Identification and characterization of the precursor of chicken matrix metalloprotease 2 (pro-MMP-2) in hen egg

Using zymography and mass spectrometry, we identified for the first time the precursor of chicken matrix metalloprotease 2 (pro-MMP-2) as a complex with TIMP-2 (tissue inhibitor of metalloproteinases) in egg white and yolk. Real-time polymerase chain reaction confirmed that MMP-2 and its inhibitors TIMP-2 and TIMP-3 were expressed all along the oviduct and in the liver of laying hens. We also demonstrated that the processing of pro-MMP-2 into mature MMP-2 by serine proteases does not occur in vivo, although purified pro-MMP-2 undergoes proteolytic maturation by these proteases in vitro. Moreover, the relative pro-MMP-2 activity assessed by gelatin zymography was shown to decrease in egg white during the storage of unfertilized or fertilized eggs. However, the mature form of 62 kDa MMP-2 could not be detected. The fact that MMP-2 is found as a proform in fresh eggs suggests that the activity of this metalloprotease is regulated under specific conditions during embryonic development.     

Conjugated linoleic acids alter the fatty acid composition and physical properties of egg yolk and albumen

Effects of dietary conjugated linoleic acids (CLAs) and docosahexaenoic acid (DHA) on the fatty acid composition of different egg compartments after storage were studied. Four dietary treatments [supplemented with safflower oil (SAFF, control group), DHA, CLAs plus DHA (CAD), and CLAs alone] were administered to Single Comb White Leghorn (SCWL) laying hens. Eggs from the different treatment groups were collected and stored for 10 weeks at 4 degrees C before analysis. Fatty acids from the yolk (yolk granules and plasma), egg albumen, and vitelline membrane were analyzed by gas chromatography. The yolk of eggs from hens given CLAs had significantly higher amounts of saturated fatty acids, typically 16:0 and 18:0, but lower amounts of polyunsaturated fatty acids (PUFAs) compared to eggs from the control group (SAFF). CLA content was highest in the yolk and present in both neutral and polar lipids, with the greatest concentrations in neutral lipids. DHA was incorporated mainly into yolk polar lipids. Lipids in yolk plasma and granules contained similar amounts of CLAs. The fatty acid compositions of vitelline membrane and egg albumen mirrored that of the egg yolk. CLA supplementation resulted in hard and rubbery yolks when compared to hard-cooked eggs from the control group. This study showed that feeding CLAs to hens led to accumulation of the isomers in polar and neutral lipids of the egg yolk and that these isomers migrated into egg albumen. Because the sensory properties of hard-cooked eggs were negatively affected by the enrichment of a mixture of CLA isomers in this study, further research should be conducted to evaluate how the different isomers alter the properties of egg yolk and albumen so that the quality of designed eggs containing CLAs and DHA can be improved.     

Proteomics analysis of egg white proteins from different egg varieties

The market of specialty eggs, such as omega-3-enriched eggs, organic eggs, and free-range eggs, is continuously growing. The nutritional composition of egg yolk can be manipulated by feed diet; however, it is not known if there is any difference in the composition of egg white proteins among different egg varieties. The purpose of the study was to compare the egg white proteins among six different egg varieties using proteomics analysis. Egg white proteins were analyzed using two-dimensional gel electrophoresis (2-DE), and 89 protein spots were subjected to LC-MS/MS. A total of 23 proteins, belonging to Gallus gallus , were identified from 72 detected protein spots. A quiescence-specific protein precursor in egg white was identified for the first time in this study. Significant differences in the abundant levels of 19 proteins (from 65 protein spots) were observed among six egg varieties. Four proteins, ovalbumin-related protein Y, cystatin, avidin, and albumin precursor, were not different among these six egg varieties. These findings suggest that the abundance, but not the composition, of egg white proteins varied among the egg varieties.     

Liquid chromatographic determination of fluoroquinolones in egg albumen and egg yolk of laying hens using fluorometric detection

A liquid chromatographic method was developed for the determination of ciprofloxacin, enrofloxacin, and sarafloxacin at 10-200 ppb in both egg yolk and egg albumen of laying hens. Egg yolk or albumen was acidified with 1 M phosphoric acid followed by deproteination with acetonitrile and centrifugation. The supernate was pipetted out, and the remaining protein pellet was extracted three times with acetonitrile. The supernates were combined and concentrated at 50 degrees C to <0.7 mL. The final volume was adjusted to 2 mL with 0.02 M potassium phosphate buffer, pH 2.5. Separation of the analytes was achieved using reversed-phase HPLC with fluorometric detection. The recoveries were >80% and coefficients of variation <20%. After validation, the method was applied for use in a national survey for fluoroquinolones in table eggs. Of the 276 eggs assayed, none was found positive for fluoroquinolones. The findings suggest that illegal use of fluoroquinolones in laying hens is not widespread.     

Distribution of total 14C residue in egg yolk, albumen, and tissues following oral [14C]sulfamethazine administration to hens

The distribution of total 14C residues was studied in egg yolk and albumen after administration of either single or multiple oral dosages of [14C]sulfamethazine (SMZ). One day after a single dose of [14C]SMZ (121 mg of sulfamethazine, 2.42 x 10(7) dpm), the 14C residue concentration peaked in egg albumen and egg yolk with the concentration in the former >4-fold greater than in the latter. Three days postdose, the 14C residue concentration in the yolk was approximately 7-fold higher than in the egg albumen. A multiple dose of [14C]SMZ containing sulfamethazine mass equivalent of an average therapeutic dose (282 mg, 2.9 x 10(7) dpm) for chickens was also administered orally for six consecutive days to hens. A significantly reduced level of egg production was observed during the medication, and most of the hens stopped laying eggs after the last dose. The 14C residue concentrations peaked on the last day (sixth) of medication in egg albumen and yolk. The 14C residue concentrations were also measured in liver, muscle, blood, and plasma of chickens sacrificed at 1, 24, 48, and 72 h after the last dose. Highest concentrations of 14C residue were accumulated in liver followed by, in decreasing order, blood, plasma, and muscle.     

利用高压静电场保鲜鸡蛋试验

为探讨高压静电场对鸡蛋贮藏保鲜的影响,该试验利用30、60、90 kV/m不同高压静电场分别对鸡蛋进行30和60 min的预处理,然后置于13℃左右的室温条件下贮藏,定期测定鸡蛋的哈夫单位、蛋黄指数、挥发性盐基氮和感官性状的变化等指标,研究高压静电场对其贮藏保鲜效果的影响。试验结果表明：高压静电场处理能很好地保持鸡蛋内部的含水量;有效地降低了哈夫单位、蛋黄指数与挥发性盐基氮的变化速率;贮藏保鲜效果明显好于对照。该试验30 kV/m、60 min剂量和60 kV/m、30 min剂量的处理,保鲜效果最好。     

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.     

基于计算机视觉的鸡蛋新鲜度无损检测

对鸡蛋的新鲜度、贮藏期进行无损检测,在鸡蛋生产、流通、加工领域具有重要意义。该文进行了鸡蛋新鲜度（哈夫值）常规指标在一定温度条件下随贮藏时间变化的试验,采用高分辨率的工业化数字摄像头,以冷光源背向照明方式（照度为10 000 Lx）获取数字图像,并提取了鸡蛋的图像特征蛋黄指数和气室指数。建立了鸡蛋新鲜度与蛋黄指数、贮藏时间与鸡蛋新鲜度、贮藏时间与蛋黄指数和气室指数的关系模型。得知鸡蛋新鲜度与蛋黄指数呈线性相关性,经检验实测值与预测值平均相对误差为6%;贮藏时间与鸡蛋新鲜度以及贮藏时间与鸡蛋蛋黄指数、鸡蛋气室指数均具有二次函数关系,经检验实测值与预测值绝对误差不超过2 d。结果表明基于计算机视觉技术、采用背向照明方式采集的鸡蛋的透射图像得到鸡蛋的蛋黄和气室的图像信息,可以预测鸡蛋的新鲜度和贮藏期。     

基于介电特性与蛋黄指数回归模型的鸡蛋新鲜度无损检测

为了更准确、快速的实现鸡蛋新鲜度品质指标蛋黄指数的无损检测,基于介电特性建立了鸡蛋新鲜度无损检测模型,获取鸡蛋的蛋黄指数信息。试验以不同新鲜度鸡蛋为研究对象,采用平行极板法测量不同新鲜度鸡蛋在温度为20℃,相对湿度为72%~89%,频率为1~200 k Hz下的介电特性参数,分析鸡蛋介电特性的变化规律,并建立鸡蛋介电特性与蛋黄指数之间的数学模型。分析了鸡蛋介电特性随测量信号频率及新鲜度指标蛋黄指数的变化曲线,发现其介电参数随频率及蛋黄指数的增大而减小。构建了蛋黄指数与相对介电常数的拟合方程,鸡蛋样品的实际蛋黄指数与模型预测的蛋黄指数间的决定系数R2为0.9115,蛋黄指数的误差为±4.2%,试验结果表明:利用拟合方程预测检验鸡蛋蛋黄指数取得了较好的预测效果,为无损检测鸡蛋新鲜度提供了一种新的可行方法。     

咸蛋黄快速腌制过程中理化性质变化规律

该研究以新鲜蛋黄为原料,利用快速腌制模具,探究在咸蛋黄的上表面添加食盐单侧腌制过程中,食盐添加量和腌制时间对咸鸡蛋黄快速腌制过程中形貌特征和理化性质变化规律的影响。借助多种仪器分析手段对蛋黄腌制过程中形貌与物性的变化、水分及盐分的迁移规律进行了表征。低场核磁及成像结果表明：在腌制过程中,蛋黄中的水分不断向外迁移,含水率显著降低,当增加食盐的添加量和延长腌制时间,会加快水分的迁移速率;原子吸收结果表明：增加食盐添加盐量越多和腌制时间越长盐分迁移速率越快,质构、色差结果共同表明咸蛋黄腌制过程中,由于水分的向外迁移和盐分的向内渗入,使得蛋黄的蛋白质发生聚集使颜色加深;同时与市售整个腌制后分离的鸡蛋黄产品相比,当腌制时间为7d,添加盐量为3%;腌制时间为3d,添加盐量为5%时,所得的样品与市面的成品咸鸡蛋黄的感官品质及量化指标差异不显著（P<0.05）,为咸蛋黄单侧腌制技术提供理论的可行性。     

Effect of selenium-enriched probiotics on laying performance, egg quality, egg selenium content, and egg glutathione peroxidase activity

A 35-day experiment was conducted to evaluate the effect of selenium-enriched probiotics (SP) on laying performance, egg quality, egg selenium (Se) content, and egg glutathione peroxidase (GPX) activity. Five hundred 58-week-old Rohman laying hens were randomly allotted to 5 dietary treatments of 100 each. Each treatment had 5 replicates, and each replicate had 5 cages with 4 hens per cage. The SP was supplemented to a corn-soybean-meal basal diet at 3 different levels that supplied total Se at 0.2, 0.5, and 1.0 mg/kg. The basal diet served as a blank control, while the basal diet with supplemental probiotics served as a probiotics control. The results showed that dietary SP supplementation not only increased (p < 0.05) the rate of egg laying, day egg weight, mean egg weight, egg Se content, and egg GPX activity but also decreased (p < 0.05) the feed:egg ratio and egg cholesterol content. The egg Se content was gradually increased (p < 0.05) along with the increasing level of dietary Se. The SP supplementation also slowed down (p < 0.05) the drop of Haugh units (HU) of eggs stored at room temperature. The egg GPX activity had a positive correlation (p < 0.01) with egg Se content and a negative correlation (p < 0.01) with egg HU drop. These results suggested that Se contents, GPX activity, and HU of eggs were affected by the dietary Se level, whereas the egg-laying performance and egg cholesterol content were affected by the dietary probiotics. It was concluded that this SP is an effective feed additive that combines the organic Se benefit for hen and human health with the probiotics benefit for laying hen production performance. It was also suggested that the eggs from hens fed this SP can serve as a nutraceutical food with high Se and low cholesterol contents for both healthy people and patients with hyperlipidemia, fatty liver, or cardiovascular disease.     

Imaging residue transfer into egg yolks

Prediction models for residue transfer into eggs are being developed. Recent results indicate that the developing egg yolk serves as an important storage depot for chemical residues. The current study was conducted to visualize incorporation and potential compartmentalization of drug residues in developing egg yolks. To this end, the drug magnevist was injected into hens to evaluate drug transfer into either early- or late-developing yolks. High-resolution magnetic resonance images (MRI) of drug residues in eggs were acquired using a 1.5 T Siemens Magnetom clinical scanner. A 10-cm circular surface coil was used for receiving the magnetic resonance signal. The eggs were positioned inside the coil cavity for an improved signal to noise ratio (SNR). Gradient-echo images were used to locate the centers of the eggs and to prescribe the position of the high-resolution image slab. The images were recorded using an inversion time (T1) weighted magnetization-prepared, rapid acquisition, gradient-recalled-echo (MPRAGE) pulse sequence. The sequence parameters used were as follows: repetition time (TR) equals 12 ms, echo time (TE) equals 5 ms, field of view (FOV) equals 200, TI = 10 ms, 1.25-mm slice thickness, and a matrix of 200 x 256. Following dosing, images of drug residues in eggs indicate that drugs can be incorporated and compartmentalized into ring structures within individual developing egg yolks. These results have significant human food safety implications because even after only a single dose, sequestered drug residues may be stored and later released to contaminate eggs for days to weeks after dosing.     

基于振动及EEMD-CMAC算法的鸭蛋散黄在线检测

针对鸭蛋长期存储以及运输过程中造成的散黄问题,构建一种基于振动信息的鸭蛋散黄在线检测流水线,可实现鸭蛋的自动触压和随动检测。通过磁致伸缩振子对鸭蛋扫频振动进行音频信息增强,对音频振动信号进行集合经验模态分解,并通过主成分分析进行降维提取主要特征,基于此,构建基于小脑神经网络的鸭蛋散黄检测模型。试验中,对320枚鸭蛋进行检测(训练集200枚,测试集120枚),结果表明,基于累积贡献率达98.14%的前5个主成分的鸭蛋散黄检测模型训练集和测试集识别率分别达98.66%和97.03%,每枚鸭蛋在线检测时间约1 s。研究表明,所研制的检测流水线基于磁致伸缩振子扫频激励未知品质鸭蛋,再结合EEMD-CMAC进行鸭蛋散黄检测是可行的,可满足流水线在线检测的要求。     

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.     

Identification of antiadhesive fraction(s) in nonimmunized egg yolk powder: in vitro study

The presence of antiadhesive component(s) in the hen egg yolk against foodborne pathogens was anticipated from results of a previous animal study conducted by the authors. The previous work showed egg yolk powder without specific antibodies is effective in controlling Salmonella enteritidis,Salmonella typhimurium, and Escherichia coli O157:H7 colonization in laying hens. Therefore, this study was necessary to locate the activity and identify the effective component(s). In vitro experiments were conducted using confluent Caco-2 cell monolayers. S. enteritidis, S. typhimurium, and E. coli O157:H7 were investigated against the various extracted granule and plasma fractions in three different assays: adhesion elimination, adhesion prevention, and antimicrobial. This study revealed original findings and identified the protective yolk fraction against the foodborne pathogens as the granule component, high-density lipoproteins (HDL). The protective activity conveyed by HDL was confirmed to remain intact despite peptic and tryptic enzymatic digestion and to have antiadhesive but not antimicrobial effect.     

Enrichment of poultry products with omega3 fatty acids by dietary supplementation with the alga Nannochloropsis and mantur oil

Experiments were conducted to evaluate the efficiency of the microalga Nannochloropsis sp. (Nanno.), as a supplement to laying hens' diet, for the production of enriched eggs and meat with omega3 fatty acids (FA). Nanno. has a unique FA composition, namely, the occurrence of a high concentration of eicosapentaenoic acid (EPA; 20:5 omega3) and the absence of other omega3 FA. The effect of supplementing diets with Nanno. on omega3 FA levels in eggs, plasma, liver, and thigh muscle was compared to that of mantur oil, high in alpha-linolenic acid (LNA; 18:3 omega3). Nanno. is rich also in carotenoids, which may be useful for egg yolk pigmentation. The observed effect of Nanno. supplementation on yolk pigmentation was dose responsive, in both the rate of coloration and the color intensity. Addition of enzyme preparations (glucanase plus cellulase or glucanase plus pectinase) slightly elevated the yolk color score. The most prominent changes in the level of omega3 FA in egg yolk were evident when the diets were supplemented with 1% Nanno. or mantur lipid extracts. Levels of dietary algal meal (0.1-1.0%) had low and inconsistent effects on the level of yolk omega3 FA. Algal EPA is not accumulated in the liver or in the egg yolk; it is apparently converted and deposited as docosahexaenoic acid (DHA). LNA from mantur oil was partially converted to DHA, and both DHA and LNA were deposited in egg yolks and livers. It is suggested that the absence of DHA and EPA from thigh muscle is due to the small amount of dietary omega3 FA used in this work, compared to other studies, and to the possibility that in laying hens the egg yolk has a priority on dietary FA over that of muscles.     

Supplementation of laying-hen feed with palm tocos and algae astaxanthin for egg yolk nutrient enrichment

Adding supplements to hen feed can increase egg nutritional value. Astaxanthin, tocotrienols, and tocopherols are potent antioxidants that provide health benefits to humans. We hypothesized that the addition of these nutrients to hen feed would result in an increased nutrient content in egg yolk with minimum changes in functional properties. Laying hens (Hy-Line W-36 breed) were fed four diets with different supplementation levels of palm toco concentrate and algae biomass containing astaxanthin for 8 weeks. Egg yolks were analyzed for physical, chemical, and functional properties. The feed with the highest nutrient concentration was also studied for stability of these antioxidants using the Arrhenius approach. No significant differences were observed in functional properties except for emulsification capacity and sensory characteristics among eggs from different diet treatments. Changes in egg yolk color reached the maximum values at day 8. Incorporation of tocopherols and tocotrienols increased until day 8, astaxanthin incorporation increased until day 10, and all decreased thereafter. Feed nutrients resulted in a dose-response relationship of these compounds in the egg yolk. The transfer efficiency ranged from 0 to 9.9% for tocotrienols and tocopherols and from 7.6 to 14.9% for astaxanthin at their peak values. Results of the Arrhenius accelerated stability study showed significant differences in the shelf life of various nutrients, and these results can be used to properly formulate and store the feed materials.     

Assessment of egg nutrient compositional changes and residue in eggs,tissues, and excreta following oral administration of atorvastatin to laying hens

Laying hens were fed a control diet alone or with 0.06 g of atorvastatin, a synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, per 100 g of diet for 20 days. Compared to controls, egg yolks from treated hens contained greater amounts of amino acids and reduced levels of total fatty acids and cholesterol. In contrast, egg albumen amino acid contents were unaffected by dietary treatments. In a residue study, seven hens each received a single oral dose of approximately 20 microCi of [(14)C]atorvastatin. Approximately 71% of the radioactivity was recovered in the excreta and liver, whereas virtually no radioactivity was detected in kidney, heart, muscle, bile, plasma, or egg albumen at 15 days postdosing. Yolk radioactivity peaked at 4 days postdosing in six of the seven birds and was absent in eggs laid after day 10. Reminiscent of that of certain antibiotic drugs, the atorvastatin egg residue pattern appeared to coincide with the physiological pattern of daily yolk accretion within the ovary.     

Liquid chromatography-tandem mass spectrometry for the determination of sphingomyelin species from calf brain, ox liver, egg yolk, and krill oil

In this study, molecular species of sphingomyelin (SM) in egg yolk, calf brain, ox liver, and krill oil were investigated. Classes of phospholipids (PLs) were purified, identified, and quantified by normal phase semipreparative high-performance liquid chromatography (HPLC) combined with evaporative light scattering detectors (ELSD). For SM molecular species identification, pure SM collected through a flow splitter was loaded to HPLC-electrospray ionization-tandem mass spectrometry (LC-ESI-MS(2)), with 100% methanol containing 5 mM ammonium formate as mobile phase. In addition to classes of PLs, the used approach allowed the determination of profiles of SM species in egg yolk, ox liver, and calf brain, whereas krill oil turned out not to contain any SM. It also allowed the separation and identification of SM subclasses, as well as tentative identification of species with the same molecular mass, including isomers. The results showed that egg yolk contained the highest proportion of (d18:1-16:0)SM (94.1%). The major SM molecular species in ox liver were (d18:1-16:0)SM (25.5%), (d18:1-23:0)SM (19.7%), (d18:1-24:0)SM (13.2%), and (d18:1-22:0)SM (12.5%). Calf brain SM was rich in species such as (d18:1-18:0)SM (40.7%), (d18:1-24:1)SM (17.1%), and (d18:1-20:0)SM (10.8%).     

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.