Isolation and characterization of immunoglobulin in yolk (IgY) specific against hen egg white lysozyme by immunoaffinity chromatography

Six hens were intramuscularly (im) immunized once a week for 3 weeks using chicken egg white lysozyme (LS) as antigen. Antibody (immunoglobulin in yolk, IgY) ELISA values of 10(3)-fold diluted yolk were almost as high as 1.879 in the sixth week and maintained a value of 0.756 in the eighth week after the initial immunization treatment. The purification efficiency (specific activity of purified IgY against LS/specific activity of antibody in yolk against LS) of IgY specific against LS isolated by laboratory-prepared LS-bound (IgY-) Sepharose 4 Fast Flow immunoaffinity column was approximately 3380. By applying various amounts (0-22 mg) of the thusly obtained IgY specific against LS to the immunoaffinity column, the binding capacity (q(m)) and dissociation constant (K(d), M(-1)) of such immunoaffinity gel for IgY against LS were found to be 0.68 mg of IgY/mL of wet gel (0.54 mg of IgY/mg of LS) and 7.13 x 10(-6) M, respectively, as determined by Langmuir-type adsorption isotherms.     

Productivity and some properties of immunoglobulin specific against Streptococcus mutans serotype c in chicken egg yolk (IgY)

Hens were immunized on thighs by using whole cells of Streptococcus mutans MT8148 serotype c strain as antigen through intramuscular (im) and subcutaneous (sc) routes to investigate the difference of immunization reactions and the changes in yolk antibody activities against antigen after initial immunization. Several properties of crude IgY were examined to evaluate the stability during food processing. Results showed that the specificity of IgY of im treated hens was nearly 10 times as high as those of sc treated antibody. IgY from the hens immunized with the serotype c strain showed significant cross-reactions against serotypes e and f, while minor reactions against serotypes a, b, d, and g were observed. In thermal stability tests, IgY activity in both yolk and crude IgY decreased with the increasing temperature, from 70 to 80 degrees C, but the thermal denaturation rates between those two samples were not significantly different. The addition of high levels sucrose, maltose, glycerol, or 2% glycine displayed effective protection against thermal denaturation of IgY. Lyophilized yolk-5% gum arabic powder showed better stability against proteases.     

Detection of gliadin in foods using a quartz crystal microbalance biosensor that incorporates gold nanoparticles

This work develops a label-free gliadin immunosensor that is based on changes in the frequency of a quartz crystal microbalance (QCM) chip. A higher sensitivity was obtained by applying 25 nm gold nanoparticles (AuNPs) to the surface of a bare QCM electrode. Subsequently, chicken anti-gliadin antibodies (IgY) were immobilized directly on the AuNP-modified surface by cross-linking amine groups in IgY with glutaraldehyde. Experimental results revealed that the change in frequency exhibited when 2 ppm gliadin was bound to the AuNP-modified electrode was 35 Hz (48%) greater than that of the bare gold electrode. The linear dynamic range in 60% ethanol was from 1 × 10(1) to 2 × 10(5) ppb gliadin, and the calculated limit of detection (LOD) was 8 ppb. The entire detection process was completed in 40 min and was highly repeatable. Additionally, the AuNP-modified QCM system generated results in the detection of gliadin in 10 commercial food products that were consistent with those obtained using an AOAC-approved gliadin kit. In conclusion, the QCM platform provides a potential alternative means of ensuring that people with wheat allergies and celiac patients have access to gliadin-free food.     

Chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY)

Chitosan-alginate microcapsules were evaluated as a method of oral delivery of IgY antibodies. Physical characteristics, encapsulation efficiency (EE%), the loading capacity for IgY (IgY loading percentage, %, w/w of microcapsules), gastro-resistance, and release characteristics of these microcapsules in vitro under varying pH were investigated. Optimum physical factors were established for preparation of homogeneous, spherical, and smooth microcapsules. IgY loading% was not significantly altered by pH of the encapsulation medium. Encapsulation efficiency was highest (73.93%) at a pH of 3.5, above which EE% decreased significantly (p < 0.05). IgY was released from microcapsules upon exposure to simulated intestinal fluid (SIF, pH 6.8), and decreasing pH increased significantly IgY release (p < 0.05). The stability of IgY in simulated gastric fluid (SGF, pH 1.2) was greatly improved by encapsulation in chitosan-alginate microcapsules, and the residual activity was not affected by pH of the encapsulation medium. Moreover, microencapsulated IgY was significantly resistant to pepsin hydrolysis. This approach may enable intact IgY to reach target microorganisms within the lower digestive tract.     

Isolation of immunoglobulin from egg yolk by anionic polysaccharides

Isolation conditions of immunoglobulin in egg yolk (IgY) were optimized by the addition of various levels of Na-alginate (Alg), lambda-carrageenan (lambda-Cg), Na-carboxymethyl cellulose (CMC), and pectin (PC) to 6-fold diluted yolk. The mixtures were then reacted at pH 4.0-6.0 for 30 min. The optimal isolation conditions of IgY for Alg, lambda-Cg, and CMC were at the 0.1% level and at pH 5.0, while those for PC were at the 0.15% level and at the same pH. The remaining lipid and remaining protein in the supernatants thus obtained was 0.5-3.8% and 10-17%, respectively, and more than 90% of lipoproteins were precipitated. The IgY recovery was determined to be 33-74% by means of single radial immunodiffusion method when IgY was isolated under the optimal conditions. PC showed the best recovery of IgY, while lambda-Cg provided the least. The interactions between polysaccharides and lipoproteins were mainly ionic bonds, hydrophobic interactions, and hydrogen bonds as determined by the addition (0-2.0 M) of NaSCN or urea to the polysaccharide-lipoprotein precipitates.     

Effects of hen egg yolk immunoglobulin in passive protection of rainbow trout against Yersinia ruckeri

Anti-Yersinia ruckeri egg yolk immunoglobulin (IgY) was transferred to egg yolk after immunization of White Leghorn hens with formalin-killed whole cells of serovar 1 (RS1154) and serovar 2 (RS1153)Y. ruckeri and its lipopolysaccharide (LPS). The IgY was specific for its homologous LPS in western immunoblot, whereas some protein bands were commonly recognized, even by IgY from eggs of unimmunized hens. Purified LPS from both Y. ruckeri serovar types 1 and 2 had a very poor immunogenicity. The IgY activity was stable when processed into pellet form by a microbial transglutaminase treatment and showed a considerable resistance against acid pepsin for at least 2 h. Feeding specific anti-serovar 1 Y. ruckeri IgY to fish either before or after immersion infection produced marginal reductions in mortalities and in intestine infection. The same IgY did passively protect rainbow trout against infection when administered by intraperitoneal injection 4 h before an immersion challenge.     

猪血清IgG干燥方式及干燥保护剂的应用

为寻求符合实际生产需要的猪血清G型免疫球蛋白（IgG）干燥技术,该文探讨了猪血清IgG喷雾干燥的可行性和热保护剂的实际作用,对不同干燥方式所得IgG产品活性和纯度进行了对比研究。结果表明：20%（m/V）的脱脂乳粉、0.5%（m/V）的甘氨酸和5%（m/V）的麦芽糖混合后作为热保护剂能够有效地减少喷雾干燥时IgG的活性损失;海藻糖能够降低IgG冷冻干燥时的活性损失。干燥方式和是否添加保护剂决定了产品中IgG的含量和活性保留率的高低。     

Development of an indirect competitive ELISA for flumequine residues in raw milk using chicken egg yolk antibodies

To detect flumequine in raw milk, an indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed. By carbodiimide conjugation, flumequine was conjugated to cationized bovine serum albumin (cBSA-flumequine) and to cationized ovalbumin (cOVA-flumequine). For the immunization of chickens, cBSA-flumequine was used, which allowed the isolation of specific chicken egg yolk immunoglobulins (IgY) for flumequine. As the coating antigen in the immunoassay, cOVA-flumequine was used. In the indirect competitive assay, standard flumequine was incubated together with the anti-flumequine antibodies. The antibody by which the lowest concentration of free flumequine that gives 50% inhibition of binding (IC50) was found in aqueous dilution was further tested for the applicability to detect flumequine in raw milk. An IC50 level in milk was reached that was about 5 times lower than in aqueous solution. So flumequine can be detected directly in raw milk at maximum residue level (50 microg/kg). No cross-reactivity was noticed with various related quinolones.     

基于海藻酸钠与鸡蛋黄静电聚集作用的低脂蛋黄酱制备

为了拓展海藻酸钠和鸡蛋黄蛋白质在低脂蛋黄酱质构设计方面的应用,该研究首先探究了海藻酸钠和鸡蛋黄分散液在不同酸性pH值条件下的聚集行为,并基于两者的静电聚集作用设计出油相比为30%(体积分数)且具有明显黏弹性和触变性的低脂蛋黄酱产品,同时以油相比为75%的蛋黄酱作对照。结果表明,当pH值低于5.0时,海藻酸钠携带负电荷,鸡蛋黄分散液携带正电荷,两者可发生明显的静电聚集作用,海藻酸钠和鸡蛋黄复合体系的结构强度增加。当白醋添加量高于2%(体积分数)时,海藻酸钠和鸡蛋黄复合体系的pH值降低至5.0以下,可诱导复合体系发生静电聚集作用,白醋添加量越高,聚集作用越明显,低脂蛋黄酱的结构化程度也越高。然而,过量的白醋添加降低了低脂蛋黄酱的热稳定性,同时也影响了产品的风味和感官接受度。综合而言,当白醋添加量为4%时(pH值4.6),制备的低脂蛋黄酱流变学特性和对照组最为接近,且感官接受度较好。该研究结果可为构建低脂食品提供理论参考。     

Ascorbic acid and ascorbic acid 6-palmitate induced oxidation in egg yolk dispersions

The oxidation in aqueous dispersions of egg yolk powder and the influence of addition of the proposed antioxidants ascorbic acid and ascorbic acid 6-palmitate indicate that both ascorbic acid and ascorbic acid 6-palmitate propagated the oxidation of egg yolk powder dispersions. Ascorbic acid 6-palmitate was found to be more prooxidative than ascorbic acid. Moreover, it was found that addition of ascorbic acid or ascorbic acid 6-palmitate gave rise to an increase in the amount of free iron Fe(II) in the egg yolk dispersions. It is proposed that ascorbic acid and ascorbic acid 6-palmitate react with the phosvitin-Fe(III) complex found in egg yolk and release Fe(II), which subsequently propagates lipid oxidation. It appears that less oxidation occurs in egg yolk dispersions exposed to high concentrations of peroxy radicals with added ascorbic acid than egg yolk dispersions with added ascorbic acid without exposure to peroxy radicals.     

Apple peels as a value-added food ingredient

There is some evidence that chronic diseases, such as cancer and cardiovascular disease, may occur as a result of oxidative stress. Apple peels have high concentrations of phenolic compounds and may assist in the prevention of chronic diseases. Millions of pounds of waste apple peels are generated in the production of applesauce and canned apples in New York State each year. We proposed that a valuable food ingredient could be made using the peels of these apples if they could be dried and ground to a powder without large losses of phytochemicals. Rome Beauty apple peels were treated with citric acid dips, ascorbic acid dips, and blanches before being oven-dried at 60 degrees C. Only blanching treatments greatly preserved the phenolic compounds, and peels blanched for 10 s had the highest total phenolic content. Rome Beauty apple peels were then blanched for 10 s and dried under various conditions (oven-dried at 40, 60, or 80 degrees C, air-dried, or freeze-dried). The air-dried and freeze-dried apple peels had the highest total phenolic, flavonoid, and anthocyanin contents. On a fresh weight basis, the total phenolic and flavonoid contents of these samples were similar to those of the fresh apple peels. Freeze-dried peels had a lower water activity than air-dried peels on a fresh weight basis. The optimal processing conditions for the ingredient were blanching for 10s and freeze-drying. The process was scaled up, and the apple peel powder ingredient was characterized. The total phenolic content was 3342 +/- 12 mg gallic acid equivalents/100 g dried peels, the flavonoid content was 2299 +/- 52 mg catechin equivalents/100 g dried peels, and the anthocyanin content was 169.7 +/- 1.6 mg cyanidin 3-glucoside equivalents/100 g dried peels. These phytochemical contents were a significantly higher than those of the fresh apple peels if calculated on a fresh weight basis (p < 0.05). The apple peel powder had a total antioxidant activity of 1251 +/- 56 micromol vitamin C equivalents/g, similar to fresh Rome Beauty peels on a fresh weight basis (p > 0.05). One gram of powder had an antioxidant activity equivalent to 220 mg of vitamin C. The freeze-dried apple peels also had a strong antiproliferative effect on HepG(2) liver cancer cells with a median effective dose (EC(50)) of 1.88 +/- 0.01 mg/mL. This was lower than the EC(50) exhibited by the fresh apple peels (p < 0.05). Apple peel powder may be used in a various food products to add phytochemicals and promote good health.     

Baking Characteristics of Chiffon Cake as Influenced by Microbial Transglutaminase

Protein modification via covalent bonds by using microbial transglutaminase (TGase) has generated many processing functionality improvements in specific food ingredients. In this study, TGase was added into different cake portions (foam and yolk batter) at levels of 0, 0.5, and 1.0% (w/w, total protein weight basis). The treatment of 0.5% TGase in the yolk batter portion significantly (P ≤ 0.05) increased its emulsion activity. The addition of 1.0% TGase in the yolk batter portion significantly increased both foam stability and emulsion activity of cake batter, whereas the addition in the foam portion only increased the emulsion activity of cake batter significantly (P ≤ 0.05). As the addition of TGase, in foam or in the yolk batter portion, rose from 0 to 1.0%, the specific volume of chiffon cake increased. Cakes with 1.0% TGase in the foam portion had the maximum specific volume, 7.078 mL/g, and the softest texture. SDS‐PAGE was used to analyze the modifications of TGase to the protein fractions from different cake portions. The effect of TGase on protein fractions from the yolk batter portion was more evident than that on protein fractions from the foam portion. However, there was no significant difference between the protein fractions of cake batters with the same level of TGase in the foam and yolk portions, which suggested that the main substrates of TGase were yolk protein and wheat protein, instead of egg white protein.     

超高压增大食品物料的导热系数

食品物料在超高压下的导热系数是研究超高压加工过程中传热与温度变化的必要参数,但有关超高压下食品物料的导热系数数据和测量方法还十分缺乏。该文基于线热源法设计了适用于超高压力环境下食品物料导热系数的测量探针和聚甲醛样品容器,利用1.5%琼脂凝胶对热探针在25℃不同压力下(0.1~400 MPa)进行标定试验,结果表明测量值与纯水导热系数的参考值非常接近且呈良好的线性相关关系(R2=0.9997),据此得到探针的标定系数为0.9944。在25℃测量了蛋清、蛋黄、火腿肠和奶油在0.1~400 MPa压力下的导热系数值。结果发现:在25℃条件下,超高压下食品物料的导热系数较常压下均有一定程度的增大(最大达到28%),且有随压力增大而增大的趋势;一定压力条件下,食品物料的导热系数随着含水量的增大而增大。建立了25℃条件食品物料在一定压力范围内(0.1~400 MPa)导热系数预测的经验公式,对研究的几种食品物料拟合得到的方程回归系数在0.91以上。     

Biochemical and functional properties of Atlantic salmon (Salmo salar) muscle proteins hydrolyzed with various alkaline proteases

Protein hydrolysates (5, 10, and 15% degrees of hydrolysis) were made from minced salmon muscle treated with one of four alkaline proteases (Alcalase 2.4L, Flavourzyme 1000L, Corolase PN-L, and Corolase 7089) or endogenous digestive proteases. Reaction conditions were controlled at pH 7.5, 40 degrees C, and 7.5% protein content, and enzymes were added on the basis of standardized activity units (Azocoll units). Proteases were heat inactivated, insoluble and unhydrolyzed material was centrifuged out, and soluble protein fractions were recovered and lyophilized. Substrate specificities for the proteases was clearly different. Protein content for the hydrolysates ranged from 71.7 to 88.4%, and lipid content was very low. Nitrogen recovery ranged from 40.6 to 79.9%. The nitrogen solubility index was comparable to that of egg albumin and ranged from 92.4 to 99.7%. Solubility was high over a wide range of pH. The water-holding capacity of fish protein hydrolysates added at 1.5% in a model food system of frozen minced salmon patties was tested. Drip loss was on average lower for the fish protein hydrolysates than for egg albumin and soy protein concentrate, especially for Alcalase hydrolysates. Emulsification capacity for fish protein hydrolysates ranged quite a bit (75-299 mL of oil emulsified per 200 mg of protein), and some were better than soy protein concentrate (180 mL of oil emulsified per 200 mg of protein), but egg albumin had the highest emulsifying capacity (417 mL of oil emulsified per 200 mg of protein). Emulsification stability for fish protein hydrolysates (50-70%) was similar to or lower than those of egg albumin (73%) or soy protein concentrate (68%). Fat absorption was greater for 5 and 10% degrees of hydrolysis fish protein hydrolysates (3.22-5.90 mL of oil/g of protein) than for 15% hydrolysates, and all had greater fat absorption than egg albumin (2. 36 mL of oil/g of protein) or soy protein concentrate (2.90 mL of oil/g of protein).     

Antioxidant‐Rich Foods Retard Lipid Oxidation in Extruded Corn

Antioxidant‐rich plant materials could provide protection against oxidation in extruded foods and feeds, but their efficacy is not well established. Degermed yellow cornmeal was mixed with 0.02% (w/w) ascorbic acid or quercetin, or with 2% (w/w) spray‐dried ginkgo extract, onion powder, potato peels, or wheat bran. The mixtures were processed in a laboratory‐scale twin‐screw extruder at a feed rate of 227 g/min. Water pump rate was 16 g/min; screw speed was 200 rpm. Mass temperature during extrusion averaged ≈170°C. Samples were cut into small spheres, dried to 5% moisture, then stored in trilaminate bags at 25°C. Ground sample headspace was assayed for hexanal and other volatile indicators of oxidation by gas chromatography. Ginkgo and potato peels significantly darkened the extrudates. Total soluble phenolics, as ferulic acid equivalents, were highest in the ginkgo sample. Volatile compounds were lower in several treatments during storage compared with the control. These findings suggest that manufacturers may be able to formulate products with improved shelf‐life through addition of antioxidant‐rich food materials.     

Plasma homocysteine and glycine are sensitive indices of folate status in a rodent model of folate depletion and repletion

The objectives of the current studies included the characterization of the temporal changes in indices of folate status and amino acid concentrations during both folate depletion and repletion phases. In trial 1, a 6 week folate depletion protocol was employed, using 60 weanling rats assigned to receive an amino acid-defined diet with or without 1 mg/kg folic acid. A 4 week folate depletion period was judged to be optimal on the basis of the development of nadirs in both plasma and hepatic folate stores and elevated (>6-fold relative to folate-adequate controls) concentrations of plasma homocysteine and glycine. In trial 2, 54 weanling rats, previously maintained on a folate-devoid diet for 4 weeks, were assigned to receive 0.25 mg/kg folate as either crystalline folic acid or folate from a folate-enriched egg yolk powder. Both forms of folate supported similar rates of gain, increases in plasma and hepatic folate stores, and reductions in plasma glycine concentrations, whereas the folate in egg yolk powder lowered plasma homocysteine concentrations further than the crystalline folic acid (P < 0.05). These data support the use of both plasma glycine and homocysteine as sensitive response criteria for folate status in a rat bioassay of folate depletion and repletion and establish appropriate temporal end-points for such studies.     

Development of an ELISA for quantifying lysozyme in hen egg white

An indirect enzyme-linked immunosorbent assay (ELISA) by inhibition was developed for quantifying lysozyme in hen egg white (HEW), a protein of value in not only the food and pharmaceutical industries but also for poultry research. Various experimental conditions (coating, antibodies dilutions, samples dilutions, preparations, blocking agents, and incubation times) were assayed to optimize this assay to the quantification of HEW in egg white samples. HEW samples were diluted 1:3000 to avoid matrix effects, possibly resulting from lysozyme interaction with other egg white proteins. Assay linearity for lysozyme ranged from 0.38 to 4.8 mug/mL, with intra- and interassay variations of 6.8% and 7.6%, respectively, and the lower detection limit was 0.264 mug/mL. We found that lysozyme concentrations in albumen from eggs laid by a hen cohort ranged from 2.2 to 4.5 mg/mL, thus underlining interhen variability. Overall, these data present an ELISA assay that is simple, quick, sensitive, accurate, and has been specifically designed to determine lysozyme concentrations in egg white samples.     

Differential incorporation of conjugated linoleic acid isomers into egg yolk lipids

The incorporation pattern of conjugated linoleic acids (CLA) isomers into the egg yolk of hens in relation to that in the diet was studied. Silver-ion high-performance liquid chromatography (Ag-HPLC) was used to separate individual CLA isomers. It was found that the isomeric distribution pattern in the egg yolk lipids was different from that in the dietary fat. Total cis/trans isomers accounted for 81.2% of total CLA incorporated into the egg yolk, which was in contrast to the value of 92.0% of total CLA in the diet. Total cis/cis isomers accounted for 3.8% total CLA in the diet but they were 6.6% of the total CLA in the egg yolk lipids. In contrast, total trans/trans isomers were 12.2% of the total CLA isomers in the egg yolk lipids, whereas they were only 4.2% of total CLA in the diet. The results showed that total trans/trans-CLA was preferentially incorporated into the egg yolk, whereas the incorporation of total cis/trans-CLA isomers was partially discriminated. Within each group, the incorporation of individual isomers into the egg yolk lipids was also selective. cis-9,trans-11/trans-9,cis-11 and cis-10,trans-12/trans-10,cis-12 were the two major isomers in the diet. Ag-HPLC analysis showed that the former was preferentially transferred into the egg yolk compared with the latter. It was observed that supplementation of CLA in the diet of laying hens decreased the concentration of oleic acid (18:1n-9), arachidonic acid (20:4n-6), and docosahexaenoic acid (22:6n-3) but increased that of linolenic acid (18:3n-3), stearic acid (18:0), and palmitic acid (16:0) in the egg yolk, suggesting that CLA may inhibit Delta6 and Delta9 desaturases.     

Enzymatic determination of cholesterol in egg yolk

The quantitative determination of cholesterol in egg yolk by using an enzymatic test kit is described. Cholesterol in the egg yolk is extracted with other lipid components by methylene chloride-methanol (2 + 1) and is enzymatically determined after saponification of the lipid extract. The method is relatively rapid, simple, and accurate and gives results which agree with those obtained by using a gas-liquid chromatographic (GLC) method. The mean cholesterol content of egg yolk determined by the enzymatic and GLC methods was 1237 and 1240 mg/100 g, respectively.     

Food matrix effects on in vitro digestion of microencapsulated tuna oil powder

Tuna oil, containing 53 mg of eicosapentaenoic acid (EPA) and 241 mg of docosahexaenoic acid (DHA) per gram of oil, delivered as a neat microencapsulated tuna oil powder (25% oil loading) or in food matrices (orange juice, yogurt, or cereal bar) fortified with microencapsulated tuna oil powder was digested in simulated gastric fluid or sequentially in simulated gastric fluid and simulated intestinal fluid. The level of fortification was equivalent to 1 g of tuna oil per recommended serving size (i.e., per 200 g of orange juice or yogurt or 60 g of cereal bar). The changes in particle size of oil droplets during digestion were influenced by the method of delivery of the microencapsulated tuna oil powder. Lipolysis in simulated gastric fluid was low, with only 4.4-6.1% EPA and ≤1.5% DHA released after digestion (as a % of total fatty acids present). After sequential exposure to simulated gastric and intestinal fluids, much higher extents of lipolysis of both glycerol-bound EPA and DHA were obtained (73.2-78.6% for the neat powder, fortified orange juice, and yogurt; 60.3-64.0% for the fortified cereal bar). This research demonstrates that the choice of food matrix may influence the lipolysis of microencapsulated tuna oil.