Pheasant Embryos Research Articles

The study of the use of specific staining in the observation of neuronal cells and myelin fibers of the cerebellum in the stages of pheasant embryos to one-day-old chicks

The study of the use of specific staining in the observation of neuronal cells and myelin fibers of the cerebellum in the stages of pheasant embryos to one-day-old chicks

Toxic interaction of glyphosate herbicide and tau-fluvalinate insecticide on pheasant embryos

Toxic interaction of glyphosate herbicide and tau-fluvalinate insecticide on pheasant embryos

Ultrastructure characteristics of primordial germ cells in stage X of pheasant (Phasianus colchicus) embryo.

Stage X is one of the formation stages in birds at which the blastoderm area is distinguished by two areas of area pellucida being responsible for formation of embryonic tissues and primordial germ cells, and area opaca forming the extra-embryonic tissues. Primordial germ cells are multi-potent stem cells giving rise to spermatogonia or oogonia. The present study was carried out to describe the characteristics of primordial germ cells in stage X of pheasants' embryo using a transmission electron microscope. The blastoderm was dissected out from embryos which were already incubated for 12 hr. Toluidine blue was used for staining semi-thin sections; lead citrate and uranyl acetate were also used to stain ultra-thin sections. Images of primordial germ cells elucidated that the nucleus was situated eccentrically and had a compact spherical structure. Moreover, the nucleolus appeared elongated and was located eccentrically. The cytoplasm was composed of yolk granules and glycogen particles. Mitochondria were observed as round structures in the cytoplasm. The most important finding was that the primordial germ cells contained yolk granules, mitochondria and small amount of glycogen at this stage.

Conservation of Migration and Differentiation Circuits in Primordial Germ Cells Between Avian Species

Germ cell differentiation in reverse-sexed reproductive organs and interspecies germ line chimeras provides insight into the mechanism of germ cell development and represents a useful tool for conservation of endangered birds. We investigated the migration and survival capacity of male chicken primordial germ cells (PGCs) in female chicken embryos and in quail and Korean ring-necked pheasant embryos of both sexes. Interestingly, the PGCs were successfully reintroduced in all cases. Furthermore, the cells survived in the recipient gonads until hatching regardless of sex and species of the recipient. In the case of male recipient chickens, PGC-derived offspring were produced. However, the reverse-sexed female chickens, quails and pheasants of both sexes did not generate any male donor PGC-derived progeny. These results suggest that migration and survival circuits in chicken PGCs are conserved in both sexes and between avian species during embryonic development.

Detection and characterization of primordial germ cells in pheasant ( Phasianus colchicus) embryos

The developmental similarity between the chicken and pheasant ( Phasianus colchicus) allows the novel biotechnologies developed in the chicken to be applied to the production of transgenic pheasants and interspecies germline chimeras. To detect pheasant primordial germ cells (PGCs) efficiently, which is important for inducing germline transmission, the ultrastructure of PGCs and their reactivity to several antibodies (2C9, QB2, anti-SSEA-1, and QCR1) and periodic acid-Schiff’s solution (PAS) were examined. To obtain PGCs, blood was taken from embryos incubated for 62–72 h or from gonads from embryos incubated for 156–216 h. The PGCs collected from both sources had the typical ultrastructure of pluripotent cells: a large nucleus with a distinct nucleolus, a high ratio of nuclear to cytoplasmic volume, and a distinct cytoplasmic membrane. In comparing the morphology of PGCs collected from different sites, more mitochondria and better-developed membrane microvilli were found in gonadal PGCs than in circulating PGCs. The nucleus of gonadal PGCs was flattened and had a large eccentrically positioned nucleolus. Of the antibodies tested, only QCR1 antibody reacted with an epitope in pheasant PGCs, and no specific signal was detected to other antibodies. The temporal change in the PGC populations in the blood and gonads of embryos was examined. In blood, the population was greater ( P < 0.0001) in embryos incubated for 64 h than in embryos incubated for 62 or 66–72 h (31.4 versus 5.6–16.2 μL −1). In embryonic gonads, the number of PGCs increased continuously from 156 to 216 h of incubation (193–2,718 cells/embryo), although the ratio of PGCs to total gonadal cells did not change significantly (0.50–0.61%). In conclusion, pheasant PGCs have typical germ cell morphology and possess the QCR1 epitope. Circulating blood and the gonads of embryos incubated for 64 and 216 h, respectively, are good sources of PGCs.

Congener-specific patterns and toxic assessment of polychlorinated biphenyls in resident and migratory birds from Southern India and Lake Baikal in Russia.

Concentrations of polychlorinated biphenyls (PCBs), including non-, mono-, and di-ortho congeners, were determined in migratory and resident birds collected from India and Lake Baikal in Russia. In the 11 different species examined, total PCBs concentrations were in the range of 11-4500 ng/g (wet wt). IUPAC 105, 118, 138, 153, and 180 were the predominant congeners in almost all the birds. White-cheeked tern collected from India and common tern collected from Lake Baikal showed high PCBs concentrations of 4400 ng/g (wet wt) and 4500 ng/g (wet wt), respectively, and accumulated relatively high ratios of penta-CBs (IUPAC 118, 105, 99). Toxic equivalents (TEQs) of non- and mono-ortho PCB congeners in birds collected from India and Lake Baikal were in the range of 1.5-56 and 2.8-370 pg/g wet wt, respectively. Toxic assessment results led by calculated TEQs of the transfer to eggs from female birds revealed that TEQs in most of migratory and resident birds were comparable to the lowest observable effect level (LOAEL) of chicken which is a highly sensitive species against dioxin-like compounds. Calculated transfer TEQs to eggs of common tern collected from Lake Baikal in autumn, however, exceeded the LOAEL of CYP1A induction in bald eagle embryos and ED50 of that in pheasant embryos, suggesting that embryo toxicity by coplanar PCBs in some avian species breeding in Lake Baikal is possible.

Intraclutch Hatch Synchronization in Pheasants and Mallard Ducks

Synchronization of hatching within clutches of precocial bird species can be achieved either by acceleration or retardation, i.e. by shortening or prolonging the incubation period. The ability of mallard (Anas platyrhynchos) and ring‐necked pheasant (Phasianus colchicus) embryos to accelerate or retard hatching was tested by incubating separate clutches, of which three eggs had 2 d longer or shorter incubation time than the others, and observing their individual time of pipping (breaking of the shell). Mallard embryos were able to delay hatching by on average 0.6 d (43% of the eggs delayed at least 1 d), but were better at acceleration (on average 1.3 d; 91% of the eggs accelerated more than 1 d). Conversely, pheasant embryos were only able to accelerate by 0.4 d (50% accelerated more than 1 d), but were better at delaying the hatching (1.2 days; 77% delayed more than 1 d). This difference between the species may depend on different degrees of relatedness within clutches in pheasants and mallards. It may also be an effect of the more developed sensory and neuromuscular systems in galliforms; a reduction of the incubation period would mean that the development of, for example, locomotion would be insufficient at hatching.

Intraclutch Hatch Synchronization in Pheasants and Mallard Ducks

Synchronization of hatching within clutches of precocial bird species can be achieved either by acceleration or retardation, i.e. by shortening or prolonging the incubation period. The ability of mallard (Anas platyrhynchos) and ring-necked pheasant (Phasianus colchicus) embryos to accelerate or retard hatching was tested by incubating separate clutches, of which three eggs had 2 d longer or shorter incubation time than the others, and observing their individual time of pipping (breaking of the shell). Mallard embryos were able to delay hatching by on average 0.6 d (43% of the eggs delayed at least 1 d), but were better at acceleration (on average 1.3 d; 91% of the eggs accelerated more than 1 d). Conversely, pheasant embryos were only able to accelerate by 0.4 d (50% accelerated more than 1 d), but were better at delaying the hatching (1.2 days; 77% delayed more than 1 d). This difference between the species may depend on different degrees of relatedness within clutches in pheasants and mallards. It may also be an effect of the more developed sensory and neuromuscular systems in galliforms; a reduction of the incubation period would mean that the development of, for example, locomotion would be insufficient at hatching.

Degradation of some pesticides in avian embryos.

On day 9 or 12 of the hatching period different pesticides (parathion, methyl-parathion, carbendazim, 2,4-D-amine Na, phosmethylane) were applied in ecotoxicological trials. The formulations were either injected into the air space of pheasant, quail or hen eggs or hen eggs were treated by the immersion technique. The residues of pesticides were measured in samples on days 13, 14 and 16 of incubation of chicken and pheasant embryos, while the Japanese quail embryos were analysed on days 10-14 of incubation. Analytical chemistry data showed a varying degradation rate of the compounds in avian embryos of the same species. The residues directly affect the embryos, disturbing their normal development and causing pathophysiological and morphological changes.

Embryonic toxicity of insecticide Sumithion 50 EC and herbicide Fusilade S in pheasants after individual or combined administration.

The purpose of this work was to determine the individual and combined effects of insecticide Sumithion 50 EC (50% fenitrothion) and herbicide Fusilade S (12.5% fluazifop-P-butyl) on the development of pheasant embryos. Eggs were treated by injection of various concentrations of pesticides into the air space on day 12 of incubation. Pathological examination of embryos was carried out on day 23 of the hatching period. Mortality rate, body weight data and morphological alterations were evaluated after the macroscopic examination. The skeletal staining method was used to detect deformities. The two pesticides used in combination moderated the toxic/teratogenic effects of individual treatment.

Embryotoxicity of 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin in the ring‐necked pheasant

AbstractFertilized eggs of ring‐necked pheasants (Phasianus colchicus) were injected into the albumin or yolk with vehicle or graded doses of 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) (0.01, 0.1, 1, 10, 100, 1,000, 10,000, or 100,000 pg TCDD/g egg) on day 0 of embryonic development and toxicity was assessed in 1‐d‐old hatchlings and 28‐d‐old chicks. The most sensitive effect of in ovo TCDD exposure was induction of hepatic ethoxyresorufin‐O‐deethylase (EROD) activity in 1‐d‐old hatchlings. The ED50 for this response was 312 pg TCDD/g egg. Embryo mortality was the most sensitive sign of toxicity. The TCDD dose that caused 50% mortality above control (LD50) when injected into the egg albumin or yolk was 1,354 and 2,182 pg TCDD/g egg, respectively. At egg TCDD doses up to and including 1,000 pg TCDD/g egg, no effect was detected in 1‐d‐old hatchlings and 28‐d‐old chicks in body growth, organ weights, carcass morphometrics, incidence of edema, or incidence of histological alterations in the liver, spleen, heart, Bursa of Fabricius, or thymus. Egg TCDD doses as high as 1,000 pg TCDD/g egg also had no effect on cardiac morphometrics or incidence of cardiac malformations in 1‐d‐old hatchlings, or on antibody‐mediated immunity in 28‐d‐old chicks. We conclude that embryo mortality is the most sensitive sign of TCDD toxicity in the ring‐necked pheasant following in ovo exposure. The ring‐necked pheasant embryo is less sensitive than the chicken (Gallus domesticus) embryo and more sensitive than the eastern bluebird (Sialia sialis) embryo to TCDD toxicity.

Differences in sensitivity of some avian species to the embryotoxicity of a PCB, 3,3′,4,4′-tetrachlorobiphenyl, injected into the eggs

Eggs of pheasant Phasianus colchicus, mallard Anas platyrhynchos, goldeneye Bucephala clangula and black-headed gull Larus ridibundus were injected with 3,3′,4,4′-tetrachlorobiphenyl (TCB) into the yolks. The highest dose given was 0·1 mg per kilogram of egg (mallard) or 1·0 mg per kilogram of egg (pheasant, goldeneye and black-headed gull). Only in the pheasants' eggs was any effect on hatchability noted. Injection of 1·0 mg TCB per kilogram of egg resulted in the death of all the pheasant embryos, but no decrease in the hatching rate occurred when 0·1 mg TCB per kilogram was injected. Embryos of the species studied appear to be considerably less sensitive than chick embryos to the toxic action of TCB. The concentration of TCB in the embryos of some of the injected eggs was determined in chickens, pheasants and goldeneyes. No difference was found in the concentration of TCB in the embryos to explain the difference in effect on hatchability.

Host range restriction of vesicular stomatitis virus on duck embryo cells

Vesicular stomatitis virus (VSV) plaque formation and replication are restricted in duck embryo cells compared to chick embryo cells. Mutants of VSV which replicate normally on both chick and duck cells were isolated. The duck-adapted mutant, when passed through chick cells, retains its ability to grow normally on duck cells. This indicates that the ability of this virus to grow on duck cells is due to mutation rather than to host-controlled modification. VSV is restricted in duck cells but not in quail or pheasant embryo cells which follows the evolutionary relationship of these species. Newcastle disease virus and Rous sarcoma virus are restricted in duck cells also. Virus-specific protein synthesis of the wild-type VSV is greatly reduced in duck cells compared to chick cells. A structural protein of the duck cell-adapted mutant is altered.

Nucleotide Sequences Derived from Pheasant DNA in the Genome of Recombinant Avian Leukosis Viruses with Subgroup F Specificity

Recombination between viral and cellular genes can give rise to new strains of retroviruses. For example, Rous-associated virus 61 (RAV-61) is a recombinant between the Bryan high-titer strain of Rous sarcoma virus (RSV) and normal pheasant DNA. Nucleic acid hybridization techniques were used to study the genome of RAV-61 and another RAV with subgroup F specificity (RAV-F) obtained by passage of RSV-RAV-0 in cells from a ring-necked pheasant embryo. The nucleotide sequences acquired by these two independent isolates of RAV-F that were not shared with the parental virus comprised 20 to 25% of the RAV-F genomes and were indistinguishable by nucleic acid hybridization. (In addition, RAV-F genomes had another set of nucleotide sequences that were homologous to some pheasant nucleotide sequences and also were present in the parental viruses.) A specific complementary DNA, containing only nucleotide sequences complementary to those acquired by RAV-61 through recombination, was prepared. These nucleotide sequences were pheasant derived and were not present in the genomes of reticuloendotheliosis viruses, pheasant viruses, and avian leukosis-sarcoma viruses of subgroups A, B, C, D, and E. They were partially endogenous, however, to avian DNA other than pheasant. The fraction of these nucleotide sequences present in other avian DNAs generally paralleled the genetic relatedness of these avian species to pheasants. However, there was a high degree of homology between these pheasant nucleotide sequences and related nucleotide sequences in the DNA of normal chickens as indicated by the identical melting profiles of the respective hybrids.

Avian leukosis viruses of different subgroups and types isolated after passage of Rous sarcoma virus-Rous-associated virus-0 in cells from different ring-necked pheasant embryos

Avian leukosis viruses of subgroups A and F (RAV-A and RAV-F) arose at a low rate after passage of Rous sarcoma virus-Rous-associated virus-0, which is subgroup E, in cells from ring-necked pheasant embryos. In cells of two embryos, all of the viruses isolated after virus passage were RAV-F. However, in cells of a third embryo, both RAV-A and RAV-F were isolated. In addition, there sometimes were type-specific differences among the different isolates of RAV-A and RAV-F from the cells of single embryos. These results indicate that the RAV-A and RAV-F probably arose by recombination of viral and cellular genes, that different ring-necked pheasant embryo may have different endogenous avian leukosis virus-related nucleotide sequences, and that recombination at different sites in these endogenous sequences might give rise to type-specific differences among the RAV-A and RAV-F.

Studies on avian erythrocyte metabolism—iv. Relationship between the major phosphorylated metabolic intermediates and oxygen affinity of whole blood in adults and embryos in several galliformes

1. 1. The major phosphorylated metabolic intermediates of the erythrocytes (RBC) and P 50 of the whole blood from the mature turkey, pea fowl, pheasant, guinea, and quail have been determined and compared with similar information from the blood of the 23-day turkey embryo, 21-day pheasant embryo, and 21-day guinea embryo. 2. 2. Inositol pentaphosphate (IPP) is the predominant organic phosphate in the RBC of the mature birds at levels of 4.05, 4.31, 3.34, 3.11, and 3.32μmoles/cm 3 RBC in the turkey, pea fowl, pheasant, guinea, and quail, respectively. 3. 3. 2,3-diphosphoglycerate (2,3-DPG) is the predominant organic phosphate in the erythrocytes of the 23-day turkey embryo, 21-day pheasant embryo, and 21-day guinea embryo at levels of 4.79, 4.29, and 5.05 μmoles/cm 3 RBC. respectively. 4. 4. The P 50's of whole blood from the mature birds ranged between 33.4 and 41.2mm Hg whereas the P 50 of whole blood from the embryos of the three species studied is between 23.5 and 22.8mm Hg. 5. 5. The relative per cent of Hb-H (hatching hemoglobin) is 13.0 in blood of embryos from the guinea eggs and approximately 0.7 in blood of embryos from pheasant eggs.

RNA tumor viruses of pheasants: Characterization of avian leukosis subgroups F and G

Endogenous leukosis-like viruses of ring-necked pheasants ( Phasianus colchicus) and golden pheasants ( Chrysolophus pictus) have been isolated and characterized. The majority of the normal pheasant embryo cultures contain helper activity for the defective Bryan high titer strain of Rous sarcoma virus. The Rous sarcoma pseudotypes produced with endogenous helper activity from ring-necked pheasants belong to subgroup F. The pseudotypes from golden pheasant cells constitute subgroup G. Subgroup F and G pseudotypes can infect all known genetic types of chicken fibroblasts as well as pheasant and Japanese quail cells, but do not plate on goose cells. Duck cells are resistant to subgroup G but not to F. The subgroup F and G helper viruses isolated from Rous sarcoma viral pseudotypes show interference with their homologous subgroup. RAV-61, a standard of subgroup F, interferes with pseudotypes produced with endogenous helper activity from ring-necked pheasant cells but not with subgroup G pseudotypes. Subgroups F and G do not cross-react with subgroup A to E in neutralization tests. Some normal ring-necked pheasant sera have anti-F activity. Subgroup F and probably also G leukosis-like viruses can undergo genetic recombination with nondefective avian sarcoma viruses.

Indications of a viral etiology for marble spleen disease in pheasants.

Evidence that marble spleen disease of pheasants (Phasianus colchicus) has a viral etiology was obtained from field cases using the following criteria: Intranuclear inclusion bodies in cells of the spleen, lungs, liver, and bone marrow; the presence of antigen in spleens of infected pheasants as detected by the agar gel precipitin test; demonstration of virus particles in the splenic intranuclear inclusions by the electron microscope; and the presence of specific fluorescence in spleen cells as shown by direct fluorescent antibody staining. Every case of marble spleen disease examined had all of these findings. Attempts to isolate a virus in cell culture using chicken and pheasant embryo fibroblasts and chick kidney cells were unsuccessful.

Measurement of endogenous leukosis virus nucleotide sequences in the DNA of normal avian embryos by RNA-DNA hybridization

Rous associated virus type O (RAV-O) is a subgroup E avian leukosis virus selected as an example of “endogenous” leukosis viruses which are associated with normal avian embryo cells. DNA sequences complementary to RAV-O 60–70S RNA genome were detected and quantitated in DNA from normal avian embryos by the technique of RNA-DNA hybridization with DNA excess. DNA from “leukosis free” white leghorn chicken embryos, as well as from line 7 chicken embryos, contained sequences related to at least 70%, if not all, of RAV-O RNA sequences. A tentative estimate of gene frequency is low, in the range of two to four copies of RAV-O per cell genome. Some difficulties in precisely calculating gene frequency by this method are discussed. Embryos of ring-necked pheasant contain only about 10% of the RAV-O genome. These findings are not affected by the presence or absence of viral group specific ( gs) antigen in chicken or pheasant embryos. Embryos of Japanese quail, a species which lacks a known leukosis virus and does not synthesize avian gs antigen, contained DNA sequences specifically related to no more than 4% of the RAV-O genome. The data document the specificity and sensitivity of this hybridization technique and provide additional firm support for the proposition that normal avian cells carry endogenous avian leukosis virus in the form of DNA proviruses.

Isolation of leukosis-type virus from pheasant embryo cells: Possible presence of viral genes in cells

Isolation of leukosis-type virus from pheasant embryo cells: Possible presence of viral genes in cells