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Comparative analysis of the prion protein open reading frame nucleotide sequences in peacock and parakeet.

The open reading frame of peacock and parakeet prion protein (PrP) genes was cloned and sequenced. The peacock and parakeet PrP genes consisted of 833 and 866 nucleotides encoding 266 and 277 amino acids, respectively (GenBank Accession numbers AY365065 and AY365066). Sequence analysis showed that the peacock and parakeet PrP genes had 93.67% homology to each other, 94.04% and 99.64% homology to the chicken PrP gene and 46.0% and 42.1% similarity to the mammalian PrP genes, respectively. The structural features of all known mammalian and avian PrPs, including N-terminal signal peptides, tandem repeats, conserved hydrophobic region, disulfide bridges and glycoinositol phospholipid anchor, were also found in peacock and parakeet PrPs. The parakeet and peacock PrPs, however, differed in the hexarepeat region, with the peacock having six and the parakeet having seven hexarepeats. The phylogenetic analysis showed that the PrP genes in 52 species were clustered into 2 distinct lineages, the avian and the mammalian. The peacock and parakeet PrP genes belonged to the same lineage but the peacock PrP was sub-classed with the pigeon PrP and the parakeet PrP was sub-classed with the duck and chicken PrPs. The present work added two more species data to the collection of the PrP genes and supported the previous findings that the PrP genes are highly conserved across species.

Amino Acid Sequence↗

Phagocytotic removal of apoptotic endocrine cells by folliculostellate cells and its functional implications in clusterin accumulation in pituitary colloids in helmeted guinea fowl (Numida meleagris).

The different cell types in the anterior pituitary behave as dynamic populations. The gland maintains a continuous renewal of cells to ensure a dynamic balance between cell division, differentiation, growth arrest and apoptosis. Apoptosis is a frequent event in the anterior pituitary in which unwanted cells are eliminated without affecting neighboring cells. We examined the link between apoptosis and the occurrence of colloids in the guinea fowl (Numida meleagris) pituitary gland and the relationship of clusterin accumulation in the colloids. S-100 positive folliculostellate (FS) cells were found surrounding colloids. Apoptotic cells detected by single stranded DNA (ssDNA) immunohistochemistry were observed in the whole anterior pituitary and preferentially near colloid masses. Clusterin protein was detected in endocrine cells, FS cells and in the colloids. In situ hybridization showed clusterin mRNA in endocrine cells and FS cells. Simultaneous localization was performed to determine whether clusterin mRNA and ssDNA within anterior pituitary was present within the same cell. Clusterin mRNA was not detected in apoptotic cells but was present in neighboring surviving cells. At the ultrastructural level, numerous endocrine cells at different stages of apoptosis were found phagocytosed by FS cells. Our results suggest that clusterin is produced by endocrine cells for cytoprotection before death. Apoptotic endocrine cells are phagocytosed by FS cells and digested by their lysosomal enzymes. In FS cells, clusterin interacts and aggregates with by-products of digestion that subsequently become stored in colloid as a residual body.

Animals↗

Effects of beta-carotene supplementation on chick growth, immune status and behaviour in the grey partridge, Perdix perdix.

Carotenoids are important for various functions during chick development. Since these pigments cannot be synthesized, they can be considered limited resources that the mother optimally allocates between herself and her offspring (maternal effect). Some studies have examined the effects of carotenoids on growth and immune function but little is known about their role in behaviour. In this study of the grey partridge, we conducted two supplementation experiments: (1) laying females were fed with beta-carotene enriched or impoverished diets; (2) chicks were fed directly with beta-carotene enriched or impoverished diets. We then evaluated the effects of this carotenoid on chick growth, immunocompetence and anti-predator behaviour (reactions to a raptor model). In the first experiment, the beta-carotene enriched diet given to mothers did not cause any difference in chick physiology. In the second experiment, beta-carotene supplementation of chicks had a significant beneficial effect on their growth and immune response, although their behavioural reactions did not differ in relation to the diet. Therefore, beta-carotene supplementation had beneficial effects on growth and immunocompetence only when directly supplied to chicks. The beneficial effect reported in other species for begging or pecking behaviours was not confirmed for the anti-predator behaviour of grey partridge chicks.

Analysis of Variance↗

The effects of fasting and refeeding on biochemical parameters in the red-legged partridge (Alectoris rufa).

Every year, about three million farm-reared red-legged partridges are released in Spain for hunting purposes, most of them for restocking where the success is very low. Some factors such as the sudden shift from abundant commercial energy-rich diet to natural food and absence of experience at looking for natural food resources could cause a rapid physical deterioration, being more vulnerable to predation and disease. To determine fasting capacity in red-legged partridges, we fasted 32 birds during 4 days and then we refed them, collecting blood samples and body masses during the experiment. Red-legged partridges entered in the third critic phase of fasting, in which body proteins became the main energetic source, after 48 h of fasting. The fasting period caused an average loss of 17% of their initial body mass and only attained a 93.6% of their initial body mass after 24 days of refeeding. The limited capacity to resist fasting in the red-legged partridge could be one of the reasons for the low success in restocking, mainly in those where neither supplementary feeding nor acclimation period is allowed for birds before they are finally released.

Adaptation, Physiological↗

Fertility and content of cadmium in pheasant (Phasianus colchicus) following cadmium intake in drinking water.

In this study, the effects of cadmium applied per os on fertility, live weight of newly hatched chicks, and cadmium concentrations in some organs of young and adult pheasants were investigated. The metal was applied at the concentration of 1.5 mg Cd(2+)/L during 3 months. After the egg laying, the numbers of eggs laid, cracked, and unfertilized were determined and the live weights of newly hatched chicks were measured. The cadmium concentrations in liver, kidney, and muscle (m. pectoralis) of young and adult pheasants were analyzed. We found that cadmium exposure of the adults did not affect the number of eggs laid but resulted in more eggs being damaged. Hatchlings were significantly heavier in the cadmium-treated group (21.36 +/- 2.28 g) compared to the control group (20.91 +/- 1.97 g) 4 weeks after the cadmium intake. Higher cadmium concentrations were observed in the muscle and kidney tissue of newly hatched pheasants after 4 weeks compared to the cadmium-exposed groups after 8 and 12 weeks. The cadmium concentrations in kidneys and liver increased significantly in adult pheasants. The metal had accumulated especially in kidneys of the adult pheasants and reached levels up to 9.64 mg/kg wet weight 3 months after the daily cadmium intake in drinking water. The concentration in liver of the adults was 3.53 mg/kg wet weight.

Animals↗

A comparative study of cadmium and copper in ruffed grouse (Bonasa umbellus) in regions with and without historic mining.

Mining activities can increase the bioavailability of metals in aquatic and terrestrial ecosystems. Following over 100 years of copper mining in portions of Michigan's Upper Peninsula (UP) terrestrial ecosystems retain vast quantities of waste rock with traces of cadmium and large concentrations of copper. We compared liver cadmium and copper concentrations in ruffed grouse (Bonasa umbellus), a popular game bird from landscapes with and without historic mining. We also used chickens (Gallus domesticus) to determine whether mine waste was a direct source of liver cadmium. Cadmium and copper levels did not differ between mining areas in Michigan and non-mining areas in Wisconsin. We found nearly significant difference between sexes in cadmium levels. Cadmium levels for all chickens were below the method detection limit of the lab (0.03 mg/kg) and copper levels did not differ in the experimental chickens. These results suggest that the historic mining in the western UP is not leading to higher cadmium or copper uptake in grouse.

Animal Feed↗

Cadmium concentrations in tissues of willow ptarmigan (Lagopus lagopus) and rock ptarmigan (Lagopus muta) in Nunavik, Northern Québec.

Willow and rock ptarmigan were obtained from Northern Québec. Willow ptarmigan were found to have mean cadmium concentrations of 179.7 microg/g (dw) in the kidneys and 25.8 microg/g (dw) in the liver; these levels were three times higher than those found in the rock ptarmigan. The cadmium levels in the ptarmigan were below the threshold above which adverse effects can be observed in birds. The difference between the two ptarmigan species in cadmium content is explained by the diet. A comparison of their diet showed that willow, which stores cadmium, is an important food resource for willow ptarmigan but not for rock ptarmigan. Because there is limited information available on the consumption of ptarmigan kidneys and liver by the Inuit, and the fact that this is a traditional way of life and provides nutritional benefits to the Inuit population, no consumption guidelines are proposed.

Air Pollutants↗

Molecular insight into a wildlife crime: the case of a peafowl slaughter.

We report a recent case in which a wildlife warden had suspected that some people had killed and cooked a peacock. Cooked meat, intestine of bird and the wooden block used for chopping were seized from the site of crime and forwarded to our laboratory for DNA testing. Mitochondrial cytochrome b sequence analysis revealed that the cooked meat and remnants of the bird were of a chicken, but the DNA obtained from the wooden block was of an Indian Peafowl (Peacock) testifying that the wooden chopping block was used to chop the meat of an endangered bird, thus bringing to light a wildlife crime.

Animals↗

The effects of flubendazole and mebendazole on cytochromes P4501A in pheasant hepatocytes.

Many benzimidazoles are known inducers of cytochromes P4501A (CYP1A) in laboratory animals and cell lines. As flubendazole and mebendazole are benzimidazole anthelmintics often used in a pheasant, in the present study an effect of these drugs in primary cultures of pheasant (Phasianus colchicus) hepatocytes was investigated. After 48 h incubation of the hepatocytes with the benzimidazoles (0.2-5 microM), CYP1A activities -- ethoxyresorufin O-deethylation (EROD) and methoxyresorufin O-demethylation (MROD) activities were measured and the CYP1A protein levels were determined by Western blotting. None of the tested benzimidazoles influenced the CYP1A protein content. No pharmacologically significant enhancement of CYP1A after exposure of the hepatocytes to flubendazole and mebendazole was found. Inhibition of the EROD/MROD activities caused by both tested substances was observed only at the highest concentration (5 microM). From a point of view of CYP1A induction or inhibition, the treatment of pheasants by both anthelmintics tested seems to be safe. Our study demonstrates the inter-species differences in CYP1A inducibility and the importance of induction/inhibition studies on target animals.

Animals↗

Accumulation of heavy metals in circumpolar willow ptarmigan (Lagopus l. lagopus) populations.

A circumpolar survey of heavy metals in willow ptarmigan liver and kidney revealed considerable variations in Cd content in Canada and Scandinavia. The Cd content in central Canada was comparable with that in Scandinavia and Russia, at least for kidney. However, in both liver and kidney the median for Canada as a whole was much higher than in the other countries. Some Canadian locations had exceptionally high levels, several birds having >50 mg kg(-1) in liver and >400 mg kg(-1) in kidney. In Norway, the Cd content was highest in central mountain areas in south Norway and inland locations in the two northernmost counties. Five locations in central and north Norway showed mean Cd levels in kidney above 100 mg kg(-1). It is difficult to evaluate regional differences in Sweden, but most locations had the same Cd level as moderately contaminated locations in Norway. Cd levels in Russia were comparable to moderately contaminated locations in the other countries. Due to a high intake of willow, naturally rich in Cd, direct comparison of the Cd level in willow ptarmigan from different locations cannot reveal the effects of long-range pollution. The Pb concentration in willow ptarmigan kidney and liver varied significantly in Norway and Canada, but not in Sweden and Russia. Levels in Sweden and Russia were comparable to those in Canada and low levels in Norway. The highest median value from all locations within countries was found in Norway, both in liver and kidney. The highest Pb content was found in south Norway, indicating an effect of long-range pollution in willow ptarmigan. The level in western Canada was significantly higher than in central Canada. The Hg content in liver varied significantly from one location to another in all the countries and in kidney everywhere except Sweden. In Scandinavia, there is no distinct regional pattern. Canada had a significantly higher Hg level in central than western regions in both tissues the opposite of that found for Cd and Pb. Cu and Zn showed significant variations from one location to another in liver and kidney in Canada and Norway, but only in kidney in samples from Sweden. Comparison between western and central Canada revealed a significant difference for Cu in liver, samples from central Canada having more. There are no significant differences from one country to another, but some localities in Canada seem to have higher Cu concentrations in kidney than are found in Scandinavia and Russia.

Air Pollutants↗

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 microL(-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.

Animals↗

Experimental toxoplasmosis in red-legged partridges (Alectoris rufa) fed Toxoplasma gondii oocysts.

Thirty red-legged partridges (Alectoris rufa), 5-month-old, were orally inoculated with oocysts of the OV-51/95 strain of Toxoplasma gondii. Birds were distributed into five groups and received, respectively, 10 (group A, 4 birds), 50 (group B, 14 birds), 10(2) (group C, 4 birds), 10(3) (group D, 4 birds) and 10(4) (group E, 4 birds) oocysts. One partridge from group B and one from group E died suddenly of acute toxoplasmosis at 7 day after inoculation (DAI) with demonstrable T. gondii in several tissues. The rest of birds remained clinically normal until killed at 44, 58, 65, 72, 79 or 100 DAI. Brain, heart, liver and skeletal muscle from these partridges were bioassayed individually in mice; T. gondii was demonstrated in all these tissues, except in heart of three birds inoculated, respectively, with 10, 50 and 10(2) oocysts. Lesions were not seen in histologic sections of tissues from surviving partridges. These results suggest that red-legged partridges are resistant to clinical toxoplasmosis.

Agglutination Tests↗

Phylogenetic analyses indicate little variation among reticuloendotheliosis viruses infecting avian species, including the endangered Attwater's prairie chicken.

Reticuloendotheliosis virus infection, which typically causes systemic lymphomas and high mortality in the endangered Attwater's prairie chicken, has been described as a major obstacle in repopulation efforts of captive breeding facilities in Texas. Although antigenic relationships among reticuloendotheliosis virus (REV) strains have been previously determined, phylogenetic relationships have not been reported. The pol and env of REV proviral DNA from prairie chickens (PC-R92 and PC-2404), from poxvirus lesions in domestic chickens, the prototype poultry derived REV-A and chick syncytial virus (CSV), and duck derived spleen necrosis virus (SNV) were PCR amplified and sequenced. The 5032bp, that included the pol and most of env genes, of the PC-R92 and REV-A were 98% identical, and nucleotide sequence identities of smaller regions within the pol and env from REV strains examined ranged from 95 to 99% and 93 to 99%, respectively. The putative amino acid sequences were 97-99% identical in the polymerase and 90-98% in the envelope. Phylogenetic analyses of the nucleotide and amino acid sequences indicated the closest relationship among the recent fowl pox-associated chicken isolates, the prairie chicken isolates and the prototype CSV while only the SNV appeared to be distinctly divergent. While the origin of the naturally occurring viruses is not known, the avian poxvirus may be a critical component of transmission of these ubiquitous oncogenic viruses.

Amino Acid Sequence↗

Full genome sequence and some biological properties of reticuloendotheliosis virus strain APC-566 isolated from endangered Attwater's prairie chickens.

Reticuloendotheliosis virus (REV) causes runting, high mortality, immunosuppression, and chronic neoplasia associated with T and/or B cell lymphomas in a variety of domestic and wild birds, including Attwater's prairie chickens (APC) (Tympanuchus cupido attwateri). The complete proviral sequence of a recent REV isolate from APC (REV APC-566) was determined. This virus was isolated from an APC maintained in captivity in a reproduction program intended to avoid its extinction. REV APC-566 was determined to be oncogenic in Japanese quail (Coturnix coturnix japonica), chickens (Gallus gallus) and turkeys (Meleagris gallopavo). Immune responses against bacteria and viruses were significantly reduced in turkeys infected with REV APC-566. The proviral genome is 8286 nucleotides in length and exhibits a genetic organization characteristic of replication-competent gammaretroviruses. The REV APC-566 provirus contains two identical long terminal repeats (LTR) and a complete set of genes including gag, gag-pol and env. As previously reported, alignments with other REV sequences showed high similarity with sequences found in the gag and pol genes from other REVs. The REV APC-566 env gene showed high nucleotide sequence homology with REV sequences inserted in fowl poxvirus (99.8%), and with spleen necrosis virus (SNV) (95.1%). Sequences coding for a previously reported immunosuppressive peptide contained in the transmembrane region of the env gene are well conserved among all REV sequences analyzed. The LTR was the most divergent region, exhibiting various deletions and insertions. REV APC-566 has a unique insertion of 23 bp in U3 and shares deletions of 19 and 5 bp with chicken syncytial virus and REV inserts in fowlpox virus.

Amino Acid Sequence↗

Testosterone and autumn territorial behavior in male red grouse Lagopus lagopus scoticus.

In many bird species, males exhibit territorial aggression outside the breeding season, when testosterone concentrations are low and may not regulate territorial behaviors. The hormonal regulation of aggression at this time of year has only been studied in passerine birds. Here, we investigated the role of testosterone in the regulation of aggression in a non-passerine bird, the red grouse Lagopus lagopus scoticus. Male red grouse are aggressive in early spring when breeding starts, in autumn when they establish territories, and sporadically through much of the winter. We first describe seasonal variations in plasma testosterone concentrations and in the size of males' sexual ornaments, their red combs, which relates to aggressiveness. Testosterone concentrations and comb size were correlated. Both increased in autumn to a peak in October, and then increased again in spring, to a greater peak in early April. Secondly, we experimentally investigated the effects of testosterone, and of an anti-androgen (flutamide) used in combination with an aromatase inhibitor (ATD), on autumn territorial behavior. Males were treated with either empty implants, as controls (C-males), testosterone implants (T-males), or with flutamide and ATD implants (FA-males). One month after implanting, both T- and FA-males had higher concentrations of testosterone than C-males. Comb size, aggressive call rate, and response to playbacks of territorial call all significantly increased in T-males. However, the increase in testosterone in FA-males did not increase comb size or aggressive behavior. In the following spring, after the content of implants was used, FA-males had significantly lower testosterone than C-males, and had a reduced seasonal increase in comb size. The results suggest that testosterone plays a significant role in regulating red grouse aggressive behavior in autumn. However, the observation that flutamide and ATD treatment did not reduce territorial behavior, suggests that estradiol may also be involved in the regulation of non-breeding aggression.

Androgen Antagonists↗

Experimental manipulation of yolk testosterone affects digit length ratios in the ring-necked pheasant (Phasianus colchicus).

In humans, most of the mammals and one bird species studied so far, the relative length of individual digits is sexually dimorphic. Most studies of humans have been concerned with the ratio between second (2D) and fourth digits (4D), whereas some studies of humans and other mammals have also investigated other digit ratios. Inter- and intra-sexual variation in 2D:4D may depend on differential exposure to androgens during embryonic life, and the genetic mechanisms linking 2D:4D to androgens may be mediated by Hox genes. Because Hox genes are conserved in vertebrates, similar patterns of variation in digit ratios might be expected across vertebrate classes. The observation of correlations between digit ratios and physiological, psychological and performance traits in humans has generated interest in exploring the possibility that digit ratios are a marker of embryonic exposure to androgens, which have diverse consequences on several phenotypic traits. However, the hypothesis that digit ratios depend on androgen effects during development has never been tested experimentally. In this study, we increased testosterone concentration in ring-necked pheasant eggs and measured length ratios between the second, third and fourth digits of both feet in fully grown offspring. Females from testosterone-injected eggs had larger 2D:3D in the left foot, whereas this was not the case in males. The other digit ratios were unaffected by hormone treatment in both sexes. However, digit ratios showed no sexual dimorphism among controls. Thus, present results are consistent with the hypothesis that variation in testosterone levels during development affects digit ratios.

Animals↗

The effect of an indirect anthelmintic treatment on parasites and breeding success of free-living pheasants Phasianus colchicus.

In Great Britain free-living common pheasants Phasianus colchicus are often managed at high densities owing to their popularity as a quarry species. They are prone to infection by a range of parasite species including Heterakis gallinarum, Capillaria spp. and Syngamus trachea. In 1995 the efficacy of an indirect anthelmintic technique for controlling parasitic worm burdens of pheasants was determined in a pilot study on a shooting estate in the south of England. Between 2000 and 2003 a large-scale field experiment was conducted on nine estates in eastern England to determine the effect of the technique on parasite burden and pheasant breeding success. In the absence of anthelmintic treatment worm burdens increased rapidly through March and April, whereas birds given anthelmintic-treated grain had lower worm burdens during the same period. The breeding success of pheasants was significantly higher on plots provided with anthelmintic treatment, although no long-term increases in population densities were observed. The burdens of the most common parasite H. gallinarum were significantly lower in pheasants from treatment plots six weeks after the anthelmintic treatment had ceased, but spring treatment did not influence parasite burden in the following winter.

Animal Feed↗