Measuring the eggshell strength of 6 different genetic strains of laying hens: techniques and comparisons.

1. Eggshell quality was compared in 6 different strains of laying hens. Three strains were commercial; the three others were experimental. 2. Four different variables describing the strength of eggshells were investigated. Three of them were the classical ones eggshell thickness, shell stiffness measured during quasi-static compression and breaking force. Dynamic stiffness, introduced by Coucke (Ph.D. Thesis, KU Leuven, 1998), was the 4th. The fact that this measurement is dynamic could be helpful in genetic selection for eggshell breakage, because forces applied to the egg in practice are dynamic, rather than static. 3. Hisex White hens produce eggs with the strongest eggshell, in terms of all 4 eggshell variables. However, their shell quality in terms of breaking force did not remain constant over the laying period, unlike 4 other strains. 4. All strains showed a decline in quasi-static stiffness over time. 5. The eggshell thickness of three strains showed a decline over time. 6. Dynamic stiffness remained constant or improved in all strain. 7. All variables describing the mechanical eggshell strength gave different information.

Measuring the mechanical stiffness of an eggshell using resonant frequency analysis.

1. A new method was introduced for the measurement of mechanical eggshell characteristics based on the analysis of the vibration response of a chicken egg excited with a non-destructive impact. Four major topics were examined. 2. The 3 dimensional vibration pattern of an intact chicken egg was described for the spherical vibration with the lowest resonant frequency. 3. The resonant frequency of this basic vibration mode and the total egg mass were used to calculate a dynamic stiffness value of the eggshell based on a mathematical mass-spring model. 4. A correlation analysis between this dynamic stiffness value and several common egg and eggshell variable was calculated. Pearson correlation coefficients between the dynamic stiffness and static stiffness, eggshell thickness at the equator, eggshell width and shape index were all significantly different from zero and were 0.71, 0.60, 0.43 and 0.51, respectively. 5. Graphical analysis of the static vs. the dynamic eggshell stiffness indicated that eggs with thick shells (>360 microm) had a higher static stiffness value than the dynamic stiffness measurement, whereas the static stiffness of eggs with thin shells (>300 microm) was lower than the dynamic stiffness. 6. The new vibration test method is a promising alternative for evaluating mechanical eggshell properties because of its non-destructive nature and brief measuring time.

The regulation of GH-dependent hormones and enzymes after feed restriction in dwarf and control chickens.

The principal objective of this study was to examine the GH-dependency of IGF-I and IGF-II changes in the chicken. To this end, the regulation of GH-dependent hormones and enzymes were studied in undernourished normal and dwarf chickens. The dwarf chickens examined exhibit a Laron-type dwarfism and have been shown to be GH receptor deficient. Thus, they provide an interesting model to determine the GH-dependency of IGF-I and IGF-II changes. Short (1 day) and long-term (7 days) feed restriction was imposed on growing normal and dwarf chickens to follow the subsequent endocrine changes. Since short-term feed restriction of dwarf chickens resulted in decreased plasma IGF-I, it appears that this is not a GH-dependent effect. However, with longer term undernutrition, IGF-I was not decreased in dwarf chickens. So, after a longer restriction period, the regulation of these factors appears to become more GH-dependent. IGF-II was not depressed at all by feed restriction in the dwarf chicken, suggesting a degree of GH-dependency.

Effect of various adjuvants on secondary immune response in chickens.

Stimulatory effects of several types of adjuvants on secondary antibody response to inactivated Newcastle disease virus (iNDV) were examined in chickens. For this purpose, animals were primed with iNDV without adjuvant resulting in a low but significant antibody response, boosted with iNDV plus adjuvant 3 weeks later, and analysed for specific antibody titres in serum 3 weeks after the booster. Water-in-mineral oil emulsion (W/O) caused significant increase in antibody titres measured in an indirect enzyme-linked immunosorbent (ELISA), haemagglutination inhibition (HI), and virus neutralisation (VN) assay. The adjuvants tested included three oil-in-water emulsions (i.e. mineral oil-in-water, sulpholipo(SL)-Ficoll400/squalane-in-water and sulpholipo-cyclodextrin/squalane-in-water), three negatively-charged polymers with high molecular weight (i.e. polyacrylate, polystyrenesulphonate and sulpho(S)-Ficoll400) and two surface-active agents (i.e. dimethyldioctadecylammonium bromide (DDA) and Quil A). These adjuvants enhanced significantly the secondary immune response but none reached the titre obtained with W/O. Combinations of adjuvants with distinct physicochemical properties, i.e. polyacrylate and DDA revealed only slight, beneficial effects. We concluded that the various types of adjuvants tested can stimulate secondary immune responses in primed animals but that W/O is superior.

Alkyl-esters of polyacrylic acid as vaccine adjuvants.

Previously, we demonstrated that polyacrylic acid (PAA) augmented significantly the immune response to inactivated Newcastle disease virus (iNDV) in chickens, but that efficacy was insufficient to replace the water-in-mineral oil (W/O) adjuvant applied for boosting primed animals. Attempting to improve its adjuvanticity, PAA with weight-average molecular weight (Mw) of 450 kDa was grafted with alkyl-chains by esterifying the carboxylic groups with octanol and butanol. The butyl-PAA and octyl-PAA esters obtained varied in degree of esterification between 10% and 92%. Adjuvant activity of water-soluble esters for humoral responses to iNDV was examined in chickens primed previously with iNDV without adjuvant. The alkyl-PAA esters exhibited significantly higher responses than unmodified PAA and titres increased with increasing dose of adjuvant. At doses of 2 mg per animal, octyl- and butyl-PAA esters with a substitution rate of 16% (octyl16-PAA and butyl16-PAA, respectively) gave similar titres as W/O. In aged animals primed with live NDV at early age, butyl16-PAA and W/O elicited comparable antibody responses. Butyl16-PAA was also more effective than PAA in stimulating primary immune responses in mice which was accompanied by stronger local reaction determined by monitoring swelling at the site of injection. Reactogenicity of butyl16-PAA was less than of W/O. We concluded that alkyl-PAA esters are strong adjuvants for primary and secondary responses and that they are promising alternatives to the mineral oil-based adjuvants presently used in various veterinary vaccines.

In ovo treatment with an aromatase inhibitor masculinizes postnatal hormone levels, abdominal fat pad content, and GH pulsatility in broiler chickens.

Vorozole, a selective aromatase inhibitor, was administered in ovo to test the specific embryonic role of estrogen in conferring the sex distinction in GH release and body phenotype in broilers. On Day 6 of incubation, eggs were injected with saline or with different concentrations of vorozole. Postnatal blood samples were analyzed for T3, T4, GH, estradiol (E2), and testosterone (T). At the age of 4 wk, control and vorozole-treated birds were cannulated, and serial blood samples were withdrawn every 10 min for 5 hr, wherein GH pulsatility characteristics were determined using deconvolution analysis. The proportional abdominal fat pad weight was reduced significantly in the treated groups, especially in female birds. The vorozole treatment increased plasma T3, E2, T, and GH concentrations, and decreased T4. The frequency of the GH pulses was lower and the interval between the bursts (min) was higher in the vorozole-treated group, as were the mass secreted per burst (ng/ml), the amplitude (ng/ml/min) and the production rate (ng/ml/5 hr). In conclusion, early in ovo treatment with a potent aromatase inhibitor is able to increase the mean serum T3 and GH concentration and masculinize the GH pulse pattern, resulting in an economically favorable decrease in abdominal fat pad content in male and female broilers at slaughter age.

Different CO2 levels during incubation interact with hatching time and ascites susceptibility in two broiler lines selected for different growth rate.

Embryos from ascites sensitive (AS) and resistant lines (AR) were subjected during the third week of incubation to a high CO2 (0.4%) (HC) or normal CO2 level of 0.2% (NC) in the incubators. The effect on embryonic and hatching parameters and growth, ascites sensitivity and related physiological parameters was followed. At NC conditions AR embryos hatch earlier than AS ones but this difference completely disappeared under HC condition, since hatching time of the AS line was reduced. Moreover, AS embryos showed slightly lower T4 and T3 levels than AR embryos. Embryos of both lines incubated at HC had higher plasma T3 concentrations than those incubated at NC. Chickens that had been incubated at HC level showed less ascites mortality than those incubated at NC. RV/TV ratios were higher in NC birds compared with HC ones. It is concluded that different ventilation levels during incubation interact with total incubation time and thereby influence ascites susceptibility.

The prophylactic effect of vitamin C supplementation on broiler ascites incidence and plasma thyroid hormone concentration.

In a two-factorial experiment, 420 1-day-old male commercial broiler chickens were randomly divided and fed a basal diet with or without vitamin C (500 parts/10(6)). Half of the birds within each group receiving the same kind of feed were fed a T(3 )supplemented diet from day 1, in order to increase the ascites incidence. Weekly body weight and feed intake were measured. Venous blood samples were taken from 10 birds per group weekly from day 14 on, for haematocrit measurements, blood gas analysis, measurement of thyroid hormones and lactate/pyruvate ratios. Ascites mortality was recorded daily. Dietary T(3) significantly increased ascites mortality. Adding vitamin C to the feed significantly reduced ascites mortality while it had no effect on performance parameters. Plasma thyroid hormone levels were significantly reduced by dietary vitamin C administration, but neither haematocrit nor blood gas levels were influenced.

Different characteristics in chick embryos of two broiler lines differing in susceptibility to ascites.

1. The relationship between the length of incubation, hatching, hypoxic condition, thyroid hormones and the occurrence of ascites were studied in embryos of 2 broiler lines differing in susceptibility to the ascites syndrome. 2. Both the time of external pipping (ep) and hatching of embryos from the ascites-resistant (AR) broiler line was earlier compared to the ascites-sensitive line (AS). The interval between internal pipping (ip) and ep was the same between the lines, but the interval between ep and hatching was shorter in the resistant line. 3. The T3 and T4 concentrations in plasma of the AS line were lower compared to the AR line. 4. Analyses of partial pressures of oxygen (pO2) and carbon dioxide (pCO2) in the air cell of the egg revealed that at day 18 the AS embryos had lower pO2 and higher pCO2 concentrations compared to the AR embryos. 5. The delay in hatching of the AS embryos might induce a more pronounced and/or extended hypoxic environment, thereby creating an environment that evokes ascites.

Partial food restriction increases hepatic inner ring deiodinating activity in the chicken and the rat.

The effects of a long term partial food restriction were studied in chickens and rats. In chickens the treatment resulted in increased plasma T4 levels while T4 levels in rats remained unchanged. Plasma T3 decreased in both species. In vitro hepatic outer ring deiodinating type I (ORD-I) activity was not influenced by the food restriction, suggesting that the amount of ORD-I enzyme present in the liver remained unchanged. In vitro hepatic inner ring deiodinating type III activity, on the contrary, was greatly increased in both species. This increase may contribute to the decreased circulating T3 levels by increasing the degradation of T3 and diverting the deiodination of T4 to rT3 instead of to T3.

Kinetic parameters of plasma thyroid hormone and thyroid hormone receptors in a dwarf and control line of chicken.

Sex-linked dwarf chickens have in plasma low triiodothyronine (T3) levels and slightly raised thyroxine (T4) concentrations and are functional hypothyroid. The kinetic parameters of T4 and T3 were investigated using 125I-labeled hormones. In addition the nuclear T3-receptors in the liver were examined using a radioreceptorassay and Scatchard analysis. Four-week-old dwarf (dw) and normal (Dw) chickens were injected with 125I-labeled T3 or T4 and blood samples taken 60, 120, 180, and 300 min after 125I-T3 injection and 120, 240, 360, and 480 minutes after 125I-T4 treatment. Labeled T4 and T3 and the degradation products were separated by paper chromatography. After the paper strips were dried, the iodinated compounds were visualized and counted in the gamma counter. The kinetic parameters, the half-life time (T1/2), the apparent distribution volume (Vd) and the metabolic clearance rate (MCR) were calculated using the natural base logarithm values of the measured radioactivity plotted against time and used for linear regression. T4 was cleared from circulation more slowly in dwarf than in control chicks and reflected a longer T1/2 (21.8%) and a reduced MCR (45%). The Vd tended to be lower (34.7%) in dwarfs. While the T1/2 of T3 was longer (28.1%) in dwarf chickens than in control animals, the MCR for T3 was considerably increased (31.8%). This results from an increased Vd (63.1%) in the dwarf chicks. The T3-receptor study in the liver of dwarf and non-dwarf chickens from Week 1 to Week 4 posthatching revealed that the total capacity and the affinity constant of the binding sites were comparable in dwarf and normal chickens. However, the occupancy of the receptors was higher in the dwarf animals.

Ontogeny of the liver nuclear T3-receptor during the last days of incubation and posthatch in the chick embryo.

The characteristics of the nuclear T3 receptor in the liver of the chick embryo were studied from incubation day 18 until day 1 posthatching. Treatment of the nuclei with 3 mol.l-1 MgCl2, which removed the endogenously bound hormone, was used in order to determine the total amount of receptors. The affinity constant Ka decreased between incubation day 18 (0.996 +/- 0.276.10(9) M-1) and day 19 (0.247 +/- 0.072.10(9) M-1), remained the same thereafter until hatching and increased again on day 1 posthatching (1.846 +/- 0.928.10(9) M-1). The total amount of receptors tended to increase from incubation day 18 to day 20 non-pipping (np) (from 4.40 to 11.55 fmol/micrograms DNA) and decreased thereafter to 2.38 fmol/micrograms DNA on day 1 posthatching. The amount of free binding sites reached a maximum on day 19 (6.91 fmol/micrograms DNA) and then decreased drastically until posthatching (0.19 fmol/micrograms DNA). The maximal specific binding was found on day 20 (np), just prior to penetration of the air chamber. During the time at which the level of T3 remains high in the plasma, a reduction in the amount of receptor was observed, which may be the consequence of a down-regulation by T3 itself.

Food intake after hatching inhibits the growth hormone induced stimulation of the thyroxine to triiodothyronine conversion in the chicken.

The effect of a single injection of 10 micrograms chicken GH on circulating thyroid hormones as well as in vitro liver 5'-monodeiodination (5'-D) activity was studied in posthatch chicks submitted to different feeding conditions. One group was normally fed after hatching, a second group was only fed after three days and a third group was food deprived after 2 days of feeding. Combination of all results indicates that the start of food intake abolishes the stimulatory effect of a GH injection on circulating T3 and liver 5'-D activity. Food deprivation after a period of food intake restores the GH effect on plasma T3 but not on liver 5'-D.

Thyrotropin-releasing hormone (TRH) is not thyrotropic but somatotropic in fed and starved adult chickens.

Adult fed and starved Warren chickens, 2 yr of age, and approaching the end of the second laying year, were injected iv with 1 of the following products: 10 micrograms of thyrotropin releasing hormone (TRH); 100 micrograms of bovine thyrotropin (bTSH); 100 micrograms of ovine growth hormone (oGH); saline. The influence on plasma concentrations of thyroxine (T4), triiodothyronine (T3) or chicken GH (cGH) were followed. Prior to injection, it was clear from the control values that starvation for 3 d decreased plasma levels of T3 and increased cGH, whereas 7 d of fasting increased T4 and cGH. The plasma levels of cGH were elevated greater than 10-fold at 15 min following the TRH challenge in food-deprived chickens compared to a less than 4-fold increase in normal fed hens. This increase was followed by a rise in T3 after 1 h, which was also more pronounced in the starved animals, whereas T4 decreased or remained unaffected. Increases in T4 can, however, be obtained with 100 micrograms TSH in normal fed (2-fold) or starved animals (greater than 3-fold). Following injection of 100 micrograms oGH, a significant increase in T3 levels was observed which in fed animals was already present at 30 min, but the higher levels persisted for 1 and 2 h in fed and starved hens. At the same time, a decrease in T4 was observed in both groups of GH-treated chickens. It is concluded that TRH at the dose used is not thyrotropic but has a somatotropic effect and is responsible for the peripheral conversion of T4 into T3.