Development of meridic and oligidic diets for rearing the aphid Myzus persicae

Meridic and oligidic diets suitable for the continuous culture of the aphid Myzus persicae were developed through modifications of a holidic diet. These included the addition of various amounts of crude nutrients and the replacement of defined nutrients by crude nutrients. The highest level of aphid growth (mean weights of 600 to 800 μg) was maintained (for 45 successive generations) on a meridic diet made by supplementing a holidic diet with 2.0% yeast extract (NBC).Continuous culture, at a level of growth (400 to 600 μg) comparable to that on the unsupplemented holidic diet (formulated with 34 defined nutrients), was supported by an oligidic diet containing only 10 weighed ingredients: 15 g sucrose, 2.5 g enzymatic casein hydrolysate (NBC), 2 g yeast extract, 123 mg MgSO₄·7H₂O, 100 mg ascorbic acid, 10 mg niacin, 5 mg Ca pantothenate, 2.5 mg pyridoxine, 2.5 mg thiamine, and appx. 1.5 gm K₂HPO₄·3H₂O (pH 6.8) per 100 ml of diet.Yeast extract at 2.0% provided adequate amounts of choline chloride, biotin, folic acid and inositol, but not of niacin, pantothenate, pyridoxine, and thiamine. Enzymatic casein hydrolyzate at 2.5% could replace the 20 defined amino acids of the holidic diet. Diets with both yeast extract and casein hydrolysate did not have to be supplemented with trace minerals (Cu, Fe, Mn and Zn). Yeast extract at 2.5% provided sufficient amounts of trace minerals, amino acids, and B-vitamins to sustain numerous successive generations, albeit at a low level of growth (200 to 300 μg). The simplicity and low cost of oligidic diets makes them attractive for aphid studies in which nutrition is not the primary consideration.     

Somatic embryogenesis and plant regeneration in an interspecific hybrid of Oryza

An embryogenic callus was obtained from immature panicle of an interspecific hybrid (Oryza sativa x O. latifolia) F₁. The medium consisted of HE salts supplemented with 2,4-D, NAA (each 2 mg/l), kinetin (3 mg/l), yeast extract (1360 mg/l) and casein hydrolyzate (300 mg/l). The callus was milk-white in colour compact and granulate in texture. Various developmental stage of embryoid, such as globular, heart-shape, scutellum-shape and mature embryoid were observed in an embryogenic callus. Plantlets were successfully regenerated from 1-month-old callus with more than 80% regenerational frequency in each subculture for 12 passages.     

Enzymatic Preparation of Crystalline Laminaribiose from Curdlan

The enzymatic preparation of laminaribiose was investigated in this research. When curdlan was hydrolyzed with the β-1,3-glucanase system of Streptomyces sp. K27-4, the hydrolyzate mainly consisted of glucose and laminaribiose in an approximate ratio of 1:1 by weight. Yeast strains selected in this study, effectively and selectively metabolized all of the glucose in the hydrolyzate, without any degradation of laminaribiose.

By successive treatment of curdlan with the glucanase system and glucose-metabolizable yeast, 31 g of crystalline laminaribiose was obtained from l00g of curdlan.     

Casein as a necessary factor in the production of stimulatory material for associative growth of lactic streptococci

Strains of Streptococcus cremoris KH and HC produced material that was stimulatory for S. cremoris R₆ in milk and in the dialyzable fraction of milk, but not in the dialysate fraction of milk, lactic acid whey, or lactose broth. The addition of casein to these latter media permitted the production of this stimulatory material to occur. Tryptone, peptone, and yeast extract could not be substituted for casein in producing the stimulatory material or in initiating associative growth in the lactic acid whey. The minimum concentration of casein required appeared to be from 2.0 to 2.5%.     

Fed-batch cultivation of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> on lignocellulosic hydrolyzate

Saccharomyces cerevisiae grows very poorly in dilute acid lignocellulosic hydrolyzate during the anaerobic fermentation for fuel ethanol production. However, yeast cells grown aerobically on the hydrolyzate have increased tolerance for the hydrolyzate. Cultivation of yeast on part of the hydrolyzate has therefore the potential of enabling increased ethanol productivity in the fermentation of the hydrolyzate. To evaluate the ability of the yeast to grow in the hydrolyzate, fed-batch cultivations were run using the ethanol concentration as input variable to control the feed-rate. The yeast then grew in an undetoxified hydrolyzate with a specific growth rate of 0.19 h⁻¹ by controlling the ethanol concentration at a low level during the cultivation. However, the biomass yield was lower for the cultivation on hydrolyzate compared to synthetic media: with an ethanol set-point of 0.25 g/l the yield was 0.46 g/g on the hydrolyzate, compared to 0.52 g/g for synthetic media. The main reason for the difference was not the ethanol production per se, but a significant production of glycerol at a high specific growth rate. The glycerol production may be attributed to an insufficient respiratory capacity.     

Proteomics as a Quality Control Tool of Pharmaceutical Probiotic Bacterial Lysate Products

Probiotic bacteria have a wide range of applications in veterinary and human therapeutics. Inactivated probiotics are complex samples and quality control (QC) should measure as many molecular features as possible. Capillary electrophoresis coupled to mass spectrometry (CE/MS) has been used as a multidimensional and high throughput method for the identification and validation of biomarkers of disease in complex biological samples such as biofluids. In this study we evaluate the suitability of CE/MS to measure the consistency of different lots of the probiotic formulation Pro-Symbioflor which is a bacterial lysate of heat-inactivated Escherichia coli and Enterococcus faecalis. Over 5000 peptides were detected by CE/MS in 5 different lots of the bacterial lysate and in a sample of culture medium. 71 to 75% of the total peptide content was identical in all lots. This percentage increased to 87–89% when allowing the absence of a peptide in one of the 5 samples. These results, based on over 2000 peptides, suggest high similarity of the 5 different lots. Sequence analysis identified peptides of both E. coli and E. faecalis and peptides originating from the culture medium, thus confirming the presence of the strains in the formulation. Ontology analysis suggested that the majority of the peptides identified for E. coli originated from the cell membrane or the fimbrium, while peptides identified for E. faecalis were enriched for peptides originating from the cytoplasm. The bacterial lysate peptides as a whole are recognised as highly conserved molecular patterns by the innate immune system as microbe associated molecular pattern (MAMP). Sequence analysis also identified the presence of soybean, yeast and casein protein fragments that are part of the formulation of the culture medium. In conclusion CE/MS seems an appropriate QC tool to analyze complex biological products such as inactivated probiotic formulations and allows determining the similarity between lots.     

New Medium for Blood Cultures

A new medium suitable for blood cultures is described. It contains dextrose, cysteine, iron, and magnesium, in a tris(hydroxymethyl)aminomethane buffer, and a mixture of peptones derived from animal tissues, casein, and yeast. In comparison with Trypticase Soy Broth, the growth rate constants of Staphylococcus aureus, Streptococcus (Viridans group), enterococcus, and Escherichia coli were higher in this medium, and growth appeared earlier in a significant number of clinical blood cultures.     

New Medium for Blood Cultures

A new medium suitable for blood cultures is described. It contains dextrose, cysteine, iron, and magnesium, in a tris(hydroxymethyl)aminomethane buffer, and a mixture of peptones derived from animal tissues, casein, and yeast. In comparison with Trypticase Soy Broth, the growth rate constants of Staphylococcus aureus, Streptococcus (Viridans group), enterococcus, and Escherichia coli were higher in this medium, and growth appeared earlier in a significant number of clinical blood cultures.     

Effects of Nitrogen Supplements on Degradation of Aspen Wood Lignin and Carbohydrate Components by Phanerochaete chrysosporium

A supplement of KH₂PO₄, MgSO₄, CaCl₂, trace elements, and thiamine accelerated the initial rate of aspen wood decay by Phanerochaete chrysosporium but did not increase the extent of lignin degradation. Asparagine, casein hydrolysate, and urea supplements (1% added N) strongly inhibited lignin degradation and weight loss. The complex nitrogen sources peptone and yeast extract stimulated lignin degradation and weight loss. Albumen and NH₄Cl had intermediate effects. Conversion of [¹⁴C]lignin to ¹⁴CO₂ and water-soluble materials underestimated lignin degradation in the presence of the complex N sources. The highest ratio of lignin degradation to total weight loss and the largest increase in cellulase digestibility occurred during the decay of unsupplemented wood. Rotting of aspen wood by P. chrysosporium gives smaller digestibility increases than have been found with some other white-rot fungi.     

MAIZE ENDOSPERM TISSUE GROWN IN VITRO III. DEVELOPMENT OF A SYNTHETIC MEDIUM

Straus , Jacob . (U. Oregon, Eugene.) Maize endosperm tissue grown in vitro. III. Development of a synthetic medium. Amer. Jour. Bot. 47(8) : 641–647. Illus. 1960.—The development of a synthetic medium for the growth of endosperm tissue cultures derived from the maize variety, ‘Black Mexican Sweet,‘ is described. Previously, these tissues required yeast extract, casein hydrolyzate, or tomato juice in the medium in order to grow. The growth-supporting activity of these complexes could be attributed to their organic nitrogen content. The effect of juice extracted from fresh tomatoes is enhanced by autoclaving under acid conditions. Presumably this treatment increases the free amino acid content of the tomato juice. One-dimensional paper chromatograms of tomato juice autoclaved under acid conditions indicated the presence of a large amount of free amino acids. Addition of 1.5 × 10^–2 M asparagine to the basal mineral-sugar-vitamin medium (White's medium plus Nitsch's trace-element solution) resulted in better growth than that supported by yeast extract, tomato juice, or casein hydrolyzate. Arginine was ineffective. Glutamine, glutamic acid and aspartic acid (all at 1.5 × 10^–2 M) supported appreciable growth of the tissue but none of them were nearly so good as asparagine in this respect. Thus, a medium containing minerals, sugar, vitamins, and asparagine is capable of supporting excellent growth of maize endosperm tissue cultures.     

Development of resistance in Saccharomyces cerevisiae against inhibitory effects of Browning reaction products

Furfural, maltol and 5-hydroxymethyl furfural, the three Browning reaction products, inhibit CO₂ production by Saccharomyces cerevisiae. The inhibition can be partially overcome by some crude extracts, such as tryptone, yeast extract and casein hydrolysate. Yeast cells show a tendency to adapt to these compounds even when exposed to them for a very short time. Sugar cane molasses can perform both the above functions simultaneously. First, it can overcome the inhibitory effect of these compounds much more effectively than the crude extracts and secondly, training of cells in molasses makes them resistant enough to withstand the lethal levels of the test compounds.     

Optimization of Low-Cost Culture Media for the Production of Biomass and Bacteriocin by a Urogenital Lactobacillus salivarius Strain

The aim of this work was to formulate a culture medium of lower cost than conventional laboratory media, in order to simultaneously obtain high amounts of both biomass and bacteriocin of vaginal Lactobacillus salivarius CRL 1328. The growth assays under different culture conditions were performed by using a 2^8?2 central composite experimental design, with a central point and sixteen additional points. The factors taken into consideration were glucose, lactose, yeast extract, tryptone, ammonium citrate, sodium acetate, MgSO₄ and MnSO₄. The simultaneous presence of a carbon source (mainly glucose), a nitrogen source (mainly yeast extract) and salts (mainly MnSO₄, MgSO₄ and sodium acetate) allowed the highest cell biomass and bacteriocin levels to be reached in the experimental design. Through the application of the desirability function, several optimal medium compositions to achieve efficient production of biomass and bacteriocin were predicted. The optimized growth media allow a cost reduction of around 25 to 40% compared with conventional broths. The results obtained represent an advance in the search of the most suitable strategies for the production of bioactive compounds for pharmaceutical products to prevent or treat female urogenital infections.     

CASEIN VISCOSITY STUDIES

1. Viscosity and pH curves of casein dissolved in NaOH, KOH, LiOH, and NH₄OH are shown and it is found that a maximum viscosity occurs at about the same pH point with each alkali; i.e., 9.1 to 9.25. The magnitude of the viscosity is largest in ammonia solutions. 2. The maximum viscosity occurs in 8 to 10 per cent solutions of casein in alkalies when about 98 x 10^–5 gram equivalents of base are combined with 1 gram of casein. 3. A maximum viscosity occurs in the same region (pH 9.1 to 9.25) when casein is dissolved in Na₂CO₃, Na₃AsO₄, Na₂SO₃, NaF, and Na₂PO₃. 4. The maximum viscosity obtained with borax solutions of casein occurs at 8.15 to 8.2 pH. It is suggested that casein acts like mannitol, glycerol, etc., in increasing the dissociation of boric acid. 5. The flattening of the viscosity curves of casein solutions, following the decline from maximum, is shown to be due to alkaline hydrolysis whence casein no longer exists as such but is cleaved into a major protein containing no phosphorus or sulfur and less nitrogen. This cleavage commences at pH 10.0 to 10.5. 6. When casein is prepared from solutions that have been subjected to high temperatures (60°C. and above) or has otherwise been heated during its preparation, it yields solutions in alkalies of high viscosity.     

Prolongation of oxidative stress by long-lived reactive protein species induced by X-ray radiation and their genotoxic action

Abstract

The formation of long-lived reactive protein species of bovine serum albumin (BSA), ovalbumin, casein and casein hydrolyzate with a half-life of 3–5 hours was shown using chemiluminescence induced by X-ray radiation. It was found that long-lived reactive protein species are capable of generating reactive oxygen species (ROS) (H₂O₂, OH^?, HO₂^{?, 1}O₂) in the aquatic environment over a long period of time in vitro. The interaction of X-ray-irradiated BSA with DNA in vitro led to the formation of 8-oxoguanine (8-oxo-7,8-dihydroguanine), a biomarker of oxidative damage to DNA. Some natural antioxidants are effective scavengers of ROS (inosine, tryptophan, methionine and ascorbate). They protect DNA from the action of long-lived reactive protein species leading to ROS generation and the formation of 8-oxoguanine. The intravenous injection of X-ray radiation-induced, long-lived reactive protein species to rats, as well as the peroral and intraperitoneal administration of these products to mice, gave rise to cytogenetic injuries in the cells of their red bone marrow through the formation of micronuclei in polychromatophilic erythrocytes. The administration of the same natural antioxidants used for in vitro experiments soon after irradiation made it possible to effectively eliminate the genotoxic action of oxidative stress caused by radiation-induced, long-lived reactive protein species. Our data represent clear evidence that the oxidative damage to proteins induced by X-rays is directly involved in the induction of a response to DNA damage in rodents.     

Detection of Type A, B, E, and F Clostridium botulinum Neurotoxins in Foods by Using an Amplified Enzyme-Linked Immunosorbent Assay with Digoxigenin-Labeled Antibodies

An amplified enzyme-linked immunosorbent assay (ELISA) for the detection of Clostridium botulinum complex neurotoxins was evaluated for its ability to detect these toxins in food. The assay was found to be suitable for detecting type A, B, E, and F botulinum neurotoxins in a variety of food matrices representing liquids, solid, and semisolid food. Specific foods included broccoli, orange juice, bottled water, cola soft drinks, vanilla extract, oregano, potato salad, apple juice, meat products, and dairy foods. The detection sensitivity of the test for these botulinum complex serotypes was found to be 60 pg/ml (1.9 50% lethal dose [LD₅₀]) for botulinum neurotoxin type A (BoNT/A), 176 pg/ml (1.58 LD₅₀) for BoNT/B, 163 pg/ml for BoNT/E (4.5 LD₅₀), and 117 pg/ml for BoNT/F (less than 1 LD₅₀) in casein buffer. The test could also readily detect 2 ng/ml of neurotoxins type A, B, E, and F in a variety of food samples. For specificity studies, the assay was also used to test a large panel of type A C. botulinum, a smaller panel of proteolytic and nonproteolytic type B, E, and F neurotoxin-producing Clostridia, and nontoxigenic organisms using an overnight incubation of toxin production medium. The assay appears to be an effective tool for large-scale screening of the food supply in the event of a botulinum neurotoxin contamination event.     

Selection of Yeasts for Breadmaking by the Frozen-Dough Method

Eleven yeast strains suitable for frozen dough were selected from over 300 Saccharomyces strains. All of these were identified as Saccharomyces cerevisiae from morphological, cultural, and physiological characteristics. The selected yeast cells accumulated a higher amount of trehalose than did commercial bakers' yeast cells.     

Optimization of high-quality dietary fiber production in submerged fermentation by Agrocybe chaxingu

The aim of this study was to determine the optimal conditions for high-quality dietary fibre (DF) production by Agrocybe chaxingu in a submerged culture fermentation system of wheat bran. The substances which may affect DF quality were first selected by Plackett–Burman design, and then the D-optimal design was applied for further optimization to obtain the highest quality of DF. The results suggest that vitamin B₁ (VB₁), MgSO₄?·?7H₂O and yeast extract are factors that have a significantly positive effect on DF production. The highest quality of DF was obtained in a solution of 50 g wheat bran and 1 L distilled water to which 0.14 g VB₁, 1.10 g MgSO₄?·?7H₂O and 2.50 g yeast extract had been added; 22.32 % soluble DF of the total DF amount was found. These results indicate that submerged culture fermentation of A. chaxingu is a novel and good method for producing high-quality DF.     

Regulation of extracellular acid phosphatase biosynthesis by culture conditions in entomopathogenic fungusMetarhizium anisopliae strain CQMa102

Metarhizium anisopliae is an imperfect entomopathogenic fungus. Once invading into its host,M. anisopliae needs to absorb basic nutrients such as phosphorus from the host haemolymph. A large number of phosphorylated compounds in haemolymph cannot be directly utilised by the fungal cell and must be hydrolysed into available form by phosphatase before ingested. Aims of this paper were to investigate optimum fermentation conditions for production of acid phosphatase and phosphatase isoenzymes byMetarhizium anisopliae. The optimum fermentation conditions were: glucose, 20 g/l; (NH₄)₂SO₄, 2 g/l; casein, 4 g/l; MgSO₄, 0.5 g; KCl, 0.5 g; microelement salt solution, 10 ml; inoculum size, 1×10⁷ spores per 100 ml medium; initial medium pH, 6.0. Under these conditions, the highest total acid phosphatase activity was 3.05 U/ml in 4 days at 27 °C and 160 rpm. Synthesis of the acid phosphatase was repressed by 0.01% inorganic phosphate in culture medium. The spectrum of isoenzymes produced byM. anisopliae varied depending on the phosphorus source employed in the culture. A specific isoform with pI 9.45 was induced by casein, and another isoform of pI 8.21 was induced by phytic acid and disodium phenyl phosphate.     

Purification and characterization of casein kinase II from lactating rabbit mammary gland

• 1.1. Highly purified 200 kDa casein kinase II from rabbit lactating mammary gland (MG-CK II) was obtained by means of a new purification procedure consisting of one phosphocellulose and three Mono Q steps.
• 2.2. Its K_m for ATP was 2.22 μM and 0.57 mg/ml and 0.13 mg/ml for partially dephosphorylated casein and phosvitin respectively. Stathmine was also suitable as substrate. 2-aminopurine and 6-dimethylaminopurine inhibited efficiently MG-CK II K_i = 5 and 1 mM respectively).
• 3.3. MG-CK II autophosphorylated on its α-, α '- and β-subunits. The β-subunit auto-phosporylation was enhanced in presence of exogenous substrate. Its modulation was highly dependent on ATP concentration.
• 4.4. The effects of basic compounds which affected dramatically the phosphorylation of dephosphorylated casein in presence of various ATP concentrations were reported.