per rat of three synthetic isoenergetic feeds containing respectively 5, 15, 25 p. 100 casein. Plasma urea was measured weekly. Hair leachable urea and soluble dry matter were measured, at the end of the experiment, on hairs regrown after an initial clipping, in a solution obtained by washing 1 g of hair in 50 ml of HCl 0,01 N under mechanical shaking. Mean plasma urea values for the whole experiment for the three groups were 0.27, 0.38, 0.45 g per liter; differences between all groups were highly significant; however differences for the 15 and 25 p. 100 casein groups were not significant for the last three weeks. Hair leachable urea and hair soluble dry matter values were respectively 0.35, 0.80, 1.02 mg/g of hair and 20.4, 24.0, 26.0 mg/g of hair. A positive correlation links hair urea to hair dry matter (r=0.76, p less than 0.001). When hair urea is expressed in per cent of soluble dry matter, the respective values for the three groups are 1.90, 3.42, 3.95. Differences between the 5 p. 100 casein group on the one hand and the 15 and 25 p. 100 casein groups on the other are highly significant (p less than 0.001).
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PMID:[Influence of protein intake on plasma and hair leachable urea in the rat (author's transl)]. 116 65

The effects of eluent composition, pH, and chaotropic agents on the recovery of T2, MS2, and indigenous coliphages from various foods were investigated. Additionally, methods of sample suspension and clarification were evaluated for coliphage recovery and application to various foods. Clarified sample suspensions were assayed for coliphages with a modified agar layer technique and appropriate Escherichia coli hosts. Centrifugation and polypropylene mesh filtration were more rapid and effective than glass wool filtration for clarification of sample suspensions and subsequent recovery of coliphages. Blending, stomaching, and shaking procedures were generally comparable for sample liquefaction and release of coliphages from foods. Complex basal eluents, EC medium and 1% casein, were generally more effective than a less complex eluent, phosphate buffer, for elution of coliphages from foods. For most foods, incorporation of sodium chloride or chaotropic agents, i.e., sodium trichloroacetate, urea, Tween 80, Triton X-100, and sodium nitrate, into basal eluents did not enhance recovery of coliphages. Indigenous coliphage recovery was not affected by sample suspension pH over a range of 6.0 to 9.0. With an optimal procedure, i.e., EC medium eluent, blending, and centrifugation, the recovery of T2 and MS2 ranged from 48 to 81% and from 58 to 100%, respectively, depending on the food type.
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PMID:Methodology for enumeration of coliphages in foods. 352 56

Pasteurella pestis, harvested after 24 to 30 hr of growth in a casein hydrolysate medium at 26 C, was resuspended and shaken in 3% lactose-0.1 m phosphate buffer for 4 hr at the same temperature. Certain characteristics of these starved cells were compared with those of control cells. No differences in the amounts of cellular carbohydrate or lipid were detected. The concentrations of the principal free amino acids were greater in the shaken cells, except that they contained no measureable arginine, and the normally large pools of intracellular tricarboxylic acid cycle intermediates were reduced. Greater viable-cell counts resulted with the cells that were shaken in lactose buffer than with the control cells when each was incubated at 5 C for several weeks. However, the reduced viabilities were apparent losses caused by the formation of aggregates of cells. The clumping of cells was caused by the polymerization of extracellular nucleic acids, principally deoxyribonucleic acid, that were excreted by the cells. Cell clumping could be partially prevented by prior shaking of the suspended cells, which removed some of the deleterious material, or by the action of crystalline deoxyribonuclease.
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PMID:Biochemical and physical changes in shaken suspensions of Pasteurella pestis. 533 54

Duerre, John A. (University of North Dakota, Grand Forks), and Patrick J. Buckley. Pigment production from tryptophan by an Achromobacter species. J. Bacteriol. 90:1686-1691. 1965.-A microorganism was isolated from the soil near the University of North Dakota. Biochemical and morphological characteristics indicated that this organism would best be classified as a member of the family Achromobacteraceae, genus Achromobacter, species unknown. The organism produced a red pigment when grown in a medium containing yeast extract and tryptophan. The pH optimum for pigment production was about 8.0 and the optimal temperature was 25 C. During a study of the nutritional requirements for growth and pigment production, it was found that the organism would grow and produce pigment in a medium containing tryptophan and nucleosides, but the rate of both growth and pigment formation in this medium was slower than that observed with tryptophan and yeast extract. The organism grew well in the presence of acid-hydrolyzed casein and nucleosides without producing pigment, indicating that the pigment is not necessary for growth. Resting-cell experiments definitely established tryptophan as the sole exogenous requirement for pigment production. The pigment was extracted from yeast extract-tryptophan medium with chloroform. Thin layer chromatographic analysis of the crude pigment extracted from this medium revealed the presence of two other pigments in addition to the major red pigment. One of these was a highly fluorescent orange pigment and the other a pink pigment. Only the red pigment was produced by resting cells in the presence of tryptophan alone. This pigment served as an electron acceptor when coupled with formic dehydrogenase, indicating its possible function as an oxidation-reduction pigment. The oxidized pigment had absorption peaks at 506 and 304 mmu. The peak at 506 mmu disappeared upon reduction with sodium sulfite. Shaking the reduced pigment in air proved to be an unsatisfactory method for returning the reduced pigment to the oxidized, colored state.
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PMID:Pigment production from tryptophan by an Achromobacter species. 585 90

Methods currently recommended for quantitative bacteriological sampling of surfaces usually suffer from either unreliable results (plain swabbing techniques and contact cultures) or high expenses in terms of material, equipment and/or labour (rinse techniques using sampling fluids). Therefore, a modified swab-rinse technique (SRT) was devised as a reasonably precise and simple alternative: A small amount of sampling fluid (1.5 ml), which can contain neutralisers, is transferred onto the flat surface under investigation; with this fluid and a pre-moistened small cotton-swab an area of 3 cm diameter is thoroughly washed for 15 s. Then 0.1 ml- and 0.5 ml-amounts of the washliquid are collected with automatic microliter pipettes and transferred and spread onto Casein-Soy-Agar for enumeration. In parallel experiments with contact cultures (rodac plates), the new SRT up to 3000 cfu per sample exhibited a linear answer to increasing inocula of E. coli on tiles (Fig. 1, rings). Rodac-plates proved to possess a rather limited span of reliable counts: above 100 colonies, increasing numbers of bacteria were prevented from forming distinguishable colonies (Fig. 1, dots and +es); thus, colony counts higher than 100 on rodac plates cannot be expected to be true estimates. In addition, in the higher count range the results of a common, simplified counting technique differed markedly from results of true counts (Fig. 2). Both methods were equally sensitive in detecting low counts of bacteria (Fig. 1, Fig. 3). Shake-rinse techniques using high volumes of sampling fluid provide lower sensitivity than contact cultures or the new SRT: The use of 100 ml of sampling fluid (15) and plating of 0.1 ml- or 1.0 ml-aliquots of sampling fluid keeps the threshold for detection of bacteria as high as 1000 and 100 per sample, respectively (Fig.3); nevertheless, sensitivity of shake-rinse techniques can be increased by filtration of the whole sample. Thus, the new swab-rinse technique combines several advantages: wide span of true estimates since washliquid can be diluted for enumeration of high counts; high sensitivity ( = ability to detect small numbers of testbacteria in sample) since about one half of the sample is plated; good recovery of testbacteria from both smooth and coarse surfaces; simplicity; the new swab-rinse technique requires basic laboratory equipment and ordinary media and no shaking- or filtration devices; option for immediate and strong neutralisation of disinfectant residues; the sampling fluid can contain any neutraliser; option for automated colony counts since any kind of petri-dishes can be used for culture.
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PMID:[A simple wash technic for the recovery of test pathogens in the evaluation of surface disinfection procedures]. 637 50

Previous morphometric or biochemical investigations have yielded different data on the distribution of free and membrane-bound ribosomes in midgut cells of Aedes aegypti. In the present paper ribosomal distribution has been morphometrically analysed to determine whether different mosquito strains, different food and different narcosis used in these previous studies, and/or methodological errors, could account for the different results. Most of the cellular parameters in the stomach epithelium of female A. aegypti, strain Rockefeller, and their changes during blood digestion, are comparable to those measured for another Aedes strain (Segemaganga, Hecker and Rudin, 1979), and ae generally similar to those of Anopheles stephensi (Hecker 1978). Proteolytic activity against casein is similar for both Aedes strains with a maximum activity being registered around 30 h after a blood meal. During digestion of human serum there is no increase in the ratio of membrane-bound to free ribosomes, and no significant increase in the surface area of the rough endoplasmic reticulum or of the number of bound or free ribosomes. Proteolytic activity is distinctly lower than during blood digestion. Immobilization of mosquitoes prior to dissection by either narcosis or by shaking in a test tube has no significant influence on cellular parameters in females fed on sugar solution and investigated 3 days after emergence. It is concluded that the differences in ribosomal parameters previously obtained by morphometrical (Hecker and Rudin 1979) and biochemical (Gander et al. 1980) methods, can only partly be explained by the selection of different food for the mosquitoes, and must also have been caused by methodological inadequacies.
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PMID:Morphometric parameters of the midgut cells of Aedes aegypti L. (Insecta, Diptera) under various conditions. 702 89

The psychrotrophs SYP-A2-3 producing the cold-adapted protease has been isolated from the bacterial samples collected from the No. 1 Glacier of China and identified as Bacillus cereus according to its morphological and physiochemical characteristics and 16s rDNA gene sequence analysis. It could grow between 0 degree C and 38 degrees C while its optimal growth temperature was 25 degrees C and the optimal temperature for its protease production was 15 degrees C. The cold-adapted protease was identified as neutral metallo-protease, the molecular weight was 34.2 kD shown by SDS-PAGE, the optimal pH and temperature for activity was 7.0-8.5 and 42 degrees C, respectively. Various fermentation conditions of its protease production were also investigated. The results showed that casein was the best nitrogen source while glucose and starch were suitable carbon source for its protease production. The initial pH of fermentation broth ranged from 6.5 to 7.0 was optimal. Under optimized conditions, the protease activity produced by SYP-A2-3 could reach 3800 U/mL and 4800 U/mL conducted in shaking flask and 5 L stirred jar experiment, respectively.
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PMID:[Identification of psychrotrophs SYP-A2-3 producing cold-adapted protease from the No. 1 Glacier of China and study on its fermentation conditions]. 1598 72


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