ZP2 cleavage blocks polyspermy by modulating the architecture of the egg coat.

Following the fertilization of an egg by a single sperm, the egg coat or zona pellucida (ZP) hardens and polyspermy is irreversibly blocked. These events are associated with the cleavage of the N-terminal region (NTR) of glycoprotein ZP2, a major subunit of ZP filaments. ZP2 processing is thought to inactivate sperm binding to the ZP, but its molecular consequences and connection with ZP hardening are unknown. Biochemical and structural studies show that cleavage of ZP2 triggers its oligomerization. Moreover, the structure of a native vertebrate egg coat filament, combined with AlphaFold predictions of human ZP polymers, reveals that two protofilaments consisting of type I (ZP3) and type II (ZP1/ZP2/ZP4) components interlock into a left-handed double helix from which the NTRs of type II subunits protrude. Together, these data suggest that oligomerization of cleaved ZP2 NTRs extensively cross-links ZP filaments, rigidifying the egg coat and making it physically impenetrable to sperm.

Effects of different selenium sources and levels on the physiological state, selenoprotein expression, and production and preservation of selenium-enriched eggs in laying hens.

Selenium (i.e., Se) is a trace element that is vital in poultry nutrition, and optimal forms and levels of Se are critical for poultry productivity and health. This study aimed to compare the effects of sodium selenite (SS), yeast selenium (SY), and methionine selenium (SM) at selenium levels of 0.15 mg/kg and 0.30 mg/kg on production performance, egg quality, egg selenium content, antioxidant capacity, immunity and selenoprotein expression in laying hens. The trial was conducted in a 3 × 2 factorial arrangement, and a total of 576 forty-three-wk-old Hyland Brown laying hens were randomly assigned into 6 treatment groups, with diets supplemented with 0.15 mg Se/kg and 0.3 mg Se/kg of SS, SY and SM for 8 wk, respectively. Results revealed that SM increased the laying rate compared to SS and SY (P < 0.05), whereas different selenium levels had no effect. Organic selenium improved egg quality, preservation performance, and selenium deposition compared to SS (P < 0.05), while SY and SM had different preferences for Se deposition in the yolk and albumen. Also, organic selenium enhanced the antioxidant capacity and immune functions of laying hens at 0.15 mg Se/kg, whereas no obvious improvement was observed at 0.30 mg Se/kg. Moreover, SY and SM increased the mRNA expression of most selenoproteins compared to SS (P < 0.05), with SM exhibiting a more pronounced effect. Correlation analysis revealed a strong positive association between glutathione peroxidase 2 (GPx2), thioredoxin reductases (TrxRs), selenoprotein K (SelK), selenoprotein S (SelS), and antioxidant and immune properties. In conclusion, the use of low-dose organic selenium is recommended as a more effective alternative to inorganic selenium, and a dosage of 0.15 mg Se/kg from SM is recommended based on the trail conditions.

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.

Egg production, egg quality, and fatty acids profiles in eggs and tissues in Lohmann LSL lite hens fed algal oils rich in docosahexaenoic acid (DHA).

Enriching eggs with omega-3 fatty acids (n-3 FA), such as docosahexaenoic acid (DHA), is a well-accepted practice that benefits the egg industry and consumers. However, issues around cost, sustainability, and product acceptance have necessitated the search for alternatives to feeding hens fish oil for DHA enrichment. The effects of feeding 2 algal oils on egg production and DHA enrichment in eggs and selected tissues were investigated. The algal oils were: 1) OmegaPro (OPAO) standardized algal oil for DHA content and 2) Crude algal oil (CAO). A total of 400, 46-wk-old Lohmann LSL lite hens were housed in enriched cages (10 birds/cage) and allocated 5 diets (n = 8) for a 12-wk trial. The iso-caloric and -nitrogenous diets were a standard corn and soybean meal diet, standard plus 0.25 or 0.76% OPAO and standard plus 0.23 or 0.69% CAO; algal oils diets supplied similar DHA at each level. Egg production indices (hen day egg production, feed intake, FCR, egg weight, egg mass, and eggshell quality) were monitored for 10 wk. Diet samples were analyzed for fatty acids (FA) on wk 1, 6, and 12 and eggs on wk 4, 5, 6, 9, and 12. At the end of the trial, one hen/cage was weighed and dissected for liver, breast and thigh for FA and long bones for ash content analyses. Concentration of omega-6 to omega-3 FA ratio was 12.9, 6.64, 3.48, 6.96, and 3.59 for standard, 0.23 and 0.76% OPAO, 0.25 and 0.69% CAO, respectively. Algal oils increased (P ≤ 0.046) eggshell thickness linearly. The concentration of DHA in the eggs from the birds fed the standard, 0.23 and 0.76% OPAO, 0.25 and 0.69% CAO was 84, 195, 286, 183, and 297 mg/100g egg, respectively, and algal oils enriched eggs with DHA linearly and quadratically (P ≤ 0.01). In conclusion, algal oils increased the concentration of DHA in eggs and had no adverse effects on egg production and eggshell quality.

Effects of supplemented multicomponent mycotoxin detoxifying agent in laying hens fed aflatoxin B1 and T2-toxin contaminated feeds.

The present study was conducted to determine the ability of multicomponent mycotoxin detoxifying agent (MMDA) in feed to prevent the gastrointestinal absorption of aflatoxin B1 (AFB1) and T2-toxin supplemented via spiked maize. For comparisons, hens were fed with uncontaminated basal diet without or with addition of MMDA at 2 g/kg feed. The trial consisted of 105 laying hens (Lohmann Brown) without obvious signs of disease allocated to 7 treatment groups in 35 pens. Responses were demonstrated on laying performance and health status throughout the 42 d experimental period. The results of laying performance indicated significantly decreased egg mass with increasing mycotoxin (AFB1 and T2-toxin) levels up to the maximum tolerated dosage, however simultaneous presence of MMDA laying performance was slightly modified linearly to increasing application. Dose-dependent pathological changes in liver and kidneys and their relative weights, changes in blood parameters and reduced eggshell weights were observed in the hens fed AFB1 and T2-toxin. The pathological changes in the hens fed with diets containing AFB1 and T2-toxin without MMDA were significantly higher as compared with the control group, but eggshell stability was not affected. The contents of AFB1, T2-toxin and their metabolites in liver and kidney tissues were significantly decreased in the hens supplemented with MMDA at 2 and 3 g/kg in feed. MMDA supplementation significantly reduced the deposition of AFB1, T2-toxin and their metabolites in liver and kidneys at the maximum tolerated dosage (2 and 3 g/kg) indicating specific binding to AFB1 and T2-toxin in the digestive tract as compared to the corresponding diets without MMDA. Exposure of AFB1 and T2-toxin indicated significantly decreased egg mass with increasing mycotoxin levels up to the maximum tolerated dosage because of the significantly reduced egg production. Therefore, in this study, MMDA could reduce negative effects of feeding AFB1 and T-2 to laying hens.

Egg residue and depletion of meloxicam in Jing Hong laying hens following multiple oral doses.

Meloxicam is a nonsteroidal anti-inflammatory drug (NSAID) commonly used in an extra-label manner in commercial laying hens for the treatment of foot lesions, which are a common issue in this species. The present study aimed to determine the depletion profiles of meloxicam in eggs with multiple oral administration under 2 different dosing regimens and to further recommend reasonable withdrawal intervals (WDIs). Meloxicam (1 mg/kg) was administered orally to laying hens under 2 dosing schedules: 10 doses at 24-h intervals and 15 doses at 12-h intervals. Eggs were collected daily after the first dosing, and meloxicam concentrations in both yolk and white were determined by a high-performance liquid chromatography (HPLC) method. The weight ratio of white to yolk in the whole egg was 1.54 (the mean of 20 eggs with repeated tests), and this value combined with the meloxicam concentrations in white and yolk were used to calculate the drug concentrations in whole eggs. Meloxicam was quickly eliminated from egg white, and its concentrations could only be quantified at 2 time points during the elimination phase. The elimination half-lives in yolk and whole egg were 3.07 ± 1.00 and 2.98 ± 0.88 d, respectively, after 10 repeated doses. And the corresponding elimination half-lives were 2.30 ± 0.83 and 2.18 ± 0.67 d, respectively, after repeated 15 doses. Considering the time when meloxicam was not detectable in eggs with the time of ovum development and maturation, a withdrawal interval (WDI) was suggested as 17 d for both dosing schedules. The current results enriched the study on the residue of meloxicam in domestic Jing Hong laying hens and provided WDIs to help ensure animal-derived food safety.

The eggshell defect as a factor affecting the egg quality after storage.

The aim of the study was to assess the influence of shell defects on the quality of eggs after storage. The study material consisted of 1,800 brown-shelled eggs from cage rearing system which were candled on the day of laying to determine the shell quality. Eggs with the 6 most common shell defects (external crack, severe stripe marks, points, wrinkled, pimples, sandy) and eggs without defects (control group) were then stored for 35 days at 14°C and 70% humidity. The weight loss of eggs was monitored every 7 days, and the quality characteristics of whole eggs (weight, specific gravity, shape), shell (defects, strength, color, weight, thickness, density), albumen (weight, height, pH) and yolk (weight, color, pH) of 30 eggs from each group were analysed at the beginning (0 days) and after 28 and 35 days of storage. The changes resulting from water loss (air cell depth, weight loss, shell permeability) were also evaluated. The study showed that all investigated shell defects significantly influenced the characteristics of the whole egg during the storage, modifying traits such as specific gravity, water loss, shell permeability, albumen height and pH, as well as proportion, index and pH of the yolk. Furthermore, an interaction between time and the shell defects presence was found.

Feed gap analysis of dual-purpose chicken production in Tanzania: feed quantity and quality limited production.

The demand for chicken meat and eggs exceeds what can be produced in Tanzania, largely due to low productivity of the sector. Feed quantity and quality are the major factors determining the potential production and productivity of chickens. The present study explored the yield gap in chicken production in Tanzania and analyses the potential of increased chicken production as a result of closing the feed gaps. The study focused on feed aspects limiting dual-purpose chicken production in semi-intensive and intensive systems. A total of 101 farmers were interviewed using a semistructured questionnaire and the amount of feed provided to chickens per day was quantified. Feed was sampled for laboratory analysis and physical assessments were made of weights of chicken bodies and eggs. The results were compared with the recommendations for improved dual-purpose crossbred chickens, exotic layers, and broilers. The results show that the feeds were offered in insufficient quantity compared with the recommendations for laying hens (125 g/chicken unit/d). Indigenous chickens were fed 111 and 67 while the improved crossbred chickens were fed 118 and 119 g/chicken unit/d under semi-intensive and intensive systems, respectively. Most feeds fed to dual-purpose chickens were of low nutritional quality, particularly lacking in crude protein and essential amino acids in both rearing systems and breeds. Maize bran, sunflower seedcake, and fishmeal were the main sources of energy and protein in the study area. The study findings show that the important feed ingredients: protein sources, essential amino acids, and premixes were expensive, and were not included in formulating compound feeds by most chicken farmers. Of all 101 respondents interviewed, only one was aware of aflatoxin contamination and its effects on animal and human health. All feed samples contained a detectable concentration of aflatoxins and 16% of them exceeded the allowed toxicity thresholds (>20 µg/kg). We highlight the need for a stronger focus on feeding strategies and ensuring the availability of suitable and safe feed formulations.

Socially transferred materials: why and how to study them.

When biological material is transferred from one individual's body to another, as in ejaculate, eggs, and milk, secondary donor-produced molecules are often transferred along with the main cargo, and influence the physiology and fitness of the receiver. Both social and solitary animals exhibit such social transfers at certain life stages. The secondary, bioactive, and transfer-supporting components in socially transferred materials have evolved convergently to the point where they are used in applications across taxa and type of transfer. The composition of these materials is typically highly dynamic and context dependent, and their components drive the physiological and behavioral evolution of many taxa. Our establishment of the concept of socially transferred materials unifies this multidisciplinary topic and will benefit both theory and applications.

The effect of adding L-carnitine to omega-3 fatty acid diets on productive performance, oxidative stability, cholesterol content, and yolk fatty acid profiles in laying hens.

In this study, different levels of Omega-3 fatty acids and L-carnitine (LC) were used in diets for laying hens. The effects of these supplements were examined on productive performance, antioxidant properties, cholesterol content, and yolk fatty acid profiles in the laying hens. A population of 120 Lohmann LSL-Lite laying hens (34-wk-old) were used in 2 × 3 factorial arrangements with 2 diets (control = 0.031 and 0.48% omega-3 fatty acids) and 3 levels of L-carnitine (0, 100, 200 mg/kg of diet) in a completely randomized design with 6 treatments. While having 5 replicates and 4 birds per replicate, the total period of the experiment lasted for 10 wk. The eggs were weighed daily, parallel to measurements of egg production, daily feed intake, feed conversion ratio, and egg mass. When the hens reached 44 wk of age, the measurements were aimed at fatty acid profiles, malondialdehyde (MDA), and cholesterol concentration in egg yolk. Feeding the hens on diets enriched by omega-3 fatty acids led to higher levels of egg production than those fed on control diets, but their daily feed intake was generally lower (P < 0.05). Egg weight decreased in birds that were fed on diets enriched with omega-3 fatty acids without L-carnitine, or with diets which contained 100 mg/kg L-carnitine, compared to control diets which contained 0 mg/kg L-carnitine (P < 0.05). Egg mass increased in birds that were fed on diets enriched with omega-3 fatty acids and which contained 200 mg/kg L-carnitine, compared to the control diet with 0 or 100 mg/kg L-carnitine (P < 0.05). The analysis of fatty acid profiles showed that L-carnitine and omega-3 fatty acids caused a significant increase in the percentage of eicosapentaenoic acid (EPA), docosahexaenoic  acid (DHA), C18: 1 (n-9), arachidonic acid (ARA) C20: 4 (n-6), and Σ n-3 in the eggs of birds (P < 0.05). Based on the results, adding L-carnitine (200 mg/kg) to diets that were already enriched with omega-3 fatty acids increased the level of production and led to a longer maintenance of fatty acids in the eggs. Also, oxidative stability was enhanced in the yolk of eggs.

Pharmacokinetics and Egg Residues of Oral Meloxicam in Bantam Cochin Chickens.

Backyard poultry are commonly treated in veterinary hospitals; however, there is limited information regarding appropriate dosing of medications and withdrawal times for eggs. Six healthy adult bantam Cochin hens were given a single oral dose of meloxicam (1 mg/kg). Meloxicam plasma concentrations and egg residues were analyzed by high-performance liquid chromatography. Noncompartmental analysis was used to calculate pharmacokinetic parameters. The apparent terminal half-life, maximum concentration, and time to maximum concentration were 5.94 ± 0.92 hours, 7.03 ± 2.68 µg/mL, and 2.83 ± 1.33 hours, respectively. Meloxicam was detected in egg whites for 4.8 ± 1.5 days and egg yolks for 9.8 ± 2.4 days. Results were compared with previous studies in white leghorn and Columbian Wyandotte hens. Bantam Cochin hens demonstrated a significantly longer mean apparent terminal half-life, greater area under the curve, smaller elimination rate constant, and longer egg residue times compared with white leghorn hens. However, the pharmacokinetic results from the bantam Cochin hens did not significantly differ from those reported for the Columbian Wyandotte hens. Until pharmacodynamic studies can be performed, dosing of oral meloxicam in bantam Cochins should follow recommendations for Columbian Wyandotte hens to reduce the likelihood of adverse effects. These results better inform appropriate dosing of meloxicam in domestic hens, as well as recommended withdrawal times for egg consumption.

Research Note: The prevalence and vertical transmission of avian hepatitis E virus novel genotypes in Tai'an city, China.

To investigate the prevalence of avian hepatitis E virus (HEV) in chickens and gather evidence of viral vertical transmission, we collected 288 cloacal swabs and 288 yolks samples from 12 farms with clinically healthy chickens in 4 different areas in Tai'an City, Shandong Province, China (i.e., Daiyue District, Xintai City, Feicheng City, and Ningyang County). We also collected 240 samples from 2 breeder farms (from each of which 30 chicks, 30 dead embryos, 30 live embryos, and 30 hatching eggs were taken). PCR detection revealed that the positive rates of cloacal swabs and yolks were 6.25% (18/288) and 4.51% (13/288), respectively. Besides, avian HEV was detected with higher positive rates in the chicks (11.67%), hatching eggs (10.00%), live embryos (13.33%), and dead embryos (26.67%) from 2 breeder farms. Sequence and genetic evolution analyses revealed that the nucleotide homology of the isolated strains was 76.4to 83.9% compared with 4 reported genotypes, but the isolated strains were located in a separate branch, indicating they were potential novel genotypes. In conclusion, those results indicate that the latent infection of avian HEV novel genotypes has been widespread in chicken farms in Tai'an City, and provide reliable evidence of the possible vertical transmission of avian HEV.

Spent fowl as a source of unintentional egg proteins exposure in Canadian food products.

The occurrence of egg proteins in products containing spent fowl manufactured under current practices was studied to assess the risk these food products may pose to egg-allergic consumers and to determine if Precautionary Allergen Labelling (PAL) was recommended. Spent fowl slaughtering and processing operations in 2 Canadian facilities were observed. Raw hen pieces (n = 134), coming from 2 facilities, and intermediate and processed products containing spent fowl (n = 57), coming from one facility, were analyzed using ELISA. All samples tested positive for egg proteins. Raw pieces were tested using a qualitative method (i.e., swabbing); estimated egg proteins concentrations suggest the presence of highly contaminated samples (>600 mg/kg in 2 hen wing samples). Swabbing was found to be efficient for rapid detection of eggs in raw hen pieces, but not for quantification. A comparison between swab and grind results showed that egg proteins concentration is underestimated by at least a factor 2 for whole carcasses and a factor 10 for breast, wings and drumsticks, when using the swab protocol. For intermediate and processed products, quantitative measurements indicate that egg protein levels were below 16 mg/kg. Additionally, 88 water samples from chiller tanks were analyzed and indicate that this step could be the cause of the global contamination observed with an increase in egg protein concentrations overtime during the production schedule. As egg contamination is not adequately controlled under the current good production practices, the use of PAL would be recommended for raw spent fowl products.

Research Note: Comparison of the texture, structure, and composition of eggs from local Chinese chickens and a highly selected line of egg-type chickens and analysis of the effects of lipids on texture.

Egg yolk texture affects consumer egg preference. The sensory characteristics of eggs are affected by not only the cooking method but also the maternal breed. In this study, we investigated the texture, structure, and nutritional differences between the cooked yolks of eggs obtained from Hetian Dahei (HTDH) and Rhode Island Red (RIR) chickens. The springiness, cohesiveness, gumminess, chewiness, and resilience of HTDH egg yolks were lower, and the hardness was higher than those of RIR egg yolks. Moreover, scanning electron microscopy revealed that HTDH egg yolk particles were smaller and that HTDH egg yolks had a denser protein network than those of RIR egg yolks. Lipid and protein levels were higher, whereas water contents were lower in uncooked HTDH egg yolks than in uncooked RIR egg yolks. Liquid chromatography tandem mass spectrometry further revealed that lower cohesiveness was associated with higher levels and greater variety of lipids in egg yolks. Moreover, increased phospholipid levels reduced egg yolk cohesiveness. Thus, the eggs of local Chinese chicken breeds were superior to those of a highly selected broiler chicken breed in terms of texture, structure, and nutritional composition, which may influence egg variety selection.

Detection of fluoroquinolone and sulfonamide residues in poultry eggs in Kunming city, southwest China.

Antibiotic residues contained in poultry eggs pose threat to human health. However, the classes and concentrations of antibiotics in poultry egg in southwestern China is unknown due to insufficient monitoring and research. A total of 513 egg samples were collected from supermarkets and farm markets in Kunming city in 2020 and the levels of 7 antibiotics were analyzed using ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. The linear correlation coefficients were above 0.990 for all antibiotics tested. The limits of detection and limits of quantification in poultry eggs were 0.002 to 0.010 µg/g and 0.007 to 0.033 µg/g, respectively. The average recoveries of the 7 analytes from poultry egg samples were 80.00 to 128.01%, with relative standard deviations of less than 13.97%. A total of 93 (18.13%) samples tested positive for antibiotics, with the highest concentration being 2.48 µg/g. The concentration range of ofloxacin, danofloxacin, difloxacin, sulfadimethoxine, sulfamonomethoxine, sulfamethoxypyridazine, and sulfamethoxazole in poultry eggs was 0.01 to 0.37 µg/g, 0.06 to 0.48 µg/g, 0.05 to 0.29 µg/g, 0.03 to 0.16 µg/g, 0.06 to 1.00 µg/g, 0.05 to 0.37, and 0.07 to 2.48 µg/g, respectively. Sulfamonomethoxine was detected from hen eggs with the highest concentration level at 1.00 µg/g. Sulfamethoxazole was detected with the highest concentration level from both duck and quail eggs, at 1.87 and 2.48 µg/g, respectively. The antibiotic with the highest residue level in pheasant eggs was danofloxacin, which was 0.37 µg/g. Sulfamethoxypyridazine was identified in 30 samples with the highest positive rate of 5.85%, sulfadimethoxine was identified in 3 samples with the lowest positive rate of 0.58%. We observed that 7 targeted antibiotic residues in quail eggs and 3 targeted antibiotic residues in pheasant eggs. We also found that there were antibiotic residues in free-range hen eggs and the concentration was not low. The antibiotic with the highest residue level in free-range eggs was sulfamonomethoxine, which was 1.00 µg/g. These findings suggest that continual antibiotic residue monitoring of poultry eggs is essential in China.

Developmental changes of free amino acids in amniotic, allantoic fluids and yolk of broiler embryo.

1. A study was conducted to evaluate the developmental changes of protein and free amino acid concentrations in amniotic, allantoic fluids and yolk during the incubation period of broiler eggs.2. A total of 120 Cobb 500 fertile eggs were individually weighed and then placed in an incubator. On incubation day: 8, 11, 13, 14, 16, and/or 18, amniotic, allantoic fluids and yolk were collected from 20 eggs for analysis of protein content and free α-amino acid concentration in allantoic and amniotic fluids and yolk.3. The total protein concentration in amniotic fluid increased from d 11 of incubation, and reached a peak at d 16 (69.85 g/l; P < 0.01), then declined at d 18 (P < 0.05). The total protein concentration in allantoic fluid increased with age of the embryo (P < 0.01). Crude protein concentration in yolk decreased (P < 0.05) from d 0 to 8, then increased gradually from d 8, and reached a peak at d 16 (P < 0.05). The concentration of most free amino acids in amniotic and allantoic fluids and yolk was related to embryo weight. Amniotic fluid amino acids gradually increased from d 13 to 18, with arginine being the most abundant at d 11 and 14. Glutamate was the most predominant amniotic fluid amino acid at d 16 and 18. From d 13 to 18, the concentrations of most α-amino acids in allantoic fluid increased, and reached a peak at d 18 (aspartate, 373 µmol/l; asparagine, 519 µmol/l; glutamine, 1230 µmol/l; threonine, 537 µmol/l; citrulline, 112 µmol/l; arginine, 2747 µmol/l; alanine, 276 µmol/l; tyrosine, 330 µmol/l; tryptophan, 212 µmol/l; valine, 140 µmol/l; phenylalanine, 102 µmol/l; isoleucine, 92.39 µmol/l; lysine, 1088 µmol/l; P < 0.05). Glutamine was the second most abundant amino acid in allantoic fluid at d 13 and 18. Glutamate was the most abundant α-amino acids at d 8, and 13 in the yolk.4. These results demonstrated that the concentration of free α-amino acids in chicken embryo fluid was related to embryo weight. Arginine, glutamine and glutamate were abundant free α-amino acids in chicken embryo fluid, to support the higher rates of tissue protein synthesis and growth for the embryo.

Genetic analysis reveals three novel QTLs underpinning a butterfly egg-induced hypersensitive response-like cell death in Brassica rapa.

BACKGROUND: Cabbage white butterflies (Pieris spp.) can be severe pests of Brassica crops such as Chinese cabbage, Pak choi (Brassica rapa) or cabbages (B. oleracea). Eggs of Pieris spp. can induce a hypersensitive response-like (HR-like) cell death which reduces egg survival in the wild black mustard (B. nigra). Unravelling the genetic basis of this egg-killing trait in Brassica crops could improve crop resistance to herbivory, reducing major crop losses and pesticides use. Here we investigated the genetic architecture of a HR-like cell death induced by P. brassicae eggs in B. rapa. RESULTS: A germplasm screening of 56 B. rapa accessions, representing the genetic and geographical diversity of a B. rapa core collection, showed phenotypic variation for cell death. An image-based phenotyping protocol was developed to accurately measure size of HR-like cell death and was then used to identify two accessions that consistently showed weak (R-o-18) or strong cell death response (L58). Screening of 160 RILs derived from these two accessions resulted in three novel QTLs for Pieris brassicae-induced cell death on chromosomes A02 (Pbc1), A03 (Pbc2), and A06 (Pbc3). The three QTLs Pbc1-3 contain cell surface receptors, intracellular receptors and other genes involved in plant immunity processes, such as ROS accumulation and cell death formation. Synteny analysis with A. thaliana suggested that Pbc1 and Pbc2 are novel QTLs associated with this trait, while Pbc3 also contains an ortholog of LecRK-I.1, a gene of A. thaliana previously associated with cell death induced by a P. brassicae egg extract. CONCLUSIONS: This study provides the first genomic regions associated with the Pieris egg-induced HR-like cell death in a Brassica crop species. It is a step closer towards unravelling the genetic basis of an egg-killing crop resistance trait, paving the way for breeders to further fine-map and validate candidate genes.

Effect of synbiotics administered in ovo on microvascularization and histopathological changes in pectoral muscle and the biochemical profile of broiler chicken blood.

The aim of this study was to investigate whether injecting synbiotics to the egg air chamber on d 12 of embryo incubation will affect the processes of angiogenesis, and thus the share of histopathological changes in superficial pectoral muscle, as well as Ca and P in blood of 42-day-old broiler chickens. The eggs containing viable embryos were injected with 0.2 mL suspension of 1/physiological saline, 2/SYN1 composed of galactooligosaccharide (GOS) (trade name: Bi2tos®, Clasado Biosciences Ltd, UK) and L. salivarius or 3/SYN2 composed of RFO and L. plantarum. All birds were fed ad libitum the standard commercial feed mixtures: starter, grower, and finisher, with a constant access to water and feed. Injecting synbiotics in ovo on d 12 of the embryonal development significantly affected the blood supply to superficial breast muscle in broiler chickens. The highest density of capillaries in the muscle area under study and per muscle fiber were identified in the group of birds the egg air chamber of which was provided with synbiotic GOS+L. salivarius. Consequently, for the muscles of the birds injected with the same synbiotic there was found the highest share of normal fibers and least necrosis and splitting, as compared with the control. The conducted research confirms the relationship between the blood supply to the muscle and the occurrence of pathological changes. We have observed a positive effect of synbiotics on the microvascularization and the size of histopathological changes in the chicken muscle, which, from a practical perspective, can affect the health status and the meat quality. Blood biochemical analyses showed that the in ovo injection of synbiotics did not significantly affect the level of parameters, except for Ca and P. A significant increase in the concentration of these minerals in the blood of chickens injected with SYN1 could have a positive effect on the angiogenesis process.

Type of hormonal treatment administered to induce vitellogenesis in European eel influences biochemical composition of eggs and yolk-sac larvae.

Egg biochemical composition is among the main factors affecting offspring quality and survival during the yolk-sac stage, when larvae depend exclusively on yolk nutrients. These nutrients are primarily embedded in the developing oocytes during vitellogenesis. In aquaculture, assisted reproduction procedures may be applied enabling gamete production. For the European eel (Anguilla anguilla), reproductive treatment involves administration of pituitary extracts from carp (CPE) or salmon (SPE) to induce and sustain vitellogenesis. In the present study, we compared the influence of CPE and SPE treatments on offspring quality and composition as well as nutrient utilization during the yolk-sac stage. Thus, dry weight, proximal composition (total lipid, total protein), free amino acids, and fatty acids were assessed in eggs and larvae throughout the yolk-sac stage, where body and oil-droplet area were measured to estimate growth rate, oil-droplet utilization, and oil-droplet utilization efficiency. The results showed that CPE females spawned eggs with higher lipid and free amino acid contents. However, SPE females produced more buoyant eggs with higher fertilization rate as well as larger larvae with more energy reserves (estimated as oil-droplet area). Overall, general patterns of nutrient utilization were detected, such as the amount of total lipid and monounsaturated fatty acids decreasing from the egg stage and throughout the yolk-sac larval stage. On the contrary, essential fatty acids and free amino acids were retained. Notably, towards the end of the yolk-sac stage, the proximal composition and biometry of surviving larvae, from both treatments, were similar.