[Simultaneous determination of 31 banned veterinary drugs during egg-laying period in poultry eggs by ultra performance liquid chromatography-tandem mass spectrometry].

The consumption of poultry eggs has increased in recent years owing to the abundance of production and improvements in living standards. Thus, the safety requirements of poultry eggs have gradually increased. At present, few reports on analytical methods to determine banned veterinary drugs during egg-laying period in poultry eggs have been published. Therefore, establishing high-throughput and efficient screening methods to monitor banned veterinary drugs during egg-laying period is imperative. In this study, an analytical method based on ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with QuEChERS-based techniques was developed for the simultaneous determination of 31 banned veterinary drugs encompassing nine drug classes (macrolides, antipyretic and analgesic drugs, sulfonamides, antibacterial synergists, anticoccidials, antinematodes, quinolones, tetracyclines, amphenicols) in different types of poultry eggs. The main factors affecting the response, recovery, and sensitivity of the method, such as the extraction solvent, purification adsorbent, LC separation conditions, and MS/MS parameters, were optimized during sample pretreatment and instrumental analysis. The 31 veterinary drug residues in 2.00 g eggs were extracted with 2 mL of 0.1 mol/L ethylene diamine tetraacetic acid disodium solution and 8 mL 3% acetic acid acetonitrile solution, and salted out with 2 g of sodium chloride. After centrifugation, 5 mL of the supernatant was cleaned-up using the QuEChERS method with 100 mg of octadecylsilane-bonded silica gel (C18), 50 mg of N-propylethylenediamine (PSA), and 50 mg of NH2-based sorbents. After nitrogen blowing and redissolution, the 31 target analytes were separated on a Waters CORTECS UPLC C18 analytical chromatographic column (150 mm×2.1 mm, 1.8 µm) at a flow rate, column temperature, and injection volume of 0.4 mL/min, 30 ℃, and 5 µL, respectively. Among these analytes, 26 analytes were acquired in dynamic multiple reaction monitoring (MRM) mode under positive electrospray ionization (ESI+) conditions using (A) 5 mmol/L ammonium acetate (pH 4.5) and (B) acetonitrile as mobile phases. The gradient elution program was as follows: 0-2.0 min, 12%B-30%B; 2.0-7.5 min, 30%B-50%B; 7.5-10.0 min, 50%B; 10.0-10.1 min, 50%B-100%B; 10.1-12.0 min, 100%B; 12.0-12.1 min, 100%B-12%B; The five other target analytes were acquired in MRM mode under negative electrospray ionization (ESI-) conditions using (A) H2O and (B) acetonitrile as mobile phases. The gradient elution program was as follows: 0-2.0 min, 12%B-40%B; 2.0-6.0 min, 40%B-80%B; 6.0-6.1 min, 80%B-100%B; 6.1-8.0 min, 100%B; 8.0-8.1 min, 100%B-12%B. Matrix-matched external standard calibration was used for quantification. The results showed that all the compounds had good linear relationships within their respective ranges, with correlation coefficients of >0.99. The limits of detection (LODs) and quantitation (LOQs) were 0.3-3.0 µg/kg and 1.0-10.0 µg/kg, respectively. The average recoveries of the 31 banned veterinary drugs spiked at three levels (LOQ, maximum residue limit (MRL), and 2MRL) in poultry eggs ranged from 61.2% to 105.7%, and the relative standard deviations (RSDs) ranged from 1.8% to 17.6%. The developed method was used to detect and analyze banned veterinary drugs in 30 commercial poultry egg samples, including 20 eggs, 5 duck eggs, and 5 goose eggs. Enrofloxacin was detected in one egg with a content of 12.3 µg/kg. The proposed method is simple, economical, practical, and capable of the simultaneous determination of multiple classes of banned veterinary drugs in poultry eggs.

Effect of Perilla seeds inclusion on the performance, egg quality characteristics, biochemical parameters and egg yolk fatty acid composition of laying hens.

This study investigates the effect of supplementation of Perilla seeds (PS) on the performance, egg quality, blood biochemical parameters, and egg yolk fatty acids composition in the diet of egg-laying chicken. A total of 1600 Lohmann laying hens were randomly assigned to four different groups with 4 replicates each (100 chickens/replicate) and were subjected to varying PS concentrations (PS0, PS6, PS12, and PS18; 0%, 6%, 12%, and 18%, respectively) for four weeks, including an acclimation period of one week. The results showed no significant differences among the groups for average egg weight (P > 0.005). The laying rate (%), feed conversion ratio (FCR) and average feed intake (AFI) decreased significantly for birds fed on 18% PS as compared to the other treatments (P < 0.005). Haugh unit, albumin height, egg-shape index and eggshell thickness among hens fed PS diets were greater averaging 80.53, 7.00, 1.29, 0.34 compared to 76.84, 6.86, 1.25 and 0.32 from Control hen eggs (P < 0.05). Serum analysis showed a trend towards elevated levels of glucose (Glu), total protein (TP) and aspartate aminotransferase (AST) among treatments. Total cholesterol (TC), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) decreased for the birds fed on 6% PS. The fatty acid composition of egg yolk showed a substantial reduction for α-linolenic acid and docosahexaenoic acid increased significantly by the incorporating PS in the diet (P < 0.001). PS incorporation in diets resulted in significant improvements in both performance indicators and greater amounts of α-linolenic acid and DHA in egg yolks. These findings indicate that PS at 6% inclusion has the potential to improve fatty acid profiles of egg yolk without any adverse effect on performance of egg quality.

Foraging animal origin food samples as passive indicators of dioxin-like POPs contamination in industry sites: Method development, characterisation and risk assessment.

Titanium dioxide (TiO2) is an important industrial chemical, and studies suggest its major production route - the chloride process could lead to the generation of unintentional dl-POPs. However, no relevant studies assessed the occurrence of dl-POPs associated with TiO2 production in the industrial zones, which is mostly due to the ultra-trace level distribution of these compounds in environmental compartments. The present study explored the novel possibility of utilising foraging animal-origin foods as sensitive indicators for addressing this challenge and generated a globally beneficial dataset by assessing the background levels of dl-POPs in the vicinity of a TiO2 production house in Southern India. Systematic sampling of foraging cow's milk and free-ranging hen's eggs was carried out from the study site, and the dl-POPs assessments were conducted utilising an in-house developed cost-effective GC-MS/MS-based analytical methodology. The median dl-POPs levels in milk and egg samples were about 3 times higher than the control samples collected from farm-fed animals and retail markets. The contaminant loads in the foraging animal-origin food samples were further traced to their presence in environmental compartments of soil and sediment and admissible degree of correlations were observed in congener fingerprints. Elevated health risks were inferred for the population in the industrial zones with weekly intakes weighing about 0.15-17 times the European Food Safety Authority-assigned levels. The consumption of foraging cow's milk was observed to have a higher contribution towards the hazard indices and cancer risk estimates and were significantly higher (p < 0.05) for children. The study also presents a critical validation of the GC-MS/MS-based method for the purpose of regulatory monitoring of dl-POPs, which could be of practical significance in economies in transition.

Colorimetry/fluorescence dual-mode detection of Salmonella typhimurium based on a "three-in-one" nanohybrid with high oxidase-like activity for AIEgen.

A colorimetry/fluorescence dual-mode assay based on the aptamer-functionalized magnetic covalent organic framework-supported CuO and Au NPs (MCOF-CuO/Au@apt) was developed for Salmonella typhimurium (S. typhimurium) biosensing. The nanohybrid combined three functions in one: good magnetic separation characteristic, excellent oxidase-mimic activity for tetrap-aminophenylethylene (TPE-4A), and target recognition capability. The attachment of MCOF-CuO/Au@apt onto the surface of S. typhimurium resulted in a significant reduction in the oxidase-mimicking activity of the nanohybrid, which could generate dual-signal of colorimetry and fluorescence through the catalytic oxidation of TPE-4A. Based on this, S. typhimurium could be specifically detected in the linear ranges of 102- 106 CFU·mL-1 and 101- 106 CFU·mL-1, with LODs of 7.6 and 2.1 CFU·mL-1, respectively in colorimetry/fluorescence modes. Moreover, the smartphone and linear discrimination analysis-based system could be used for on-site and portable testing. In addition, this platform showed applicability in detecting S. typhimurium in milk, egg liquid and chicken samples.

Effects of the dose of administration, co-antioxidants, food matrix, and digestion-related factors on the in vitro bioaccessibility of rosmarinic acid - A model study.

This study aimed to determine the effect of the administration dose, combinations with co-antioxidants (vitamin C, caffeic acid, chlorogenic acid, catechin, rutin), and different food matrices (cooked and lyophilized hen eggs, chicken breast, soybean seeds, potatoes) on the potential bioaccessibility of rosmarinic acid (RA) in simulated digestion conditions, depending on the digestion stage (gastric and intestinal) and the contribution of physicochemical and biochemical digestion factors. The in vitro bioaccessibility of RA depended on the digestion stage and conditions. The physicochemical factors were mainly responsible for the bioaccessibility of RA applied alone. The higher RA doses improved its bioaccessibility, especially at the intestinal stage of digestion. Furthermore, the addition of vitamin C and protein-rich food matrices resulted in enhanced intestinal bioaccessibility of RA. In the future, the knowledge of factors influencing the bioaccessibility of RA can help enhance its favorable biological effects and therapeutic potential.

Albumen and Yolk Plasma Peptidomics for the Identification and Quantitation of Bioactive Molecules and the Quality Control of Hen Egg Products.

Hen eggs and the corresponding food products are essential components of human diet. In addition to supplying basic nutrients, they contain functional peptides that are released in vivo within the intact raw material following physiological proteolytic events affecting specific proteins or derive from technological processing of albumen and yolk fractions as a result of the dedicated use of proteases from plant and microbial sources. Besides their potential importance for functional applications, peptides released under physiological conditions in intact egg can be used as markers of product storage and deterioration. Therefore, characterization and quantitation of peptides in egg and egg-derived products can be used to implement evaluation of potential bioactivities as well as to assess food product qualitative characteristics. Here, we provide dedicated information on extraction, identification, and quantitative analysis of peptides from albumen and yolk plasma; nano-liquid chromatography-mass spectrometry combined with bioinformatic analysis of resulting raw data by different software tools allowed to assign molecules based on database searching and to evaluate their relative quantity in different samples.

Effects of replacing Na selenite in laying hen feed with selenized glucose on production performance, egg quality, egg selenium content, microbial population, immunological response, antioxidant enzymes, and fatty acid composition.

This study aimed to explore the effects of selenized glucose (SeGlu) and Na selenite supplementation on various aspects of laying hens such as production performance, egg quality, egg Se concentration, microbial population, antioxidant enzymes activity, immunological response, and yolk fatty acid profile. Using a 2 × 2 factorial design, 168 laying hens at 27-wk of age were randomly divided into 4 treatment groups with 7 replications. Se source (Na selenite and SeGlu) and Se level (0.3 and 0.6 mg/kg) were used as treatments. When 0.3 mg SeGlu/kg was compared to 0.3 mg Na selenite/kg, the interaction findings revealed that 0.3 mg SeGlu/kg increased egg production percent and shell ash (P < 0.05). When compared to 0.3 mg Na selenite/kg, dietary supplementation with 0.3 and 0.6 mg SeGlu/kg resulted in an increase in albumen height, Haugh unit, and yolk color of fresh eggs (P < 0.05). SeGlu enhanced albumen height, Haugh unit, shell thickness (P < 0.01), albumen index, yolk share, specific gravity, shell ash (P < 0.05) of fresh eggs and shell thickness (P < 0.05) of stored eggs as compared to Na selenite. The interaction showed that 0.6 mg SeGlu/kg enhanced yolk Se concentration while decreasing malondialdehyde levels in fresh egg yolk (P < 0.05). SeGlu enhanced Se concentration in albumen and glutathione peroxidase activity in plasma (P < 0.05) as compared to Na selenite. 0.6 mg Se/kg increased lactic acid bacteria, antibody response to sheep red blood cells, and lowered ∑n-6 PUFA/ ∑n-3 PUFA ratio (P < 0.05). As a result, adding SeGlu to the feed of laying hens enhanced egg production, egg quality, egg Se concentration, fresh yolk lipid oxidation, and glutathione peroxidase enzyme activity.

Comparative Metabolomic Profiling of Eggs from 3 Diverse Chicken Breeds Using GC-MS Analysis.

Eggs, as a crucial source of essential nutrients for consumers, possess a high nutritional value owing to their rich composition of vital components essential for human health. While previous research has extensively investigated genetic factors influencing egg quality, there has been a limited focus on exploring the impact of specific strains, particularly within the African context, on the polar metabolite profile of eggs. In this extensive study, we conducted an untargeted analysis of the chemical composition of both albumen and yolk from 3 distinct strains of hens-Blue Holland, Sasso, and Wassache-raised under identical feeding conditions. Utilizing gas chromatography coupled with mass spectrometry (GC-MS), we meticulously examined amino acids, carbohydrates, fatty acids, and other small polar metabolites. In total, 38 and 44 metabolites were identified in the whites and yolk, respectively, of the 3 studied strains. The application of chemometric analysis revealed notable differences in metabolite profiles with 8 relevant metabolites in each egg part. These metabolites include amino acids (N-α-Acetyl-L-lysine, lysine, L-valine, L-Tryptophan), fatty acids (oleic acid, linoleic acid, palmitic acid and stearic acid), and carbohydrates (d-glucose, maltose, lactose). These findings shed light on strain-specific metabolic nuances within eggs, emphasizing potential nutritional implications. The ensuing discussion delves into the diverse metabolic pathways influenced by the identified metabolites, offering insights that contribute to a broader understanding of egg composition and its significance in tailoring nutritional strategies for diverse populations.

Quality characteristics, lysozyme activity, and albumen viscosity of fresh hatching duck eggs after a week's storage at various temperatures.

The study aimed to analyze the qualitative features of Cherry Valley duck' hatching eggs during storage at different temperatures. Eggs were divided into 3 equal groups with 30 eggs each: fresh egg and stored at 7 °C and 17 °C within one week. Qualitative analyses of duck eggs were carried out, considering the morphological composition, physicochemical characteristics, lysozyme activity, and albumen viscosity. The highest weight of yolk and its percentage was found in the 17 °C group. The weight and percentage of albumen were significantly the highest in the group of fresh eggs. Higher egg weight loss was observed in the group stored at higher temperatures. Higher thick albumen height and Haugh units were found in fresh eggs and eggs stored at 7 °C. Different temperatures of egg storage did not affect lysozyme activity in thick and thin albumen. Stored eggs were characterized by lower albumen viscosity only at a shear rate of 10 rpm. The higher viscosity of thick albumen compared to thin ones was demonstrated at 10 and 20 rpm shear rates. The presented research results indicate a large diversity of selected qualitative indicators of hatching duck eggs, which may affect their storage and suitability for incubation.

Testing adulterated liquid-egg: developing rapid detection techniques based on colorimetry, electrochemistry, and interfacial fingerprinting.

To develop rapid detection techniques for liquid eggs' adulteration, three types of adulterations were considered: water dilution, manipulation of yolk ratio in whole egg, and blending different varieties of egg white or yolk. Objective: Establish detection techniques utilizing colorimetry, electrochemistry, and interfacial fingerprinting for these adulterations, respectively. Results: Colorimetry allows for detection (1 min·sample-1) of water dilution through linear (R2 ≥ 0.984) and exponential fitting (R2 ≥ 0.992); Electrochemistry enables detection (6 min·sample-1, R2 ≥ 0.979) of the adulteration of yolk ratio in whole egg; Interfacial fingerprinting technique effectively detects (detection duration: 10 min·sample-1, detection limit: 1.0-10.0 wt%) the adulteration of different varieties of egg white. Subsequently, through 3D-fluorescence microscopy (interface height variation: 22.49-573.45 µm), interfacial tension variation (65.54-35.48 mN·m-1), contact angle variation (89.7°-32.9°), particle size range (free water: 0.94-14.29 µm; protein aggregation: 6.57-10.76 µm), and etc., interfacial fingerprinting mechanism was elucidated. This research contributes novel insights into the detection of adulteration in liquid eggs.

The shellac and shellac nanocomposite coatings on enhanced the storage stability of fresh eggs for sustainable packaging.

Shellac bio-coatings can enhance to improve quality and storage stability of fresh egg qualities with improved shell strength therefore minimizing the reduction the egg losses. Shellac bio-chitosan at 3 concentrations (1 %, 4 % and 8 % w/w) and shellac-1 % montmorillonite nanocomposites were applied as biocoatings to improve storage stability. Shellac-8 % (SH-8 %) coated eggs exhibited the lowest weight loss (1.28 %), significantly. The weight loss of shellac 1 % + MMT and 4 % shellac (SH-4 %) coated eggs was similar each other and had lower weight loss than 1 % shellac (SH-1 %). The Haugh Unit (HU) of eggs with SH-8 % (63.75) had the significantly the highest HU. The SH-4 % (60.24) and SH-1 %/MMT-1 % (58.04) were similar, and the control was the lowest one. The albumin pH of SH-8 % (9.15) coated exhibited a significantly lower than SH-4 % (9.21) and SH-1 %/MMT-1 % (9.24), while the control (9.39) was the highest value at end of storage. For the shellac coated group, total soluble values of albumen reached 12.87 (initial) to 16.331 (SH-1 %), 15.96 (SH-4 %), 15.60 (SH-8 %) and 16.15 (SH-%1-MMT-1 %) at the end of storage. The RWC and foam stability of SH-8 %, SH-4 % and SH-1 % MMT-1 % were similar and higher than 1 % SH and uncoated egg samples. The rheology behaviors were maintained with increasing shellac concentration through the storage. SH-8 % biocoatings were very most effective in filling and sealing the porous in the eggshell and protecting the storage stability and enhancing the strength of the eggshell. Shellac bio-coatings acted as a tiny layer for an effective protective barrier to gas permeability for enhancing the storage stability of the fresh eggs. Higher shellac concentrations (4 and 8 %) and 1 %-MMT were enhanced the storage stability and can be vital solutions for improving shell strength, so it decreases breakage rates.

Influence of the genotype of the hen (Gallus gallus domesticus) on main parameters of egg quality, chemical composition of the eggs under uniform environmental conditions.

The objective of this study was to identify and compare the quality characteristics and concentrations of various compounds in eggs from several pure breeds and lines of hens reared under the same environmental conditions and fed a commercial feed. A total of 280 hens aged 52 to 56 wk belonging to 14 different breeds or lines of hens worldwide were included in this study. Their eggs were characterized by wide differences in various egg quality parameters. Breeds and lines of hens with a higher lutein content in eggs were characterized by a lower beta-carotene content (e.g. Hy line brown, Cochin miniature, Ayam Cemani) (P < 0.001). Additionally, vitamin D, cholesterol, and fatty acid contents were also different between eggs, from 1.51 to 1.79 µg/100g; from 14.1 to 15.4 mg/g fat, PUFA from 19.6 to 22.8 g/100g fat, and SFA from 32.8 to 37.8 g/100g fat respectively (P < 0.001). Lysozyme content also exhibited significant variation among breeds, with some showing a 2-fold higher content in eggs compared to others (0.31% - cochin miniature, 0.66% Faverolle) (P < 0.001). Our study demonstrated that intensively selected hen breeds like Hy-line Brown Hybrid had an improved egg quality seen by the increase in many parameters (e.g., egg weight, Haugh unit, Lutein, vitamins D, MUFA) compared to pure breed hens. In conclusion, genetic differences between breeds and lines of hens have a significant impact on the quality of eggs.

In-house validation of an LC-MS method for the multiplexed quantitative determination of total allergenic food in chocolate.

Mass spectrometry has been widely accepted as a confirmatory tool for the sensitive detection of undeclared presence of allergenic ingredients. Multiple methods have been developed so far, achieving different levels of sensitivity and robustness, still lacking harmonization of the analytical validation and impairing comparability of results. In this investigation, a quantitative method has been validated in-house for the determination of six allergenic ingredients (cow's milk, hen's egg, peanut, soybean, hazelnut, and almond) in a chocolate-based matrix. The latter has been produced in a food pilot plant to provide a real and well-characterized matrix for proper assessment of method performance characteristics according to official guidelines. In particular, recent considerations issued by the European Committee for Standardization have been followed to guide a rigorous single-laboratory validation and to feature the main method performance, such as selectivity, linearity, and sensitivity. Synthetic surrogates of the peptide markers have been used both in native and labelled forms in matrix-matched calibration curves as external calibrants and internal standards, respectively. A two-order of magnitude range was investigated, focusing on the low concentration range for proper assessment of the detection and quantification limits (LOD and LOQ) by rigorous calibration approach. Conversion factors for all six allergenic ingredients have been determined for the first time to report the final quantitative information as fraction of total allergenic food protein (TAFP) per mass of food (µgTAFP/gfood), since such a reporting unit is exploitable in allergenic risk assessment plans. The method achieved good sensitivity with LOD values ranging between 0.08 and 0.2 µgTAFP/gfood, for all ingredients besides egg and soybean, whose quantitative markers reported a slightly higher limit (1.1 and 1.2 µgTAFP/gfood, respectively). Different samples of chocolate bar incurred at four defined concentration levels close to the currently available threshold doses have been analyzed to test the quantitative performance of the analytical method, with a proper estimate of the measurement uncertainty from different sources of variability. The sensitivity achieved resulted in compliance with the various threshold doses issued or recommended worldwide.

Oral bioavailability and egg drug residue of lincomycin in laying hens after different treatment.

Lincomycin (LCM) is an antibiotic used to treat severe bacterial infections in livestock and companion animals. In this study, we aimed to investigate the oral bioavailability of LCM with PK data after IV and PO administration and to compare differences in drug residue patterns in eggs. To ensure food safety, an additional study on egg residue was conducted using 3 different commercial LCM drugs. For bioavailability study, laying hens were divided into oral and intravenous (n = 8/group) groups and received single dose (10 mg/kg) of LCM. The limits of quantification for LCM were 0.729 µg/mL and 0.009 mg/kg in plasma and eggs, respectively. The oral group exhibited a significantly lower average serum drug concentration than the IV group, with a bioavailability of 2.6%. Furthermore, the egg residue profiles confirmed reduced systemic drug exposure after oral administration. For the commercial LCM drug egg residue experiment, laying hens were divided into low- and high-dose groups (n = 12/group) for each drug and treated with the recommended dosage and administration method for each respective drug. The eggs were collected and analyzed until 14 d after the last drug treatment. Despite differences in the LCM content and formulation among commercial drugs, all the tested commercial drugs showed average concentrations below the MRL in eggs within approximately 3 d after the last drug treatment. In this study, we have confirmed that LCM has a low oral absorption rate in laying hens, and this was consistent with the findings from the egg residue profiles. Further studies are requested to elucidate the exact reasons for evidently low oral drug absorption in laying hens.

Per- and poly-fluoroalkyl substances in commercial organic eggs via fishmeal in feed.

Chicken eggs can be a significant source of human PFAS exposure. A survey of PFAS in commercial eggs from larger farms across Denmark showed the absence or low contents of PFAS in free-range and barn eggs. However, organic eggs from eight farms collected in September 2022 had a similar profile of nine PFASs with a predominance of odd over even carbon length PFCAs. Farm 11-13 e.g. had egg yolk ng/g concentrations of PFOA 0.07 ± 0.02; PFNA 0.37 ± 0.04; PFDA 0.13 ± 0.00; PFUnDA 0.22 ± 0.04; PFDoDA 0.06 ± 0.02; PFTrDA 0.15 ± 0.04; PFTeDA 0.02 ± 0.02; PFHxS 0.10 ± 0.04; PFOS 2.62 ± 0.11. Normalised to PFOS, the relative sum of other PFAS showed no difference between the eight organic egg samples, but significant differences between mean individual PFASs (p = 1.4E-25), reflecting a similar profile. The PFAS found in two fishmeal samples with the same origin as the fishmeal used for the organic feed production, could account for the contents in the eggs via estimated transfer from the feed. Furthermore, the estimated transfer from concentration in feed to concentration in egg increased with the carbon length of the PFCA. Exposure (95th percentile) of ∑4PFAS (PFOA, PFNA, PFHxS, PFOS) solely from consumption of 311 g âˆ¼ 5-6 organic eggs/week was for children 4-9 years 10.4 ng/kg bw, i.e. a significant exceedance of the tolerable weekly intake of 4.4 ng/kg bw established by the European Food Safety Authority. Based on the PFAS exposures from organic egg consumption, the organic egg producers decided voluntarily to cease adding fishmeal to the feed. Since the feed-to-egg half-lives are ≤1 week for PFOA, PFOS, and PFHxS, the removal of fishmeal as a feed ingredient should eliminate PFAS after 1-2 months. This was demonstrated in analyses of ten organic egg samples collected by the authorities without PFAS in eight and with 0.1 and 0.4 ng/g ∑4PFAS in two samples.

Development of an enhanced analytical method utilizing pepper matrix as an analyte protectant for sensitive GC‒MS/MS detection of dimethipin in animal-based food products.

Herein, an analytical method using gas chromatography-tandem mass spectrometry (GC‒MS/MS) was devised to detect the presence of the troublesome pesticide dimethipin in various animal-based food products, including chicken, pork, beef, eggs, and milk. The injection port was primed with a matrix derived from pepper leaves that acts as an analyte protectant (AP) to safeguard the target compound from thermal degradation during gas chromatography. The presence of AP resulted in a remarkable limit of quantification of 0.005 mg/kg for dimethipin in five matrices. Three different versions (original, EN, and AOAC) of the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method were compared for dimethipin extraction, with a double-layer solid-phase extraction (SPE) cartridge utilized for matrix purification. A seven-point external calibration curve was established for dimethipin in the five matrices, demonstrating excellent linearity with determination coefficients (R2) ≥ 0.998. The developed quantitative method was validated by fortifying each matrix with three different concentrations of standard dimethipin, and the average recovery fell within the acceptable range outlined in the CODEX guidelines (ranging from 88.8% to 110.0%), with a relative standard deviation (RSD) of ≤ 11.97%. This method effectively addresses the challenge of analyzing dimethipin and can therefore be used as a routine monitoring tool for dimethipin across various matrices.

Transcriptome and histological analyses on the uterus of freckle egg laying hens.

BACKGROUND: In this study, we explored the characteristics and causes of freckle formation. We collected 15 normal and freckled eggs each for eggshell index testing and hypothesized that the structure and function of the uterus would have a direct effect on freckled egg production given that eggshells are formed in the uterus. To test this hypothesis, we collected uterine tissue from laying hens (418 days of age) that laid normal (Group C, n = 13) and freckled (Group T, n = 16) eggs for 7 consecutive days. RESULTS: When we examined the eggshell quality, we found that the L value was significantly lower (P < 0.05) in the freckled site group of freckled eggs compared to the normal egg group during the detection of blunt pole, equator, and sharp pole of the eggshell color. The a-values of the three positions were significantly higher (P < 0.05) in the freckled site group of freckled eggs, and the a-values of the blunt pole were significantly lower (P < 0.05) in the background site group of freckled eggs, compared to the normal egg group. The b-values were significantly higher (P < 0.05) at three locations in the freckled site group of freckled eggs compared to the normal egg group. During the detection of eggshell thickness, the blunt pole was significantly higher (P < 0.05) in the freckled egg site group of freckled eggs compared to the normal egg group, and there was no significant difference between the other groups (P > 0.05). There was no significant difference (P > 0.05) between the transverse and longitudinal diameters of the eggs in each group.We then performed histopathology and transcriptome analyses on the collected tissue. When compared with group C, uterine junctional epithelial cells in group T showed significant defects and cilia loss, and epithelial tissue was poorly intact. From transcriptomics, genes that met (|log2FC|) ≥ 1 and P < 0.05 criteria were screened as differentially expressed genes (DEGs). We identified a total of 136 DEGs, with 101 up- and 35 down-regulated genes from our RNA-seq data. DEGs identified by enrichment analyses, which were potentially associated with freckled egg production were: IFI6, CCL19, AvBD10, AvBD11, S100A12, POMC, and UCN3. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses showed that pathways were associated with immunoreaction and stress stimulation, e.g., complement activation, interleukin-1 cell reactions, viral responses, cell reactions stimulated by corticotropin releasing hormone, steroid hormone mediated signaling pathways, staphylococcal infections, B cell receptor signaling pathways, and natural killer cell mediated cytotoxicity. CONCLUSIONS: From these data, freckled areas deepen freckled eggshell color, but background areas are not affected. At the same time,we reasoned that freckle eggs may result from abnormal immune responses and impaired uterine functions induced by stress. Therefore, the uterus of laying hens in a state of stress and abnormal immune function can cause the appearance of freckled eggs.

Fluorescence Polarization Assay Based on a New Recognition Motif QepA for the One-Step Detection of Fluoroquinolones in Eggs.

A recognition motif is vital in determining the specificity and sensitivity of the fluorescence polarization assay (FPA) for detecting chemical contaminants in food. Four candidates (Gyrase, GyrBA, TopIV, and QepA) were prepared for this study. The applicability of QepA was confirmed through DNA cleavage assay, inhibition effects, and mechanism investigations using molecular docking, compared to other counterparts. Finally, a novel FPA based on QepA and a CIP-FITC tracer for the detection of fluoroquinolones (FQs) in eggs was developed. The limits of detection (LODs) for eight fluoroquinolones ranged from 2.2 to 5.1 ng g-1, with enrofloxacin, danofloxacin, and difloxacin meeting the maximum residue limits (MRLs). The spiked recoveries ranged from 65.8 to 103.6% with coefficients of variation (CVs) of 5.4-12.8%. Therefore, a new recognition motif for FQs that did not belong to conventional antibodies was identified, and QepA-based FPA could be a potential tool for rapid, homogeneous, and sensitive monitoring of the residue of FQs in eggs.

Pharmacokinetic profiles and egg residue patterns of levamisole in laying hens at two dosing rates and two routes of administration.

The levamisole maximum residue limit for edible fat, kidney, and muscle of chickens is 0.01 mg/kg. However, no maximum residue limit has been established for eggs. In the present study, the pharmacokinetic profile and levamisole residue in the eggs from laying hens were investigated using ultra-performance liquid chromatography-tandem mass spectrometry. A single dose of levamisole (30 mg/kg) was administered via the intramuscular or oral route, and an additional egg residue study was performed with 300 or 600 mg/kg commercial LEV drug (30 or 60 mg/kg as levamisole) orally. The limit of quantification was 0.0056 µg/mL and 0.0015 mg/kg for plasma and eggs, respectively. The plasma concentration was below the limit of quantification 10 and 12 h after intramuscular and oral administration, respectively. The half-life of the absorption phase was comparable between the intramuscular and oral routes, which was approximately 1 h, and the mean maximum concentration value was significantly higher in intramuscular (2.29 ± 0.30 µg/mL) than in oral (1.45 ± 0.38 µg/mL) route. The relative oral bioavailability after intramuscular administration was 92.3%. In the egg residue study, dose-dependent area under concentration and maximum concentration were observed after single oral administration of 30 and 60 mg/kg egg residue, and the calculated withdrawal period for both 30 and 60 mg/kg groups based on the positive list system standard (0.01 mg/kg) was 7 d after the treatment.

The physicochemical features of eggshell, thick albumen, amniotic fluid, and yolk during chicken embryogenesis.

The study aimed to analyze the hatching egg and physiochemical features of eggshells, thick albumen, amniotic fluid, and yolk during the incubation of Ross 308 chicken eggs. Eggs (n = 755) were incubated for 21 d. Quality analysis of fresh eggs was performed. Eggshells, albumen, and yolk were collected from fresh eggs and incubation d 1, 7, and 14. Eggshell thickness and strength, pH, vitelline membrane strength, fatty acid (FA) in the yolk, pH, viscosity, lysozyme activity, and crude protein content in thick albumen and amniotic fluid were analyzed. Hatching parameters were calculated. Egg weight loss was constant (8.04% overall). Lower egg surface temperature was found on d 7 compared to d 4, 14, and 18. A lower thickness of posthatch eggshells was found. The strength of the vitelline membrane significantly decreased within 24 h (by over 58%). During incubation, there was a decrease in thick albumen/amniotic fluid pH; an opposite trend was found in yolk pH. The vitelline membrane strength was negatively correlated with the albumen pH. Lysozyme activity was higher in fresh thick albumen and up to 2 wk of incubation. On d 7, the lowest activity was found in the amniotic fluid. On d 14, lysozyme activity increased in amniotic fluid. The higher viscosity of the thick albumen was demonstrated on d 7 and 14 of incubation. The lowest viscosity in amniotic fluid was found on the same days. Crude protein content was higher in thick albumen (d 7 and 14) and lowest in amniotic fluid on d 7. The FA content changed between d 0 and 14. The results indicate different use of FA, where PUFA decreased. Eggshell is used in the last week of incubation. The thick albumen is reduced, while the biological value of amniotic fluid is increasing. Lysozyme activity, viscosity, and crude protein content may be interdependent. It may indicate the flow of substances and the transfer of functions from the thick albumen to the amniotic fluid during chicken embryogenesis.