Effect of incubation temperature on sex-dependent embryo mortality and morphological traits in Mallard.

Although birds have genetically determined sex, the sex ratio has been reported to deviate from parity in several studies. Temperature-dependent sex determination, which is common in reptiles, is absent in birds. However, females are able to adjust their investment into eggs according to the sex of the embryo, which may cause sex-specific embryonic mortality. Incubation temperature may also cause sex-biased embryonic mortality, and it may differentially affect the phenotype of male and female hatchlings. We aimed to investigate differences between male and female Mallard embryos regarding their egg size, mortality during incubation and hatchling phenotype in relation to incubation temperature. Mallard eggs were incubated under six constant incubation temperatures (ranging from 35.0 to 38.0 °C). Hatchlings were weighed, and their morphological traits were measured. We determined the sex of hatchlings and unhatched embryos by genetic analysis and found higher male embryonic mortality at 35.5 °C (44 males vs. 28 females) and a higher proportion of female hatchlings at 38 °C (24 males vs. 38 females); however, these results were not statistically significant. Our results suggest that Mallard females do not differentiate quantitatively between sexes during egg production. Male hatchlings were significantly larger but not heavier than females. The size difference between sexes was most pronounced at temperatures around 36 °C, which is the mean temperature of naturally incubated Mallard eggs.

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.

The effect of gaseous ozone treatment on egg components.

BACKGROUND: Because of the salmonella problem in poultry, disinfection technologies are necessary. Ozone is a strong oxidant used for the disinfection of surfaces, drinking water and foods. However, since ozone not only destroys bacteria but may also damage eggs, it is necessary to clarify the effects of ozone treatment on hatching egg components. In this study, doses of gaseous ozone ranging from 10 to 50 mL L(-1) were tested. The vitamin A and E contents and fatty acid composition of the egg yolk were determined. To detect possible damage to the DNA of the germ disc, single-cell gel electrophoresis was used. Moreover, free SH groups were measured in the egg white. The soluble cuticula proteins were analysed by polyacrylamide gel electrophoresis. RESULTS: The yolk was not significantly affected by ozone treatment. However, the DNA of the germ disc was attacked and a significant decrease in free SH groups in the egg white was recorded at 50 mL L(-1) ozone. Even at low ozone doses the soluble cuticula proteins were completely destroyed. CONCLUSION: Significant alterations of egg components were caused by 50 mL L(-1) ozone. At lower ozone doses the oxidative processes occurred mainly at the egg surface and are therefore probably harmless to the developing embryo.

The eggshell: strength, structure and function.

In making a journey through the literature of the last 50 years one can easily highlight a sequence of seminal works-but the route has not been direct and to avoid the many profitable diversions and detours that have enriched and deepened our collective understanding of the subject of eggshell structure and function is to do the subject a serious disservice. This is a route march of science enabled by advances in technology.

An appraisal of nuclear organisation in interphase embryonic fibroblasts of chicken, turkey and duck.

Determining the nuclear 'addresses' of chromosome territories is a well-documented means of assaying for nuclear organisation in many cell types and species. Data in avian species are however limited at best, despite the pivotal role played by birds (particularly chickens) in agriculture, and as model organisms in developmental biology. That is, studies have hitherto focussed mostly on mammals (especially humans) and have demonstrated the importance of chromosome territory positioning in embryology, disease and evolution. Thus a detailed study of nuclear organisation in many species, many cell types and many developmental stages in birds is warranted, however, before this is achieved, 'baseline' needs to be established to determine precisely the relative locations of chromosome territories in at least 1 cell type of at least 1 bird. With this in mind we hybridised FISH probes from chicken chromosomes 1-28 to embryonic fibroblast nuclei, determining nuclear addresses using a newly developed plug-in to the image analysis package ImageJ. In our experience, evenly spaced representative BAC clones yielded more consistent results than hybridisation of chromosome paints. Results suggested that chromosome territory distribution best fitted a chromosome size-based (rather than gene density-based) pattern. Identical BAC clones were then hybridised to turkey and duck in a comparative genomic strategy. Observations were consistent with those seen in chicken (although, less well-defined in duck), providing preliminary evidence of conservation throughout evolution.

On the histogenetic potency of the tailbud mesoderm.

The caudalmost part of the tailbud mesoderm (terminal paraxial tailbud mesoderm) does not develop into somites. It is not clear whether this terminal paraxial tailbud mesoderm can be considered to be a part of the segmental plate. To elucidate the nature of the tailbud mesoderm, grafts containing caudal somites, caudal prospective somitic mesoderm and the terminal paraxial tailbud mesoderm were grafted from quail embryos into the wing bud mesoderm of chick embryos. The distinct nuclear difference between quail and chick allows the identification of the grafts on a cellular level. The grafts containing caudalmost somites and the prospective somitic mesoderm differentiate into muscle and cartilage. The terminal paraxial tailbud mesoderm, on the other hand, did not give rise to either of these tissues. From this it can be concluded that the terminal paraxial tailbud mesoderm cannot be considered to be a part of the segmental plate.

On the origin of cells determined to form skeletal muscle in avian embryos.

Pieces of quail embryos from various developmental stages ranging from unincubated blastoderms (before the appearance of a primitive streak) to embryos having formed somites were grafted to the wing buds or into the coelomic cavity of chicken embryos. The grafts, which can be identified on a cellular level by virtue of the prominent nucleolus-associated chromatin, present in the quail and absent in the chicken, were screened after suitable periods of reincubation for the presence or absence of skeletal myotubes containing quail nuclei. Grafts having contributed to such skeletal myotubes were considered as having contained determined myogenic cells at the time of the grafting procedure. Determined myogenic cells appeared first in the primitive streak and in the mesodermal cells formed by the invagination (gastrulation) of epiblastic cells through the primitive streak. This is true for both the head process and the paraxial mesoderm. Epiblastic cells never gave rise to skeletal myotubes. Therefore it can be said, that the onset of myogenic determination coincides with gastrulation. It remains, however, to be established, whether these two events are causally related to one another.

On the differentiation and migration of some non-neuronal neural crest derived cell types.

Neural tubes containing premigratory neural crest cells from head and trunk levels as well as somites containing neural crest cells that have migrated away from the neural crest were grafted orthotopically and heterotopically from quail embryos to chicken embryos. Schwann cells and melanocytes of donor origin developed after all grafting procedures. Cartilage developed only from neural crest cells of head levels. No skeletal muscle was ever observed to develop from the neural crest. The development of these different cell types from heterotopically grafted premigratory neural crest cells indicates that the neural crest is not a population of pluripotent undeterminated cells, but that at least some determinated cells are present within it before the onset of emigration of neural crest cells from the neural crest. Different neural-crest-derived cell populations exhibit different migratory behaviour: After heterotopically grafting quail neural crest cells to the wing buds of chicken embryos, Schwann cells and non-epidermal melanocytes were found to have migrated proximally and distally away from the grafts. Epidermal melanocytes of donor origin were found to have migrated in a distal direction essentially.

Fertilization and early embryonic development in poultry.

There are two major controllers of development in the early stages of bird embryos. These are: 1) gravity, probably acting through the distribution of yolk and its components, which lays down the initial plans for polarity that are later established firmly through the genes; and 2) the primitive streak, which controls the orderly ingression of the cells and imposes a pattern on the developing tissues.

25-hydroxycholecalciferol in poultry nutrition.

Vitamin D is a complex of secosteroids that must undergo metabolic alterations to reach optimal biological activity. The parent compounds 1) ergocalciferol (D2) and 2) cholecalciferol (D3) can be synthesized in the leaves of many plants or in the skin of most animals, respectively. Transport of vitamin D steroids after absorption is associated with vitamin D binding proteins (DBP). In general, the relative binding affinities of the vitamin D steroids are: 25-hydroxy vitamin D3 [25-(OH)D3] = 24,25-dihydroxy vitamin D3 [24,25-(OH)2D3] = 25,26-dihydroxy vitamin D3 [25,26-(OH)2D3] > 25-hydroxy vitamin D2 (25-(OH)D2) > 1,25-dihydroxy vitamin D3 [1,25-(OH)2D3] > vitamin D3. The DBP in poultry does not bind D2 forms effectively, and therefore poultry can not use this form of vitamin D adequately. The concentration of 25-(OH)D3 in blood seems to be well correlated with dietary vitamin D intake or exposure to ultraviolet light. The 1 alpha hydroxylase enzyme in the kidney is subject to negative feedback regulation and is critical for formation of the active metabolite 1,25-(OH)2D3. The intracellular vitamin D receptor (VDR) specifically binds 1,25-(OH)2D3 and is necessary for cellular action. Increased levels of two to three orders of magnitude are required for 25-(OH)D3 to compete with 1,25-(OH)2D3 for binding on VDR. Feeding studies with 25-(OH)D3 suggest it has nearly twice the activity of vitamin D3. Hatchability studies have shown that 25-(OH)D3 supports good fertility and hatchability, whereas hens fed only 1,25-(OH)2D3 did not have normal hatchability. Likewise, 1,25-(OH)2D3 seems to reach toxic levels at dietary concentrations only two to three times optimal dietary levels whereas feeding 25-(OH)D3 for extended periods at levels 8 to 10 times requirement seems to have no adverse effects. It seems that 25-(OH)D3 is the most active metabolite of vitamin D3, ultimately capable of supporting both cellular functions and embryonic development in chickens and turkeys when fed as the sole source of vitamin D3.

Genetic control of cellular infection by subgroups A and C RNA tumour viruses in guinea fowl.

An investigation was carried out in guinea fowl to determine their susceptibility to infection by Rous sarcoma viruses of subgroups A and C. A standard dose of each subgroup virus was inoculated into 14-day-old embryos via the chorioallantoic membrane (CAM). On the 10th day after inoculation, 50% of the embryonic chorioallantoic membranes were harvested to assess their infection status (CAM(+) or (-)), while the rest were allowed to hatch. The hatchabilities of the embryos inoculated with subgroups A and C were about 50% and 57%, respectively. The relative sensitivities of guinea fowl to infection by viruses of subgroups A and C were observed to be 0.220 and 0.003, respectively, as compared to chickens (1.00). Mortality due to subgroup A virus-induced liver tumours (LT) was 54% and four phenotypic subclasses, namely CAM(+) LT(+), CAM(+) LT(-), CAM(-) LT(+) and CAM(-) LT(-), were observed in guinea fowl as in chickens. However, a higher incidence (31%) of conversely associated phenotypes, i.e. CAM(+) LT(-) and CAM(-) LT(+), were observed in guinea fowl. Mortality caused by subgroup A virus-induced liver tumours was first observed in inoculated guinea fowl keets during the 3rd week after hatching, and 93% of the mortality occurred within 6 weeks. The peak mortality occurred in the 4th week after hatching. The target organs for transformation were considered to be the liver and spleen because of the equal incidence of tumours in these organs. Males and females were equally likely to die from liver tumours.(ABSTRACT TRUNCATED AT 250 WORDS)

Posture in ovo as a precursor of footedness in ostriches (Struthio camelus).

Two categories of behaviour involving lateralized posture were observed in semi-natural conditions in ostriches (Struthio camelus). Observing preferences for left or right foot, both in the forward foot posture (the foot standing in front at rest) and the starting foot used to initiate locomotion, a population-level right-foot preference was shown for the whole group and for each of the three age ranges considered (chick, young and adult). Ostriches are known to rely upon a lateralized behaviour during hatching (using their right foot to break the egg shell) suggesting the hypothesis that the precocious motor laterality observed at hatching might stand as a precursor of limb preference later in development, as already observed in other avian species.

Efeito do período e temperatura de armazenamento de ovos férteis sobre o rendimento de incubação e características de qualidade de codornas neonatas / Effect of period and storage temperature of hatching eggs from breeder quails on hatching results and quality characteristics of neonate quails

Foram estudados parâmetros de incubação e de qualidade física de codornas japonesas neonatas oriundas de ovos submetidos a diferentes condições de armazenamento. Os tratamentos constituíram-se de combinações entre temperaturas e períodos de armazenamento dos ovos férteis. O delineamento foi em blocos casualizados em arranjo fatorial 2x5, temperaturas (ambiente (28±1,5ºC) e refrigerada (14ºC)) x períodos de armazenamento (um, três, seis, nove ou doze dias), totalizando 10 tratamentos com 105 repetições, sendo cada ovo uma unidade experimental. Os ovos foram incubados nas condições de 37,9ºC e 60% de umidade relativa (UR). Houve aumento de perda de peso do ovo com o período de armazenamento, com efeito linear positivo. Observou-se maior taxa de eclosão/ovos férteis para os ovos armazenados sob refrigeração. Houve efeito linear negativo para o período de armazenamento sobre a taxa de eclosão somente para os ovos armazenados em 28ºC. Para o peso da codorna neonata, houve efeito linear negativo, independentemente da temperatura de armazenamento. O comprimento do intestino delgado em temperatura ambiente sofreu influência do período de armazenamento, sendo que o período de 12 dias apresentou pior resultado. O pior escore de qualidade física foi encontrado em codornas neonatas oriundas de ovos armazenados em temperatura ambiente por 12 dias. Pode-se concluir que, para melhorar a eclosão e a qualidade física da progênie, ovos de codorna devem ser armazenados por até nove dias e em temperatura de 14ºC.

Incubation parameters and physical quality of neonate Japanese quails hatched from eggs that were subject to different storage conditions were evaluated. The treatments consisted of a combination of temperatures and storage periods of hatching eggs from breeder quails. The experiments were conducted in a randomized block design in a factorial arrangement 2x5, with two storage temperatures (ambient temperature ± 28°C and refrigerated temperature 14°C) and five storage periods (one, three, six, nine and twelve days), totaling 10 treatments with 105 repetitions, each egg considered to be an experimental unit. The eggs were incubated at 37.9°C with 60% of relative humidity (RH). There was a positive linear effect between egg weight loss and storage period. Higher rates of hatching/fertile eggs were observed for eggs stored in refrigerated conditions. A negative linear effect was observed between storage period and hatching rate when the eggs where stored at a temperature of 28ºC. There was a negative effect for neonate quail weight, independent of storage temperatures. The intestine length was influenced by storage period and 12 days of storage showed the shortest intestine length. Lower scores of physical quality were observed in quails that hatched from eggs stored in ambient temperatures during 12 days. In this context, to improve hatching rates and physical quality of neonate quails the eggs should be stored up to nine days at a temperature of 14°C.

Influência do aquecimento artificial de ovos de matrizes pesadas sobre o rendimento de incubação / Influence of artificial heating on hatchability of broiler breeder eggs

O desenvolvimento embrionário nas aves tem início antes mesmo da postura. A variabilidade do estágio de desenvolvimento embrionário no momento da oviposição é conhecida e influencia a taxa de eclosão, uma vez que estágios muito avançados ou muito precoces são prejudiciais por tornarem os embriões mais sensíveis ao estresse do armazenamento. O aquecimento de ovos férteis no período entre a postura e o armazenamento vem sendo estudado como forma de reduzir os efeitos negativos do armazenamento sobre o rendimento de incubação por permitir que os embriões progridam até um estágio em que são mais aptos a suportar o estresse do armazenamento. Este experimento teve como objetivo avaliar os efeitos do aquecimento artificial de ovos de matrizes pesadas no período entre a coleta e o armazenamento sobre o rendimento de incubação e o peso do pinto ao nascimento. Foram utilizados 5.760 ovos de matrizes pesadas Cobb(r) com 57 semanas de idade. O delineamento experimental foi inteiramente ao acaso, constituído por quatro tratamentos definidos com base no tempo de aquecimento dos ovos (zero, três, seis e nove horas). O aquecimento foi feito em câmara de fumigação a 30°C, e os ovos foram armazenados por três dias. O aquecimento artificial no período entre a coleta e o armazenamento não influenciou a eclodibilidade, a mortalidade embrionária e o peso do pinto ao nascimento, tendo sido, nessas circunstâncias, uma prática injustificada...

In birds, embryonic development begins before laying. The embryonic development variability at the time of egg laying is known and influences hatching rate, since very early or advanced stages are detrimental for embryos because they become more sensitive to stress storage. The heating of fertile eggs in the period between posture and storage has been studied as a way to reduce the negative effects of storage on hatchability since it allows embryos to progress to a stage where they are more able to survive during storage. This experiment aimed to evaluate the effects of artificial heating of fertile broiler breeder eggs in the period between the collection and storage on hatchability and chick weight at birth. For this, 5760 eggs from Cobb(r) broiler breeders, 57 weeks old, were used. The experimental design was completely randomized. It consisted of four defined treatments based on the heating time of eggs (zero, three, six and nine hours). Heating was done in a fumigation chamber at 30°C, and eggs were stored for three days. The artificial heating in the period between collection and storage didn't affect hatchability, embryo mortality and chick weight at birth, being, in this case, an unjustified practice...

The guinea fowl spleen at embryonic and post-hatch periods.

The spleen of the guinea fowl was bean-shaped but without a dented hilus. It is supplied by three short arteries that came from the ventral surface, two on the cranial end and one at the caudal end of the organ. The whole organ had a thin but tough capsule covering the outer surface except at the point of entry of the blood vessels. By day 18 of incubation, the spleen had a thin but well-defined capsule and internal to this been complete network of sinusoids filled with erythrocytes, lymphocytes and granulocytes. By day 19, dark and light staining zones, which could be termed red and white pulps, had appeared. By day 20, the granulocytes with a lot of granules within their cytoplasm, had become the biggest-sized cells in the spleen. At day 21, arteries and veins were noticed clearly in the spleen and many lymphocytes, few granulocytes and reticular cells surrounded these. Red pulp with its sinusoids was now distinct. A giant cell containing three nuclei was seen within the red pulp. At day 1 post-hatch, the capsule was at its greatest thickness so far and muscle cells were seen at the inner most part of the capsule. Granulocytes that had been a constant feature suddenly disappeared. At day 5, the small lymphocytes had dominated the large and medium-sized ones. By 2 weeks, the red and white pulps were virtually equal in distribution but by 3 weeks, the red pulp was convincingly greater. By 7 weeks, plasma cells had appeared in the peripheral splenic cords. Monocytes were observed in the sinusoids. Two germinal centres were identified for the first time in week 13 post-hatch.

The structure of the harderian gland of the guinea fowl at embryonic and post embryonic stages.

The Harderian gland of the guinea fowl is a bright, pink and relatively large orbital organ situated at the ventromedial aspect of the orbit. It possesses a single duct that has its exit on the convex medial surface. The outline is irregular with its caudal half narrower than the rostral half, and possessing a slight constriction about the mid point. Histologically, the gland had been outlined with the existence of a large contorted lumen by day 18 of incubation. The surface epithelial lining showed two layers of cells all through and lymphocytes were also present. By day 19 of incubation, certain zones of the surface epithelium had become pseudostratified and some of these cells contained vacuoles indicative of secretory materials within their cytoplasm. By day 21, few definitive acini with lumina had been observed and at day 23, the epithelium had assumed only a single layer of cells that were tall columnar cells, except at the junctions of the folds. By day 24, the tunica propria seemed to have completely disappeared with the acini occupying every available space. Plasma cells were seen three days after hatching.

The hatching sequence of ostrich (Struthio camelus) embryos with notes on development as observed by candling.

1. The hatching sequence of the ostrich (Struthio camelus) was determined by observing embryos during candling, those in the process of hatching and in dead-in-shell eggs. 2. Candling showed that there was a progressive increase in dark shadowing within the egg but fine details were not easily seen. Nevertheless candling is considered to be an important method of assessing development of ostrich eggs during incubation. 3. The ostrich embryo adopts a hatching position and follows a hatching sequence different from that of the fowl. During internal pipping, the air space is pulled towards the beak in the former but the beak moves to the air space in the latter. The right foot has an important role in breaking the shell during hatching. 4. The hatching process is considered to reflect an adaptation to the egg having a hard, brittle shell. 5. Malpositions in the ostrich may have been misinterpreted in the past because of the unusual hatching position. The commonest malposition described here was head-in-the-small-end.