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PubMed2006

Menstrum for culture preservation and medium for seed preparation in a tetanus toxin production process containing no animal or dairy products.

AIMS: To completely eliminate animal and dairy products from the lyophilization menstrum and the seed medium used to produce tetanus toxin with Clostridium tetani. METHODS AND RESULTS: Tetanus toxin production in a recently developed fermentation medium lacking animal and dairy products was studied with different seed media. It was found that soy peptone could completely replace the beef heart infusion plus animal peptone previously used as seed medium. In addition, we found that cells lyophilized in soy milk could replace the usual type of cells lyophilized in cow's milk. CONCLUSIONS: We have now developed a complete tetanus toxin production process containing no animal and dairy products. SIGNIFICANCE AND IMPACT OF THE STUDY: Toxoid preparations made from toxin produced with animal and dairy products can contain undesirable contaminants such as the prion causing bovine spongiform encephalopathy (Mad Cow's Disease) or antigenic peptides that stimulate anaphylactic reactions and other undesirable immune reactions in immunized hosts. The new vegetable-based process described here avoids such unfortunate possibilities.

Animals↗
PubMed1977

In vitro studies on normal, stimulated and immunologically activated mouse macrophages. I. Oxidation of 1-14C glucose by macrophages in monolayer cultures.

Oxidation of 1-14C glucose of BCG/listeria activated, proteose-peptone stimulated and typhoid-parathypoid vaccine (T.A.B.) stimulated peritoneal and spleen macrophages from C3H, CBA and C57Bl mice was determined at various times after treatment. The study showed no major differences between the oxidation of stimulated and activated macrophages, but a variation was seen in C3H mice in the kinetics of glucose oxidation of BCG/listeria activated and proteose-peptone stimulated peritoneal macrophages. It was also observed, in these mice, that the response appeared and disappeared earlier in the spleen than in the peritoneal macrophages.

Animals↗
PubMed1977

In vitro studies on normal, stimulated and immunologically activated mouse macrophages. II. Degradation of radioactive antigen/antibody complexes.

Radioactive antigen/antibody complexes are degraded to a greater extent by PE macrophages from C3H, CBA and C57Bl mice injected intraperitoneally with proteose-peptone (stimulated macrophages) and T.A.B. vaccine than by PE macrophages from normal mice and mice immunized with BCG (activated macrophages). On the other hand, spleen macrophages from mice immunized with BCG had a greater digestive capacity than macrophages from mice injected with proteose-peptone or T.A.B. vaccine or from normal mice. Thus, activated macrophages are capable of degrading antigen/antibody complexes, provided that the cells are harvested from one of the foci of BCG infection.

Animals↗
PubMed1974

The role of vagal innervation in gastric and pancreatic responses to meals varying in pH.

In three dogs provided with chronic gastric and pancreatic fistulae, gastric acid response to a peptone meal was measured by a method of intragastric titration at pre-selected pH values ranging from 6.0 to 2.0 or at a constant pH during intravenous infusion of secretin in graded doses ranging from 0.25 to 4.0 u./kg.hr. Simultaneously, pancreatic secretion was determined by the standard collection technique and serum gastrin response was measured by radio-immunoassay. In this way the pH profile for the inhibition of gastric response and stimulation of pancreatic secretion as well as serum gastrin response to a peptone meal have been determined and compared with that obtained with exogenous secretin or feeding a liver meal. Truncal vagotomy resulted in a significant decrease of gastric acid response to the meal at all pH levels or at all doses of secretin but the inhibitory effect of gastric acidification or secretin on serum gastrin and gastric acid response to a meal remained unchanged. Vagotomy caused a decrease of pancreatic secretory response to meals at all pH levels and at all doses of secretin except the largest one. Pancreatic secretion stimulated by secretin alone was not affected by vagotomy, whereas that induced by feeding a liver meal was markedly decreased by this procedure. From these studies we have tentatively concluded that the vagus nerve facilitates the release of intestinal hormones and that vagotomy causes a decrease of gastric and pancreatic responses to endogenous but not to exogenous stimuli.

Animals↗
PubMed1976

Production of an extracellular maltase by thermophilic Bacillus sp. KP 1035.

Production of extracellular maltase was studied with thermophilic Bacillus sp. KP 1035, which was selected as the organism producing the highest levels of maltase. The final enzyme yield was increased by maltose, peptone, and yeast extract but reduced by succinate and fumarate. Maximum enzyme production was achieved at 55 degrees C and at an initial pH of 6.2 to 7.0 on a medium containing 0.3% maltose, 1% peptone, 0.1% meat extract, 0.3% yeast extract, 0.3% KH2PO4, and 0.1% KH2PO4. Maltase was synthesized in cytoplasm and accumulated as a large pool during the logarithmic growth phase, which preceded sporulation. At the end of this phase, the enzyme appeared in the culture broth, and its accumulation increased in parallel with a rise in the extracellular protein level. Maltase was stable for 24 h at 60 degrees C over a pH range of 5.6 to 9.0 and retained 95% of the original activity after treatment for 20 min at 70 degrees C at pH 6.8.

Bacillus↗
PubMed1977

Procedures involving lipid media for detection of bacterial contamination in breweries.

The liquid equivalent of universal beer agar, designated universal beer liquid medium, and its beer-free equivalent, universal liquid medium (UL), were equally effective in demonstrating bacterial contamination in 120 of 200 samples from different stages of commercial brewing process. Growth of the contaminants after 3 days was consistently more luxuriant in the UL medium. A yeast-water substrate medium failed to reveal many contaminants detected with UL in 392 samples from three breweries and revealed only a few not detected with UL. The use of UL and a lactose-peptone medium, with microscope examination of the media for bacterial growth, permitted detection of 93% of the known contaminants compared to 87%, detected with UL alone; this combination or universal beer liquid medium plus lactose-peptone medium can therefore be recommended for the detection of bacterial contaminants in brewery samples. Bacterial contamination of pitching yeasts appeared to be a particular problem in the breweries investigated.

Bacteria↗
PubMed1977

Diluent sensitivity in thermally stressed cells of pseudomonas fluorescens.

Thermally injured cells of Pseudomonas fluorescens were unable to produce colonies on Trypticase soy agar (TSA) after dilution with 0.1% peptone. Nutritional exigency could not be used as the criterion for this injury, since varying the composition of the plating medium had little effect on the number of colonies that developed. The injured cells had no requirement for compounds known to leak out during the heat treatment in order to recover. The cells did not exhibit injury if dilution preceded heat treatment on the plating medium, demonstrating that the heat treatment sensitized the cells to the trauma of dilution. Substitution of 0.1% peptone with growth medium as the diluent largely offset the previously observed drop in TSA count. Little difference in survival was observed when monosodium glutamate or the balance of the defined medium was used as the diluent. The diluent effect was ionic rather than osmotic. The presence of cations was important in maintaining the integrity of the injured cell, and divalent cations enhanced this protective effect. The role of these cations at the level of the cell envelope is discussed.

Cations, Divalent↗
PubMed1981

Culture medium for selective isolation and enumeration of Gram-negative bacteria from ground meats.

We developed a new medium, designated peptone bile amphotericin cycloheximide (PBAC) agar, which contains (per liter) 10 g of peptone, 300 mg of bile salts, 1 mg of amphotericin B, 1 g of cycloheximide, and 15 g of agar. When 21 samples of fresh ground beef were studied and plate count agar counts were used as references, we obtained a mean recovery of 28% of total counts with violet red bile agar overlay, whereas we obtained 48% recovery with PBAC agar. With 12 samples of frozen ground beef, recovery on violet red bile agar overlay was 29% of the recovery on plate count agar, whereas the corresponding value on PBAC agar was 45%. PBAC agar allowed the enumeration of 1.4 times as many gram-negative bacteria as violet red bile agar overlay. None of eight strains of gram-positive bacteria and none of eight strains of yeasts grew on PBAC agar. Of 158 colonies randomly selected from pour plates of eight fresh ground meat samples, 95% stained gram negative. In comparison, only 70% of 151 colonies selected from corresponding plate count agar plates were gram negative. The lack of background color, turbidity, and ease of use make PBAC agar easier to handle than other media used for gram-negative bacteria, such as violet red bile agar, violet red bile agar overlay, and crystal violet tetrazolium agar. In the preparation PBAC agar, all ingredients are autoclaved together except amphotericin B, which is filter sterilized and added before the plates are poured.

Amphotericin B↗
PubMed1983

Broth medium for enrichment of Vibrio fluvialis from the environment.

A medium was designed for the enrichment and enumeration of Vibrio fluvialis from environmental samples. The medium contains 1% peptone plus 4% sodium chloride and 5 micrograms of novobiocin per ml, pH 8.5. This V. fluvialis enrichment medium (FEM) was tested, in comparison with alkaline peptone (AP), in field samplings. A total of 177 samples (estuarine waters and sediment, sewage, and crabs) collected over a 14-month period were examined with FEM and with AP broth. Results showed that FEM was more effective than AP in detecting V. fluvialis, particularly from water and sewage samples with low salinities (less than 6%). The best recovery of V. fluvialis occurred when both enrichment media were used simultaneously.

Brachyura↗
PubMed1998

Enhanced broth media for selective growth of Vibrio vulnificus.

Rapid detection of Vibrio vulnificus can be enhanced by optimizing the components of enrichment broth. PNC (5% peptone, 1% NaCl, and 0.08% cellobiose [pH 8.0]) enhanced the growth of V. vulnificus compared to alkaline peptone broth. PNCC (PNC with 1.0 to 4.1 U of colistin methanesulfonate per ml) increased the growth of low levels of V. vulnificus while suppressing non-target bacteria.

Anti-Bacterial Agents↗
PubMed1970

Bacteriology of manganese nodules. IV. Induction of an MnO2-reductase system in a marine bacillus.

Bacillus 29, isolated from a ferromanganese nodule from the Atlantic Ocean, was shown to possess an MnO(2)-reductase system which is induced in the presence of manganous ion. Maximal activity of the enzyme system was induced in about 5 hr in the presence of 4.35 mm MnSO(4) and was minimally dependent on the presence of either glucose or peptone and oxygen. Induction of optimal activity required the simultaneous presence of glucose and peptone. At least 30% of maximal activity was induced in 5 hr in the presence of 0.4 mum MnSO(4). Actinomycin D (5 mug/ml) or chloramphenicol (35 mug/ml), when added to the induction medium, inhibited approximately 90% of MnO(2)-reductase synthesis and incorporation of uracil-2-(14)C or leucine-1-(14)C. Cell-free extracts having MnO(2)-reductase activity were prepared by sonic disruption of cell suspensions of induced Bacillus 29. Such extracts used glucose metabolism as a source of electrons. They had an average specific activity of 1.15 nmoles of Mn(II) produced per mg of protein per hr at 25 C. They had a temperature optimum of 18 C for reductase activity and retained 50% of their activity at 4 C, the approximate temperature of the natural habitat of the organism. Extracts were stable for several days at 4 C but rapidly lost over 50% of their activity on freezing and thawing. Over 90% of the activity of the extract could be destroyed by heating in a boiling-water bath for 5 min. At a concentration of 1 mm, HgCl(2) and atebrine dihydrochloride inhibited MnO(2)-reductase activity by at least 50%, but sodium azide was ineffective. The MnO(2)-reductase activity of induced cells and extracts from them was no greater in the absence of oxygen than in its presence, confirming an earlier observation that MnO(2) and O(2) do not compete as terminal electron acceptors in the respiratory activity of this organism.

Bacillus↗
PubMed1973

Growth and cloning of Tetrahymena pyriformis on solid meduium.

A method of obtaining clones of Tetrahymena pyriformis on solid medium has been developed. The medium consists of a basal layer of 1.5% agar topped with 2 ml of 0.3% agar in sterile, plastic petri plates (100 by 15 mm). Both agar layers contain either 2% proteose peptone and 0.1% liver extract (complex medium) or defined medium supplemented with proteose peptone. After drying, 0.5 ml of liquid culture is spread evenly over the top agar, and the plates are then sprinkled lightly and evenly with autoclaved dry Sephadex G-25 (fine). Cell colonies can be observed after 5 days of incubation either by viewing with a microscope or without the aid of a microscope after staining. Plating efficiency is high on either complex or defined medium with a number of strains of Tetrahymena, both micronucleate and amicronucleate. Colonies can be picked and transferred to liquid culture for further growth. The existence of clones was demonstrated by plating a mixture of two different drug-resistant mutants. The method should prove useful in selective procedures for the isolation of mutants and for determining survival after treatments such as ultraviolet irradiation.

Agar↗
PubMed1996

High-affinity maltose/trehalose transport system in the hyperthermophilic archaeon Thermococcus litoralis.

The hyperthermophilic marine archaeon Thermococcus litoralis exhibits high-affinity transport activity for maltose and trehalose at 85 degrees C. The K(m) for maltose transport was 22 nM, and that for trehalose was 17 nM. In cells that had been grown on peptone plus yeast extract, the Vmax for maltose uptake ranged from 3.2 to 7.5 nmol/min/mg of protein in different cell cultures. Cells grown in peptone without yeast extract did not show significant maltose or trehalose uptake. We found that the compound in yeast extract responsible for the induction of the maltose and trehalose transport system was trehalose. [14C]maltose uptake at 100 nM was not significantly inhibited by glucose, sucrose, or maltotriose at a 100 microM concentration but was completely inhibited by trehalose and maltose. The inhibitor constant, Ki, of trehalose for inhibiting maltose uptake was 21 nM. In contrast, the ability of maltose to inhibit the uptake of trehalose was not equally strong. With 20 nM [14C]trehalose as the substrate, a 10-fold excess of maltose was necessary to inhibit uptake to 50%. However, full inhibition was observed at 2 microM maltose. The detergent-solubilized membranes of trehalose-induced cells contained a high-affinity binding protein for maltose and trehalose, with an M(r) of 48,000, that exhibited the same substrate specificity as the transport system found in whole cells. We conclude that maltose and trehalose are transported by the same high-affinity membrane-associated system. This represents the first report on sugar transport in any hyperthermophilic archaeon.

Archaea↗
PubMed1962

Constancy of characteristics in the streptomycetes.

Sanchez-Marroquin, A. (University of Mexico, México D.F.). Constancy of characteristics in the streptomycetes. J. Bacteriol. 83:1183-1192. 1962.-A total of 150 Streptomyces strains isolated from soil was studied during a 3-year period in regard to constancy and variation of the following characteristics: sporophore micromorphology, color of the aerial and substrate mycelium, surface configuration of the spores, assimilation of carbon compounds, production of H(2)S and melanoid pigment, and reduction of nitrates. A remarkable constancy in the following characteristics was found: (i) sporophore micromorphology when only three of the seven morphological series of Pridham et al. were developed on tomato paste-oatmeal agar or yeast extract-malt extract agar; (ii) color of the aerial mycelium if only four fundamental colors are distinguished (white to cream or buff shades; yellow to orange or brown; pink to cinnamon, red or pinkish tan to lavender; and green to gray or blue); (iii) surface configuration of the spores divided in two types (smooth and warty to spinous or hairy); (iv) assimilation of five carbon compounds (arabinose, xylose, rhamnose, raffinose, and mannitol); (v) production of H(2)S on Difco peptone-iron agar supplemented with 0.1% Difco yeast extract; and (vi) production of melanoid pigment on peptone agar, giving similar results to those of the H(2)S test. Color of the substrate mycelium, size and shape of the spores, and reduction of nitrates should be used only as complementary data in the species descriptions, owing to their inconsistency and unreliability.

Actinomycetales↗
PubMed1964

Proteolytic activity and general characteristics of a marine bacterium, Aeromonas proteolytica sp. N.

Merkel, Joseph R. (Fort Johnson Marine Biological Laboratory, College of Charleston, Charleston, S.C.), Eugene D. Traganza, Barid B. Mukherjee, Travis B. Griffin, and J. M. Prescott. Proteolytic activity and general characteristics of a marine bacterium, Aeromonas proteolytica sp. n. J. Bacteriol. 87:1227-1233. 1964.-A highly proteolytic bacterial species was isolated from the alimentary canal of the marine borer, Limnoria. The morphological and biochemical characteristics of the organism indicated that it was a new Aeromonas species, for which the name A. proteolytica is proposed. When freshly isolated, the organism required seawater for growth; but, upon prolonged culture in the laboratory, it was able to grow in media of greatly reduced salt concentration, provided that relatively large amounts of peptone were supplied. Peptone or hydrolysates of casein were capable of supplying all organic nutrients required for growth and proteinase production. Certain individual amino acids were also able to furnish all energy, carbon, and nitrogen requirements. Inorganic nitrogen was utilized in the presence of citrate, but could not serve as the only source of nitrogen in the presence of glucose. The organism was facultatively anaerobic, but best growth and proteinase production occurred only with vigorous aeration. The amount of growth obtained in 24 hr increased rapidly as the incubation temperature was increased up to a maximum of 40 C, but no growth occurred at 42 C.

Aeromonas↗
PubMed1968

Spore germination in strains of Dictyostelium discoideum and other members of the Dictyosteliaceae.

Spores of all strains of Dictyostelium discoideum tested in this study germinated after a heat shock of 45 C for 30 min. Whereas the strains differed in their rates of germination, the rate for each strain was constant. A correlation existed between the rate of germination and the rate of vegetative growth when spores were inoculated into bacterial streaks. Heat shock clearly increased spore germination in D. purpureum, but the response was less dramatic than in D. discoideum. Enhancement also occurred in D. rosarium, but only in media containing peptone. Strains of D. mucoroides gave varied responses, and these could be divided into those which required mutrients for spore germination and those which did not. The spores of Polysphondylium pallidum were resistant to mild heat (45 C), but were not activated; peptone was required for germination. In contrast, the microcysts of this species were heat-labile and required no added nutrients for excystment.

Culture Media↗
PubMed1990

Assessment of conventional and commercial methods for identification of clinical isolates of cysteine-requiring strains of Escherichia coli and Klebsiella species.

Cysteine-requiring strains of the family Enterobacteriaceae that are auxotrophic for this amino acid because of defects in the sulfur assimilatory pathway account for about 1.5% of urinary tract isolates of Escherichia coli and Klebsiella species. Forty Escherichia and eight Klebsiella cysteine-requiring strains were used to test the ease with which various test systems identified clinical isolates of cysteine auxotrophs. In a preliminary experiment, the growth yield of 10 cysteine-requiring E. coli in 10 solutions of commercially available peptones was in each instance less than that of prototrophic control and showed that these sources of nutrients were suboptimal for these strains. A significant proportion of the cysteine-requiring strains were not adequately identified by growth-dependent tests which used various peptones as a nutrient source. Problems were encountered with all test systems examined, which were as follows: conventional methods; the API 20E, Microbact, and Vitek systems; and two rapid methods for the identification of E. coli, the Rapidec coli and the beta-D-glucuronidase tests. The performance of the test systems was only partly improved when inocula were derived from appropriately supplemented media. However, the problems of the growth-dependent tests were resolved when a cysteine-supplemented suspension was used to inoculate each test system.

Bacteriological Techniques↗
PubMed1976

Medium-dependent inhibition of Peptostreptococcus anaerobius by sodium polyanetholsulfonate in blood culture media.

Of 13 species of anaerobic cocci, Peptostreptococcus anaerobius was the only species tested that was sensitive to 0.1% sodium polyanetholsulfonate (SPS). However, the sensitivity of P. anaerobius to SPS varied according to the media in which the cultures were grown. In supplemented peptone (B-D) and brain heart infusion media, most strain of P. anaerobius were not inhibited by SPS. Gelatin and proteose peptone were the medium components which were protective. The minimal inhibitory concentration of SPS for P. anaerobius was approximately 60-fold higher in media. However, the concentration of SPS required to neutralize the bactericidal properties of human serum was only four fold higher in media containing geltain. In a commerical medium containing SPS (0.03%) and gelatin (1.2%), SPS-sensitive strains of P. anaerobius were not inhibited by SPS, and the bactericdal action of human blood on Escherichia coli C and Serratia marcescens SM 29 was eliminated.

Anaerobiosis↗