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PubMedDate not supplied

Ultrastructural location of the cross-protection factor on in vivo and in vitro grown Pasteurella multocida.

Pasteurella multocida from infected turkey tissues expresses a unique immunogen called cross-protection factor (CPF) that induces immunity to challenge by both homologous and heterologous serotypes. In this study, we used a monoclonal antibody (AMP MAb) to CPF and protein A-colloidal gold (PACG) to locate CPF on P. multocida. After incubation with AMP MAb and PACG, CPF was detected at the bacterial surface and cell periphery of P. multocida in infected turkey liver and P. multocida isolated from infected turkey blood. CPF was not detected on P. multocida incubated with control monoclonal antibody. Pasteurella multocida isolated from infected turkey blood and cultivated in the peptone-based medium did not express CPF consistently, and some cells contained more CPF than others. The location of CPF also varied, and CPF was detected both intracellular and extracellular on the cell surface. In the latter cells, CPF was heavily concentrated to a specific lateral site or detected sloughing from the cell surface. These results correlate with laboratory observations that CPF detected on P. multocida from infected turkey tissues, P. multocida isolated from infected turkey blood, and P. multocida cultivated in peptone-based medium is associated with outer membrane fractions.

Animals↗

PubMed2001

Cultural requirements for in vitro seed germination, protocorm growth and seedling development of Geodorum densiflorum (Lam.) Schltr.

Effects of different nutrient solutions, organic supplements and plant growth regulators on in vitro seed germination and protocorm development of Geodorum densiflorum (Lam.) Schltr. were studied. Seed germination was very high (up to 96%) in all the basal media, with Knudson's C and half-strength Murashige & Skoog being slightly more productive than Vacin & Went. Application of organic supplements and NAA had little effect on germination, but BAP proved inhibitory. After germination, protocorms exhibited a clear preference for peptone and NAA for much faster growth, while BAP resulted in stunted growth. Beside normal development, disorganisation of protocorms, followed by callusing occurred in presence of peptone and NAA. The calli were compact with limited growth and frequently regenerated protocorm like bodies. Development of seedlings was preceded by an intermediary rhizome phase. Growth of rhizomes was slow in the plant growth regulator free medium and about 15 months of culture was required for seedling formation. However, it was possible to hasten the process by 8-10 months with the employment of NAA, which also enhanced the number of seedlings per protocorm through axillary branching. Combined application of high BAP and low NAA was also useful for high rate of seedling formation.

Culture Media↗

PubMedDate not supplied

[Effect of mineral fertilizers on the microflora of corn roots].

The paper describes studies of the density of microorganisms grown on meat-peptone agar, fungi, nitrifiers, cellulose degraders, actinomycetes and azotobacter in the soil of the radical area of corn grown in field experiments with various doses and ratios of mineral fertilizers. In the forest-steppe zone (leached chernozem) of the Krasnoyarsk region mineral fertilizers activated proliferation of radical microorganisms grown on meat-peptone agar, fungi, nitrifiers and cellulose degraders. The effect of the tested fertilizers was dissimilar during plant vegetation. Under the influence of fertilizers proliferation of actinomycetes increased at the phase of 3--4 leaves. At later phases of the plant development a similar tendency was manifest upon soil application of N120P60K60, N60P120K60 and N120P120K120. The azotobacter density increased only after application of N60P120K60, N60P60K120 and N30P60K30.

Chemistry, Agricultural↗

PubMed1975

Studies on the cultivation of Clostridium oncolyticum M 55. 5th communication: The influence of iron, zinc, and cobaltous ions on the growth and the kininase activity of clostridium oncolyticum M 55 ATCC 13.732.

The precipitation or iron sulfide by hydrogen sulfide excreting Clostridium oncolyticum M 55 led to further examinations in order to avoid this disturbing factor. The method should render the cultivation of Cl. oncolyticum M 55 under definite conditions and should also intensify the formation and the enrichment of the clostridial kininases. Cobaltous ions and zinc granules were found useful for cultivation, because no disturbances increased from both during sterilization and incubation. The determination of the growth cycles and the replication rates showed the usefulness of zinc for substitution of iron. Cobaltous ions did only allow a low replication rate. These results confirmed those showing the influence of chelate-forming agents on the kininase activity. Cobaltous ions were not able to exchange the central atom of the metallo-proteide. The influence of the peptone quality on the kininase activity in presence of zinc was also examined, since examinations with iron media have shown a direct connection between peptone quality and enzyme activity. The results with zinc-containing media were nearly 5-10% lower than those with iron. The results allowed also the conclusion that the kininase excreted by Cl. oncolyticum M 55 is an iron-containing metallo-proteide, whose central atom is exchangeable for zinc. The consequences of the loss of enzyme activity for the oncolytic therapy with Cl. oncolyticum M 55 will be shown in further examinations.

Acetates↗

PubMed2000

Crayfish broth for bacterial culture.

OBJECTIVE: To formulate a new growth medium for bacteriological studies. METHODS: Sun dried white shrimps, Nematopalaemon hastatus, were ground into fine powder and infused in distilled water at 7+/-1 degrees C for 24 h. The pH of the infusion was adjusted to 7.2 with NaOH (0.1 m) and HCl (0.1 m) and evaporated to dryness. Crayfish broth (CB) was produced by reconstituting the concentrate in distilled water and sodium chloride was added at 0.9% w/v. Crayfish infusion agar (CIA) and crayfish starch agar (CSA) were prepared with CB as nutrient base. The crayfish broth and agar were evaluated alongside with nutrient broth and agar (Oxoid) for ability to support growth of pathogens such as Escherichia coli, Klebsiella aerogenes, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis. Crayfish broth was tested for suitability for sugar fermentation and indole test and results compared with those obtained with peptone water. Its suitability as a sensitivity test medium was evaluated using ampicillin (10 microg) and gentamicin (10 microg) discs and results compared with those obtained using Mueller-Hinton agar (Oxoid). RESULTS: Crayfish broth comprises crayfish infusion concentrate 3.5 g and sodium chloride 9.0 g dissolved in one litre of distilled water. The crayfish broth, CSA supported the growth of the test pathogens with cultural characteristics similar to those seen on nutrient agar. The pH of the spent crayfish broth was near neutral (pH 6.9). The broth was as important as peptone water for sugar fermentation test but was not suitable for indole test. The results of antibiotic susceptibility test on CIA were comparable with those obtained on Mueller-Hinton agar with difference in zone diameters of not more than 1 mm. CONCLUSION: Crayfish broth possesses enormous potential as a new growth medium for bacterial studies.

Animals↗

PubMed2003

Evaluation of germ tube test in various media.

One of the most valuable tests for the rapid presumptive identification of Candida albicans is germ tube production. Previously researchers have concluded superior results with serum in germ tube test. However, handling of pooled human serum has danger of acquisition of hepatitis and HIV infections. Therefore, in the present study we compared various media--pooled human serum, sterile horse serum, peptone water and tryptic soy broth. It was observed that human serum remains to be the best with 100% positivity for identification of C. albicans followed by horse serum (76.3%), peptone water (61.8%) and tryptic soy broth (61.8%).

Candida albicans↗

PubMed2004

Effect of combined nutrients on biosurfactant produced by Pseudomonas putida.

This work investigated biosurfactant production by Pseudomonas putida in combined C/P, C/Ninorganic, C/Fe, C/Mg nutrient ratios and peptone concentration. Analysis of the 2(5-1) fractional factorial experimental design showed that only the C/Fe ratio had a significant (p<0.02) effect on biosurfactant production. The highest amount of biosurfactant was obtained at low C/Fe ratios, but net surface tension did not show significant differences. In addition, low amounts of peptone and the C/P-C/Mg nutrient ratios interaction significantly (p < 0.05) enhanced the biomass produced by P. putida. Analysis of biosurfactant by gas chromatography (GC) showed that the hydrophilic fraction was composed by rhamnose and the hydrophobic fraction, mainly by palmitic (C16), stearic (C18:0), oleic (C18:1) and linoleic (C18:2) fatty acids.

Bioreactors↗

PubMed2001

Three-factor response surface optimization of the production of dextran dextrinase by Gluconobacter oxydans.

Response surface methodology (RSM) and a five-level three-factor central composite rotatable design (CCRD) were used to evaluate the effect of glycerol and peptone concentration and initial pH on dextran dextrinase (DDase) production by Gluconobacter oxydans. Optimal fermentation conditions were 20.59 g/l of glycerol, 6.67 g/l of mycological peptone and an initial pH of 6.14. The predicted DDase yield of the optimised fermentation was 0.207 U/ml, whereas an actual experimental yield of 0.208 +/- 0.025 U/ml was obtained.

Fermentation↗

PubMed2001

Biosynthesis of exopolysaccharides by two strains of Lactobacillus bulgaricus in whey-based media.

Exopolysaccharides /EPS/ produced by lactic acid bacteria /LAB/ play a role in the rheology and texture of fermented milks (Cerning 1990) and could also provide a new source of safe additives for use in various food products. The physical properties of EPS, such as rheology and behaviour, depend on several factors, including sugar composition, type of sugar linkages, the presence of organic or inorganic substituents, the degree of polymerization and the length of the side chains. On the other hand certain physiological properties of lactic bacteria are of particular importance in the mechanism of their probiotic functioning: metabolism leading to accumulation of organic acids and other fermentation products in the media; resistance to those metabolites; competitive assimilation of the major nutrients of the media. The significant antimicrobial effect of probiotics towards pathogens and potentially pathogenic microorganisms is outlined in this context. This study was conducted on LAB strains with experimentally established preventive effect in control of chemical carcinogenesis induced by dimethylhydrazine, heavy metals and nitrites. The present study reports the biosynthesis of EPS by strains with proved anticancer effect Lactobacillus bulgaricus LB3 and Lactobacillus bulgaricus LBs on whey-based media. A study on the action of different nitrogen sources (yeast extract and peptone) on the productivity of EPS and the rheological properties of medium was done. When whey-based medium was used the highest production of EPS was determined at 1% peptone--145 mg/g(-1). The carbohydrate substrate influenced the EPS composition. Not only the concentration but also the structure of the EPS is important for its thickening effect. The functional properties of the EPS produced could be modified by influencing the growth conditions. The influence of the different content of glucose on the fermentation process and the viscosity of the whey was performed.

Culture Media↗

PubMed2003

[Effects of nutrient conditions and fed-batch culture on CoQ10 production by Rhizobium radiobacter WSH2601].

The effects of the variables, including the concentrations of glucose, sucrose, corn steep liquor (CSL) and peptone, and the conditions of fed-batch culture, on CoQ10 fermentation by Rhizobium radiobacter WSH2601 were assessed. The results showed that the optimum concentrations of glucose, sucrose, CSL and peptone were 30 g/L, 40 g/L, 11 g/L and 16 g/L respectively. Addition of CSL and tomato juice stimulated the cell growth. CSL, L-methionine and isopentyl alcohol efficiently increased the biosynthesis of CoQ10. In a 7L fermentor, the fed-batch culture improved both cell growth and CoQ10 production compared to a batch culture; and suitable mixed feeding of CSL and sucrose enhanced CoQ10 yield to 52.4 mg/L. The DCW reached 26.4 g/L, an increase of 53% in comparison to that without feeding, and an increase of 24% to that feeding with sucrose only. The C/B value reached 2.38 mg/g(DCW), representing an increase of 33% to that of without feeding, and an increase of 26% to that of feeding with sucrose only.

Agrobacterium tumefaciens↗

PubMed2002

High-level production of prourokinase-annexin V chimeras in the methylotrophic yeast Pichia pastoris.

Prourokinase (proUK)-annexin V chimeras expressed by the methylotrophic yeast Pichia pastoris in a synthetic medium as part of a system designed to yield a novel thrombolytic agent are degraded, as it is thought, by various yeast proteases present in the culture supernatant. Minimization of proteolysis was therefore investigated to increase the yield of intact proUK-annexin V. Protease inhibitor screening study indicated several proteases including at least serine protease like chymotrypsin were involved in the proteolysis. Addition of more than 10% of peptone or more than 0.2 mol l(-1) of arginine to the medium was effective in minimizing proteolysis in shake-flask culture. Culture condition of higher pH was also effective, however, induced a cell death. Cell improvement by increasing the methanol utilization ability yielded greater tolerance to high pH. As a result, the culture condition with highly concentrated peptone solution fed under controlled conditions of pH 8.0 was established, which greatly reduced proteolytic degradation in fed-batch fermentation. These optimal conditions, which enabled fibrinolytic activity to reach 7800 IU ml(-1), could easily be applied in industrial scale production.

Journal Article↗

PubMedDate not supplied

[Effect of cultivation conditions of Streptomyces globisporus 3-1 on landomycin E production].

The producing of antitumor angucycline antibiotic landomycin E has been studied in dynamics of the surface and deep growth of Streptomyces globisporus 3-1 on different full-value media--soya, maize and peptone-yeast media. It is shown that the highest yield of landomycin E is observed on the maize, a bit lower--on soya, and the lowest--on peptone-yeast media independent of the method of growing. The antibiotic stability is affected by alkali pH of the medium. Growth dynamics of the culture in the maize and soya media in the laboratory 2-litre fermenter is characterized by intensive assimilation of glucose, absorption of dissolved oxygen, accumulation of biomass and landomycin E from the 24th to 48th hour of fermentation. The antibiotic is synthesized as primary metabolite reaching its maximum (180-200 mg/l) for the 48th hour of the culture growth. A hypothesis is put forward on the inhibition of the two-component system of signals transduktion by isoflavone genistein, which stimulates synthesis of landomycin E.

Aminoglycosides↗

PubMed1992

[Studies of bacterial indicators for water pollution--growth of Escherichia coli and enterococci in limited nutrient conditions].

Bacteria, that would be an ideal indicator for fecal contamination in environmental water, should not proliferate but at the same time should survive a little longer than enteropathogenic bacteria in the environmental water. While conventional methods have recommended Escherichia coli as an indicator, enterococci is recommended as a preferable indicator for the estimation of fecal contamination based on the following observation. The relationship between the specific growth rate of organism and the concentration of peptone was determined and the value of Ks, a parameter analogous to Michaelis-Menten constant, was estimated. Ks of Escherichia coli, Enterococcus faecalis and Enterococcus faecium were 3.8, 11.1 and 12.5 mg/ml respectively, indicating that Escherichia coli will grow at lower concentration of peptone than enterococci. Escherichia coli also grew in human fecal suspension and in filtrated (0.22 microns) river water, and while in those conditions enterococci did not grow or survive longer than Escherichia coli, survival time was longer than for Salmonella typhi and Shigella sonnei. It appears that enterococci reflect more accurately actual fecal contamination.

Enterococcus faecalis↗

PubMed1976

The fermentative production of acetone-butanol by Clostridium acetobutylicum.

Fourteen different media were used in the fermentative production of acetone-butanol. The highest total yields were achieved in medium I. Potato starch and soluble starch were suitable as carbon sources. The best concentrations of potato starch and soluble starch were 500.0 and 10.0 g/l, respectively. Peptone was the most favourable nitrogen source. The best concentration of peptone was 4.0 g/l. Calcium carbonate in 3.6 g/l acted as buffering agent in the fermentation process. The best initial pH value of the fermentation medium was 6.0. The optimum temperature was 32--33degreesC. The fermentation process required 120 h to obtain maximum yields of acetone-butanol.

Acetone↗

PubMedDate not supplied

[Effect of cultivation conditions of Bacillus subtilis IMV B-7023 on surface properties of cells].

It has been established that the cells of Bacillus subtilis IMV B-7023 grown on the medium where peptone or calcium glycerophosphate served as the only source of carbon and energy have similar acid type of the surface and are characterized by a high negative charge even at low values of pH of the dispersion medium. This may be determined by the presence of phosphate groups, teichoic acids being their carriers in Gram-positive bacteria. Glucose addition to peptone or glycerophosphate leads to the change of the bacteria surface type, i.e., at low values of pH the cells have total positive charge, that allows to suppose availability of protein nature components in the surface layers of the cell wall. Bacteria grown in these conditions are also characterized by the higher degree of adhesion to inert carrier (glass).

Bacillus subtilis↗

PubMed1991

Effect of corn-steep liquor on growth and mosquito larvicidal activity of Bacillus thuringiensis var israelensis de Barjac 1978 and B. sphaericus Neide 1904.

Influence of corn steep liquor on the cell yield and toxicity of three strains of B. thuringiensis var israelensis and two strains of B. sphaericus was studied and compared with peptone-yeast extract using a laboratory fermentor. Large increase in the cell yield of all the three strains of B. thuringiensis var israelensis was observed when cornsteep liquor was used as the sole nitrogen source. Significant increase in toxicity was also observed in B. thuringiensis var israelensis strains B17 and B113. Among the two B. sphaericus strains tested, the strain 1593 showed no significant change in cell yield and toxicity, whereas the strain VCRC B42 showed increased cell yield and toxicity in this medium. The results indicate that cornsteep liquor can effectively replace both peptone and yeast extract in the media presently used for large scale multiplication of the two larvicidal bacilli.

Alcohols↗

PubMed1975

[A new butyric acid-producing bacteroides species: B. splanchnicus n. sp. (author's transl)].

Three butyric acid-producing saccharolytic Bacteroides cultures (1651/6, BM 158, and IPP 3751) were described by WERNER and REICHERTZ in 1971 (Zbl.Bakt.Hyg., I. Abt. Orig. A 217,206-216). Since then, 6 strains closely resembling 1651/6 were isolated from stool specimens and surgically removed appendices. In the present communication, strains 1651/6, S2/34, S3/38, S4/28, S6/6, A5/2 are described as members of a new species, Bacteroides splanchnicus n.sp. The strains were morphologically very similar (Gram negative non-sporing non-motile rods, 1-2.5 mu in length and 0.7 mu in width) and fermented glucose, fructose, galactose, mannose, lactose, and arabinose (pH values of 4.6-5.4, moderate gas formation). Negative reactions (pH values of 5.8-7.2) were observed with 20 other carbohydrates. The strains were positive in the glutamic acid decarboxylase test and formed indole and H2S. In peptone-yeast extract broth and peptone-yeast extract-glucose broth acetic, propionic, isobutyric, butyric, and isovaleric acids were produced. Washed cells of strains 1651/6 and S4/28 incubated anaerobically in sterile solutions of single amino acids produced butyrate from lysine only. Abundant butyric acid was also produced from glucose. The in vitro activity of 15 antibiotics on 5 strains was studied by broth dilution tests. Uniformly, the strains showed resistance to aminoglycosides and polymyxins (MIC values, 60-500 mug/ml) and susceptibility to tetracyclines, lincomycin, clindamycin, rifampicin, and erythromycin (MIC values, 0.05-0.5 mug/ml). Chloramphenicol, penicillins, and cephalosporins showed bacteriostatic activity at concentrations of 5-40 mug/ml. The serological behaviour of 5 strains was studied in cross-agglutination and gel-diffusion experiments. Cross-reactivity was pronounced in gel-diffusion tests using rabbit antisera and autoclaved extracts and extracts prepared by repeated deep-freezing and thawing of whole cell suspensions as antigens. However, antisera against the B. splanchnicus strains did not react with antigens of B. fragilis 6869, B. thetaiotaomicron AS 126, B. vulgatus AM 45 a, B. distasonis AII 104 and butyric acid-producing strains BM 158 and IPP 3751 in agglutination and gel-diffusion experiments, and there were no cross-reactions between B. splanchnicus antigens and antisera against B. fragilis 6869 and the other aforementioned strains. The new species has been confirmed by members of the ICSB Taxonomic Subcommittee for Gram negative anaerobic rods. Strains 1651/6 and S4/28 have been deposited in The National Collection of Type Cultures, London (NCTC numbers 10825 and 10826). Strain NCTC 10825 (= 1651/6) is the type of the species, B. splanchnicus.

Acetates↗

PubMed1990

Enteric pathogen screening: a multipoint technique with a simple numerical profile.

The use of peptone water sugars to screen non-lactose fermenting enteric isolates is expensive and has limited potential for identification. The multipoint system uses seven media from a commercial system containing either single sugars or biochemical substrates. Up to 20 organisms can be tested on each set of plates, the results being used to generate a four figure profile from which a presumptive identity can be obtained. This identity must still be confirmed, but the number of organisms requiring further investigation can be as low as one third of those following use of peptone water sugars. The multipoint system represents a rapid, more accurate and considerably cheaper alternative.

Bacteriological Techniques↗