Ammonia volatilization from tropical legume mulches and green manures on unlimed and limed soils

In the humid tropics, legumes are harvested and surface applied as mulch or incorporated as green manure. Studies on N dynamics and budgets from these systems report unaccounted losses of N. Ammonia volatilization may account for a significant percentage of these unexplained N deficits. The main objectives of this study were to: 1) determine the rate and amount of ammonia volatilization from organic amendments, both incorporated (green manure) and unincorporated (mulch), 2) compare ammonia volatilization of organic amendments on both acid (unlimed) and limed soils, and 3) correlate quality, i.e. polyphenolic and lignin concentration and carbon-to-nitrogen ratio, of the organic amendments with ammonia volatilization and net N mineralization. In an incubation experiment, ammonia volatilization losses and net N mineralization were measured from fresh leaflets of 10 legumes over a three-week period. Ammonia volatilization losses for the 10 species ranged from 3.4 to 11.8% of the total N applied in the organic amendment. Lignin content was negatively correlated to ammonia volatilization. Ammonia volatilized from mulches but not green manures, on both unlimed and limed soils, suggesting that ammonia volatilization is a surface phenomenon and not affected by soil pH. Net N mineralization was affected by species and soil pH, but was unaffected by placement (green manure or mulch). For the farmer in low-input agriculture where N tends to be limiting, volatilization losses of N from legume mulch systems could be on the same order of magnitude as crop removal.     

Characterization of Phosphorus in Animal Manures Collected from Three (Dairy,Swine, and Broiler) Farms in China

In order to identify the phosphorus species and concentration in animal manure, we comparatively characterized phosphorus in dairy manure, swine manure, and broiler litter, using a sequential procedure, a simplified two-step procedure (NaHCO₃/NaOH+EDTA), and a solution Phosphorus-31 Nuclear Magnetic Resonance (³¹P-NMR) spectroscopy procedure. In the sequential procedure, deionized water extracted 39, 22, and 32%; NaHCO₃ extracted 48, 26, and 37%; NaOH extracted 8, 9, and 13.8%; and HCl extracted 3, 42.8, and 17% of the total phosphorus in dairy manure, swine manure and broiler litter, respectively. Total phosphorus extracted by the NaHCO₃/NaOH+EDTA procedure was 7.5, 32.4, and 15.8 g P kg⁻¹ for dairy manure, swine manure, and broiler litter, respectively. The solution ³¹P-NMR procedure detected that 9, 34, and 29% of total phosphorus was phytic acid in dairy manure, swine manure, and broiler litter, respectively. These results show that phosphorus forms, availability, and quantities differ between animal manures, which provides valuable information for P characterization of animal manures in China.     

In situ measurement of ammonia and greenhouse gas emissions from broiler houses in France

An experimental technique was developed for measuring gaseous emissions including ammonia (NH(3)), nitrous oxide (N(2)O) and methane (CH(4)) from broiler houses. This technique included the monitoring of the air flow rate and the gaseous concentrations. NH(3) was determined using acid trap while N(2)O and CH(4) were determined continuously by infrared gas analyser and sequentially by gas chromatography. Moreover, N(2)O and CH(4) emissions were monitored above the litter using closed flux chambers at the end of the experiment. No emissions of N(2)O and CH(4) were observed neither during the growth of the broiler nor above the litter at the end of the experiment. Ammonia concentration varied between 0.8 and 32 ppm in the building. Total ammonia emissions were estimated to 5.74 g N animal(-1) during this experiment. According to this result, ammonia emissions from broiler houses could be estimated to 5.3 kt of N per year in France.     

几种控释氮肥减少氨挥发的效果及影响因素研究

采用“静态吸收法”和“土柱淋溶法”、室内模拟试验,研究几种控释氮肥施入土壤后的氨挥发损失情况、N溶出速率、土壤脲酶活性及pH值变化的关系．结果表明,施氮450mg·kg^-1土时,3种控释氮肥氨挥发损失氮总量分别比普通尿素减少49．7％、28．0％和71．2％;施氮600mg·kg^-1土时,3种控释氮肥氨挥发损失氮总量分别比普通尿素减少34．6％、12．3％和69．9％．控释氮肥能显著降低土壤氨挥发量,减少因施肥而引起的大气环境污染．控释氮肥氨挥发量与不同氮肥引起的土壤脲酶活性、pH值、土壤中氮溶出速率密切相关．土壤的氨挥发总量与肥料在土壤中溶出总量的相关系数达到0．9533,在肥料施入的前期土壤氨挥发量同土壤脲酶活性、pH值的相关系数达到0．9533和0．9908。     

双水相体系萃取分离杜仲叶中桃叶珊瑚甙的研究

建立了由高分子化合物聚乙二醇(PEG4000)与葡聚糖40000(D40)形成的双水相体系萃取分离杜仲叶中桃叶珊瑚甙的新方法.考察了萃取体系相图,研究了PEG4000/D40质量分数、样品溶液加入量、pH值和温度等因素对双水相成相及桃叶珊瑚甙萃取率的影响.结果表明:PEG4000的质量分数为11%,D40质量分数为8%、样品溶液加入量为8 g,温度为60℃,溶液pH为7时,双水相体系对桃叶珊瑚甙有较高的萃取率,重复三次可达到66.32%,而且萃取得到的桃叶珊瑚甙产品的纯度达到48.67%,远远高于粗提物中的8.750%.     

Extraction of starches from tuber crops using ammonia

Ammonia solution (0·03 ) was used to extract starch from various tuber crops by the conventional settling method. It was found that there was noticeable improvement in the yield of starch from Colocasia (6–16%), while it fell for sweet potato starch and remained almost the same for the other starches. The various properties of starch, thus extracted, were compared with those for starch obtained by water extraction. It was found that total amylose of all starches were unaffected while the ‘soluble amylose’ was slightly suppressed for Colocasia starch extracted with ammonia solution. Peak viscosity was found to be increased to a large extent for Colocasia and Dioscorea esculenta starches by ammonia extraction, while it was lowered for sweet potato starch. The swelling volume of Colocasia starch extracted with ammonia was similarly enhanced by 25%, but the Dioscorea esculenta starch did not show such a tendency. Sweet Potato starch suffered a reduction in swelling volume. Phosphorus content was found to be independent of the extraction medium.     

Managing ammonia emissions from dairy cows by amending slurry with alum or zeolite or by diet modification

Animal agriculture is a significant source of atmospheric ammonia. Ammonia (NH3) volatilization represents a loss of plant available N to the farmer and a potential contributor to eutrophication in low-nitrogen input ecosystems. This research evaluated on-farm slurry treatments of alum or zeolite and compared three diets for lactating dairy cows in their effectiveness to reduce NH3 emissions. NH3 emissions were compared using a group of mobile wind tunnels. The addition of 2.5% alum or 6.25% zeolite to barn-stored dairy slurry reduced NH3 volatilization by 60% and 55%, respectively, compared to untreated slurry. The alum conserved NH3 by acidifying the slurry to below pH 5, while the zeolite conserved ammonia by lowering the solution-phase nitrogen through cation exchange. The use of alum or zeolite also reduced soluble phosphorus in the slurry. NH3 loss from fresh manure collected from lactating dairy cows was not affected by three diets containing the same level of crude protein but differing in forage source (orchardgrass silage vs. alfalfa silage) or neutral detergent fiber (NDF) content (30% vs. 35% NDF). NH3 losses from the freshly excreted manures occurred very rapidly and included the urea component plus some unidentified labile organic nitrogen sources. NH3 conservation strategies for fresh manures will have to be active within the first few hours after excretion in order to be most effective. The use of alum or zeolites as an on-farm amendment to dairy slurry offers the potential for significantly reducing NH3 emissions.     

Yellow Mealworm Protein for Food Purposes - Extraction and Functional Properties

A protocol for extraction of yellow mealworm larvae proteins was established, conditions were evaluated and the resulting protein extract was characterised. The freeze-dried yellow mealworm larvae contained around 33% fat, 51% crude protein and 43% true protein on a dry matter basis. The true protein content of the protein extract was about 75%, with an extraction rate of 70% under optimised extraction conditions using 0.25 M NaOH, a NaOH solution:ethanol defatted worm ratio of 15:1 mL/g, 40°C for 1 h and extraction twice. The protein extract was a good source of essential amino acids. The lowest protein solubility in distilled water solution was found between pH 4 and 5, and increased with either increasing or decreasing pH. Lower solubility was observed in 0.5 M NaCl solution compared with distilled water. The rheological tests indicated that temperature, sample concentration, addition of salt and enzyme, incubation time and pH alterations influenced the elastic modulus of yellow mealworm protein extract (YMPE). These results demonstrate that the functional properties of YMPE can be modified for different food applications.     

豆腐柴叶提取低酯果胶的研究

豆腐柴叶含有丰富的果胶。文中结合盐酸浸提法的基础上,采用醇氨降酯法提取豆腐柴叶的果胶并检测,提取率为部颁19.5%,pH值为2.9,水分和灰分分别为5.13%和8.67%,半乳糖含量为65%,酯化度为38.5%,所提取的低酯果胶的基本性质与国标检测标准接近。果胶成品经AB-8树脂柱纯化后,所提果胶的成分经硅胶板薄层层析初步测定,果胶多糖成分为葡萄糖,果糖,D-甘露糖3种,果胶的溶解度与温度呈正相关性,pH值对其溶解度影响不大。     

Effective extraction of astaxanthin pigment from shrimp using proteolytic enzymes

The present study was to investigate efficient extraction conditions of astaxanthin from shrimp wastes for utilizing it as a functional food additive. In order to enhance the stability of pigments, proteolytic enzymes were applied to extract astaxanthin as carotenoprotein. Also, various conditions such as acid ensilaging of samples, using EDTA solution and adding various enzymes were examined to optimize extraction processing. After extraction, all of the extracts were partitioned in a separate funnel and each astaxanthin content was analyzed by using UV/VIS spectrophotometer. Carotenoprotein was effectively extracted from non-acid ensilaged shrimp wastes by using EDTA medium and a proteolytic enzyme. In that case, the reddish top layer showed 91.9% of recovery and the blackish bottom layer did 2.3% and its separation ratio was about 0.2 (v/v); therefore concentration and purification of reddish top layer were more desirable than those of whole extract.     

响应面法优化白头翁总黄酮提取工艺

采用无水乙醇C2H5OH/硫酸铵(NH4)2SO4双水相体系分离白头翁中的黄酮。确定双水相体系组成为21%C2H5OH/22%(NH4)2SO4,通过单因素试验和Box-Benhnken实验设计探讨黄酮粗提液质量分数、NaCl质量分数和pH值对萃取效果的影响。确定最佳萃取条件为:黄酮粗提液质量分数12.5%,NaCl质量分数1.5%,pH 5.99,在此条件下,白头翁总黄酮主要分布在上相,萃取率可达73.6%。     

Odour and ammonia emissions from intensive poultry units in Ireland

Odour and ammonia emissions were measured from three broiler, two layer and two turkey houses in Ireland. The broiler units gave a large range of odour and ammonia emission rates depending on the age of the birds and the season. A considerable variation between the odour and ammonia emission rates was evident for the two layer units which may have been due to the different manure handling systems utilised in the houses. There was relatively little difference in the odour and ammonia emissions from the two turkey houses. As a precautionary principle, odour emission rates utilised in atmospheric dispersion models should use the maximum values for broilers and turkeys (1.22 and 10.5 ou(E) s(-1) bird(-1) respectively) and the mean value for the layers depending on the manure handling system used (0.47 or 1.35 ou(E) s(-1) bird(-1)).     

双水相萃取法分离竹叶黄酮的工艺研究

以竹叶黄酮水提溶液为原料,采用PEG(聚乙二醇)/(NH4)2SO4双水相体系对竹叶黄酮进行萃取,考察了PEG平均相对分子质量、PEG质量分数、(NH4)2SO4质量分数、pH值、NaCl质量分数、原液质量分数、萃取温度等对双水相及竹叶黄酮萃取效果的影响。双水相萃取法提取竹叶黄酮的最优条件为:PEG 400 31%,(NH4)2SO411%,pH 3.9,NaCl 0.7%,原液51.5%,萃取温度20℃,在此条件下得到的竹叶黄酮萃取率为97.8%。结果说明,双水相萃取法操作简单方便,成本低,不会引起生物质失活或变性,适合于黄酮类化合物的萃取分离。     

Use of Solid-Phase Extraction To Determine Ergosterol Concentrations in Plant Tissue Colonized by Fungi

At present, the ergosterol and acetate-to-ergosterol techniques are generally considered to be the methods of choice to quantify fungal biomass, growth rate, and productivity under natural conditions. Both methods rely on the accurate isolation and quantification of ergosterol, a major membrane component of eumycotic fungi. Taking advantage of the solid-phase extraction (SPE) technique, we present a novel method to determine the ergosterol concentration in lipid extracts derived from plant tissues and dead organic matter colonized by fungi. In this method, a primary alkaline extract is acidified and passed through a reversed-phase (C(inf18)) SPE column. The column is then washed with an alkaline methanol-water solution to eliminate interfering substances and increase pH and is thoroughly dried in air. Ergosterol is eluted with alkaline isopropanol. This eluting solvent was chosen to produce a strongly basic pH of the final extract and thus confer stability on the ergosterol molecule before high-performance liquid chromatography analysis. The recovery of ergosterol from plant tissues and the O(infhf) horizon of a woodland soil ranged from 85 to 98%, and the overall extraction efficiency was similar to that obtained by a conventional procedure involving liquid-liquid extraction. Potential pitfalls of ergosterol analysis by SPE include (i) insufficient acidification before sample loading on the extraction column, resulting in a poor affinity of ergosterol for the sorbent; (ii) incomplete drying of the sorbent bed before the elution step; and (iii) chemical breakdown of ergosterol after elution, which was found to be related to a low pH of the final extract and a high concentration of matrix compounds. When these pitfalls are avoided, SPE is an attractive alternative to existing methods of ergosterol analysis of environmental samples.     

Spore toxin complex recovery from solid-state fermentation of some mosquitocidal Bacilli

The efficiency of different methods for recovery of spore toxin complex (STC) from solid-state fermentation (SSF) extracts of Bacillus thuringiensis israelensis (Bti) and Lysinibacillus sphaericus 14N1(Ls14N1) was studied. Drying at 30–50°C, lyophilisation, flocculation by ferric chloride at 0.1–0.2% and encapsulation by entrapment in calcium alginate beads, agar, or polyacrylamide gel were efficient methods for recovery of STC of Ls14N1 and Bti from SSF extracts. Also, STC was precipitated with remained the mosquitocidal activity through adjusting the pH of the SSF liquid extract before centrifugation at pH 2–5 or after addition of acetone (200–400%), ethanol (400%) or acetic acid (2.5–4%) to SSF liquid extract of Ls14N1. On the other hand, STC of Bti showed a good recovery by centrifugation after adjusting SSF liquid extract at pH 4 followed by pH 3 and pH 5 or after the addition of ethanol (300%) or acetone (200–300%). This study contains simple and multiple methods for efficient recovery of STC of Bti and Ls14N1 grown under SSF. The choice of the best method depends on the availability of the necessary equipment as well as the cost of the treatment.     

Detection of Staphylococcal Enterotoxin in Foods

A short procedure for the extraction of staphylococcal enterotoxins from food materials has been developed. The procedure involves extraction of the food at pH 4.5, centrifugation, extraction of the supernatant with CHCl(3) (pH 7.5), extraction of the enterotoxin from the water layer with CG-50 ion exchange resin (pH 5.4 to 5.9), and treatment of the eluate with agar and concentration with Carbowax 20-M. The concentrate was extracted with CHCl(3), and the water layer was lyophilized. The dried material was dissolved in a 1% trypsin solution and placed on microslides, which were incubated 24 h at 37 C. The time required for enterotoxin analysis was 3 days with microslides and 1 day with the reversed passive hemagglutination technique.     

Characterization of organic matter from animal manures after digestion by earthworms

The humification index (HI) values of three different manures and earthworm casts were calculated for three different extractant solutions (0.5M sodium hydroxide, 0.1M sodium pyrophosphate pH 7 and 0.1M sodium pyrophosphate plus 0.1M NaOH). The alkaline sodium pyrophosphate solution was found to be the most suitable because of both its extraction efficiency and the quality of the organic matter extracted which allows a good characterization of the stabilization degree attained by composting. Neutral sodium pyrophosphate extracts also show characteristic HI values for different samples but lower extraction efficiencies. The HI values for sodium hydroxide extracts show only little differences between manures and composts. The good correspondence found between HI data and isoelectric focusing (IEF) patterns confirmed on one side that humification indexes give a quantitative measure of the humification degree, on the other side that IEF is a suitable technique in order to obtain qualitative informations on organic matter stabilization in earthworm casts.     

A procedure for purifying jack bean urease for clinical use

Urease with a purity meeting the requirements of analytical use was purified from jack bean meal through steps consisting of 20% acetone extraction, heat treatment, acid precipitation, and lyophilization. For extraction of urease, one part of bean meal was mixed with 5 parts of 20% acetone containing 1 mM EDTA and 1 mM 2-mercaptoethanol, and stirred at 20 degrees C for 5 min. Milky substances in the extract were removed by heat treatment. Urease in the clear yellow supernatant was precipitated by adjusting the pH of the solution to 5.4 with citric acid. The acid precipitated urease was neutralized by dissolving in 0.015 M phosphate buffer, pH 8.5 (final pH 6.8 to 7.0) and then lyophilized. By this procedure, the purity of the enzyme was increase 14.7 fold, the recovery of activity was 63%, and the yield was 6.75 g from 1 kg of bean seeds. The specific activity of the preparation was 411 units/mg protein (240 units/mg solid), and the free ammonia content was less than 0.01 microgram per unit. Some other proteins were present in the urease preparation as examined by gel filtration and gradient polyacrylamide gel electrophoresis. The molecular weight of the enzyme estimated by gel filtration was 480,000. However, two urease activity bands with molecular weight of 230,000 and 480,000 were observed in the polyacrylamide gel electrophoregram. From the result of determination of blood urea nitrogen (BUN), this simple purification procedure could be used for practical preparation of urease from jack bean meal for clinical analysis.     

Bacterial action of fertilization envelope extract from eggs of the fish Cyprinus carpio and Plecoglossus altivelis

The vitelline envelope (VE) and fertilization envelope (FE) in eggs of the fish Cyprinus carpio and Plecoglossus altivelis were purified by homogenization of eggs or embryos in 5 mM Tris-HCl buffer, pH 7.0, containing 2 mM ethylenediamine tetraacetic acid disodium salt (EDTA), except for processing of VEs in Plecoglossus eggs, and by repeated washing wih the same buffer. To extract the outermost layer material, the purified VEs and FEs were processed overnight at 4 degrees C in 5 mM Tris-HCl buffer, pH 7.0, containing 8 mM 2-mercaptoethanol, 2 mM EDTA, 0.3 M alpha-lactose, 0.3 M glucose, and 0.9% NaCl. Since extraction of the outermost layer of the VEs of Cyprinus eggs in this solution was found to be ultrastructurally incomplete, further sonication in the same buffer was necessary. The solution extracted from purified VEs or FEs was dialyzed against 5 mM Tris-HCl buffer, pH 7.0, followed by lyophilization. The extracts from the FEs from both fish species contained two kinds of lectins, one agglutinated human B-type erythrocytes and the other nonspecifically agglutinated fish spermatozoa, and both extracts had a strong bactericidal effect on Vibrio anguillarum that was isolated from diseased cultured fish, but not on Aeromonas hydrophila and Escherichia coli. The extracts of purified VEs from eggs of both fish had no bactericidal effect on the bacteria examined, nor any agglutination effect on human erythrocytes and fish spermatozoa.     

Improved method of lipopolysaccharide isolation from gram-negative bacteria

A modification of the traditional method for lipoplysaccharide isolation from the cells of grammnegative bacteria was elaborated on the basis of extraction by the hot water solution of phenol (the method of Westfahl). To make the method simpler and to raise the yield of the product it was proposed to use the water-phenol extract without its division for plases. The nucleic acids are eliminated by precipitation from dialyzed extract at pH 3,2-3,4 achieved by addition of acetic acid. The comparative isolation of lipopolysaccharides by the classic and modified methods has confirmed the advantages of a new technique.