Protein synthesis measured in the whole body of chick embryos cultured in vitro

1. Whole body protein synthesis was measured in chick embryos cultured in vitro. On day 7 of incubation chick embryos were cultured for 60 min in synthetic serum‐free medium containing 4‐[³H]phenylalanine. Specific radioactivities in free and protein‐bound phenylaline in the whole embryo were measured, starting 2 min after commencement of the culture process.

2. The values for fractional synthesis rate (FSR) estimated in vitro at 20, 30, 45 and 60 min during the embryo culture agreed well, ranging from 35 to 40%/d, suggesting that the method would serve as a useful model for studying the effect of growth promoters in chick embryos.

3. Bovine insulin in the synthetic medium did not affect FSR of protein in chick embryos cultured in vitro.     

Hormone-mediated maternal stress affects embryonic development during incubation without adverse effect on chick weight and body composition

1. The objective of this study was to evaluate the effect of maternal stress (MS) induced by supplementing the hen’s diet with 2 mg/hen/d dietary corticosterone (CORT) on embryonic development, biochemical blood parameters and hatching performance of broiler chicks.

2. A total of 200 Ross broiler breeder hens at 42 weeks of age were randomly divided into two groups: MS or control. Hens in the MS were fed 2 mg/hen/d CORT for 14 d. Eggs (648 and 635 eggs for MS and control, respectively) were collected from d 3 to 14 of dietary CORT supplementation and incubated. Weights of embryo, chicks and organs and body composition were determined during incubation and at hatch. Biochemical blood parameters were measured at internal pipping stage and day of hatch. Hatching performance and embryonic mortalities were recorded.

3. Hens fed a diet supplemented with CORT had lighter body weight and produced less eggs at the end of the 14-d treatment period. Although MS embryos were heavier than control from 12 to 18 d of incubation, chick weight was similar at the day of hatch. Lower relative weights for yolk sac and bursa were observed at 12 d of incubation for MS chicks compared to control. Chicks from both groups had similar body content in spite of higher fat content of MS embryos on d 18 of incubation.

4. MS had no effect on the duration of incubation or hatching performance but increased mortality at the pipping stage.

5. The results suggest that hormone-mediated MS might affect embryonic development during incubation without adverse effect on chick weight and body composition.     

Injection of porcine growth hormone releasing hormone gene plasmid in skeletal muscle increases piglets' growth and whole body protein turnover

The aim of the current study was to investigate the effects of a porcine growth hormone releasing hormone (pGHRH) gene plasmid injection in piglets on growth performance and whole body protein turnover. Sixty male Canadian Landrace × Chinese Taihu piglets were assigned to an intramuscular injection of 0 (control), 0.25, 0.5, 1 and 2 mg. All pigs were fed with the same diet (crude protein: 239.8 g/kg, digestible energy: 14.28 MJ/kg) at ad libitum intake. Protein turnover was determined on the 22nd day with a three-pool model by using a single-dosage, end-product analysis method with ¹⁵ N-glycine as a tracer. Injection of the pGHRH gene plasmid increased the piglets' growth rate, altered feed intake and decreased feed conversion ratio. It increased plasma growth hormone releasing hormone (GHRH), growth hormone (GH), insulin-like growth factor-I (IGF-I) and somatostatin but reduced serum urea and triglyceride. It reduced the urinary nitrogen excretion and led to higher nitrogen retention as well as the efficiencies of nitrogen retention and digestible N utilization. It increased the rates of protein synthesis, protein breakdown and net protein gain. Excretion of endogenous urinary nitrogen was reduced and nitrogen reutilization rate was improved. Conclusions: Injection of the pGHRH gene plasmid in skeletal muscle stimulated GHRH, GH and IGF-I excretion in piglets. Protein deposition was increased by an increase in protein synthesis and a smaller increase in protein breakdown, which was accompanied by reducing amino acid oxidation and increasing nitrogen reutilization.     

Eye variability in chick embryos

1. In two strains of Barred Plymouth Rock and one strain of Rhode Island Red birds a highly significant effect of strain was found on the size of eye cups and pupils in 6‐d‐old chick embryos. The average values of these eye characteristics and their variations were also influenced by the composition and energy content of the diet fed to the dams.

2. A strong positive phenotypical relationship was found between the size of eye cups and pupils, especially between the size of the left and the right eye cups. No relationship was found between the eye characteristics and the weight of egg.

3. An asymmetry of the left and the right eye was recorded; the left eye cups and pupils were smaller than the right on average by 2.2 and 3.9%, respectively.     

Macromolecular composition of fast, slow and cardiac muscles of chick during incubation.

Total muscle mass increases about five-fold in fast (anterior latissimus dorsi: ALD) and slow (posterior latissimus dorsi: PLD) muscles and 30-fold in cardiac muscle between the 12th and 20th d of incubation. 2. Increase in muscle protein does not parallel the rise in muscle RNA. 3. The DNA concentration of the ALD muscle decreases while that of the heart and PLD muscle show an increase between 8 and 12 d and a steady decrease thereafter. 4. The increase in glycogen concentration is greater in heart than in ALD and PLD up to the 20th d; 1 d after hatching, ALD has more glycogen than PLD.     

Higher levels of CO2 during late incubation alter the hatch time of chicken embryos

1. It has been reported that the increasing CO₂ tension triggers the embryo to pip the air cell and emerge from the egg. However, the mechanism by which higher CO₂ concentrations during the last few days of incubation affect chick physiology and the hatching process is unclear. This study investigated the effect of CO₂ concentrations up to 1% during pipping, on the onset and length of the hatch window (HW) and chick quality.

2. Four batches of Ross 308 broiler eggs (600 eggs per batch) were incubated in two small-scale custom-built incubators (Petersime NV). During the final 3 d of incubation, control eggs were exposed to a lower CO₂ concentration (0.3%), while the test eggs experienced a higher CO₂ concentration programme (peak of 1%).

3. There were no significant differences in blood values, organ weight and body weight. There was also no difference in hatchability between control and test groups. However, a small increase in the chick weight and the percentage of first class chicks was found in the test groups. Furthermore, plasma corticosterone profiles during hatching were altered in embryos exposed to higher CO_2; however, they dropped to normal levels at d 21 of incubation. Importantly, the hatching process was delayed and synchronised in the test group, resulting in a narrowed HW which was 2.7 h shorter and 5.3 h later than the control group.

4. These results showed that exposing chicks to 1% CO₂ concentration during pipping did not have negative impacts on physiological status of newly hatched chicks. In addition, it may have a significant impact on the physiological mechanisms controlling hatching and have benefits for the health and welfare of chickens by reducing the waiting time after hatching.     

鸡包涵体肝炎鸡胚肝细胞包涵体特性的研究

应用鸡包涵体肝炎病毒 FAV- Hb株试验感染 SPF鸡胚 ,通过透射电镜对感染鸡胚肝脏的观察 ,表明 FAV- Hb株可引起鸡胚肝细胞内形成三种类型的包涵体 ,即非病毒性中等电子密度包涵体 ,病毒性包涵体和非病毒性高电子密度包涵体 ,各型包涵体均可见于胞核或胞浆内 ,但胞核是包涵体首先形成的部位 ,核内包涵体通过核膜进入胞浆。各型包涵体在其形成过程中有较密切的关系。     

Whole body protein turnover of growing rats in response to different dietary proteins--soy protein or casein.

Whole body protein turnover was studied in growing rats fed restrictively on isoenergetic (GE 17.6 MJ/kg DM) and isonitrogenous (104 g CP/kg DM) diets based on soy protein isolate or casein supplemented with D,L-methionine. During each of the three separate experiments six male Fischer rats per group were housed individually in metabolic cages at 24 degrees C. Prefeeding of both dietary groups up to similar body weights at the start of the main experimental periods (105-134 g) lasted up to 16 d for casein-fed rats and up to 30 d for the soy protein-fed rats. Following the energy and nitrogen balance periods whole-body protein synthesis was estimated by the end-product method using a single tracer dose of a mixture of 15N-labelled amino acids. Fractional protein accretion rate [% of the protein pool accreted per day] was significantly lower in soy protein-rats than in casein-fed rats in all three experiments whereas fractional synthesis rate was not significantly lower. Therefore, protein breakdown subsequently calculated as the difference between synthesis and accretion showed a tendency towards higher values in this group. In soy protein-fed rats also a tendency towards higher excretion of 3-methylhistidine as a marker of myofibrillar protein breakdown was observed. It is concluded that increase in lean tissue growth resulting from improved protein quality is brought about by changes of both rates, by small increase of protein synthesis and by reduced rate of body protein breakdown.     

Investigation of mRNA expression for secreted frizzled-related protein 2 (sFRP2) in chick embryos

The roles of secreted frizzled-related protein 2 (sFRP2) in organ development of vertebrate animals are not well understood. We investigated expression of sFRP2 during embryogenesis of Arbor Acre broiler chicken eggs. Expression of sFRP2 was detected in the folds and lateral layer of developing brains. The sFRP2 signals in the developing eye were marked as a circle along the orbit. In younger embryos on days 3-6, the sFRP2 signals were consistent with growth of the sclerotome, suggesting that sFRP2 may be associated with somite development. Furthermore, with the exception of bones, sFRP2 mRNA was detectable in the interdigital tissue of embryos older than eight days as the limbs matured. This revealed that sFRP2 might play a role in myogenesis. In situ hybridization was also used to analyze the expression of sFRP2 in day 3-10 chick embryos. Signals were expressed in the gray matter of the developing brain coelom, including the optic lobe, metencephalon, myelencephalon, mesencephalon and diencephalon. The developing eyes contained an intercellular distribution of sFRP2 in the pigmented layer of the retina and photoreceptors. Furthermore, sFRP2 was expressed in the mantle layer of the neural tube and notochord. Based on these findings, it seems reasonable to suggest that sFRP2 may play an active role in embryogenesis, especially in development of the neural system, eyes, muscles and limbs.     

胚胎肌细胞生成期间提高和降低孵化温度影响肉鸡胚胎肌肉的生长和能量代谢

正正如一些论文中所报道的那样,肉鸡种蛋的胚胎发育可通过改变外部因子来进行调节,如孵化温度。肉鸡胚胎肌细胞的形成分别以胚胎发育期(Embryonic Day,ED)的第3~8天(ED 3~ED 8)和第8~14天(ED 8~ED 14)期间形成的初级和次级肌小管为特征。研究表明,在此阶段提高孵化温度,会影响胚胎和出壳后的动物生长发育(Maltby等,2004;Hammond等,2007;     

Pulmonary ventilation in a population of hatching chick embryos

Two experiments were made to study pulmonary ventilation in the hatching chick embryo with particular reference to the part played by the air cell.

The hatchability of embryos without access to the air cell was the same as for normally positioned embryos. The lungs of about 35 per cent of the normally hatching embryos were inflated in the air cell, but 65 per cent were inflated after breathing atmospheric air. The shell was cracked but not pipped before the outer membrane was penetrated. Comparisons between different groups of normally and abnormally positioned unhatched embryos revealed that the air cell had an insignificant respiratory value, but that the mechanical advantages of the large end seemed apparent.

The time relationships between lung inflation, inner membrane perforation, pipping, yolk sac retraction, outer membrane perforation, lung discoloration and hatchability were studied. Equations were derived for the regression on time of each of the seven variables studied. Statistically insignificant differences in hatchability were observed between the 2 groups of embryos which hatched from the large or small end of the egg.

It is suggested that lung inflation occurs when a certain threshold in the respiratory movements of thoracic muscles is reached. The threshold is attained by the rising anoxia, and probably other stimuli. The chick embryo's lungs, which lack elastic recoil, respond to the threshold and start utilising air. It is doubtful whether the embryo needs the very high partial pressure of carbon dioxide in the air cell. It is feasible to believe that CO₂ is an inevitable metabolic by‐product, and that it is stored in the air cell. The air cell also serves as a resting place for the chick while retracting the yolk sac. Through evolution, however, the chick embryo may have built up a high physiological tolerance to the CO₂ which it encounters upon entering the air cell.     

Gastrointestinal emptying in the final days of incubation of the chick embryo

1. The gastrointestinal (GI) emptying of the chick embryo was studied in days 19 and 20 of incubation using radioactive (14C-PEG-4000) and coloured (Evans Blue dye) tracer markers. 2. Following administration by oral cannula, the percentages of tracer marker recovered in the yolk sac and the GI tract showed the existence of a GI motor pattern in both 19 d and 20 d chick embryos and suggested that the fluids drunk by the chick embryo during the final days of incubation could have reached the yolk sac from the GI tract via the vitelline diverticulum.     

Methodic recommendations for the determination of whole body protein synthesis rates in tracer studies

Methodical recommendations are suggested--predominantly for laboratory and small animals (rats and young chickens)--for the determination of parameters of the protein metabolism of the whole body after single doses of a mixture of 15N labelled amino acids by means of the determination of the temporal course of cumulative 15N excretion in urine and the assessment of the tracer kinetic data in a compartment model. These recommendations are to make it possible to carry out purposefully such experiments under comparable conditions. The advantages of this method are: the non-invasive character of the method; the possibility of repeating the experiment with the same animal; the adaptability to other methods of investigation (e.g. measuring energy metabolism); the relatively low expenditure of labour and requirement of test animals; the relatively good reproducibility of the method. Thus this method is a good supplement to the flooding and permanent infusion methods and should be used wherever the determination of parameters of the protein metabolism of the total body is sufficient.