Growth of Salmonella enteritidis in yolk from eggs laid by immunized hens

After hyperimmunization of laying hens with Salmonella enteritidis, antibodies can be found in egg yolks. This study was conducted to ascertain whether the growth of S. enteritidis would be suppressed in the presence of antibodies contained in egg yolk. Specifically pathogen-free (SPF)-laying hens were immunized with S. enteritidis; eggs were collected, the yolk was separated and the concentration of S. enteritidis antibodies was determined quantitatively by using the enzyme-linked immunosorbent assay (ELISA), the radial immunodiffusion and the bicinchoninic acid protein assay. Then, the yolk was inoculated with approximately 10, 100 or 1000 S. enteritidis cells/ml and incubated at 15, 20 and 30 degrees C for 0, 2, 6 and 24 h. The growth of organisms in each yolk was examined, and the generation times were calculated. The egg yolk from nonimmunized hens served as negative control. The highest level of antibody concentration was found in the hyperimmunized group. There was no difference in the generation times of S. enteritidis between the antibody-positive yolk and the negative yolk at the three different incubation temperatures. The results suggest that antibodies in the yolk do not influence the growth of S. enteritidis, even if the hens are highly immunized.     

Antimicrobial susceptibilities of Salmonella isolates obtained from layer chicken houses on a commercial egg-producing farm in Japan, 1997 to 2002

Susceptibility to antimicrobial agents was examined for 325 isolates of Salmonella enterica serotypes Cerro, Infantis, Livingstone, and Montevideo isolated from layer houses on a commercial egg-production farm in the western region of Japan between 1997 and 2002. No antimicrobials were used for therapeutic purposes on the farm during this period. From 1.8 to 3.1% of the isolates were resistant to ampicillin, chloramphenicol, and tetracycline. Resistance to streptomycin and sulfisoxazole was found in 52.9 and 65.5%, respectively, of Salmonella Montevideo isolates and in 0 to 13.2% of the isolates of the other serotypes. All the streptomycin-resistant isolates of Salmonella Montevideo also exhibited resistance to sulfisoxazole. Salmonella Montevideo isolates were first isolated in 1998, and 80.0% of the isolates obtained in this year were resistant to streptomycin and sulfisoxazole. The results suggest that Salmonella Montevideo isolates that had already acquired resistance may have been introduced into the layer houses, although the route and vehicle of transmission were uncertain. The proportion of Salmonella Montevideo isolates resistant to streptomycin and sulfisoxazole significantly decreased (P < 0.01) from 79.5% for 1998 through 1999 to 37.3% for 2000 through 2002. This decrease probably was due to the fact that no antimicrobials were used on the farm. Among 10 isolates from different serotypes obtained from 1997 to 2002 that were resistant to ampicillin, two and five isolates harbored 42- and 63-kb R plasmids and identical DraI restriction enzyme digest patterns, respectively, and carried the blaTEM gene. The results suggest that the ampicillin resistance determinants were transferred among different serotypes of Salmonella in the layer houses.     

Aflatoxin B1 residues in eggs of laying hens fed a diet containing different levels of the mycotoxin.

The present study was carried out to evaluate the excretion of aflatoxin B1 residues in eggs of young laying hens fed aflatoxin B1-contaminated rations for 8 weeks. To this end, 96 twenty-week-old hens were randomly distributed into four experimental groups (24 birds per group) and given rations containing either 0 (controls), 100 micrograms, 300 micrograms or 500 micrograms aflatoxin B1/kg feed. Egg aflatoxin B1 residues were determined by thin layer chromatography; two samples per treatment per week were used for analysis. Egg production and average egg weights were not affected (p < 0.05) in the groups receiving aflatoxin B1-contaminated rations. Residues of aflatoxin B1 were detected only in the eggs of hens given 500 micrograms/kg feed, at levels that ranged from 0.05 to 0.16 microgram/kg (average: 0.10 microgram/kg). The results indicate that the feed to eggs aflatoxin B1 transmission ratio was approximately 5000:1, emphasizing the importance of controlling aflatoxin levels in rations for laying hens.     

Deposition and depletion of maduramicin residues in eggs after oral administration to laying hens determined by LC-MS

The coccidiostat maduramicin has been approved as a feed additive for chickens and turkeys, although it is prohibited for use in laying hens. In the present study, laying hens were divided into three groups and fed for 14 days with medicated feed containing maduramicin, at three different concentrations: 50, 100 and 500 µg kg^?1. Eggs were collected during treatment and for 26 days after the end of feeding with medicated feed. Maduramicin residues were found exclusively in egg yolk, with the highest concentration in egg yolk of 459 µg kg^?1 for the highest dose. The maximum concentration of maduramicin in whole egg was 16.6 µg kg^?1 for the group receiving feed containing the maximum permitted level of maduramicin in feed (50 µg kg^?1). The half-life of elimination of maduramicin, calculated for post-treatment days 1–10, was 6.5 days. Twelve days after drug administration, the concentration of the maduramicin in egg yolk for Group 3 (fed with 500 µg kg^?1 maduramicin) still exceeded 20 µg kg^?1, while the concentrations for Groups 1 and 2 were 1.2 and 2.7 µg kg^?1, respectively.     

Effects of L‐carnitine supplementation to diets with different fat sources and energy levels on fatty acid composition of egg yolk of laying hens

BACKGROUND: The present study was carried out to determine the effects of feeding diets with two different levels of metabolisable energy (ME) (11.51 or 10.88 MJ ME kg^?1 diet) and three different sources of fat (palm oil, sunflower oil or fish oil) with or without supplemental L ‐carnitine (0 or 500 mg kg^?1 diet) on the fatty acid (FA) composition of egg yolk and the passage of n3 polyunsaturated FAs to egg yolk in laying hens. RESULTS: The ∑n3, particularly C22:6n‐3, FA contents of egg yolk were significantly reduced by adding of L ‐carnitine (C2) to different fat sources (P < 0.01). The ratio of n6/n3 was reduced from 53.77 to 17.72 in eggs yolks when ME was lowered in the diet with C2‐sunflower oil (SFO) whereas it was enhanced from 2.19 to 9.31 in C2‐E2 (low energy) diet with fish oil (FO) (P < 0.001). The diet with E2 or C2 containing FO resulted in a decrease of the C22:5n‐3, C22:6n‐3 and ∑n3 FA contents of egg yolk (P < 0.001). On the other hand, supplementation of C2 to diets with SFO or palm oil (PO) caused to a decrease in the C22:6n‐3 and ∑n3 FA contents of egg yolk (P < 0.01). A significant increase of the ratio of n6/n3 in egg yolk can be seen by feeding with E2 diet by adding of C2 to all fat sources like in E1 (normal energy) diet (P < 0.001). CONCLUSION: Dietary treatments resulted in major changes in FA composition of egg yolk. The supplemental C2 in diet decreased the C22:5n‐3, C22:6n‐3 and ∑n3FA contents in egg yolk. The use of FO in diets with E2 significantly reduced the passage rate of C22:6n‐3 FA to egg yolk. Copyright © 2008 Society of Chemical Industry     

Effects of dietary microalgae,other lipid sources,inorganic selenium and iodine on yolk n‐3 fatty acid composition,selenium content and quality of eggs in laying hens

BACKGROUND: This aim of this study was to investigate the effects of supplementing laying hen diets with different n‐3 fatty acid sources (linseed oil, fish oil and microalgae) on egg quality and fatty acid profile. The addition of inorganic selenium (Se) and iodine (I) and their effects on egg characteristics and Se concentration were also evaluated. RESULTS: A significantly lower egg weight was found in the control group and in groups fed a diet containing fish oil or a diet containing microalgae plus Se and I. Yolks from all supplemented diet groups exhibited a significant reduction in n‐6 fatty acid content compared with the control group. The docosahexaenoic acid (DHA) content of yolks was higher (P < 0.01) in all supplemented diet groups. Selenium supplementation increased the Se concentration (P < 0.001) in eggs. CONCLUSION: The results of this study provide evidence that an improvement in egg weight and in the DHA content of yolks can be achieved by feeding hens a microalgae‐rich diet, which avoids the unpleasant flavours associated with fish oil supplementation. Dietary inorganic Se, especially in combination with I, can enhance the concentration of Se in eggs. Copyright © 2009 Society of Chemical Industry     

Effects of heating procedures on deoxynivalenol, nivalenol and zearalenone levels in naturally contaminated barley and wheat.

The influence of heating temperature and time on deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEA) contents in naturally co-contaminated barley and wheat was investigated intending to establish the basis for a decontamination model of Fusarium mycotoxins in cereals. The standard toxins and whole barley powder samples were heated in a convection oven at 140, 160, 180, 200, or 220 degrees C, and kernel subsamples (200 g each) were roasted in an experimental rotary gas-fired roaster at 150, 180 or 220 degrees C. All treatments resulted in a time-temperature-dependent decomposition of the toxins; the logarithm of the toxin remaining % presented a linear relationship with heating time. The lines equations were used to estimate the half (H) and decimal (D) decomposition times (time required to destroy 50 or 90% of the toxin, respectively). DON and NIV H and D decomposition times were similar and 50% shorter for heated standards than for whole barley powder. ZEA standard values were considerably longer, while whole barley powder values were comparable with those of DON and NIV. At 220 degrees C, D decomposition times of DON, NIV and ZEA heated standards were 11, 10 and 85 min, respectively, while the values obtained in whole barley powder were the same for the three toxins (25 min). The determination of H and D decomposition values constitutes a basis to understand the heating stability nature of each toxin.     

Distribution and elimination of levamisole in eggs and tissues after oral administration to laying hens,determined by LC-MS/MS

Levamisole was administered to laying hens, and concentrations in eggs and tissues (thigh muscle, breast muscle, liver and kidney) were determined by a newly developed liquid chromatography tandem mass spectrometry method, which allowed trace level quantification of levamisole. The adopted analytical method showed good sensitivity, repeatability and percentage of recovery from spiked matrices. Maximum concentrations of levamisole were found on the first day after the administration (531.1 μg/kg in liver, 164.3 μg/kg in egg yolk, 130.7 μg/kg in kidney, 78.0 μg/kg in breast muscle, 70.7 μg/kg in thigh muscle and 64.0 μg/kg in egg white), after which there is a decline. The compound was rapidly eliminated from eggs, with a half-life of 1.3 days. Elimination appeared to be slower in thigh muscle (3.5 days), breast muscle (3.4 days) and liver (3.3 days). According to this experiment, the levamisole withdrawal periods calculated for eggs, liver, kidney, breast muscle and thigh muscle in laying hens were 14.1, 6.1, >4.0, 14.5 and 13.0 days, respectively. The longest time for levamisole residues to be completely released from tissues was seen in liver samples (37.4 days), followed by thigh muscle, breast muscle and kidney. Elimination from eggs was fastest (16.4 days for levamisole residues to drop below the method quantification limit).     

The effect of a hypocrellin A enriched diet on egg yolk quality and hypocrellin A distributions in the meat of laying hens

Hypocrellins, including hypocrellin A and hypocrellin B, are natural red pigments isolated from a traditional Chinese medicine. The present work was designed to investigate the influence of dietary HA supplementations on the colour and emulsifying properties of egg yolk and HA distributions in tissues of laying hens. A Chinese hen breed (black-bone silky fowl) was used. Ninety-six layers were assigned randomly to diets containing 0, 10, 30 and 50 mg HA per kilogram for 21 days. HA concentrations in egg yolk and tissues of laying hens such as ovary, liver, lung, heart, gizzard and breast muscle were determined by HPLC. Dietary HA supplementations improved the colour of egg yolk (P < 0.05), whereas no significant differences (P > 0.05) in the emulsifying properties of egg yolk were observed. There was a dose-dependent manner increase in egg yolk HA content in response to dietary HA supplementations (P < 0.05). The highest HA concentration was measured in ovary followed by liver and egg yolk.     

The effect of pre-enrichment protocol on the sensitivity and specificity of PCR for detection of naturally contaminated Salmonella in raw poultry compared to conventional culture

Salmonella spp. are the leading cause of foodborne illness worldwide. Conventional culture techniques for the detection of Salmonella spp. are labor intensive and time consuming. Several rapid detection methods have been developed over the past few years. However, standard methods for sample handling and preparation have not been established and limited data are available on the sensitivity and specificity of these methods for detection of Salmonella in naturally contaminated retail meat. Using culture as the gold standard for Salmonella detection in naturally contaminated raw poultry products, the sensitivity and specificity of a polymerase chain reaction (PCR) detection method was determined under varying enrichment protocols. Chicken meat samples (ground, boneless/skinless breast meat, and bone-in breast meat with skin) from retail grocery stores were pre-enriched in buffered peptone water (BPW) and Salmonella specific primers ST 11 and ST 15 were used to amplify a 429 bp region of random fragment target specific to all Salmonella spp. There was a significant decrease (P-value<0.001) in the sensitivity of the PCR test when BPW pre-enrichment alone (85%) was used compared to the sensitivity achieved after both BPW enrichment and selective enrichment with RV and TT-H (100%). PCR failed to detect any positive samples when no pre-enrichment was conducted. A minimum of 12h pre-enrichment was required for detection of Salmonella by PCR at a limit of 100 colony forming unit (cfu)/1 ml of sample. No detectable amplification product was seen in those naturally contaminated meat samples testing negative by culture methods.     

Study on the transmission of deoxynivalenol and de-epoxy-deoxynivalenol into eggs of laying hens using a high-performance liquid chromatography-ultraviolet method with clean-up by immunoaffinity columns

The transmission of deoxynivalenol (DON) and of its metabolite de-epoxy-DON into eggs has not been sufficiently elucidated until now. This question was addressed within the scope of a 16-week experiment with laying hens which were fed a maize-based diet with a DON concentration of 11.9 mg x kg(-1 )dry matter. Eggs were collected during weeks 2, 4, 8, and 16 of the experiment, and DON and its metabolite de-epoxy-DON were analyzed in freeze-dried yolk and albumen. In order to cover possible conjugates, all samples were incubated with beta-glucuronidase prior to extraction. Yolk and albumen were extracted with acetonitrile-water, and the extracts were purified with immunoaffinity columns (IACs) after a precleaning step. The toxins were determined by high-performance liquid chromatography (HPLC) with UV detection. The detection limits of both toxins were 5 and 8 microg x kg(-1) in freeze-dried yolk and albumen, respectively, corresponding to approximately 2.5 and 1 microg x kg(-1) in fresh samples. The recovery of DON and de-epoxy-DON in yolk was 80% and 78%, respectively, and in albumen 77 and 72%. Neither DON nor de-epoxy-DON or glucuronide conjugates of both substances could be detected in any of the samples. These results indicate that eggs do not contribute significantly to the dietary DON intake of humans.     

Comparison of a real-time PCR method with a culture method for the detection of Salmonella enterica serotype enteritidis in naturally contaminated environmental samples from integrated poultry houses

Conventional culture methods have traditionally been considered the "gold standard" for the isolation and identification of foodborne bacterial pathogens. However, culture methods are labor-intensive and time-consuming. A Salmonella enterica serotype Enteritidis-specific real-time PCR assay that recently received interim approval by the National Poultry Improvement Plan for the detection of Salmonella Enteritidis was evaluated against a culture method that had also received interim National Poultry Improvement Plan approval for the analysis of environmental samples from integrated poultry houses. The method was validated with 422 field samples collected by either the boot sock or drag swab method. The samples were cultured by selective enrichment in tetrathionate broth followed by transfer onto a modified semisolid Rappaport-Vassiliadis medium and then plating onto brilliant green with novobiocin and xylose lysine brilliant Tergitol 4 plates. One-milliliter aliquots of the selective enrichment broths from each sample were collected for DNA extraction by the commercial PrepSEQ nucleic acid extraction assay and analysis by the Salmonella Enteritidis-specific real-time PCR assay. The real-time PCR assay detected no significant differences between the boot sock and drag swab samples. In contrast, the culture method detected a significantly higher number of positive samples from boot socks. The diagnostic sensitivity of the real-time PCR assay for the field samples was significantly higher than that of the culture method. The kappa value obtained was 0.46, indicating moderate agreement between the real-time PCR assay and the culture method. In addition, the real-time PCR method had a turnaround time of 2 days compared with 4 to 8 days for the culture method. The higher sensitivity as well as the reduction in time and labor makes this real-time PCR assay an excellent alternative to conventional culture methods for diagnostic purposes, surveillance, and research studies to improve food safety.     

Effects of dietary black cumin seed (Nigella sativa L.) on performance,egg traits,egg cholesterol content and egg yolk fatty acid composition in laying hens

BACKGROUND: The objective of this study was to determine the effects of dietary black cumin seed on performance, egg traits, egg cholesterol content and egg yolk fatty acid composition in laying hens during a 12 week period. For this purpose a total of 160 Lohmann Brown laying hens 36 weeks of age were allocated to four dietary treatments with one control group and three treatment groups. Black cumin seed (Nigella sativa L.) was used at the level of 5, 10 and 15 g kg^?1 in the diets of the first, second and third treatment groups, respectively. RESULTS: Dietary treatments did not significantly affect body weight, feed intake, egg production, egg quality characteristics and blood parameters. Diets containing 10 and 15 g kg^?1 black cumin seed increased egg weight (P < 0.01), improved feed efficiency (P < 0.01) and decreased egg yolk cholesterol, saturated fatty acids (% of total fatty acid methyl esters) and the ratio of saturated to unsaturated fatty acids (P < 0.05) compared to the diet of control group. CONCLUSION: Dietary black cumin seed at the level of 10 and 15 g kg^?1 had beneficial effects on egg weight, feed efficiency, egg cholesterol content and egg yolk fatty acid composition. Therefore it can be used at the level of 10 and 15 g kg^?1 in the diets of laying hens. Copyright © 2009 Society of Chemical Industry     

两种不同检测方法分析不同类型天然和人工染菌食品中菌落总数的比较研究

目的：验证分析测试片检测不同类型食品菌落总数计数的适用性。 方法：与传统国标平板计数方法平行比较,应用测试片检测肉制品、面食、冷饮、奶制品、豆制品、果蔬和坚果7大类70份天然食品,以及以最常见的两种食源性微生物金黄色葡萄球菌和大肠埃希菌作为标准菌株,制备22的份不同食品基质的人工染菌样品。所有检测结果使用两种不同数据方法进行处理分析,以此探讨不同方法检测结果的相关性。 结果：依据ISO 16140-2 2016推荐的数据处理分析方法发现测试片和国标平板计数方法检测不同类型天然和人工染菌食品样品的菌落总数结果对数值差值的平均值均小于0.5;同时、单因素方差分析结果也证实两种方法检测结果无显著性差异（P>0.05）,相关系数高达0.938,不同方法检测结果之间呈正相关。以此发现本研究随机选择食品中菌落总数最高的分别为豆沙春卷、京酱肉丝和鲜猪皮。 结论：本研究证实测试片对不同类型食品菌落总数检测结果与国标平板计数方法结果一致性好,并且具有简单、快捷、易于标准化的优点,适用于不同层级食品承检机构。     

Reduction of aflatoxins by Korean soybean paste and its effect on cytotoxicity and reproductive toxicity--Part 3. Inhibitory effects of Korean soybean paste (doen-jang) on aflatoxin toxicity in laying hens and aflatoxin accumulation in their eggs

This study was conducted to determine the effects of Korean soybean paste (doen-jang [dwen-jahng]) (at concentrations of 0.5, 1, and 5%) on the toxicity of 500 ppb of aflatoxin in the diets of 60 laying hens (Isa Brown) divided into five groups and treated from week 15 to week 67. The aflatoxin-treated hens exhibited many deleterious effects, including reduced body weight; increased relative organ weights; decreased egg production; aflatoxin accumulation in eggs; decreased serum calcium, phosphorus, and alanino amonotransferase (ALT) levels; increased serum gammaglutamil transferase and lactic dehydrogenase levels; and, most significantly, severely altered cell foci and sinusoid dilatation in the liver, relative to control hens. The feeding of 1% soybean paste to hens reduced the adverse effects of aflatoxin on body weight, relative organ weights, egg production, and aflatoxin accumulation in eggs and improved serum calcium and ALT levels and the histopathological lesions of the liver. The feeding of 5% soybean paste to hens resulted in higher levels of the same types of improvements, especially with regard to the histopathological findings for the liver. On the basis of these results, it was suggested that a diet including 5% (and in some cases only 1%) Korean soybean paste protected laying hens and their eggs from the major deleterious effects of 500 microg of aflatoxin per kg of diet and from aflatoxin accumulation. These results indicate that dietary supplementation with Korean soybean paste reduces aflatoxin toxicity in laying hens that ultimately produce human foods such as eggs and poultry.     

Effects of log position in the stem and commercial thinning on jack pine chip dimensions produced by a chipper-canter

Fifteen stems of jack pine (Pinus banksiana Lamb.) coming from three commercial thinned (CT) plots (control, moderate, and intensive thinning) in the Abitibi-Témiscamingue region, Canada, were cross-cut into three 2.4 m length sections: bottom, middle, and top logs. Logs were processed with a chipper-canter at three cutting widths (CW 12.7, 19.1, and 25.4 mm), producing chips and a three faced cant. Chips were assessed by thickness, width, and length. Knot characteristics [total knot number (TKN) and area (TKA)] were assessed in the three cant faces. Growth ring attributes [earlywood density, latewood density (LWD), ring density (RD), earlywood proportion (EWP), ring width, and rings per mm], mechanical properties (shear, splitting, modulus of elasticity and modulus of rupture in bending), and basic density were evaluated on samples obtained within each CW area. The weighted mean chip thickness (WCT) was significantly affected by the log position in the stem (LPS) and CW. WCT increased as CW increased. Jack pine produce thicker chips than black spruce, when processed under the same cutting parameters. Thickness of chips coming from bottom and middle logs was similar due to a greater taper of the bottom log, which slightly increased this dimension. Thickest chips were produced in the top log due to a higher TKA and TKN towards the top of the stem. Multiple linear regressions showed that TKA, CW, and RD were significant predictors of WCT. Chip size distributions were significantly affected by the CT, LPS, and CW. Thinned stands logs appeared to produce smaller chips than natural stand logs. Moreover, chip thickness distribution was affected primarily by TKA and EWP, while the width and length distribution was mainly affected by EWP, LWD, and TKN. Chip size in jack pine is to some degree determined by knot attributes, growth ring characteristics, and wood density of the raw material. These results suggest the potential advantage of sorting logs prior to chipping, either by species, LPS and/or provenance (thinned or natural stands).     

Rapid detection of Salmonella in milk by a nuclear magnetic resonance biosensor based on the streptavidin-biotin system and O-carboxymethyl chitosan target gadolinium probe

Rapid and sensitive detection of foodborne pathogens is of great importance for food safety. Here, a set of nuclear magnetic resonance (NMR) biosensors based on a O-carboxymethyl chitosan target gadolinium (Gd) probe was developed to quickly detect Salmonella in milk by combining NMR technology and bioimmunotechnology with membrane filtration technology. First, O-carboxymethyl chitosan (O-CMC) was biotinylated to prepare biotinylated O-carboxymethyl chitosan (biotin–O-CMC) through amide reaction, and biotinylated magnetic complexes (biotin–O-CMC–Gd) were obtained by using O-CMC, which has strong chelating adsorption on Gd. The target probe was obtained by combining biotin–O-CMC–Gd with the biotinylated antibody (biotin–antibody) via streptavidin (SA) by introducing the SA–biotin system. Then, Salmonella was captured by the target probe through antigen–antibody interaction. Finally, NMR was used to measure the longitudinal relaxation time (T₁) of the filtrate collected by membrane filtration. This NMR biosensor with good specificity and high efficiency can detect Salmonella with the sensitivity of 1.8 × 10³ cfu/mL within 2 h; in addition, it can realize the detection of complex samples because of its strong anti-interference capability and may open up a new method for rapid detection of Salmonella, which has a great application potential.     

Effects of different watering methods,soil weights,soil diluents and soil coverings on the yield and nutrient uptake by ryegrass grown in a controlled environment

A pot experiment was made to test the effects of various experimental techniques on yield and nutrient uptake by ryegrass grown in a controlled environment. Plants grown in 200 g soil yielded more per unit weight of soil than plants grown in the larger weight of soil (600 g). Supplying water to the saucers in which the plants stood gave significantly larger yields than watering to 80% water holding capacity (WHC) either once or twice daily. %N, %K and %Mg were larger in plants grown in the most soil, %P was unaffected and %Ca was less. With either soil weight, watering and amount of soil diluent had more effect on %P and %Ca than on the other nutrients.