Evaluation of an early granulocytic response of chick embryos inoculated with herpesvirus of turkeys

1. Early granulocytic response was evaluated in chick embryos inoculated with herpesvirus of turkeys (HVT). 2. Fifty 10-d-old specific pathogen-free embryos were divided into two groups, inoculated via yolk sac. Group 1 were inoculated with a complete dose of HVT and group 2 with vaccinediluent only. 3. Samples were taken for histological evaluation of yolk sac, liver, chorioallantoic membrane, brain and heart from 5 embryos per group on days 12, 14, 16, 18 and 20 of embryonic life. 4. Increases in numbers of granulocytes were detected on days 14 and 16 in the yolk sac, and on d 14 and 20 in the liver of in embryos, which received HVT. In addition, the chorioallantoic membrane was infiltrated with granulocytes. 5. The results confirm that granulopoiesis in the yolk sac is stimulated in the early stages of incubation if a viral antigen is present. The virus also appears to trigger the presence of granulocytes in embryonic liver and chorioallantoic membranes.     

Tissue-specific changes in the activities of antioxidant enzymes during the development of the chicken embryo.

1. Tissue-specific profiles of the expression of the antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) and the concentrations of reduced glutathione (GSH) during the development of the chick embryo were investigated. 2. The liver, brain, yolk sac membrane (YSM), kidney, lung, heart and skeletal muscles were collected at the following days of embryo development: 10, 11, 13, 15, 17, 19, 21 and 22 (day-old chicks). 3. The different tissues of the embryo displayed distinct development strategies with regard to the acquisition of antioxidant capacity. In the liver the specific activity of SOD increased between days 10 and 11 of development, then significantly decreased up to day 15 and remained at the same value during the rest of the developmental period. GSH-Px specific activity increased through the time of development. CAT had 2 peaks of specific activity at day 10 of the development and in day-old chicks. 4. The brain was characterised by comparatively high SOD-specific activity especially during the last days of incubation. The specific activities of GSH-Px and CAT were low throughout development. 5. In the YSM maximal GSH-Px and CAT-specific activities were found on day 15 of incubation. In the kidney and heart GSH-Px-specific activity increased at hatching time. CAT-specific activity in the kidney increased just after hatching. 6. It is concluded that each tissue studied expressed a profile of antioxidant defence mechanisms to deal with oxidative stress at hatching time.     

Effects of breeder hen age and dietary L-carnitine on progeny embryogenesis

1. Ross 308 broiler breeder hens were given diets containing 0 or 25 mg L-carnitine/kg (8 replications per treatment) from 21 weeks of age. 2. Hens were inseminated with semen from Ross broiler breeder males. In a common facility, subsequent progeny hatchability and embryonic mortality at 25, 30, 32, and 38 weeks of breeder age were evaluated. 3. Subsequent egg component weights, incubational egg water loss, progeny embryo growth, and embryo, yolk sac and liver composition through 18 d of incubation at 27, 32, and 38 weeks of breeder age were evaluated. 4. Calculated additions of L-carnitine were in agreement with analysed contents of 3.5 and 31.1 mg free L-carnitine/kg of diet, respectively, and total L-carnitine concentrations increased by 48.6, 21.7, and 10.0% in 0-d yolk, 18-d yolk sac, and 18-d liver samples, respectively, due to the addition of dietary L-carnitine. 5. Supplemental L-carnitine resulted in increased (0.6%) relative 0-d egg yolk weight across weeks 27, 32, and 38, and reduced (0.38%) 18-d yolk sac palmitoleic acid concentration at week 27 without altering embryogenesis. 6. In conclusion, dietary L-carnitine (25 mg/kg of the diet) was deposited in the yolks of broiler breeder hens and was subsequently transferred to the embryonic liver via yolk sac absorption through 18 d of incubation. Furthermore, dietary L-carnitine supplementation increased ovarian follicle yolk deposition in 27-, 32-, and 38-week-old breeder hens, and influenced yolk sac fatty acid beta-oxidation in embryos from 27-week-old breeder hens causing yolk sac palmitoleic acid concentrations to be reduced by 18 d of incubation.     

Comparative evaluation of the effect of two maternal diets on fatty acids, vitamin E and carotenoids in the chick embryo.

1. The fatty acid profile of egg yolk and vitamin E and carotenoid accumulation in the egg yolk and embryonic tissues were investigated in relation to the maternal diet. 2. Two hundred fertile eggs (Ross 308 Broiler Breeder), obtained from hens fed on a maize-based (M-diet) or a wheat-based diet (W-diet), were incubated using standard conditions. 3. The egg yolk and embryo tissues (residual yolk, yolk sac membrane, liver, kidney, lung, muscles, adipose tissue and plasma) were collected on d 18 of incubation and on d 21 (newly-hatched chicks) and analysed for fatty acids, vitamin E and carotenoids. 4. The diets did not differ in terms of fatty acid or alpha-tocopherol concentrations. The concentration of carotenoids in the M-diet was 11.8 mg/kg and in the W-diet was 5.6 mg/kg with lutein and zeaxanthin being major carotenoids. 5. Eggs from the M-group contained higher (P<0.01) concentrations of beta+gamma-tocopherols, total carotenoids, lutein and zeaxanthin. Chickens hatched from those eggs were characterised by the increased concentrations of total carotenoids and zeaxanthin in all the tissues studied. The concentration of beta+gamma-tocopherol was enhanced only in the liver and yolk sac membrane. 6. It is concluded that the maternal diet plays an important role in antioxidant systems formation during chick embryonic development; the M-diet can increase the antioxidant potential of the egg yolk and embryonic tissues compared to the antioxidant potential provided by parent birds fed the W-diet.     

Effects of coated hatching eggs obtained from old broiler breeders with chitosan on embryonic growth,hatching results and chick quality

The aim of this research was to determine the effects of coating broiler hatching eggs with chitosan on egg quality, embryonic growth, hatching results and chick quality. Eggs obtained from old broiler breeder, aged 59 weeks, were used. Eggs were divided into two groups (coated with chitosan and uncoated) groups. Each group was divided into three groups according to the storage period. Eggs were stored for periods of 1, 4 and 7 days at 15°C and 80% humidity. Then, they were incubated. Storage period of eggs up to 7 days did not affect the egg quality, egg weight loss, embryo growth, chick properties and hatchability of fertile eggs. However, embryo development, relative residual yolk sac weight and relative chick weight were affected by coating eggs with chitosan. Interaction was not found between storage lengths and coating of eggs with chitosan for all examined parameters. The results of the present study suggested that hatching eggs obtained from old broiler breeder flocks were immersed into chitosan solution once and quickly have negative effect on the embryo weight, yolk sac absorption and relative chick weight. It has been observed that covering the hatching eggs with chitosan is not suitable in this respect even if immersion takes place once and in a short time.     

Pathogenicity of Mycoplasma synoviae in chicken embryos.

Pathogenicity of Mycoplasma synoviae (MS) was examined in specific-pathogen-free (SPF) white leghorn chicken embryos. Six isolates of MS were inoculated into 7-day embryos via the yolk sac. Isolates were evaluated for gross and microscopic lesions through 19 days' incubation and for embryo lethality through 20 days' incubation. Isolates in decreasing order of lethality, from lowest to highest 50% embryo lethal dose, were WVU 1853, K1968, K1858, FMT, 92D8034, and F10-2AS. Embryo lethality was consistent with lesion incidence and severity. Embryo lethality did not correlate with previous results regarding pathogenicity of these same six isolates in SPF broiler chickens.     

Factors affecting the weight and water status of the chick at hatch

1. Variation in chick weight at the moment of escape from the shell is accounted for by the fresh weight of the egg, the weight lost from the egg during the incubation period and the weight of the shell and residues at hatch.

2. Over 97% of the variation in chick weight at hatch can be explained by two factors, fresh egg weight and weight loss during incubation. These two factors still account for a high (87%) proportion of the variation in chick weight at 21 d after setting when chicks would normally be removed from the hatcher.

3. The water content of the true chick and the residual yolk sac are both affected by water loss from the egg during incubation and account for the differences in the weight of the whole chick at hatch from eggs of the same size.

4. When the fluid reserves within the egg become exhausted in the few days before hatch, the water: dry weight ratio of the yolk rather than embryo is maintained despite variation in water losses between eggs.     

卵黄中脂肪酸含量的变化对鸡胚组成的影响

对孵化过程中,卵黄囊中各种脂肪酸的含量变化和鸡胚或出壳雏鸡机体的水分、脂肪和蛋白质的含量变化关系进行了两次试验。试验1于入孵的第6,9,12,15,18和21 d测定胚胎或出壳雏的湿重和干重、水分、粗蛋白质和粗脂肪的含量。试验2于入孵的第6,9,1 2,15和18 d测定胚胎的干重、湿重、卵黄脂类和脂肪酸的含量。结果表明试验1:胚体水分含量和胚体蛋白含量呈正相关,但和干胚重、体脂肪含量呈负相关。试验2卵黄中脂类含量和湿胚重、胚胎水分含量、卵黄中亚油酸含量呈正相关。而卵黄中脂类含量和干胚重、相对卵黄重呈负相关。相对干胚重(以入孵蛋重为基础)表现出和硬脂酸呈负相关,孵化过程中胚胎的水分、粗脂肪、粗蛋白含量受来自卵黄脂肪的影响是不同的。胚胎摄取的硬脂酸和亚油酸对胚胎的干物质组成和卵黄的花生四烯酸浓度的影响是不同的。     

Effect of maternal corticosterone on utilisation of residual yolk sac fatty acids by developing broiler embryo

1. Increased concentrations of maternal corticosterone are deposited into egg yolk under stress conditions. This experiment investigated the effect of maternal corticosterone on yolk and yolk sac fatty acid profiles of eggs and developing broiler embryos. 2. At 42 weeks of age, 200 broiler breeder females were randomly divided into two groups: maternal corticosterone (MC): 2 mg/hen/d of corticosterone dissolved in 1 mL of 99% ethanol and mixed in the individual daily feed for a 14 d period, and control: 1 mL of ethanol added to the individual daily feed. Hens were inseminated, and eggs were collected daily from d 3 of corticosterone feeding to d 14 and incubated. Fatty acid profiles of egg yolk and yolk sac were analysed before incubation and at 12, 14, 18 and 21 d of incubation. 3. Yolk corticosterone concentrations were greater in MC eggs. Before incubation, eggs from MC had lower DHA (22 : 6 n-3) but higher stearic (18 : 0) and dihomo-gamma linolenic (20 : 3 n-6 Cis 8,11,14) acid content compared with the control. 4. Higher concentrations of stearic (18 : 0), elaidic (18 : 1 tr n-9), eicosenoic (20 : 1 n-9), eicosadienoic (20 : 2 n-6), dihomo-gamma linoleic (20 : 3 n-6) and arachidonic (20 : 4 n-6) acids and lower concentrations of myristic (14 : 0), palmitoleic (16 : 1 n-7), linoleic (18 : 2 n-6) and DHA (22 : 6 n-3) were found at different periods of incubation in the residual yolk sac of MC than from controls. 5. The results suggest that the preferentially selective absorption of fatty acids is related to specific tissue requirements.     

孵化期间鸡胚、肝脏、卵黄及蛋白组成的变化及其相关关系

将1188枚种蛋随机码放在6个蛋盘内，同机人孵，每盘为1个试验重复。于人孵的第6、9、12、15、18和21天从各盘取出36个胚蛋用于测定蛋清葡萄糖的含量；胚质量及其水分、蛋白质和脂肪含量；肝脏质量、水分和脂肪含量；卵黄囊质量、卵黄脂类及其脂肪酸的含量。结果表明，随着孵化时间的增加，肝脏的脂肪含量不断增加，但胚胎的水分含量、蛋清的葡萄糖含量和卵黄囊质量则不断减少，脂肪在孵化的第12天含量最低，蛋白质在孵化的第6、12和15天含量较高。相对肝质量与胚胎的水分含量正相关，与孵化第15天的卵黄棕榈酸和油酸分别为正相关和负相关。卵黄硬脂酸含量与孵化第6天的亚油酸和孵化第12天的油酸均为负相关，但与孵化第6和15天的花生四烯酸均正相关。     

Experimental transmission of turkey viral hepatitis to day-old poults and identification of associated viral particles resembling picornaviruses.

Turkey viral hepatitis (TVH) was experimentally reproduced in two experiments in 1-day-old poults. In the first experiment, an infectious inoculum was prepared from filtered yolk materials harvested from dead embryonating chicken eggs (ECE) previously inoculated with suspensions of liver and pancreas tissues collected from TVH-affected birds in commercial turkey flocks. One-day-old poults given a yolk-sac inoculation or oral gavage with this preparation developed lesions in the liver and pancreas characteristic of TVH at 20 days postinoculation (PI) in 60% and 14% of the experimentally infected birds, respectively. With the identical inoculum, embryo mortality occurred at 8 and 10 days PI in embryonating turkey eggs (ETE) inoculated into the yolk sac. In the second experiment, an infectious inoculum was prepared from filtered yolk materials from dead ETE harvested in the first experiment. One-day-old poults given a yolk-sac inoculation with this filtered yolk material developed lesions in the liver and pancreas within 5 days PI. At 20 days PI, 67% of the experimentally infected birds had similar lesions. With the inoculum given to these poults, embryo mortality occurred at 6, 8, and 10 days PI in ETE inoculated into the yolk sac. Virus particles 26-28 nm in diameter with icosahedral morphology typical of picornaviruses were identified by EM in the yolk sacs of ETE that died in both experiments, and inoculated ETE that died following passage of filtered suspensions of pancreatic tissues collected from affected birds in the first experiment.     

Twin embryos in mares. II: Post fixation embryo reduction

Recent findings on the development and natural outcome of twins from Day 17 (immediately after fixation) to Day 40 are reviewed. Incidence of embryo reduction was increased significantly when the vesicles became fixed unilaterally, rather than bilaterally, and when the vesicles were unequal in diameter. Of 68 mares with twins on the day of fixation, post fixation embryo reduction occurred in 41 (60 per cent). The incidence of reduction was 41 of 48 (85 per cent) following unilateral fixation; reduction occurred in all of 22 mares with vesicles of dissimilar size (4 mm or more difference in diameter) and in 19 (73 per cent) of 26 mares with vesicles of similar size (0 to 3 mm difference). Embryo reductions were complete (vesicle no longer visible ultrasonically) by Day 20 (59 per cent of the reductions), during Days 21 and 30 (27 per cent), or Days 31 to 38 (14 per cent). In 80 per cent of the early reductions (by Day 20) the eliminated vesicle disappeared within one day. Reductions that occurred after Day 20 were preceded by a gradual decrease in size. As the number of days after Day 17 increased, the frequency of reduction decreased and the time required for completion of reduction increased. When the twins were dissimilar in diameter (4 mm or more), they were more likely to undergo reduction by Day 20. In summary, dissimilarity in diameter increased the likelihood of unilateral fixation, increased the incidence of reduction for unilaterally fixed vesicles, hastened the day of occurrence of reduction and shortened the interval from initiation to completion of reduction. The deprivation hypothesis proposes that embryo reduction occurs when a major portion of the three walled area of the yolk sac or the vascularised wall of the yolk sac or allantoic sac is in apposition with the wall of the adjacent vesicle rather than with the endometrium; the vesicle is deprived of adequate embryonal-maternal exchange and therefore regresses.     

Yolk sac utilization in ostrich (Struthio camelus) chicks

The mass of residual yolk sac expressed as a percentage of initial mass of the egg from which the chick hatched decreased sharply in the first 2 days post-hatching. A gradual reduction occurred between 3 and 10 days after which a sharp decline was noted between 11 and 13 days post-hatching. The highest number of chicks with unabsorbed yolk sac was noted on day 5 post-hatching followed by days 6 and 7. Chick mortality followed the same pattern. The dynamics, causes and clinical consequences of yolk sac utilization are discussed.     

Antioxidant systems of the avian embryo: tissue-specific accumulation and distribution of vitamin E in the turkey embryo during development

Tissue-specific accumulation of tocopherols and tocotrienols in turkey tissues during embryonic development and their susceptibility to lipid peroxidation were investigated. Fertile turkey eggs were incubated using standard commercial conditions. Embryonic tissues were collected at 16, 22, 25 d of incubation and from day-old poults (referred to as day 29) and alpha-; beta- + gamma- and delta-tocopherols and respective tocotrienols were analysed by HPLC. A turkey diet provided to the parent hens contained the complete range of tocopherols and tocotrienols. Between days 16 and 22 of embryo development, the alpha-tocopherol concentration in the liver remained constant and then increased significantly (P<0.01) reaching a maximum just after hatching. Similar changes were observed for the other tocopherols and tocotrienols. The accumulation of alpha-tocopherol in the yolk sac membrane (YSM) started after day 20 of development and at hatching the alpha-tocopherol concentration in the YSM was twice that of beta- + gamma-tocopherols and 15 times greater than that of alpha-tocotrienol. In the kidney, heart, lung, muscle and adipose tissues a gradual increase in tocopherol and tocotrienol concentrations took place between days 20 and 25 of development with a sharp increase in particular of alpha-tocopherol between days 25 and 29. There was a discrimination between tocopherols and tocotrienols during their assimilation from the diet by the parent hen and during metabolism by the developing turkey embryo. Tissue-specific features in the susceptibility to lipid peroxidation were found with the brain being the most susceptible to lipid peroxidation at day 25 and in day-old poults.     

Effect of BioSentry 904 and ethylenediaminetetraacetic acid-tris disinfecting during incubation of chicken eggs on microbial levels and productivity of poultry

Proper sanitation practices and the use of efficacious disinfectants in a hatchery have an effect on chick quality. Aerosol bacterial counts, egg moisture loss, hatchability, chick quality, and broiler productivity were evaluated when egg surfaces were contaminated by immersion of each egg into a broth medium containing a field isolate of Pseudomonas aeruginosa and incubated with exposure to one of three disinfectant treatments administered by fine spray: distilled water, BioSentry 904 (904), and a 1:1 ratio of 904 and ethylenediaminetetraacetic acid (EDTA)-Tris. The aerosol bacteria levels were statistically greater on day 21 of incubation in the group treated with distilled water than in those receiving disinfectants. Overall hatch of fertile eggs and egg moisture loss were comparable among all three treatments. At 1 day of age, the chicks incubated with 904 had a statistically lower yolk sac contamination rate than those incubated with 904+EDTA-Tris or distilled water. The 2-wk mortality rates, body weights, feed conversion ratios, yolk sac weights, and yolk sac contamination rates were all similar among the three treatments.     

Importance of sub-embryonic fluid and albumen in the embryo's response to turning of the egg during incubation

1. The effects of removal of sub-embryonic fluid and albumen on the development of the fowl embryo were investigated. 2. A deficiency of sub-embryonic fluid on day 7 of incubation resulted in a reduction in the volume of the yolk sac and of allantoic fluid, and a reduction in the mass of the embryo later in incubation. 3. A deficiency of albumen on day 3 of incubation led to a reduction in allantoic fluid and embryo mass after day 12 incubation. 4. The results suggest that the physiology of embryos in unturned eggs is better modelled by eggs deprived of albumen than by eggs deprived of sub-embryonic fluid.     

Pathogenesis of Brucella abortus in chicken embryos

Chicken embryos inoculated with Brucella abortus at 6, 10, and 12 days of incubation were examined by light and electron microscopy. B. abortus was identified by avidin-biotin immunoperoxidase and immunogold techniques. Death occurred from 2 to 5 days post-inoculation, depending on age of the embryo and route of inoculation. B. abortus was recovered from all infected eggs. Brucellae had spread throughout all tissues and localized preferentially within cells of mesodermal derivation. Organ distribution and degree of bacterial replication varied with age of the embryo at time of inoculation. In 6-day-old embryos, B. abortus localized preferentially in endoderm and mesoderm of yolk sac wall, extra- and intraembryonic serosal epithelia, and glomeruli of the mesonephros. In 10- and 12-day-old embryos, B. abortus spread to all tissues; renal glomeruli, liver, spleen, and heart were most severely infected. Intracellular B. abortus was within the rough endoplasmic reticulum of mesenchymal, mesothelial, yolk endodermal, and hepatic cells. In mononuclear phagocytes, endothelial cells, and granulocytes, bacteria were within membrane-bound vacuoles. Intracellular replication of B. abortus in embryonic tissues, especially yolk endoderm, closely resembled that in experimental infections of trophoblasts.     

Chicken embryo propagation of type I avian adenoviruses

Forty-two clone-purified, cell-culture-propagated type I avian adenoviruses (AAV) representing 11 serotypes and two intermediate strains were evaluated for virus replication (evidenced by embryo death and lesions) resulting from the inoculation of specific-pathogen-free chicken embryos via the chorioallantoic sac or yolk sac. Commonly observed embryonic changes were death, stunting and curling, hepatitis, splenomegaly, congestion and hemorrhage of body parts, and urate formation in the kidneys. Basophilic or eosinophilic intranuclear inclusion bodies characteristic of fowl adenoviruses were observed in hepatocytes. The magnitude and relative uniformity of intra- and interserotypic embryo mortality, gross lesions, and virus titers was greater in embryos inoculated via the yolk sac. This work identifies the yolk sac as a practical and sensitive chicken embryo inoculation route for poultry diagnosticians to employ. It is suggested that the yolk sac may be a reliable alternative to cell culture for the successful isolation of all type I avian adenoviruses.     

Ontogeny of 5-aminolevulinic dehydratase and porphobilinogen deaminase activities in the yolk sac membrane and liver of chick embryos

1. Chick embryos of 7, 9, 11, 12, 13, 14, 15, 17 and 19 d of embryonic development were examined to determine the activities of 5-aminolevulinic dehydratase (ALA-D, EC 4.2.1.24) and porphobilinogen deaminase (PBG-D, EC 4.3.1.8). 2. Liver and yolk sac membrane ALA-D specific activities showed a maximum between 12 and 13 d of embryonic development, yolk sac membrane PBG-ase activity a maximum at 9 d and at 7 d in liver. Total activities of ALA-D and PBG-D were not constant during the course of embryonic development but probably related to the changes of intensity of haem synthesis. 3. ALA-D and PBG-ase activities were higher in yolk sac membrane than in liver, showing the importance of the yolk sac membrane as erythropoietic tissue. PBG-D catalysed the rate-limiting reaction of the cytosolic steps in the biosynthetic pathway in both tissues.     

Dynamics of yolk sac resorption and post-hatching development of the gastrointestinal tract in chickens, ducks and geese

Under similar environmental conditions, 100 male Shaver-Starbro hybrid chickens, 100 male Mulard cross-line ducklings and 100 male Landaise goslings kept in 30 metabolic cages from the first day of life (10 birds of each species separately per cage) were chosen for the study. The animals were fed semi ad libitum diets with crude protein content and energy density similar to that used for poultry species, avoiding feed residues. During the first 21 days of life they received starter diets and from day 22 to 42 grower diets. Maize accounted for 23-40% and barley constituted 10-18%. Wheat accounted for 20.0% in starter diets and 10-18% in grower diets. Resorption of yolk sac residues, performance, development of the intestinal tract in young chickens, ducklings and goslings were assessed. During the first 5 days of life intensive absorption of yolk sac ingredients was observed. On day 7, residues of the yolk sac were found in more than 30% of chickens compared with approximately 10% in geese. In ducklings residues of the yolk sac were not found. On day 16 unabsorbed yolk sacs were found in approximately 10% of chickens. Liver and pancreas weighed the highest in geese and in ducks; in chickens the weight of these organs was significantly lower (p < 0.01). The relative values calculated to 100 g of metabolic body weight show that the average small intestine in chickens was significantly longer (p < 0.01) than in ducks or geese. The intestinal tract developed earlier in the chicken than in the water fowl.