鸡胚发育过程中胚胎液中几种游离氨基酸的动态变化

本实验旨在研究鸡胚发育过程中尿囊液和羊水中天冬氨酸、谷氨酸、精氨酸和亮氨酸浓度的动态变化。选用72枚6胚龄的三黄鸡鸡胚,按重量相近原则分为6组,每组12个重复,分别在孵化第7、8、11、13、14、18天采集尿囊液和羊水,并利用高效液相色谱法分析其氨基酸浓度。结果表明:尿囊液中,除8胚龄外,其他各胚龄精氨酸浓度均高于另外3种氨基酸(P<0.05),13胚龄精氨酸浓度显著低于其他胚龄(P<0.05),而18胚龄显则著高于其他胚龄(P<0.05)。羊水中,各胚龄精氨酸浓度均高于其他3种氨基酸;除11胚龄外,13胚龄精氨酸浓度显著高于其他胚龄(P<0.05);7胚龄天门冬氨酸浓度显著高于13、14、18胚龄(P<0.05);但18胚龄谷氨酸及亮氨酸浓度则显著升高(P<0.05)。以上结果说明,在尿囊液及羊水中,所测4种氨基酸浓度变化不尽相同,但总体精氨酸含量最丰富,这可能与鸡胚发育的营养与代谢有关。     

Importance of albumen content in whole-body protein synthesis of the chicken embryo during incubation.

The importance of egg albumen content in whole-body protein synthesis was investigated in developing chicken embryos by using lines genetically selected for high and low albumen contents and by removing albumen from eggs before incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine intravenously on day 12 of incubation. 3. Embryos from high albumen eggs had higher whole-body protein synthesis rates than those from low albumen eggs. 4. Whole-body protein synthesis was reduced by the removal of albumen from eggs before incubation. 5. It was concluded that albumen content per se was of crucial importance in regulating whole-body protein synthesis in chicken embryos during incubation.     

Characteristics of unturned eggs: critical period, retarded embryonic growth and poor albumen utilisation

1. The physiological effects of egg turning during incubation are described. 2. There was a critical period for turning, from days 3 to 7 of incubation, which affected hatchability, embryo growth and utilisation of extra-embryonic fluids. 3. Failure to turn eggs retarded formation of allantoic and amniotic fluids, restricted alumen uptake and retarded growth of the embryo, but only after day 12 of incubation.     

Failure to turn eggs during incubation: effects on embryo weight, development of the chorioallantois and absorption of albumen

Turning eggs during incubation is essential for good hatchability. In the present paper additional effects on the development of the chorioallantois, absorption of albumen and growth of the embryo are recorded. The ability of an unturned egg to hatch was not affected by egg weight, egg shell porosity or water loss during incubation. The ability of the chorioallantois to spread around the inner surface of the inner shell membrane and the degree of absorption of the residual albumen affected the growth of the embryo and its ability to hatch. Unturned eggs hatched later than eggs which were turned throughout incubation.     

Effect of immersion fluid temperature on the chicken embryo in teratogenicity tests: short communication

The influence of immersion fluid temperature on the development of the chicken embryo was studied on the day most commonly used for treating incubated eggs in teratological trials. Embryonated eggs were immersed in tap water for 30 min on the 12th day of incubation at 22-25 degrees C or at incubation temperature without a waiting time or after 30 min. The incubation was then continued and the eggs were processed on day 19 of the incubation period. Treatment of eggs at 22-25 degrees C caused a significant increase in embryonic mortality, while the 30-min waiting time did not exert an influence on embryogenesis.     

种毒接种剂量与鸡胚尿囊液量的相关性研究

对分成3组而同机孵化的180个9日龄罗曼蛋鸡鸡胚分别尿囊腔接种NDVLasota种毒液0.1mL／个,0.15mL／个和0.20mL／个。结果：尿囊液的收集量与种毒接种量呈显著负相关,而血凝价受接种剂量的影响不显著。     

Effects of electrical field on hatchability performance of eggs from a layer-type breeder

1. Eggs from a layer-type breeder flock (Baladi, King Saud University) between 50 and 63 weeks of age were used in three trials to study the effects of electrical field (EF) during incubation on albumen and yolk heights, incubation temperature, egg weight loss and hatchability traits. The effects of egg size and eggshell characteristics on hatchability traits of eggs incubated under EF were investigated. 2. Eggs were weighed and graded into three weight classes (small, medium, and large). The physical dimensions, eggshell characteristics, and conductance of eggs were examined. The incubator was divided into two compartments for the control and EF treatments. Two aluminium plates were fitted on the inside walls of the EF compartment, face to face, and connected to a step up electric transformer. Eggs were exposed constantly to the EF during the first 18 d of incubation at the level of 30 kV/m, 60 Hz. 3. Egg size influenced the physical dimensions and eggshell characteristics of eggs. Large eggs had higher egg weight, egg surface area, egg volume, eggshell conductance, and eggshell weight and lower yolk weight percentage than medium or small size eggs. Small eggs had lower egg length and higher egg density than large or medium size eggs. Large eggs had higher eggshell thickness than small size eggs. 4. EF incubation of eggs raised incubation temperature by 0.06 degrees C, and increased the percentage of egg weight loss, hatchability, and weight of hatching chicks and reduced the early embryo deaths, and length of incubation by approximately 9.8, 19.6, 1.7, 62.1 and 2.1%, respectively. 5. There was no significant difference between the two incubation treatments in the heights of albumen and yolk of incubated eggs, percentages of late embryo deaths, and pips with live and dead embryos. Hatchability traits were not significantly influenced by egg size. 6. It was concluded that EF incubation of eggs increased hatchability, chick-hatching weight, and reduced the length of incubation of Baladi eggs. Differences in the physical dimensions and eggshell characteristics of eggs did not influence hatchability traits of eggs under EF incubation.     

用异源蛋白和蛋壳体外培养鹌鹑胚胎的研究

将50个鹌鹑胚胎在鸡蛋壳中用鸡稀蛋白培养,对种蛋产出体外阶段裸黄状态的鹌鹑胚胎进行培养的方法进行了探讨。1～2.5d、2.5～14d和15～17d胚胎培养温度分别是38.0℃、37.8℃和37.5℃,相对湿度分别是60%、55%和70%,1～14d每小时翻蛋1次,翻蛋角度1～2.5d为90°,2.5～14d为50°,15d后静止落盘。2.5d和15d胚胎存活率以及孵化率分别为92%、78%和28%。结果表明裸黄状态的鹌鹑胚胎不排斥鸡稀蛋白,用鸡稀蛋白培养鹌鹑胚胎可行。     

Detection of early embryonic development in chicken eggs using visible light transmission

1. In two separate experiments, the possibility of detecting embryonic development in chicken eggs was assessed using the same spectrophotometric method used to detect blood in Table eggs, using a combination of two wavelengths (577 and 610 nm) of the transmission spectrum. 2. In the first experiment, during the first 10 d of incubation, transmission spectra of 30 Hisex White eggs and 30 Hybro eggs were measured daily. 3. In the second experiment, 292 Hisex White eggs were incubated. Seven groups were randomly assigned. Six received an injection of sodium azide (NaN3) at different times during incubation in order to stop embryonic development, and during the first 12 d of incubation the transmission spectrum was measured daily. The acoustic resonance analysis method was also used on a group of uninjected eggs. 4. In the first experiment, it was possible to detect embryonic development from 120 h of incubation onwards in fertile eggs. In the second experiment changes in light transmission due to embryonic development were detected from 108 h of incubation. Detection of embryonic development using the acoustic resonance analysis method in the second experiment was possible only from 120 h of incubation. 5. It was concluded that the detection of embryonic development using visible light transmission is not directly linked with the formation of blood, but with the formation of sub-embryonic fluid, which takes place from 72 h of incubation onwards. This fluid makes the yolk sac translucent so that absorption of light at 577 nm can be detected.     

Effect of egg turning and incubation time on carbonic anhydrase gene expression in the blastoderm of the Japanese quail (Coturnix c. japonica)

1. The gene expression of carbonic anhydrase, a key enzyme for the production of sub-embryonic fluid (SEF), was assessed in turned and unturned eggs of the Japanese quail. The plasma membrane-associated isoforms CA IV, CA IX, CA XII, CA XIV, and the cytoplasmic isoform CA II, were investigated in the extra-embryonic tissue of the blastoderm and in embryonic blood. 2. Eggs were incubated at 37.6 degrees C, c.60% RH, and turned hourly (90 degrees ) or left unturned. From 48 to 96 h of incubation mRNA was extracted from blastoderm tissue, reverse-transcribed to cDNA and quantified by real-time qPCR using gene-specific primers. Blood collected at 96 h was processed identically. 3. Blastoderm CA IV gene expression increased with the period of incubation only in turned eggs, with maxima at 84 and 96 h of incubation. Only very low levels were found in blood. 4. Blastoderm CA II gene expression was greatest at 48 and 54 h of incubation, subsequently declining to much lower levels and unaffected by turning. Blood CA II gene expression was about 25-fold greater than in the blastoderm. 5. The expression of CA IX in the blastoderm was the highest of all isoforms, yet unaffected by turning. CA XII did not amplify and CA XIV was present at unquantifiable low levels. 6. It is concluded that only gene expression for CA IV is sensitive to egg turning, and that increased CA IV gene expression could account for the additional SEF mass found at 84 to 96 h of incubation in embryos of turned eggs.     

Allantoic fluid electrolytes and urate of embryonic Japanese quail subject to differential water loss

1. The composition of allantoic fluid from Japanese quail was measured from days 9 to 14 of incubation.

2. The effect of the inhibitor of urate synthesis, allopurinol, upon weight, electrolytes and urate of allantoic fluid was investigated in eggs subject to different rates of water loss. The composition of plasma and the solid fraction of allantoic fluid was also investigated.

3. During the second half of incubation allantoic fluid water weight decreased, as did pH, and sodium, chloride and bicarbonate concentrations. Fluid urate and potassium concentrations increased, but osmolality was unchanged.

4. Allantoic urate and electrolyte concentrations were greatest, and fluid weight lowest, in eggs subject to high water loss.

5. Allopurinol reduced the total urate of allantoic fluid, and increased osmolality and electrolyte concentrations. Allopurinol had no effect on plasma osmolality.

6. It is concluded that in eggs of high water loss allantoic fluid osmolality was reduced by sequestration of sodium by urate in the fluid solid fraction. In eggs subject to low water loss fluid osmolality was reduced by the replacement of chloride by colloidal urate.     

Implantation of chicken embryonic tissue and cells into unfertilised eggs

1. Chick embryo cells and halved embryos were successfully implanted into unfertilised eggs. Yolks containing implants were placed in recipient eggshells, covered by transparent vacuum-formed plastic cones and incubated for 72 h. 2. Dispersed cells were obtained from eggs expelled from the uterus or from eggs that had been laid. Implantation of these cells often resulted in aggregation and epithelial growth, in several cases with axial development. 3. Growth of implanted halved embryos of different ages was often observed, including one 10-somite embryo. Non-axial epithelia, sometimes with a central hole, a central fluid-filled cellular vesicle or a vesicle only, were also observed. 4. In another culture system, whole and halved embryos obtained from laid eggs were cultured on a vitelline membrane stretched across semi-solid egg albumen. During the 72 h incubation, axial development was observed only in whole embryos, while halved embryos grew either into epithelia containing fluid-filled cellular vesicles or into vesicles only. 5. It was found that daily drainage of the accumulating fluid from the embryo compartment encouraged axial development in halved embryos, and almost abolished vesicle formation. Holes were formed in half the embryos cultured on a vitelline membrane. 6. It appeared that physical and biological conditions could inflict serious malformations on the implants.     

Effect of turning upon the sub‐embryonic fluid and albumen of the egg of the Japanese quail

1. The effects of egg turning upon the formation of sub‐embryonic fluid (SEF), and albumen weight and composition are described up to day 8 of incubation.

2. The density of albumen increased, and the density of yolk sac contents decreased, during the first 5 d of incubation. Failure to turn eggs diminished these changes.

3. The rate of formation of SEF and its maximum weight were lower if eggs were not turned, as was the rate of decrease in albumen weight.

4. The concentration of sodium and chloride, as well as osmolality, were higher in SEF than in thin albumen, and were affected by a failure to turn eggs.

5. Static incubation altered the concentrations of yolk nutrients in SEF.

6. It is concluded that the formation of SEF is primarily dependent upon the transfer of sodium and chloride from albumen to SEF so creating an osmotic force for water movement in the same direction. Turning the eggs promotes this process by ensuring an adequate supply of ions in thin albumen adjacent to the blastoderm.     

Effect of egg turning and fertility upon the sodium concentration of albumen of the Japanese quail

1. The effects of egg turning and fertility upon sodium concentration of albumen of the Japanese quail is described for up to 72 h incubation.

2. For incubated eggs the sodium concentration of albumen adjacent to the yolk sac was lower than that from albumen next to the shell. Static incubation increased the magnitude of this difference, such that albumen adjacent to the yolk sac was substantially depleted of sodium. This was found at the yolk equator and the yolk vegetal pole of both fertilised and unfertilised eggs.

3. Unincubated eggs also had a lower sodium concentration of albumen adjacent to the yolk sac compared with albumen next to the shell.

4. After 48 h of incubation yolk sodium concentration was substantially lower than albumen sodium concentration in both fertilised and unfertilised eggs, whether eggs were turned or not.

5. It is concluded that in unturned eggs the depletion of sodium from albumen adjacent to the vitelline membrane is not produced by ion transport processes but results from a passive movement of sodium into the yolk. Egg turning reduces the magnitude of the depletion of sodium from the albumen adjacent to the yolk sac by stirring the albumen, so permitting the full expression of ion and water transport across the blastoderm into the yolk sac.     

Effect of in ovo vaccine delivery route on herpesvirus of turkeys/SB-1 efficacy and viremia

A study was designed to ascertain the influence of in ovo site of inoculation and embryonic fluid type on the development of Marek's disease (MD) vaccine viremia and efficacy against MD challenge. The experiments were divided into in vitro and in vivo phases. In the in vitro phase, herpesvirus of turkeys/SB-1 vaccine was combined with basal medium eagle (BME) medium (control), amniotic fluid, or allantoic fluid and subsequently titrated on secondary chick embryo fibroblast cultures. There were no significant differences in titer between the virus inoculum carried in BME and the virus inoculum combined with either the allantoic fluid or the amniotic fluid. In the in vivo phase, five routes of inoculation, amniotic, intraembryonic, allantoic, air cell, and subcutaneous at hatch, were compared for generation of protection against virulent MD challenge. Comparisons were made in both specific-pathogen-free and commercial broiler embryos/chicks and, for the amniotic and allantoic routes, injection at either day 17 or day 18 of embryonation. Reisolation of the vaccine virus at day 3 of age was also done for all routes with the exception of the air cell route. Vaccine virus was recovered from all birds tested that were injected in ovo via the amniotic and intraembryonic routes and the subcutaneously at hatch route but was isolated only sporadically from birds inoculated via the allantoic route. Vaccination protective efficacy against virulent MD for all birds vaccinated in ovo via the amniotic or intraembryonic routes and birds vaccinated subcutaneously at hatch was over 90% regardless of day of in ovo injection or bird type. Protective efficacy for vaccines delivered in ovo by either the allantoic or the air cell routes was less than 50% regardless of day of injection or bird type. Therefore, in ovo MD vaccines must be injected either via the amniotic route or the intraembryonic route for optimal performance.     

雏半番鸭呼肠孤病毒的致病性

从鸭体、禽胚和细胞 3个方面探讨了雏半番鸭呼肠孤病毒 FZ2 株的致病性。经试验表明 ,在实验室条件下 ,该株病毒可导致雏番鸭、雏半番鸭发病、死亡 ,对雏番鸭的致死率为 14 .3%～ 4 1.7% ,对雏半番鸭的致死率高达 38.5 % ,且死亡鸭表现出与自然感染呼肠孤病毒病死的雏番鸭、雏半番鸭相同的病变 ;人工感染幸存鸭大多生长发育明显受阻。该株病毒经尿囊腔途径接种 ,对番鸭胚、半番鸭胚、北京鸭胚、SPF鸡胚的致死率分别为 10 0 %、96 %、2 8%和 0 ,致死鸭胚的肝脏、脾脏表面见白色坏死点。经蛋传试验表明 ,该株病毒有可能经胚蛋垂直传染。以该株病毒在番鸭胚成纤维细胞 (MDEF)上连续传接 10代 ,结果细胞病变 (CPE)仍不明显 ,表明其不易适应 MDEF。由此可见 ,该株雏半番鸭源呼肠孤病毒具有较强的致病力     

Estimations of repeatability and heritability of egg albumen antimicrobial activity and of lysozyme and ovotransferrin concentrations

1. The repeatability and heritability of growth inhibition by egg albumen of two major pathogenic bacteria, a Gram-negative (Salmonella Enteritidis) and a Gram-positive (Staphyloccocus aureus) and of two antimicrobial albumen proteins, lysozyme and ovotransferrin, were estimated in commercial pedigree hens. 2. Repeatability was evaluated in 100 egg-type hens at the beginning, middle and end of the laying cycle on eggs collected for 3 weeks. Heritabilities were estimated at 36 to 40 weeks of age on 400 pedigree hens (2 eggs/hen), which were the offspring of 25 sires each mated with 4 dams. Ovotransferrin and lysozyme were quantified by ELISA. Salmonella Enteritidis (S.E.) and Staphyloccocus aureus (S.A.) were inoculated into a sample of sterilised albumen and enumerated after incubation. 3. Total protein content in albumen decreased with age of laying hens, whereas there were increases in lysozyme or ovotransferrin concentrations and in the bacteriostatic effect of albumen. 4. Repeatability for bacterial growth in albumen ranged from 0.29 to 0.39 for the number of S.E. (log cfu/ml) one day post inoculation (p.i.) but was lower and more variable at 5 d p.i. or for S.A. number. It ranged from 0.27 to 0.38 for S.E. and S.A. number at the mid period of the laying cycle. Repeatabilities were low and variable for total egg albumen protein or lysozyme and ovotranferrin concentrations (0 to 0.22). 5. Negative phenotypic correlations were observed between lysozyme concentrations and S.E. number but that between lysozyme and S.A. number was not significant. 6. Heritabilities were low (0.01 to 0.09) for protein traits. They were 0.11 for S.A. number and 0.16 for S.E. number one day p.i. 7. It appears to be more efficient to select on global bacterial growth than on specific antimicrobial proteins. The most promising trait is the number of S.E. one day p.i.     

Double-yolked pheasant eggs provide an insight into the control of albumen secretion in bird eggs

1. The possible role of the presence of the yolk in stimulating secretion of albumen was investigated. 2. Double-yolked and single-yolked pheasant (Phasianus colchinus) eggs were opened to determine the masses of the shell, albumen and yolk(s). 3. In double-yolked eggs, the two yolk masses were not significantly different. Albumen mass was increased above that expected from an egg with a single-yolk of comparable size but below that expected from an egg having a mass of the combined yolks. The mass of shell per unit area reflected the mass of the initial mass of the egg irrespective of the number of yolks. 4. The additional mass of albumen is unrelated to yolk or initial egg mass. It is postulated that in double-yolked eggs the oviduct is mechanically stimulated by the presence of both yolks, which empties the stores of water-soluble albumen proteins in the magnum wall. Such stores are insufficient to provide the same amount of protein for the two yolks.