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Extraction of Clostridium perfringens spores from bottom sediment samples.

Two extraction-separation procedures were developed and evaluated for use in conjunction with the mCP membrane filter method for the enumeration of Clostridium perfringens spores in bottom sediments. In the more facile of the two procedures, a distilled-water suspension of the sediment sample is pulse sonicated for 10 s and allowed to settle. Portions of the supernatant are then removed for membrane filtration. This procedure is recommended for general use. The more complicated procedure is recommended for situations in which the presence of high levels of toxic materials is suspected or in which relatively low spore densities are present in fine silts. In this procedure, sonication is followed by a distilled water wash. The centrifuged sediment is resuspended in distilled water and mixed with the components of a two-phase separation system (50% polyethylene glycol in distilled water and 25% sucrose in 3 M phosphate buffer [pH 7.1]). After equilibration of the system and low-speed centrifugation, the top phase and interphase are removed, mixed, and membrane filtered. The recoveries of C. perfringens spores by the two procedures, when used in conjunction with the mCP method, were comparable to each other and significantly greater than those by the British most-probable-number method. It was estimated that more than 85% of the spores were recovered by the procedures. The precision of the sonicate-and-settle-mCP procedure was markedly better than that obtained theoretically by the most-probable-number method and approached that theoretically attributable to counting an average of 85 colonies on each of two plates.

Centrifugation↗

Fate of Fumonisin B(1) in Naturally Contaminated Corn during Ethanol Fermentation.

Two lots of corn naturally contaminated with fumonisin B(1) (15 and 36 ppm) and a control lot (no fumonisin B(1) detected) were used as substrates for ethanol production in replicate 8.5-liter yeast fermentations. Ethanol yields were 8.8% for both the control and low-fumonisin corn, while the high-fumonisin corn contained less starch and produced 7.2% ethanol. Little degradation of fumonisin occurred during fermentation, and most was recovered in the distillers' grains, thin stillage, and distillers' solubles fractions. No toxin was detected in the distilled alcohol or centrifuge solids. Ethanol fermentation of fumonisin-contaminated corn coupled with effective detoxification of distillers' grains and aqueous stillage is suggested as a practical process strategy for salvaging contaminated corn.

Journal Article↗

Volatile Flavor Compounds Produced by Molds of Aspergillus, Penicillium, and Fungi imperfecti.

Strains of molds Aspergillus niger, A. ochraceus, A. oryzae, A. parasiticus, Penicillium chrysogenum, P. citrinum, P. funiculosum, P. raistrickii, P. viridicatum, Alternaria, Cephalosporium, and Fusarium sp. were grown on sterile coarse wheat meal at 26 to 28 C for 120 h. The volatiles from mature cultures were distilled at low temperature under reduced pressure. The distillates from traps -40 and -78 C were extracted with methylene chloride and subsequently concentrated. All the concentrates thus obtained were analyzed by gas-liquid chromatography, mass spectrometry, chemical reactions of functional groups, and olfactory evaluation. Six components detected in the culture distillates were identified positively: 3-methylbutanol, 3-octanone, 3-octanol, 1-octen-3-ol, 1-octanol, and 2-octen-1-ol. They represented 67 to 97% of all the volatiles occurring in the concentrated distillate. The following 14 components were identified tentatively: octane, isobutyl alcohol, butyl alcohol, butyl acetate, amyl acetate, octyl acetate, pyridine, hexanol, nonanone, dimethylpyrazine, tetramethylpyrazine, benzaldehyde, propylbenzene, and phenethyl alcohol. Among the volatiles produced by molds, 1-octen-3-ol yielding a characteristic fungal odor was found predominant.

Journal Article↗

Antagonistic effect of monovalent cations in maintenance of cellular integrity of a marine bacterium.

The susceptibility of a marine bacterium, designated isolate c-A1, to lysis in distilled water and in salt solutions has been found to be a function of Na(+) concentration. Optical densities of cells pre-exposed to 0.05 m MgCl(2) were maintained in 1.0 m KCl, whereas those of cells pre-exposed to 1.0 m NaCl were not maintained at any KCl concentration tested. Cells transferred from MgCl(2) to low concentrations of NaCl underwent more extensive lysis than did those transferred to distilled water. The degree of disruption of cells transferred to distilled water from mixtures of 0.05 m MgCl(2) and NaCl (0 to 1.0 m) was dependent on the concentration of NaCl; similar results were obtained with LiCl, but not with KCl. In electron micrographs of thin sections, c-A1 cell envelopes consisted of two double-track layers which fractured and peeled apart on lysis after pre-exposure to NaCl-MgCl(2) mixtures. Envelope eruptions or "hernias" occurred only in lysed cells pre-exposed to NaCl alone. No evidence for a functional lytic enzyme was found. Comparative studies on a terrestrial pseudomonad with a multilayered envelope indicated that preexposure to NaCl did not enhance the susceptibility of this cell to lysis in distilled water. The lytic susceptibility of the marine bacterium is considered to be the consequence of competition between specific monovalent cations and Mg(++) for electrostatic interactions with components of the cell envelope of this organism.

Bacteria↗

Laryngeal chemoreflexes induced by acid, water, and saline in nonsedated newborn lambs during quiet sleep.

Laryngeal chemoreflexes (LCR) are triggered by the contact of assorted liquids with the laryngeal mucosa. In the neonatal period, the immature LCR consist primarily of apnea and bradycardia, which at times can be life threatening. The aim of this study was to assess LCR induction in nonsedated, newborn full-term lambs by several acid solutions, compared with distilled water and saline. Twelve lambs were instrumented for recording of glottal adductor and diaphragm EMG, EEG, eye movements, heart rate, systemic arterial pressure, and respiratory movements. LCR were induced during quiet sleep by the injection (0.5 ml) of saline, distilled water or two acid solutions (HCl and citric acid, pH 2, diluted in either water or saline). A chronic supraglottal catheter was used to inject the solutions in a random order. Distilled water and acid solutions did not induce any significant decrease in heart rate or respiratory rate. However, significant lower airway protective responses (swallowing, cough, and arousal) were observed after distilled water and especially acid solution administration. In conclusion, LCR in full-term lambs, particularly with acid solutions, are merely characterized by lower airway protective responses resembling mature LCR reported in adult mammals.

Acids↗

Effect of Tween 80 on adriamycin cytotoxicity in murine P388 leukemia.

Studies on mice bearing P388 adriamycin sensitive and resistant lymphocytic leukemia were carried out to compare antitumor activity of adriamycin dissolved in Tween 80 (10% in distilled water) with that of adriamycin dissolved in distilled water alone. The antitumor activity of adriamycin dissolved in acqueous solutions of Tween 80 was higher than that of adriamycin dissolved in distilled water against P388 leukemia sensitive to adriamycin. However, no change in the activity was observed in P388 leukemia resistant to adriamycin when the drug was administered either in aqueous solution of Tween 80 or in distilled water alone.

Animals↗

Absence of hydrocarbon-induced nephropathy in rats exposed subchronically to volatile hydrocarbon mixtures pertinent to gasoline.

Ninety-day inhalation studies were conducted on 50:50 weight percent (wt %) mixtures of n-butane:n-pentane and isobutane:isopentane, respectively, and on a distillation cut boiling below 145 degrees F of a reference unleaded gasoline blend to assess the nephrotoxicity of these volatile mixtures. The mixtures of butanes and pentanes were selected because these four hydrocarbons are the most prevalent components of gasoline vapors encountered under typical occupational exposures. The 0-145 degrees F gasoline distillation fraction was tested because it reasonably approximates the composition of gasoline vapors measured under occupational settings. Male and female F-344 rats were exposed to 2 levels of each mixture, 6 hours per day, 5 days per week, for 13 weeks. The target concentrations for the butane:pentane mixtures were 4500 and 1000 parts per million (ppm), while 5200 and 1200 ppm were set for the gasoline distillation fraction. An interim sacrifice was conducted after 28 days. The rats were not significantly affected by the exposures, and there was no evidence of hydrocarbon-induced nephropathy in either sex at the termination of each study. However, at the 28-day interim sacrifice period for both butane:pentane mixtures, mild, transient treatment-related but not exposure-related kidney effects were observed in the male rats. These perturbations were absent at the interim sacrifice period for the gasoline distillation fraction.

Animals↗

Influence of pH and temperature on kinetics of ceftiofur degradation in aqueous solutions.

The objective of this study was to evaluate the stability of ceftiofur (1 mg mL(-1)) in aqueous solutions at various pH (1, 3, 5, 7.4 and 10) and temperature (0, 8, 25, 37 and 60 degrees C) conditions. The ionic strength of all these solutions was maintained at 0.5 M. Ceftiofur solutions at pH 5 and 7.4 and in distilled water (pH = 6.8) were tested at all the above temperatures. All other solutions were tested at 60 degrees C. Over a period of 84 h, the stability was evaluated by quantifying ceftiofur and its degradation product, desfuroylceftiofur, in the incubation solutions. HPLC was used to analyse these compounds. At 60 degrees C, the rate of degradation was significantly higher at pH 7.4 compared with pH 1, 3, 5 and distilled water. At both 60 degrees C and 25 degrees C, degradation in pH 10 buffer was rapid, with no detectable ceftiofur levels present at the end of 10 min incubation. Degradation rate constants of ceftiofur were 0.79+/-0.21, 0.61+/-0.03, 0.44+/-0.05, 1.27+/-0.04 and 0.39+/-0.01 day(-1) at pH 1, 3, 5, 74 and in distilled water, respectively. Formation of desfuroylceftiofur was the highest (65%) at pH 10. The rate of degradation increased in all aqueous solutions with an increase in the incubation temperature. At pH 7.4 the degradation rate constants were 0.06+/-0.01, 0.06+/-0.01, 0.65+/-0.17, and 1.27+/-0.05 day(-1) at 0, 8, 25, 37 and 67 degrees C, respectively. The energy of activation for ceftiofur degradation was 25, 42 and 28 kcal mol(-1) at pH 5, 7.4 and in distilled water, respectively. Desfurylceftiofur formation was the greatest at alkaline pH compared with acidic pH. Ceftiofur degradation accelerated the most at pH 7.4 and was most rapid at pH 10. The results of this study are consistent with rapid clearance of ceftiofur at physiological pH.

Cephalosporins↗

Experimental polyacrylamide-induced acute injury in rat lung.

We recently reported the first case of accidental aspiration of polyacrylamide occurring in a 26-year-old man. The patient developed severe airway obstruction and parenchymal lung damage and died. Autopsy revealed numerous polyacrylamide particles in his lungs, as well as extensive bronchiolar and alveolar damage. Gas chromatographic and mass spectrometric assessment of the lung tissue failed to reveal polyacrylamide activity, although assessment of the suspending solvent of the polyacrylamide showed a pattern characteristic of an aliphatic hydrocarbon mixture with a prominent dodecane peak. This experimental study was performed to determine the nature and extent of damage to rat bronchial and alveolar epithelia following endotracheal instillation of polyacrylamide, hydrocarbon mixture (petroleum distillate), dodecane (C12H26), or normal saline. The rat lungs were examined grossly and microscopically 10 min and 24, 72, and 96 h after endotracheal instillation, following inflation and fixation with 10 percent buffered formaldehyde. Gross examination revealed congested, mottled visceral pleural surfaces in the rats treated with polyacrylamide and dodecane. There were no pleural exudates or effusions. Microscopically, vascular engorgement, bronchiolitis, and focal pneumonia were observed. Vascular engorgement was most pronounced at 72 to 96 h in rat lungs treated with polyacrylamide and dodecane and was moderate at 24 h in rats treated with petroleum distillate. Focal organizing pneumonia was marked at 96 h in rats treated with petroleum distillate, at 72 h in those treated with polyacrylamide, and at 24 h in those treated with dodecane. The saline-treated control animals showed no change. Our findings suggest that polyacrylamide, dodecane, and petroleum distillate are strong irritants to the airways. However, a direct obstructive/mechanical effect of the polyacrylamide upon the airway has not been excluded. Airway exposure to polyacrylamide may result in lung injury secondary to the polyacrylamide itself, its suspending agents, or both.

Acrylic Resins↗

Recovery of volatile products from dilute high-fouling process streams.

As biomass hydrolysis and fermentation technologies approach commercial viability, advancements in product recovery technologies will be required. For cases in which fermentation products are more volatile than water, recovery by distillation is often the technology of choice. Distillation technologies that will allow the economic recovery of dilute volatile products from streams containing a variety of impurities have been developed and commercially demonstrated. Distillation tower and tray designs, along with specialized heat-exchanger designs, allowing for extended processing intervals on solutions containing lignocellulosic residues, organic acids, and inorganic salts concentrations > 100 g/L are in commercial operation. In the case of ethanol, distillation energy consumption efficiencies for processing solutions containing < 40 g/L of desired product can approach demonstrated energy consumption efficiencies for solutions containing concentrations > 120 g/L. These proprietary technologies have been applied individually at commercial scale, and designs have been developed that incorporate the combined technologies with only a marginal increase in capital investment compared to traditional methods.

Biomass↗

Effects of intranasal administration of atrial natriuretic hormone on spontaneously hypertensive rats.

The effects of the intranasal administration of synthetic alpha-human atrial natriuretic polypeptide (alpha-hANP) were investigated in 14 anesthetized spontaneously hypertensive rats (SHR; Okamoto-Aoki strain). They were given intranasally synthetic alpha-hANP in distilled water at doses of 10 micrograms/kg, 50 micrograms/kg and 100 micrograms/kg. Intranasal application of 200 microliter of distilled water as a control was also performed in 3 anesthetized SHR. Sixteen anesthetized SHR were examined for the effects of intravenous administration of alpha-hANP at doses of 4 micrograms/kg, 10 micrograms/kg, 20 micrograms/kg and 40 micrograms/kg. Urinary volume and the urinary excretion of sodium increased 2- to 3-fold during the 50 minutes following intranasal administration of a single dose of 50 micrograms/kg or 100 micrograms/kg, although neither the urinary volume nor the urinary excretion of sodium increased after intranasal administration of 10 micrograms/kg of alpha-hANP or 200 microliter of distilled water. There were no significant changes in arterial pressure or heart rate after the intranasal administration of synthetic alpha-hANP or distilled water. In contrast, arterial pressure was decreased and urinary volume and urinary excretion of sodium were increased, in a dose dependent manner, within 5 minutes after intravenous bolus-injection of alpha-hANP and returned to their baseline levels within 20 minutes. These results indicate that intranasal administration of synthetic alpha-hANP exerts its diuretic effect without concomitant changes in arterial pressure or heart rate in SHR.

Administration, Intranasal↗

Improvement of silver impregnation technique (protargol) to obtain morphological features of protists ciliates, flagellates and opalinates.

The research on ciliates, flagelates and opalinates have been widespread by the utilization of techniques employing silver impregnation (protargol), modified by several authors. However, these are time consuming and its results are variable. The present work is a variant of the technique described by Tuffrau (1964, 1967) showing some adaptations made in our laboratory. The organisms can be preserved by different fixatives (alcoholic Bouin, Stieve's fluid, 2.5% glutaraldehyde and others) and then rinsed in destilled water followed by a fast clarification by 3% sodium hypochloride. If the organism is very sensitive to hypochloride, 4% sodium lauryl sulfate may be used and then washed 3 times in distilled water. The protista can be adhered to the glass slides with Mayer's glycerinated-albumin (1 glycerin vol. to 1 or 2 albumin vol.), diluted in water at a proportion of 1:10 Cv/v., or with 1% polylysine followed by fast washes with distilled water. After the slide preparation, they were covered with a layer of 0,8% Silver proteinate. Right after that, the slide has to be placed in a glass tray lined with moist tissue and covered to prevent the proteinate to dry. The tray was placed in a incubator at 40 degrees - 50 degrees C for 30 minutes. The slides are rinsed for 1 minute. with warm (35 degrees C) distilled water. The development of the material should be done with 0.4% hydroquinone with a maximum incubation time of 1 minute. It should be developed gradually, controlling the silver impregnation intensity by observation under optical microscope. Next, rinse in distilled water for 1 minute, and then, fix in 2,5% Sodium thiosulfate. Rinse the slide for two minutes before dehydrating it in an alcoholic serial 50-100 degrees. Finally rinse the slides in xylene. Mount the slides with Entellan MerckTM or Canada balsam.

Animals↗

[Changes in chewing rhythm during repetitive series of mastication in normal adults as a function of time elapsed and the four basic taste stimuli].

Surface EMG activities in the masseter muscle were recorded from 8 healthy men with normal permanent dentition during habitual chewing of a small cube of synthetic cellulose sponge which had absorbed distilled water. Three parameters of chewing rhythm, viz., cycle time, duration, and interval, were shown as the respective averages of 40 measurements of the EMG traces of a masticatory series of 42 consecutive chewing strokes. Twenty-five recording sessions, in each of which the above masticatory series were repeated 10 times in a day with a 5 minutes' recess between each series, were performed on a subject during continuous 25 days. The amount of scatter in the distribution of 25 cycle time values was great at the 1st masticatory series, gradually decreased to reach a node relatively free from the dispersion at the 5th series, and then gradually increased from the 6th to 10th series. The same phenomenon was observed in all 8 subjects, although there was a small difference among them in the number of the series where the node was apparent. The average of 5 cycle time values was used to compare in the same subject the effect of chewing the cellulose sponge which had absorbed one of the 4 basic taste solutions with the effect of chewing that which had absorbed distilled water. The average value while chewing the sponge which had absorbed 1 M saccharose was significantly lower in the subjects who obtained a relatively large average value during distilled water sponge-chewing, whereas it was significantly greater in those who obtained a relatively small average value while chewing the sponge which had absorbed distilled water. The other 3 taste stimuli induced a significant increase in the average value in most subjects.

Adult↗

Reproducibility of seasonal effects in antibody formation and host resistance in the outbred mouse.

The purpose of this study was to assess the reproducibility of any seasonal effects in the outbred CD1 mouse of antibody production to sheep red blood cells (SRBCs) and host resistance to the bacterium Listeria monocytogenes. A marked seasonal effect on antibody production was seen when 5- to 6-week-old female CD1 mice were studied on a weekly basis for a period of 2 years. Maintained on a 12:12 h light:dark schedule, animals were held for 12 days prior to experiment to insure physical condition and acclimatization to the lighting regimen. Beginning at 4 h after lights on (HALO) for day 1 and 2 HALO thereafter, groups of mice were (a) not treated, (b) administered a vehicle (corn oil, 1% methylcellulose, or distilled water) by oral gavage for 5 days, or (c) not treated, but given an intraperitoneal injection of cyclophosphamide 24 h prior to assay. On the fifth day, mice were injected with SRBCs intravenously. Four days later, antibody formation against SRBCs was measured using spleen cells. Circannual and seasonal rhythms were displayed by each group of animals, with greatest antibody production, indicated by the number of plaque-forming cells (PFCs)/million viable cells, in the Spring (range of double amplitude = 36-108%). The timing of these rhythms was reproducible from one year to the next. In contrast, the magnitude of the response in year 1 was significantly different from year 2 for animals given vehicle or not treated. Cyclophosphamide-treated mice had consistently low numbers of plaque-forming cells. Host resistance was studied in separate mice treated with vehicles at the same time as the antibody assay. On the third day of dosing, mice were injected intravenously with Listeria monocytogenes (LM) and monitored for death for 10 days. When analyzed by Kruskal-Wallis life table analysis, there was no overall effect of vehicle on survival for 1987 but there was an effect for 1988 and when data from both years were combined. Distilled water-treated mice had lower survival rates than the other two vehicles. Mice treated with distilled water displayed a circannual rhythm for survival for 1988 and for both years combined, in contrast to the other two vehicles. When each vehicle was analyzed separately for seasonal effect, a significant effect of season occurred for corn oil- and distilled water-treated animals. The greatest survival rate and longest survival time generally occurred in the months between July and December.

Analysis of Variance↗

A new method for easy demonstration of argyrophil cells.

A new method for silver impregnation of endocrine cells of the gastrointestinal mucosa is described. It offers great reliability, eveness of impregnation, and, since it can be used on batches of slides, is also suitable for histology class and investigation material. The procedure for paraffin sections of formalin-fixed material is as follows: dewax and transfer to distilled water, leave in 0.5% silver nitrate solution for 2 hours at 60 C. Rinse in distilled water, then treat in Bodian developer (hydroquinone, 1 g; sodium sulphite, 5 g; distilled water, 100 ml) previously heated to 60 C. Rinse in running tap water, distilled water, and then re-impregnate for 10 minutes at 60 C in the same silver solution and reduce in Bodian's solution. Since the background is not impregnated by this method, sections may be counterstained by any basic anilin dye to bring out nuclei. A 0.1% kernechtrot solution was found very satisfactory in this respect. The granulations of argyrophil cells stand out sharply black against a red background.

Chromaffin System↗

Substitution of neutral detergent fiber from forage with neutral detergent fiber from by-products in the diets of lactating cows.

Four lactating dairy cows that were ruminally and duodenally cannulated were used in an experiment with a 4 x 4 Latin square design to determine the effects of the substitution of neutral detergent fiber (NDF) from forage with NDF from wheat middlings, corn gluten feed, or a blend of distillers dried grains and hominy. Dietary crude protein and NDF averaged 18 and 31%, respectively, for the diet with 71.2% of the NDF from forage (control diet) and for diets with 55% of the NDF from forage (by-product diets). The substitution of NDF from these by-products for forage NDF did not affect dry matter intake (20.1 kg/d) or digestibility of organic matter. Total tract digestibility of acid detergent fiber was lower for cows fed the diet containing a blend of distillers dried grains and hominy than for cows fed the diet containing corn gluten feed. Microbial crude protein synthesis, milk production (23.9 kg/d), and milk fat percentage were similar for all cows, regardless of diet. Cows fed the diets containing wheat middlings or a blend of distillers dried grains and hominy had reduced ruminal pH compared with that of cows fed the diet containing corn gluten feed or the control diet. Diets containing 55% of total NDF from forage with 31% of total NDF from corn gluten feed, wheat middlings, or a blend of distillers dried grains and hominy can supply sufficient effective fiber to maintain normal ruminal function.

Ammonia↗

Modified extraction 2-thiobarbituric acid method for measuring lipid oxidation in poultry.

A simple and accurate objective test for monitoring lipid oxidation in poultry products is needed. Thiobarbituric acid (TBA) values of cooked poultry meat were determined by a distillation method and by a modified extraction method. Perchloric acid was used in the extraction method to release TBA reactive substances from the meat. The maximum absorption wavelength of the TBA-malonaldehyde complex was 531 nm. The TBA values for cooked chicken and turkey were highly correlated (r = .91) for both the extraction and distillation methods. Sensory scores of warmed-over flavor in precooked poultry were highly correlated with TBA values in both the distillation (r = .83) and extraction (r = .85) methods. Results indicate that the extraction method is faster, easier to perform, and as accurate as the distillation method for monitoring lipid oxidation in poultry.

Animals↗

Characterization of enamel and dentin surfaces after removal of temporary cement--study on removal of temporary cement.

Enamel and dentin surfaces of bovine teeth were characterized after mechanical and chemical removal of temporary cements. The surfaces were examined by scanning electron microscopy and X-ray microanalysis. Contact angles of distilled water were measured to monitor surface wettability. Mechanical removal with a dental probe did not completely remove all of the temporary cement from both enamel and dentin surfaces. These surfaces usually produced contact angles of distilled water greater than that on polished surfaces. Etching with thirty-seven percent phosphoric acid effectively removed the temporary cement that remained on enamel surfaces after mechanical removal but the etchant was not effective on dentin surfaces. Acid etching after mechanical removal of cement significantly decreased the contact angles of distilled water on enamel surfaces but the etchant only slightly improved contact angles of distilled water on dentin surfaces.

Acid Etching, Dental↗