Deoxynivalenol and zearalenone residues in eggs of laying hens fed with a naturally contaminated diet: effects on egg production and estimation of transmission rates from feed to eggs

The potential for the Fusarium mycotoxins 4-deoxynivalenol (DON) and zearalenone (ZON) to enter the human food chain through contaminated eggs was assessed using a controlled feed study. Four groups of laying hens (eight in each group) were fed a diet that included differing amounts of naturally contaminated wheat containing DON ( approximately 20 mg kg(-1)) and ZON (0.5 mg kg(-1)). Eggs were collected and pooled from each group on a daily basis. Pooled samples were analyzed by liquid chromatography with mass spectrometry detection (LC-MS/MS). The method allowed DON, other type B trichothecenes, ZON, and its metabolites to be determined in a single multi-residue analysis. The selectivity of the MS/MS procedure allowed cleanup to be minimized (for DON, cleanup by immunoaffinity column was used) or eliminated (for ZON). The limits of detection of 0.01 microg kg(-1) for DON and 0.1 microg kg(-1) for ZON in eggs were lower than previously published methods. None of the samples analyzed had detectable levels of ZON or its metabolites. Although maximum levels of DON contamination (10 mg kg(-1) feed) were relatively high, no adverse effects were observed on egg production. On the basis of the determined DON levels in the hen's diet and the determined levels of DON in the corresponding eggs, transmission rates of 15 000:1, 18 000:1, and 29 000:1 for treatment levels 5, 7.5, and 10 mg DON kg(-1) feed, respectively, were found. These results show that, although eggs could be a human exposure route for DON, the levels are insignificant compared to the other sources, although the presence of metabolites of DON was not studied.     

Determination of total 14C residues of sarafloxacin in eggs of laying hens

[14C]sarafloxacin was orally administered to six laying hens for five consecutive days. Eggs were collected for 15 days after the initial drug treatment. Egg yolk and egg albumen were separated and assayed for total radioactive residues (TRR) using a combustion oxidizer and scintillation counting techniques. Radioactivity was detected in egg yolk and egg albumen on the second day of dosing and reached a maximum at 24 h after drug withdrawal. Thereafter, the sarafloxacin TRR levels in egg albumen declined rapidly and were undetectable 2 days after the last dose, whereas the levels in egg yolk declined at a much slower rate and were undetectable 7 days after drug withdrawal. In both the egg albumen and yolk, HPLC analysis indicated that the parent sarafloxacin was the major component.     

Enantioselective determination of chiral toxaphene congeners in laying hens and eggs using multidimensional high-resolution gas chromatography

A total of 22 chiral toxaphene congeners were analyzed in organ tissues and eggs of laying hens after they had been fed with food spiked with technical toxaphene. For the analysis, multidimensional high-resolution gas chromatography using a chiral column coated with randomly silylated heptakis(O-tert-butyldimethylsilyl)-beta-cyclodextrin, electron capture detection, and valveless "live column switching" technique was applied. The analytical results were additionally confirmed with mass spectral data, recorded in electron-capture negative ionization mode with selected-ion monitoring mass spectrometry. During both the feeding period of the laying hens with toxaphene-contaminated food (38 weeks, accumulation phase) and the following subsiding period without toxaphenes (another 14 weeks, decontamination phase), organs (liver, kidney, skin/fat), blood, meat, and eggs of the hens served as model matrices for toxaphene uptake. The enantiomeric ratios (ERs) of congeners 26, 31, 32, 40, 41, 42(a+b), 44, 50, and 62--known as the most important components of technical toxaphene occurring in the environment--could be analytically determined. Significant differences were observed with respect to their initial racemic ratios. On the basis of their chemical structures, the metabolic pathways of some congeners could be explained. Astonishingly, some of the toxaphenes applied as racemates could merely be found as single enantiomers at the end of the feeding program, for example, congener 32 in blood and meat samples or congener 44, especially in organ tissues, which showed ERs of zero or infinity. The findings of this study impressively emphasize that it is essential to isolate and analyze individual toxaphene enantiomers in food and biota tissues to be capable of evaluating their toxicity and metabolization more specifically.     

Effect of short-term UVB exposure on vitamin D concentration of eggs and vitamin D status of laying hens

Vitamin D deficiency in humans is widespread, and only a few food items are important natural sources of vitamin D. This study investigated the effect of UVB exposure of laying hens on the vitamin D content in egg yolk. In a two-factorial design, hens fed a vitamin D-deficient (-D) or -adequate (+D) diet were nonexposed or exposed to UVB light over a period of 4 weeks. UVB exposure of the -D group caused nearly normal egg production rate and egg shell quality; exposure of the +D group did not further improve these parameters. UVB exposure tended to improve the concentration of plasma 25-hydroxycholecalciferol (25(OH)D(3)), but had no effect on 1,25-dihydroxycholecalciferol in plasma or on cholecalciferol and 25(OH)D(3) in egg yolk. The present study shows that a short-term exposure of laying hens to UVB light is not an appropriate way to improve the vitamin D content of egg yolk.     

Effect of red mold rice supplements on serum and egg yolk cholesterol levels of laying hens

Monacolin K is a secondary metabolite of Monascus species and reduces cholesterol levels. This research focuses on the effect of adding red mold rice to hens' diet on cholesterol level in egg yolk and on cholesterol, triglyceride, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) levels in serum. Forty-eight Hy-line laying hens of 48 weeks of age were studied by dividing them into four groups. Except for the control group, the feed for three other groups contained 2.0, 5.0, and 8.0% red mold rice (monacolin K concentrations were 0.0145, 0.035, and 0.056%, respectively). The experiment lasted 6 weeks. During this period, egg weight and egg production were recorded every day, and cholesterol, triglyceride, HDL, and LDL in serum were measured weekly as well. The result showed that the cholesterol in eggs produced by experimental groups was lower than that of the control group (0%, 194.14 +/- 8.30; 2%, 167.17 +/- 4.34; 5%, 168.93 +/- 9.38; 8%, 183.02 +/- 7.63 mg/egg; p < 0.05), and the triglyceride (0%, 1494 +/- 178; 2%, 1280 +/- 174; 5%, 1189 +/- 248; 8%, 1381 +/- 218 mg/dL; p < 0.05) and LDL levels (0%, 36.81 +/- 5.53; 2%, 32.25 +/- 7.93; 5%, 30.06 +/- 4.39; 8%, 28.81 +/- 4.16 mg/dL; p < 0.05) were also significantly lowered in the experimental groups. However, the HDL level did not show significant change for either control or experimental groups (0%, 36.06 +/- 3.96; 2%, 36.25 +/- 5.39; 5%, 33.13 +/- 3.68; 8%, 31.44 +/- 4.29 mg/dL; p > 0.05). Besides, the addition of red mold rice also helps to inhibit production of malondialdehyde (MDA) in serum lipid oxidation (0%, 27.42 +/- 0.53; 2%, 25.62 +/- 0.76; 5%, 24.35 +/- 0.59; 8%, 23.63 +/- 0.48 microM; p < 0.05).     

Study of enrofloxacin depletion in the eggs of laying hens using diphasic dialysis extraction/purification and determinative HPLC-MS analysis

A study of the depletion of enrofloxacin residues in eggs was carried out using a diphasic dialysis procedure for the extraction of fluoroquinolone residues from the matrix. Enrofloxacin was administered to laying hens through the intramuscular route (15 mg/day) and orally (12 mg/day). After daily collection, the egg albumen and the egg yolk were separated, and the residue levels were determined using an HPLC-MS (API-ESI) method. The enrofloxacin and ciprofloxacin peaks gradually increased until the fifth day, because the drug was employed for 5 days. However, differences were observed in the depletion curves of enrofloxacin and its metabolite when both parts of the egg and the mode of administration were considered.     

Quantitative analysis of tylosin in eggs by high performance liquid chromatography with electrospray ionization tandem mass spectrometry: residue depletion kinetics after administration via feed and drinking water in laying hens

Maximum residue limits (MRLs) have been established by the European Union when tylosin is used therapeutically. They are fixed at 200 microg/kg for eggs. A highly sensitive and selective quantitative liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI/MS/MS) method suitable for monitoring tylosin residues in eggs to determine its depletion kinetics was developed and validated. For sample pretreatment all samples were liquid-liquid extracted with citrate buffer (pH 5.0) and acetonitrile. Liquid chromatographic separation was carried out on a reversed phase C18 column employing a 0.5% formic acid/acetonitrile gradient system. The tylosin recovery in eggs at a concentration range from 1.0-400 microg/kg was >82% with relative standard deviations between 1.5 and 11.0%. In two experimental studies administrating tylosin via feed (final dosage: 1.5 g/kg) or drinking water (final dosage: 0.5 g/L), no residues above the MRL were found during and after treatment. Moreover, all samples were well below the actual MRL of 200 microg/kg. Therefore, our residue data suggest that a withholding period for eggs is not required when laying hens are treated with tylosin in recommended dosages via feed or drinking water. Keywords: Tylosin; residue; depletion; laying hen; withholding period; mass spectrometry.     

Effect of dietary lipid-lowering drugs upon plasma lipids and egg yolk cholesterol levels of laying hens.

To evaluate the effect of lipid-lowering agents upon egg quality, reproductive performance, plasma lipids, and egg yolk cholesterol levels, 30-week-old Shaver laying hens were fed a basal diet (commercial ration) supplemented with 0.1% probucol (PROB), 0.025% gemfibrozil (GEMF), or lovastatin at 0.0005% (LOV1), 0.001% (LOV2), or 0.0015% (LOV3) for a 12-week experimental period. It was observed that the supplementation of the drugs did not impair albumen and shell quality. Hen performance was not adversely affected. The depression in triglyceride concentrations approached statistical significance only in LOV2 (38.5%), and total cholesterol was significantly depressed in LOV2 (36.0%), LOV3 (36.8%), PROB (29.6%), and GEMF (30.4%) treatments. Egg cholesterol content, expressed per gram of yolk, was significant lowered in LOV1 (7.5%) and LOV3 (12. 7%).     

蛋鸡发声音频数据库的构建与应用

蛋鸡发声含有丰富的机体信息,充分挖掘其声学特性,并利用其无接触、无应激的优点,为建立基于发声信息的蛋鸡养殖远程监测平台提供基础依据。该研究借助音频数字化处理技术和数据库管理平台,以海兰褐蛋鸡为例,搭建系统分别采集其在小规模(5只)饲养条件下的叫声信息及其体态行为。运用音频处理软件Adobe Auditionv1.0和音频分析软件Praat5.3提取蛋鸡发声特征参数,包括持续时间、基音频率、频谱质心、共振峰及其衍生的统计值,以此构建出蛋鸡发声音频数据库,在此基础上分别选取蛋鸡产蛋行为发声、鸣唱声和鸣叫声等典型发声行为对比分析。结果表明,蛋鸡产蛋行为发声与鸣唱声均为多次重复的、有节奏的、短促的音节所构成(称其为句子),前者先抑后扬、后者先扬后抑,句子的音节个数分别是7.8±2.0、15.2±7.7,但其时频域特征间存在着显著差异(P<0.05),与鸣叫声相比,其发声特征参数如频谱质心、共振峰等有着显著差异。研究表明,掌握蛋鸡发声的含义,有助于了解其行为特性、机体状态以及种群间的信息传递,并为蛋鸡行为特征识别与数字化监测平台的构建提供数据支持。     

笼养和栖架养殖模式下蛋鸡的发声特征

为了分析不同饲养模式和阶段对蛋鸡发声的影响,并为构建基于蛋鸡声音信息的健康养殖评价系统提供参考,该研究对蛋鸡的发声进行了监测,通过声音预处理、特征提取、数据挖掘和统计分析等方法,研究笼养和栖架饲养模式下、育成期和产蛋期蛋鸡的声学特征.结果表明,典型蛋鸡声音可分为产蛋叫声、鸣唱声、鸣叫声和争斗尖叫声等四类.产蛋期蛋鸡发声...     

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.     

Perchlorate, nitrate, thiocyanate, and iodide levels in chicken feed, water, and eggs from three farms

Perchlorate is an inhibitor of iodide uptake that is found widely in the environment. Given the potential for perchlorate accumulation during egg formation and the widespread consumption of eggs, it is important to examine eggs as a source of exposure to perchlorate and other potential inhibitors of iodide uptake (nitrate and thiocyanate). This study was conducted to determine potential human exposure to perchlorate from eggs produced by chicken flocks consuming differing amounts of perchlorate. The mean concentrations of perchlorate (7.16 ( 1.99 microg/kg of dry weight), nitrate (2820 ( 2100 microg/kg of dry weight), thiocyanate (574 +/- 433 microg/kg of dry weight), and iodide (2980 ( 1490 microg/kg of dry weight) in eggs (n = 180) from 15 chicken houses on 3 U.S. farms were determined. Chickens secreted into eggs an average of 23% of the perchlorate ingested from feed and water. Perchlorate levels in eggs were positively correlated with perchlorate intake (p < 0.001). Increased intake of perchlorate, nitrate, and thiocyanate was associated with decreased iodide levels in eggs, possibly indicating a competitive transport mechanism, such as sodium-iodide symporter. It was estimated that egg consumption contributes minimal perchlorate (approximately 0.040 microg) compared to the average total intake of approximately 10.5 microg for U.S. adults. Additionally, it was found that egg consumption was not associated with increased perchlorate exposure in 2820 individuals from the National Health and Nutrition Examination Survey (p value for the difference of least-squares means, pDiff = 0.225). From these findings it was concluded that, although chickens secrete perchlorate in eggs, eggs do not appear to be a significant source of perchlorate exposure for adults in the United States.     

Metabolism of 2,4-dichlorophenoxyacetic acid in laying hens and lactating goats

2,4-Dichlorophenoxyacetic acid (2,4-D) labeled with (14)C was found to be rapidly eliminated by laying hens and lactating goats dosed orally for 7 consecutive days at 18 mg/kg of food intake and for 3 consecutive days at 483 mg/kg of food intake, respectively. Excreta of hens and goats contained >90% of the total dose within 24 h after the final dose. Tissue residues were low and accounted for <0.1% of the dose in these animals. For hens, the residues in muscle, liver, and eggs (0.006-0.030 ppm) were lower than those found in fat and kidney (0.028-0.714 ppm), 2,4-D equivalents. The tissue with highest residue in goat was the kidney at 1.44 ppm, 2,4-D equivalents. Milk, liver, composite fat, and composite muscle had significantly lower residue levels of 0.202, 0.224, 0.088, and 0.037 ppm, respectively. The most abundant tissue residue was 2,4-D and acid/base releasable residues of 2,4-D. A minor metabolite was identified as 2,4-dichlorophenol.     

Determination of dimetridazole and ipronidazole in feeds at cross-contamination levels

A rapid method for the determination of dimetridazole and ipronidazole in feeds is described. The compounds are extracted from a borate buffer (pH 8.65) with benzene, partitioned into 1N HCl, and then partitioned back into benzene from a basic aqueous phase. The benzene extract is concentrated and injected onto a nonpolar (Apiezon L) gas chromatographic column for determination by 63Ni electron-capture detection. Recoveries from feeds of various composition, spiked at 0.2 ppm with both dimetridazole and ipronidazole, ranged from 70 to 115%; for the same feeds spiked at 1 ppm or more, the recoveries were greater than 80%. Carbadox, furazolidone, levamisole, oxytetracycline, chlortetracycline, sulfamethazine, sulfaquinoxaline, arsanilic acid, piperazine, penicillin, and commonly added vitamins and minerals do not interfere. A 2-dimensional thin layer chromatographic system is presented as a means of additional identification.