Fertilizer potential of some commercially available forms of keratin and microbial biomass

The mineralization of nitrogen from citric acid waste mycelium (Aspergillus niger) and Pruteen (Methylophilus methylotrophus), and nitrogen and sulphur mineralization from keratin (feather meal) have been studied in a fertile loam soil. Ammonium nitrogen was rapidly released from the organic nitrogen sources and was subsequently nitrified. Sulphate release from feather meal was also demonstrated. Both the advantages and disadvantages of using these commercially available forms of microbial biomass and keratin as fertilizers are discussed.     

Production of keratinolytic enzyme by a newly isolated feather-degrading Stenotrophomonas maltophilia that produces plant growth-promoting activity

A novel feather-degrading Stenotrophomonas maltophilia R13 was isolated from rhizospheric soil of reed. The strain R13 produces keratinolytic enzyme using chicken feather as the sole carbon and nitrogen source. Addition of 0.1% glucose and 0.12% polypeptone to the feather medium increased the enzyme production. The optimum temperature and initial pH for the enzyme production were 30 °C and 7.0. The maximum yield of the enzyme was 82.3 ± 1.0 U/ml in the optimal feather medium; this value was about 5.5-fold higher than the yield in the basal feather medium. S. maltophilia R13 possessed disulfide reductase activity along with keratinolytic activity. As a result of feather degradation, 18 free amino acids were produced in the culture; the concentration of total amino acid was 2298.8 μM. The strain R13 produced IAA in the optimal feather medium without l-tryptophan supplementation, indicating simultaneous production of keratinolytic activity and IAA by S. maltophilia R13. The strain R13 grown in the optimal feather medium also inhibited mycelial growth of some phytopathogenic fungi. This result suggests that antifungal activity of the strain R13 could be produced in the same conditions observed for keratinolytic activity. Thus, S. maltophilia R13 could be not only used to enhance the nutritional value of feather meal but is also a potential bioinoculant in agricultural environments.     

A biotechnological process for treatment and recycling poultry feathers as a feed ingredient

A strain of Kocuria rosea with keratinolytic capacity was cultured aerobically on submerged feathers to obtain a fermented feather meal (FFM). This FFM enriched with cells of K. rosea mainly contains crude protein (71%). The pepsin digestibility of the fermented product (88%) was similar to the value of the commercial feather meal and more than 70% greater that untreated feathers. The bacterial biomass improved the content of amino acids lysine (3.46%), histidine (0.94%) and methionine (0.69%). Additionally, the amino acid availability tested by in vivo assay was greater than commercial feather meal. The microbial cells also supplied carotenoid pigments to FFM (68 ppm). These results suggest that feather meal enriched with K. rosea may be useful in animal feeding as protein and pigment source.     

6种石斛属植物氨基酸组成及营养价值评价

为比较不同种石斛的氨基酸组成差异,作出营养评价,使用氨基酸自动分析仪检测6种石斛的氨基酸含量,分析其必需氨基酸组成成分,并通过计算氨基酸评分(AAS)、化学评分(CS)及氨基酸比值系数分(SRCAA)等非生物指标进行营养评价。结果表明:石斛中氨基酸种类齐全、营养丰富,均含有被检测的17种氨基酸。6种石斛氨基酸总量存在差异,介于3.58%～8.09%。必需氨基酸组成总含量介于42.62%～47.23%之间,明显高于WHO/FAO模式值(35.00%),其中,化学评分显示其限制氨基酸均为半胱氨酸和蛋氨酸。6种石斛药用氨基酸含量相差较大(2.24%～5.37%),其中蜻蜓石斛含量最高,为含量最低细叶石斛的2.40倍,但在总氨基酸中所占的比例基本一致(60.96%～66.47%)。对6种石斛进行聚类分析可分为4类,其中杓唇石斛、叠鞘石斛、细叶石斛可视为高品质蛋白质种。综上所述,石斛氨基酸具有重要的营养价值,6种石斛氨基酸差异显著。该结果可为石斛营养价值评价、品种选育及药用开发提供理论指导。     

Yield,composition and nutritive value of a leaf protein concentrate obtained by fractionating the shelled aerial part of Pisum sativum L.

A leaf protein concentrate (LPC) extracted from shelled plants of pea (Pisum sativum L.) yielded 108 kg DM ha^?1. Dry matter (DM) was analysed for ether extract (13.59%), neutral detergent fibre (NDF, 1.79%), acid detergent fibre (ADF, 0.81%) and total ash (10.57%). The main nitrogen fraction was represented by protein-N (92% of total N). Mineral composition showed an adequate ratio of Ca to P (3). Lysine was present in nutritionally suitable amounts (7.42 g per 100 g recovered amino acids), but methionine was not (1.78). Apparent digestibility (AD; 75.5), true digestibility (TD; 87.6), biological value (BV; 68.8) and net protein utilisation (NPU; 59.7) were determined with Wistar albino rats. Methionine addition improved AD (81.3), BV (80.6) and NPU (70.5), although not TD (87.5) which was already very high.     

Nitrogen partitioning and blood meal utilization by Aedes aegypti (Diptera Culicidae)

The utilization of the blood meal by mosquitoes was investigated by first feeding females quantities of blood ranging from 1 to 5 mg, and then analyzing the faeces for the various by-products of protein catabolism that were subsequently eliminated. The nitrogeneous waste products in order of importance were uric acid, histidine, ammonia and arginine. Only traces of the other amino acids were excreted.The total amount of each faecal substance varied linearly with the quantity of blood ingested, however their relative proportions did not change. Regardless of blood meal size the quantily of uric acid and ammonia produced indicates that about 80% of the non-histidine and arginine amino acids are deaminated and utilized for metabolic purposes other than egg protein synthesis.Most of the histidine and about one half of the arginine content of the blood were excreted as free amino acids, but the other amino acids were lost in trace amounts.Nineteen per cent of the total ingested amino acids was incorporated into soluble yolk proteins and this proportion was constant even for small blood meals that result in a reduction in the numbers of eggs produced.The comparative aspects of nitrogen partitioning and blood meal utilization by haematophagous insects, as well as the factors that affect blood meal utilization and fecundity in A. aegypti are discussed.     

Mosquito reproduction: Incomplete utilization of the blood meal protein for oögenesis

Fecundity, measured by utilization of dietary protein for the production ofyolk protein was examined in several mosquito species (A. aegypti, A. simpsoni, A. atropalpus, A. triseriatus) using blood from eight vertebrate hosts over a whole range of biological blood mean volumes. All experimental blood meals were injected by enema in measured amounts. The total ovarian and excretory nitrogen produced during each gonotrophic cycle was quantified.Blood proteins are utilized for oögenesis only to a limited extent: two thirds, at the most, are detectable in the mature ovaries. This efficiency decreases with increasing blood meal size to about one fifth. The nitrogen content of each mature egg is constant and species-specific, irrespective of blood meal size or source.There are no significant differences in the efficiency of conversion of protein from different types of vertebrate blood into oöcyte protein except for human blood which led to a drastically reduced efficiency. This is shown to be due to a lack of isoleucine which is essential for reproduction. Hominid primates lack this amino acid in their haemoglobin molecule and it exists at a low titre in human plasma. Apparently, in other vertebrates the amino acids are better balanced in the haemoglobin or, can be mobilized in sufficient amounts from various non-haemoglobin protein fractions.All blood protein is catabolized and the excess nitrogen excreted quantitatively; excretory nitrogen therefore reflects the quantitative demands of a gonotrophic cycle. These processes are not dependent upon receiving a mating stimulus.     

The role of hemolymph proline as a nitrogen sink during blood meal digestion by the mosquito Aedes aegypti

Mosquitoes utilize the amino acids derived from blood meal protein to produce egg proteins. But the amino acids can also be used to produce egg lipid or can be oxidized for energy production. These latter two processes result in the release of nitrogen as toxic ammonia. Therefore, amino acids must be processed in such a way that amino acid nitrogen can be incorporated into non-toxic waste products. Proline is the predominant amino acid in the hemolymph of the adult female mosquito Aedes aegypti. After feeding on albumin meal, hemolymph proline levels increased five-fold over unfed levels, reached maximal levels in the first hours after feeding and remained high through oviposition. Hemolymph proline levels increased as the concentration of protein in the meal increased. When starved of sugar for 24 h prior to feeding on an albumin meal, hemolymph proline levels increased four-fold over the proline levels of non-starved mosquitoes. Proline levels after feeding on a protein deficient in essential amino acids, pike parvalbumin, increased to twice the levels of albumin fed mosquitoes. Based on these observations, we propose that mosquitoes utilize proline as a temporary nitrogen sink to store ammonia arising from deamination of blood meal amino acid.     

Feed formulations to reduce N excretion and ammonia emission from poultry manure

This summary focuses on reducing nitrogen (N) and ammonia emissions from poultry manure through the use of improved amino acid digestibilities and enzyme supplementation. Proper feed processing techniques, phase feeding, and the minimization of feed and water waste can contribute to additional minor reductions in these emissions. Reductions in environmental pollution can be achieved through improved diet formulation based on available nutrients in the ingredients, reducing crude protein (CP) levels and adding synthetic amino acids. Use of amino acid and CP digestibilities can reduce N excretion up to 40% and a 25% increase in N digestibility can be achieved with enzyme supplementation in broiler diets. Digestibilities can be measured by two methods: the excreta and ileal amino acid digestibilities. Both methods allow amino acid levels to be reduced by 10% or more. Enzyme supplementation decreases intestinal viscosity, improves metabolizable energy levels, and increases amino acid digestibilities. Many feed manufacturers still use total amino acid content to formulate feeds. To meet amino acid requirements, crystalline amino acids are needed. The use of feather, meat and bone meal must not be overestimated or underestimated and the limiting amino acids such as cystine, tryptophan, and threonine must be carefully analyzed.     

Lupin-seed meal (Lupinus angustifolius cv. Uniwhite) as a protein supplement to barley-based diets for growing pigs

The apparent digestibility coefficients of the energy and nitrogen of diets containing lupin-seed meal (LSM) were similar to those of diets containing fish meal, despite the comparatively high fibre content of the former.In pigs of 25–85 kg, barley-based diets containing LSM as the sole protein concentrate supported similar rates of gain and efficiency of gain as those containing mixtures of LSM and either fish meal or meat and bone meal, and at least as fast and efficient gains as those containing fish meal or meat and bone meal alone as the source of supplementary protein. However, they depressed dressing percentage. Pigs growing from 12 to 25 kg on barley diets in which up to 45% of the supplementary lysine was supplied in the form of LSM grew as well as those given diets containing various mixtures of dried blood, fish meal and skim-milk powder.     

黄伞菌丝蛋白质营养价值评价

本文测定了黄伞Pholiotaadiposa菌丝粗蛋白含量和氨基酸组成,采用国际上通用的营养价值评价方法,对其蛋白质的营养价值进行全面评价,并与营养价值较高的白灵侧耳Pleurotusnebrodensis、刺芹侧耳Pleurotuseryngii和金顶侧耳Pleurotrscitrinopileatus进行比较。分析结果表明,黄伞菌丝的必需氨基酸含量最高,占其氨基酸总量的43.9%,蛋白质的氨基酸评分(AAS)、化学评分(CS)、必需氨基酸指数(EAAI)、生物价(BV)、营养指数(NI)和氨基酸比值系数分(SRCAA)分别为92.0、71.3、88.5、84.8、40.4和78.8,六项指标均比参比食用菌高。结果说明黄伞菌丝具有很高的营养价值。     

Production of feather hydrolysate by <Emphasis Type="Italic">Elizabethkingia meningoseptica</Emphasis> KB042 (MTCC 8360) in submerged fermentation

A keratinolytic bacterium Elizabethkingia meningoseptica KB042 was isolated from dropped off feathers. The bacterium showed 82.50 ± 0.3% feather degradation when grown on medium containing 10 g/l chicken feathers with initial pH 7.0 at 37°C, 150 rpm in 6 days. The pH of the medium was increased up to 10.02 ± 0.10 during 6 days of incubation. Soluble protein and amino acids concentration in the culture fluid was also found increased until the end of incubation. During the cultivation of strain KB042 on feather as sole source of carbon and nitrogen, the maximum cysteine release was noted on the 3rd day. Varying feather concentration 1.0–2.0% in basal medium resulted in soluble protein release between 1814.42 and 1954.61 μg/ml. The amino acid concentration was found to be maximum, i.e. 937.85 ± 11.9 μg/ml in the cultures grown with 2% feather. The hydrolysate was also found rich in essential amino acids valine, tryptophan, threonine, leucine and cysteine and contains minor amount of methionine and arginine. These data indicate a potential biotechnology for biotransformation and utilization of feather keratin as a source of protein which can be used as animal feed after successful animal trials.     

一株高效羽毛降解菌株的分离与鉴定

采用以羽毛粉为唯一碳源和氮源的培养基,从自然界中分离到一株能够高效降解羽毛角蛋白的细菌,经形态学观察,生理生化实验和16SrRNA基因鉴定,初步确定该菌株为短小芽孢杆菌(Bacillus pumilus),且命名为短小芽孢杆菌WHK4。发酵48 h时,羽毛粉降解率达到85.76%。本研究为微生物降解羽毛角蛋白提供了优良的菌株,在蛋白饲料生产中具有潜在的广泛的应用前景。     

Composition of amino acids in feed ingredients for animal diets

Dietary amino acids (AA) are crucial for animal growth, development, reproduction, lactation, and health. However, there is a scarcity of information regarding complete composition of “nutritionally nonessential AA” (NEAA; those AA which can be synthesized by animals) in diets. To provide a much-needed database, we quantified NEAA (including glutamate, glutamine, aspartate, and asparagine) in feed ingredients for comparison with “nutritionally essential AA” (EAA; those AA whose carbon skeletons cannot be formed by animals). Except for gelatin and feather meal, animal and plant ingredients contained high percentages of glutamate plus glutamine, branched-chain AA, and aspartate plus asparagine, which were 10–32, 15–25, and 8–14% of total protein, respectively. In particular, leucine and glutamine were most abundant in blood meal and casein (13% of total protein), respectively. Notably, gelatin, feather meal, fish meal, meat and bone meal, and poultry byproduct had high percentages of glycine, proline plus hydroxyproline, and arginine, which were 10–35, 9.6–35, and 7.2–7.9% of total protein, respectively. Among plant products, arginine was most abundant in peanut meal and cottonseed meal (14–16% of total protein), whereas corn and sorghum had low percentages of cysteine, lysine, methionine, and tryptophan (0.9–3% of total protein). Overall, feed ingredients of animal origin (except for gelatin) are excellent sources of NEAA and EAA for livestock, avian, and aquatic species, whereas gelatin provides highest amounts of arginine, glycine, and proline plus hydroxyproline. Because casein, corn, soybean, peanut, fish, and gelatin are consumed by children and adults, our findings also have important implications for human nutrition.     

作物节木代料香菇与纯木屑代料香菇蛋白质营养比较研究

采用国际通用的蛋白质营养评价方法,对2种作物节木代料和纯木屑代料(CK)香菇子实体蛋白质的营养价值进行了比较研究。结果表明,2种作物节木代料香菇子实体蛋白质的NI指标低于纯木屑代料香菇,而CS、AAS、EAAI、BV、SRCAA等指标均高于纯木屑代料香菇。结论:作物节木代料香菇子实体和纯木屑代料香菇子实体蛋白质均具有较高营养价值,且二者之间无显著差异。     

Compartmental modeling of postprandial dietary nitrogen distribution in humans

A linear 11-compartment model was developed to describe and simulate the postprandial distribution of dietary nitrogen. The values of its 15 constant diffusion coefficients were estimated from the experimental measurement of (15)N nitrogen kinetics in the intestine, blood, and urine after the oral administration of (15)N-labeled milk protein in humans. Model structure development, parameter estimation, and sensibility analysis were achieved using SAAM II and SIMUSOLV softwares. The model was validated at each stage of its development by testing successively its a priori and a posteriori identifiability. The model predicted that, 8 h after a meal, the dietary nitrogen retained in the body comprised 28% free amino acids and 72% protein, approximately 30% being recovered in the splanchnic bed vs. 70% in the peripheral area. Twelve hours after the meal, these values had decreased to 18 and 23% for the free amino acid fraction and splanchnic nitrogen, respectively. Such a model constitutes a useful, explanatory tool to describe the processes involved in the metabolic utilization of dietary proteins.     

蛋白核小球藻粉中氨基酸含量及饲用价值分析

测定了蛋白核小球藻粉的蛋白质含量及18种氨基酸组分,用氨基酸分(AAS)、必需氨基酸指数(EAAI)等指标对蛋白核小球藻粉氨基酸营养进行评价。结果表明：蛋白核小球藻粉粗蛋白和必需氨基酸含量高,是一种优良的饲料蛋白源。     

Optimal protease production condition for Prevotella ruminicola 23 and characterization of its extracellular crude protease

In this study, Prevotella ruminicola 23 (ATCC 19189), a ruminal proteolytic bacterium, was used as protease producer to examine the optimal condition for protease production. The best carbon and nitrogen sources for the maximum growth were glucose with peptone. Both sucrose and glucose could stimulate high protease production. Casein and peptone are better nitrogen sources for protease production than other choice in this study. The best enzyme production condition was 18-20 h incubation which was at late log phase in the broth of 5% glucose or sucrose as carbon source with 0.1% ammonium chloride and 0.2% peptone as nitrogen sources. Most of the protease activity was secreted into broth (65%) and on cell surface (18%). The optimal temperature and pH for protease reaction were 40 degrees C and pH 6.8, respectively. After incubation for 6h, the crude extract maintained 50% of original protease activity at 30 and 50 degrees C, and protease activity was stable between pH 6 and 8. The protease inhibitor test showed that serine, aspartic acid and metallo-protease inhibitors could cause inhibition of proteolysis. Protein feedstuff degradation experiments suggested that protease in crude extract had higher degradation ability on fish meal, whey, and feather meal (2.39, 2.60 and 1.76 micromol aminoacid/mg enzyme/h) in comparison to soybean meal and blood meal (1.11 and 1.09 micromol aminoacid/mg enzyme/h). The protease in the crude extract should have application potential in term of improving utilization of fish meal and feather meal for monogastric animals.     

Studies on Amino Acid Contents of Processed Soybean

Effect of heating on the nutritive value of defatted soybean flour has been investigated by animal experiments. Loss due to heat degradation was evaluated in two ways. In the first method, the amino acids lost during overheating were supplemented by cystine and mixture of lysine, arginine, tryptophan, and serine at dietary levels of 1.6% nitrogen, and cystine and mixture of those amino acids plus histidine at dietary levels of 3.2% nitrogen. The other procedure adopted was the absorbent test used with amino acid mixtures based on the pattern of amino acids released by pancreatic hydrolysis of unheated, properly heated, and overheated defatted soybean flour at 6 and 120 hr hydrolysis.

At 1.6% dietary nitrogen level, the nutritive value of overheated soybean flour increased by supplementation with cystine and amino acid mixture, but at the 3.2% nitrogen level only cystine was effective. Supplementation of lost amino acids to overheated flour did not restore the nutritive value to that of the properly heated flour. Based on the amino acids released by pancreatic hydrolysis of unheated, properly heated, and overheated soybean flour after 6 and 120 hr reaction, amino acid mixtures were prepared and tested for their nutritive value. While the nutritive value of amino acid mixture prepared based on the pattern of amino acid liberated by 6 hr digestion of unheated, properly heated, and overheated flour did not show similar trend to that of 3 kinds of flour itself, the nutritive value of the amino acid mixture prepared after the data obtained by 120 hr digestion agreed well with the trend of unheated or heated soybean flour.

The nutritive value was also measured by the nitrogen balance of test animals.