Changes in metabolism and cell size of the anaerobic bacterium Selenomonas ruminantium 0078A at the onset of growth in continuous culture

Initial metabolism of Selenomonas ruminantium 0078A in continuous culture was characterized by a high lactate and low volatile fatty acid production; this was associated with poor growth as determined by bacterial dry weight production, yet individual cells were considerably larger than those of the inoculum. Biomass production increased, cell size decreased and the fermentation pattern reverted to the characteristic low lactate and high volatile fatty acid production after approximately 90 h growth.     

Determination and quantification of monoterpenoids secreted into the medium of cell cultures of thuja occidentalis

Cell cultures of Thuja occidentalis excrete monoterpenoids into the culture medium. Highly volatile monoterpene hydrocarbons were only detectable in the presence of Miglyol, a water-insoluble non-toxic trap for lipophilic compounds. In the presence of Miglyol the levels of oxygenated monoterpenoids were two-fold higher than in dichloromethane-extracts of culture media. The cells excrete up to 3 mg regular monoterpenoids/g dry wt/day. Continuous extraction of lipophilic compounds by Miglyol also allowed quantification of a Tris—thujaplicinato—(Fe III)-complex.     

Effects of Lavandula officinalis and Equisetum arvense dry extracts and isoquercitrin on the fermentation of diets varying in forage contents by rumen microorganisms in batch culture

The short-term actions of Lavandula officinalis and Equisetum arvense dry extracts, and of isoquercitrin, flavonoid present in Equisetum arvense, on in vitro fermentation by rumen microbes were studied in batch culture. The orchard grass hay:barley ratios in the three experimental diets were 100:0, 75:25, 50:50 on a DM basis. The production rates of all volatile fatty acids except isobutyrate were strongly influenced by the composition of the diet and to a lesser extent, by plant extracts, with significant interactions between both factors. When hay was the only substrate, the addition of L. officinalis and E. arvense enhanced the fermentation rate by 50%, through an increased release of acetate and propionate. On the contrary, with the two other diets, the fermentation rate was strongly lowered by isoquercitrin. Gas outputs were not significantly influenced by plant extracts.     

Changes in metabolism and cell size of the anaerobic bacterium Selenomonas ruminantium 0078A at the onset of growth in continuous culture

S illey , P. & A rmstrong , D.G. 1984. Changes in metabolism and cell size of the anaerobic bacterium Selenomonas ruminantium 0078A at the onset of growth in continuous culture. Journal of Applied Bacteriology 56 , 487–492.
Initial metabolism of Selenomonas ruminantium 0078A in continuous culture was characterized by a high lactate and low volatile fatty acid production; this was associated with poor growth as determined by bacterial dry weight production, yet individual cells were considerably larger than those of the inoculum. Biomass production increased, cell size decreased and the fermentation pattern reverted to the characteristic low lactate and high volatile fatty acid production after approximately 90 h growth.     

The physiological activity of volatile substances of plants in air and water media

Physiological effects of volatile substances released by the overground as well as by the underground organs of higher plants were studied. The activity of the volatile substances was tested both when these substances were allowed to act directly in the air and when they were dissolved in water in the form of solutions. Plants which do not contain essential oils or which are not rich in them as well as those abounding in essential oils and other volatiles were used in the experiments. The physiological activity of the volatile substances was tested on rye seedlings. The overground as well as underground mature organs of the tested plants were found to release volatile substances causing, when acting directly, in the majority of cases an inhibition of the growth in length and of the formation of dry matter in rye seedlings. A pronounced inhibition of the growth of rye seedlings was brought about especially by the volatile substances of “aromatic” plants such as common dill, wild thyme, yarrow milfoil, garden thyme, marjoram, etc. The volatile substances released by the organs of “non-aromatic” plants like sugar-beet, common sunflower, quackgrass, etc., were found to bring about a significant inhibition of the growth of rye seedlings, too. The volatile substances released by the plant organs were found to be altogether absorbable in water and physiologically active also in the form of water solutions. With the exception of volatile substances from hemp and quackgrass leaves, which brought about a mild stimulation of the dry matter formation in rye seedlings, inhibitory effects of these solutions were found to prevail in all cases. Most effective were the solutions of the volatiles from some of the “aromatic plants”. An assay for olefines in the atmosphere of the experimental vessels demonstrated that in almost all cases ethylene is being released by the plant organs.     

Explant type and natural ventilation systems influence growth and content of carvacrol and thymol of Lippia gracilis Schauer

The objective of this study was to evaluate the effects of different natural ventilation systems and explant types on the growth and volatile compound content of Lippia gracilis cultured in vitro. The treatments consisted of four membrane systems (without membrane, with one, two, and four porous membranes) and two explant types (nodal segments with and without a pair of leaves). The evaluation of growth, photosynthetic pigments and chemical analysis of the volatile fraction were performed at 35 days of cultivation in half strength MS basal medium. Natural ventilation systems significantly influenced the in vitro growth and volatile fraction of L. gracilis. Explants with a pair of leaves obtained the best experimental responses. The natural ventilation system with four membranes provided the best growth parameters and leaf area response of L. gracilis explants with leaves. The photosynthetic pigments decreased with an increase in the number of porous membranes in the culture flask. Variations in the number, content, and profile of volatile compounds under the influence of natural ventilation systems were observed. Major constituents such as ρ-cymene, γ-terpinene, thymol, carvacrol, and E-caryophyllene, regardless of experimental conditions, were identified. The highest carvacrol and thymol contents were observed in plantlets grown in culture flasks with four porous membranes. To maximize the content of carvacrol and thymol from the in vitro culture of L. gracilis, explants with a pair of leaves and four porous membranes in culture flasks are recommended for use.

     

Application of poly(butyl methacrylate-co-ethylene glycol dimethacrylate) monolith microextraction coupled with high performance liquid chromatography to the determination of polycyclic aromatic hydrocarbons in smoked meat products

This paper presents a study of the synthesis of a polymer monolith column and its application to the analysis of PAHs in smoked meat products. A poly(butyl methacrylate-co-ethylene glycol dimethacrylate) monolith capillary has been successfully prepared with in situ polymerization method. The polymer monolith microextraction combined with HPLC determinations is employed for the analysis of naphthalene, biphenyl, phenanthrene, and anthracene. Various parameters affecting the extraction efficiency have been investigated and optimized. Under the optimum experimental conditions, the method provides an acceptable linearity (2-10,000 μg/L), low limits of detection (1.4-2.0 μg/L), and good precision (intraday relative standard deviations<4.1%, interday relative standard deviations<5.7%). When applied to the determination of the four PAHs in smoked meat samples, recoveries are obtained in the range of 86.6-101.5%.     

Mathematical modeling of ethanol production in solid-state fermentation based on solid medium’ dry weight variation

In this work, mathematical modeling of ethanol production in solid-state fermentation (SSF) has been done based on the variation in the dry weight of solid medium. This method was previously used for mathematical modeling of enzyme production; however, the model should be modified to predict the production of a volatile compound like ethanol. The experimental results of bioethanol production from the mixture of carob pods and wheat bran by Zymomonas mobilis in SSF were used for the model validation. Exponential and logistic kinetic models were used for modeling the growth of microorganism. In both cases, the model predictions matched well with the experimental results during the exponential growth phase, indicating the good ability of solid medium weight variation method for modeling a volatile product formation in solid-state fermentation. In addition, using logistic model, better predictions were obtained.     

Characterization of technological features of dry yeast (strain I-7-43) preparation, product of electrofusion between Saccharomyces cerevisiae and Saccharomyces diastaticus, in industrial application

The aim of the study was to verify the technological usability and stability of biotechnological features of active dry distillery yeast preparation (strain I-7-43 with amylolytic abilities) applied to full-scale production of agricultural distillery. Various reduced doses of glucoamylase preparation (San-Extra L) were used for starch saccharification, from 90% to 70% in relation to the full standard dose of preparation. The dry distillery yeast I-7-43 were assessed positively in respect to fermentation activity and yield of ethanol production. Application of the dry yeast I-7-43 preparation in distillery practice lowers the costs of spirit production by saving the glucoamylase preparation (up to 30%) used in the process of mash saccharification. Concentrations of the volatile fermentation by-products in raw spirits obtained from fermentations with application of I-7-43 strain were on the levels guaranteeing good organoleptic properties of distillates.     

Growth and sesquiterpenoid production by Calypogeia granulata inoue cells in suspension culture

Growth and the production of volatile sesquiterpenoids by a chlorophyllous cell suspension culture from gametophytes of C. granulata, a leafy liverwort, were examined. Glucose was more effective than 2,4-dichlorophenoxyacetic acid for callus induction, and elimination of glucose from the medium resulted in prompt redifferentiation of plantlets. The cells grew photoheterotrophically, but not in the dark. 1,4-Dimethylazulene, a trinorsesquiterpenoid, was produced as the major volatile sesquiterpenoid in the cultured cells; bicyclogermacrene, compound II, an indene-type aldehyde (a trinorsesquiterpenoid aldehyde), compound I and tetrahydro-1,4-dimethylazulene (a trinorsesquiterpenoid) followed in decreasing order. The azulene was produced both in light and the dark, and its yield was proportional to the growth in light. The yield in light was four times higher than that in the dark. The content of 1,4-dimethylazulene was 0.9–10.% and that of total essential oils was 2.0–3.3% of the dry werght of the cultured cells. The quantity, quality, and proportions of the volatile sesquiterpenoids of the cell culture were almost equal to those of intact (original) plants and redifferentiated plantlets. Previous name: Institute of Food Chemistry     

Quantitative Method for the Gas Chromatographic Analysis of Short-Chain Monocarboxylic and Dicarboxylic Acids in Fermentation Media

A method for the preparation and gas chromatographic analysis of the butyl esters of volatile (C-1-C-7) and nonvolatile (lactic, succinic, and fumaric) acids in microbial fermentation media is presented. Butyl esters were prepared from the dry salts of the acids. The esters were separated by temperature programming on a column of Chromosorb W coated with Dexsil 300 GC liquid phase and analyzed with a flame ionization detector. Apparent recoveries with butanol-HCl or butanol-H2SO4 as butylating agents were 80 to 90% for most acids. Chromatographic profiles of the butyl esters demonstrated that both volatile and nonvolatile acids can be detected and separated in 24 min on a single column. Standard calibration curves (peak area versus concentration) of the butyl esters were linear in the range of 5 to 40 mumol of acid per ml. The advantages of using an internal standard (heptanoic acid) for quantitating fatty acids in a mixture are given. Chromatograms of butylated fermentation media in which rumen anaerobic bacteria were grown illustrated that this method is useful for determining short-chain volatile and nonvolatile acids of toxonomic significance.     

The composition of volatile components of dry cepe and oyster mushroom

The composition of aroma compounds in dry cepe mushroom (Boletis edulis Fr.) and oyster mushroom (Pleurotus ostreatus Fr.) was studied using capillary gas chromatography and chromatography—mass spectrometry. In dry cepe, 53 volatile compounds were identified, and in dry oyster mushroom 41 compounds were identified. Volatile organic substances with various functional groups formed the flavor of dry mushrooms. Unsaturated alcohols and ketones with eight carbon atoms were responsible for the mushroom notes of products. Their content in dry cepe was much higher than in dry oyster mushroom. The specific aroma of dry cepe was formed by the complex mixture of methional, substituted furans, pyrazines, and pyrroles. The content of these compounds was higher in dry cepe than in dry oyster mushroom. The content of aromatic and aliphatic aldehydes with six, nine, and ten carbon atoms was higher in dry oyster mushroom. The differences in the qualitative and quantitative composition of volatile compounds are responsible for more intensive and pleasant aroma of dry cepe in comparison to that of dry oyster mushroom.     

Influence of cysteamine supplementation and culture in portable dry-incubator on the in vitro maturation, fertilization and subsequent development of mouse oocytes

The need to transport oocytes and embryos between two laboratories have prompted us to evaluate the effects of in vitro maturation of immature mouse oocytes in a CO2-deficient dry heat portable incubator and subsequent in vitro development of these fertilized mouse oocytes in a standard CO2 incubator. In addition, the effects of cysteamine supplementation on maturation rate and embryonic development during in vitro maturation (IVM) and culture of embryos in the portable incubator were also investigated. Germinal vesicle stage mouse oocytes, recovered at 40-h post-FSH from 6- to 8-week-old C57BL/6xCBA F1 healthy female mice, were matured in vitro in a modified TCM-199 supplemented with or without 100 microM cysteamine in a standard incubator (5% CO2; 37 degrees C) or cultured in a CO2-deficient dry heat portable incubator for 5 h at 37 degrees C and thereafter transferred to a standard incubator for further culture. The addition of cysteamine in the IVM medium significantly improved maturation rates of the GV mouse oocytes to metaphase II stage. However, cysteamine supplementation in the culture medium did not significantly improve fertilization and blastocyst formation rates of IVM and ovulated oocytes, and in vivo-derived zygotes. Culture conditions in a CO2-deficient dry heat portable incubator did not adversely affect the developmental competence of in vivo-derived zygotes and in vitro matured mouse oocytes after IVF or parthenogenetic activation. Cysteamine supplement in the IVM medium could enhance nuclear maturation of these immature oocytes during shipment.     

Volatile compounds from the symbiotic system Azolla filiculoides‐Anabaena azollae bacteria

Abstract

Azolla filiculoides is an aquatic pteridophyte that may be used as animal food, biofertilizer and phytoremediation. Its volatile composition was never studied although several phytochemical analyses were performed. The volatile composition of A. filiculoides grown outdoors in a pond at the Botanical Garden of Lisbon University (BGLU) or in culture conditions as well as the effect of different harvesting times and the storage type were evaluated. The volatiles isolated by hydrodistillation and distillation‐extraction were analysed by gas chromatography and gas chromatography‐mass spectrometry. The oil of all the A. filiculoides samples studied affords a yellowish colour and an unpleasant odour in a yield of 0.01% (v/fw). Alcohols, aldehydes, alkanes and ketones dominated the culture samples, while aldehydes, alcohols, terpenoids and alkanes represented the main volatiles of the BGLU samples. Some quantitative differences were detected in seasonal and type of storage (fresh, dry or frozen at ?20°C) studies of A. filiculoides from the BGLU. The BGLU and culture volatiles showed qualitative differences: 2‐ethyl‐1‐hexanol was only identified in the fern culture, whereas acetophenone, pentylfuran, acetylpyridine and 2‐octanone were only detected in BGLU samples. The dendrogram showed two distinct clusters (culture and BGLU samples). The possible biological origin and bioactivity of some of the volatile compounds is discussed.

Abbreviations: FID, flame ionization detector; i.d., internal diameter; v/fw, volume by fresh weight; GC, gas chromatography; GC‐MS, gas chromatography‐mass spectrometry; u, unified atomic mass unit     

Detection of endogenous ethanol and other compounds in the breath by gas chromatography with on-column concentration of sample

A new method is described for collecting and concentrating volatile compounds in the breath, in order to facilitate their assay by gas chromatography. Breath was collected into sealed Mylar bags containing an internal standard (isopropyl alcohol). The sample was pumped through a cooled gas chromatograph column, where the volatile compounds were concentrated by adsorption onto the resin packing (Porapak Q) at 35 degrees C. The column was then heated, and the volatilized sample was separated for assay by flame ionization detection. The assay was highly sensitive for ethanol (detecting at least 4.0 nmol) and linear up to 20 nmol (r2 = 0.98). Accuracy and precision were determined by assaying nine replicates of a sample containing 12.0 nmol ethanol; a mean value of 12.18 nmol ethanol was obtained with a coefficient of variation of 10.26%. In a group of normal volunteers, endogenous breath ethanol concentrations ranged from 2.23 to 6.51 nmol/liter. This assay provided a number of advantages over previously described methods: The use of breath collection bags enabled the collection of samples outside the laboratory. The use of an internal standard in the collection bag reduced errors that might have resulted from leakage of the specimen. An on-column concentration of the sample in the gas chromatograph eliminated the need for an additional preconcentration device, such as a cryogenic or adsorptive trapping apparatus.     

Formation of flavor of dry champignons (Agaricus bisporus L.)

The composition of aroma compounds of dry champignons (Agaricus bisporus L.) were identified using capillary gas chromatography and chromatography-mass spectrometry. In total, 56 compounds were identified. It was found that the flavor of dry mushrooms was formed by the volatile compounds produced as a result of enzymatic and oxidative conversion of unsaturated fatty acids as well as in the Maillard reaction. Unsaturated alcohols and ketones containing eight carbon atoms determined the mushroom note of the product. The specific aroma of dry mushrooms was determined by a complex composition of substituted sul- fur-, oxygen-, and nitrogen-containing heterocyclic compounds as well as by aliphatic carbonyl compounds and methional. It was found that the concentrations of volatile carbonylic and heterocyclic compounds increased after the addition of a mixture of amino acids to mushrooms before drying. As a result, the intensity of the aroma of dry mushrooms increased.     

Optimization of culture medium for aroma production byCeratocystis fimbriata

Summary The fungusCeratocystis fimbriata is known to produce among others a banana-like aroma. With a fractional factorial experimental design a culture medium was optimized to produce characteristic volatile metabolites of this aroma. HPLC was used to separate and quantify some of the metabolites. This showed the influence of the composition of the basal culture medium. It is not yet possible to correlate these results with sensorial evaluation.     

Incorporation of cigarette butts into nests reduces nest ectoparasite load in urban birds: new ingredients for an old recipe?

Birds are known to respond to nest-dwelling parasites by altering behaviours. Some bird species, for example, bring fresh plants to the nest, which contain volatile compounds that repel parasites. There is evidence that some birds living in cities incorporate cigarette butts into their nests, but the effect (if any) of this behaviour remains unclear. Butts from smoked cigarettes retain substantial amounts of nicotine and other compounds that may also act as arthropod repellents. We provide the first evidence that smoked cigarette butts may function as a parasite repellent in urban bird nests. The amount of cellulose acetate from butts in nests of two widely distributed urban birds was negatively associated with the number of nest-dwelling parasites. Moreover, when parasites were attracted to heat traps containing smoked or non-smoked cigarette butts, fewer parasites reached the former, presumably due to the presence of nicotine. Because urbanization changes the abundance and type of resources upon which birds depend, including nesting materials and plants involved in self-medication, our results are consistent with the view that urbanization imposes new challenges on birds that are dealt with using adaptations evolved elsewhere.     

Study of the effect of volatile substances from cereal roots

We traced the liberation and biological effect of volatile substances released from the roots of cereals,i. e. barley, wheat, rye and oats, on seedlings of the same and other plant species. Experiments were carried out in a closed glass apparatus with a static or circulating atmosphere in which the CO₂ and O₂ were permanently absorbed and supplemented, respectively. In some experiments the air was bubbled through water or through solutions of boric acid, barium hydroxide and potassium permanganate. The roots of all four cereals tested released volatile substances with a biological activity which appeared to be non-specific with respect to plant species. The effect of volatile substances was partially decreased by bubbling through water, barium hydroxide and boric acid and was completely removed after passing through the solution of potassium permanganate. Volatile substances liberated from roots of barley inhibited elongation of roots and coleoptile, decreased SH-group content and caused excessive formation of root hairs as well as inhibition of both dry matter production and respiration of roots of rye seedlings. Ethylene was found in the atmosphere of experimental vessels.     

A gas chromatographic investigation of the influence of different carbon sources on the production of volatile compounds by Dipodascus aggregatus

Summary The yeast fungus Dipodascus aggregatus was cultivated aerobically on different carbon sources. The growth was measured turbidimetrically and related to the simultaneus production of volatile compounds, which was determined by a gas chromatographic head-space technique. The same culture could be analyzed many times and up to 15 components were detected in a chromatogram recorded in 18 min. The main peaks were identified and the chromatograms quantitatively evaluated by peak height measurements.All the carbon sources tested were utilized for growth even if the lag phase was prolonged on xylose and ethanol. The production of volatile compounds from the different carbon sources decreased as follows; ethanol>glucose>fructose>glycerol >xylose and succinic acid. A good carbon source for growth could be unsuitable for the formation of volatile products and vice versa.The time course production of volatile components was recorded. On each separate carbon source the formation of volatiles was correlated to growth until the end of the exponential phase of growth.An extraction procedure including the addition of an internal standard was used to determine the exact concentrations of 9 components at near optimum production. The amounts varied between 0.1–32 mg/l medium when the fungus was grown on 5% glucose.