Gene Set Enrichment Analysis of Selenium-Deficient and High-Selenium Rat Liver Transcript Expression and Comparison With Turkey Liver Expression.

BACKGROUND: Better biomarkers of selenium (Se) status and a better understanding of toxic Se biochemistry are needed to set safe dietary upper limits. In previous studies, differential expression (DE) of individual liver transcripts in rats and turkeys failed to identify a single transcript that was consistently and significantly (q < 0.05) altered by high Se. OBJECTIVES: To evaluate the effect of Se status on rat liver transcript expression data at the level of gene sets, and to compare transcript expression in rats with that in turkeys to identify common regulated transcripts. METHODS: Gene set enrichment analysis (GSEA) was conducted on liver from weanling rats fed an Se-deficient basal diet (0.005 µg Se/g) supplemented with 0, 0.24 (Se-adequate), 2, or 5 µg Se/g diet as selenite for 28 d. In addition, transcript expression was compared with liver expression in turkeys fed 0, 0.4, 2, or 5 µg Se/g diet as selenite. RESULTS: Se deficiency significantly downregulated the rat selenoprotein gene set but also upregulated gene sets for a variety of pathways, processes, and disease states. GSEA of 2 compared with 0.24 µg Se/g found no significantly up- or downregulated gene sets, showing that 2 µg Se/g is not particularly toxic to the rat. GSEA analysis of 5 compared with 0.24 µg Se/g transcripts, however, found 27 significantly upregulated gene sets for a wide variety of conditions. Cross-species GSEA comparison of transcript expression, however, identified no common gene sets significantly and consistently regulated by high Se in rats and turkeys. In addition, comparison of individual marginally significant (unadjusted P < 0.05) DE transcripts between rats and turkeys also failed to find common transcripts. CONCLUSIONS: The dramatic increase in significant liver transcript DE and GSEA gene sets in rats fed 5 compared with 2 µg Se/g clearly appears to be a biomarker for Se toxicity, albeit not Se-specific. These analyses, however, failed to identify specific transcripts or pathways, biological states, or processes that were directly linked with high Se status, strongly indicating that adaptation to high Se lies outside transcriptional regulation.

Transcriptome analysis of the hypothalamus and pituitary of turkey hens with low and high egg production.

BACKGROUND: High egg producing hens (HEPH) show increased hypothalamic and pituitary gene expression related to hypothalamo-pituitary-gonadal (HPG) axis stimulation as well as increased in vitro responsiveness to gonadotropin releasing hormone (GnRH) stimulation in the pituitary when compared to low egg producing hens (LEPH). Transcriptome analysis was performed on hypothalamus and pituitary samples from LEPH and HEPH to identify novel regulators of HPG axis function. RESULTS: In the hypothalamus and pituitary, 4644 differentially expressed genes (DEGs) were identified between LEPH and HEPH, with 2021 genes up-regulated in LEPH and 2623 genes up-regulated in HEPH. In LEPH, up-regulated genes showed enrichment of the hypothalamo-pituitary-thyroid (HPT) axis. Beta-estradiol was identified as an upstream regulator regardless of tissue. When LEPH and HEPH samples were compared, beta-estradiol was activated in HEPH in 3 of the 4 comparisons, which correlated to the number of beta-estradiol target genes up-regulated in HEPH. In in vitro pituitary cell cultures from LEPH and HEPH, thyroid hormone pretreatment negatively impacted gonadotropin subunit mRNA levels in cells from both LEPH and HEPH, with the effect being more prominent in HEPH cells. Additionally, the effect of estradiol pretreatment on gonadotropin subunit mRNA levels in HEPH cells was negative, whereas estradiol pretreatment increased gonadotropin subunit mRNA levels in LEPH cells. CONCLUSIONS: Up-regulation of the HPT axis in LEPH and upstream beta-estradiol activation in HEPH may play a role in regulating HPG axis function, and ultimately ovulation rates. Thyroid hormone and estradiol pretreatment impacted gonadotropin mRNA levels following GnRH stimulation, with the inhibitory effects of thyroid hormone more detrimental in HEPH and estradiol stimulatory effects more prominent in LEPH. Responsiveness to thyroid hormone and estradiol may be due to desensitization to thyroid hormone and estradiol in LEPH and HEPH, respectively, due to up-regulation of the HPT axis in LEPH and of the HPG axis in HEPH. Further studies will be necessary to identify possible target gene desensitization mechanisms and elicit the regulatory role of the HPT axis and beta-estradiol on ovulation rates in turkey hens.

The hepatic transcriptome of the turkey poult (Meleagris gallopavo) is minimally altered by high inorganic dietary selenium.

There is interest in supplementing animals and humans with selenium (Se) above Se-adequate levels, but the only good biomarker for toxicity is tissue Se. We targeted liver because turkeys fed 5 µg Se/g have hepatic Se concentrations 6-fold above Se-adequate (0.4 µg Se/g) levels without effects on growth or health. Our objectives were (i) to identify transcript biomarkers for high Se status, which in turn would (ii) suggest proteins and pathways used by animals to adapt to high Se. Turkey poults were fed 0, 0.025, 0.4, 0.75 and 1.0 µg Se/g diet in experiment 1, and fed 0.4, 2.0 and 5.0 µg Se/g in experiment 2, as selenite, and the full liver transcriptome determined by RNA-Seq. The major effect of Se-deficiency was to down-regulate expression of a subset of selenoprotein transcripts, with little significant effect on general transcript expression. In response to high Se intake (2 and 5 µg Se/g) relative to Se-adequate turkeys, there were only a limited number of significant differentially expressed transcripts, all with only relatively small fold-changes. No transcript showed a consistent pattern of altered expression in response to high Se intakes across the 1, 2 and 5 µg Se/g treatments, and there were no associated metabolic pathways and biological functions that were significant and consistently found with high Se supplementation. Gene set enrichment analysis also found no gene sets that were consistently altered by high-Se and supernutritional-Se. A comparison of differentially expressed transcript sets with high Se transcript sets identified in mice provided high Se (~3 µg Se/g) also failed to identify common differentially expressed transcript sets between these two species. Collectively, this study indicates that turkeys do not alter gene expression in the liver as a homeostatic mechanism to adapt to high Se.

Ascorbic Acid Differentially Affects Stress Response and Resistance to Colibacillosis in Turkeys from Genetic Lines Differing in Growth Rate.

Genetic selection for fast growth can affect the ability of male turkeys to cope with stressors, resulting in decreased immunity to opportunistic bacterial infection. The purpose of the current study was to compare the effects of ascorbic acid (AA) on the stress response and resistance to Escherichia coli challenge of birds selected for increased 16-wk body weight (BW; F-line) with their random-bred parent line (RBC2). Male turkeys were raised in duplicate floor pens in a two line×two AA treatment×two stress challenge (SC) design. At 5 wk of age, AA (1200 ppm) was provided in drinking water for a 24-hr period, during which all birds were weighed. After AA treatment, the SC group was subjected to a transport stress protocol. Six hours after the start of transport, SC birds were also inoculated in the thoracic air sac with 1×10(4) colony-forming units of E. coli. The following morning four birds from each pen were bled, and all birds were weighed and necropsied 2 days later. BW and gain after SC were decreased in the F-line but not the RBC2 line, and there were no AA effects on BW. The weight of the bursa of Fabricius relative to BW was higher in the RBC2 line than in the F-line, was decreased by SC, and was not affected by AA. The heterophil∶lymphocyte ratio was higher in the SC F-line as compared to the SC RBC2 and was decreased by AA only in the SC F-line. Corticosterone (C) levels were increased by SC only in the F-line, and AA decreased C levels only in the RBC2 line. Airsacculitis scores were increased in the F-line SC birds. The challenge strain of E. coli was only detected in the air sac and liver of the AA-treated F-line SC birds and in the liver of the no-AA F-line birds. These results suggest that SC at 5 wk of age had a more deleterious effect on the fast-growing F-line than on its parent line and that AA may have increased susceptibility to colibacillosis in the SC F-line birds.

Tuberal hypothalamic expression of the glial intermediate filaments, glial fibrillary acidic protein and vimentin across the turkey hen (Meleagris gallopavo) reproductive cycle: Further evidence for a role of glial structural plasticity in seasonal reproduction.

Glia regulate the hypothalamic-pituitary-gonadal (HPG) axis in birds and mammals. This is accomplished mechanically by ensheathing gonadotrophin-releasing hormone I (GnRH) nerve terminals thereby blocking access to the pituitary blood supply, or chemically in a paracrine manner. Such regulation requires appropriate spatial associations between glia and nerve terminals. Female turkeys (Meleagris gallopavo) use day length as a primary breeding cue. Long days activate the HPG-axis until the hen enters a photorefractory state when previously stimulatory day lengths no longer support HPG-axis activity. Hens must then be exposed to short days before reactivation of the reproductive axis occurs. As adult hens have discrete inactive reproductive states in addition to a fertile state, they are useful for examining the glial contribution to reproductive function. We immunostained tuberal hypothalami from short and long-day photosensitive hens, plus long-day photorefractory hens to examine expression of two intermediate filaments that affect glial morphology: glial fibrillary acidic protein (GFAP) and vimentin. GFAP expression was drastically reduced in the central median eminence of long day photosensitive hens, especially within the internal zone. Vimentin expression was similar among groups. However, vimentin-immunoreactive fibers abutting the portal vasculature were significantly negatively correlated with GFAP expression in the median eminence, which is consistent with our hypothesis for a reciprocal relationship between GFAP and vimentin expression. It appears that up-regulation of GFAP expression in the central median eminence of turkey hens is associated with periods of reproductive quiescence and that photofractoriness is associated with the lack of a glial cytoskeletal response to long days.

Phospholipid hydroperoxide glutathione peroxidase (Gpx4) is highly regulated in male turkey poults and can be used to determine dietary selenium requirements.

The dietary selenium recommendation for turkeys of 0.2 microg Se/g is higher than for many other species. Liver glutathione peroxidase-1 (Gpx1) activity levels determined using hydrogen peroxide (H(2)O(2)) in previous studies suggest that 0.2 microg Se/g may still be too low and that some of this Gpx1 activity might be due to phospholipid hydroperoxide Gpx (Gpx4). Thus we separated Gpx1 from Gpx4 by chromatography, demonstrated that 47% of the H(2)O(2) activity in Se-adequate turkey liver was due to Gpx4, and determined a factor for calculation of each activity. Day-old male poults were fed an Se-deficient torula diet (0.007 microg Se/g) supplemented with graded levels of Se (0-0.5 microg Se/g) for 27 days. Final body weights indicated a minimum Se requirement for growth of 0.05 microg Se/g. The liver had the highest Gpx4 activity in Se-adequate poults, and Gpx4 activity in Se-deficient liver decreased to 5% of Se-adequate levels, with an Se requirement of 0.29 microg Se/g. Liver Gpx1, gizzard Gpx1 and gizzard Gpx4 activities also had Se requirements of 0.28-0.30 microg Se/g, collectively yielding an Se requirement of 0.3 microg Se/g, which is three times higher than the requirements found in comparable rodent studies. We also sequenced partial cDNA clones for turkey Gpx1 (GQ502186) and Gpx4 (GQ502187), and found >60% identity with rodents and humans and >90% identity with chickens. Ribonuclease protection analysis showed that Gpx4 mRNA levels decrease substantially in Se-deficient turkey liver, unlike in rodents. These underlying differences in selenoprotein molecular biology may explain the elevated dietary Se requirements of turkeys.

Effects of Escherichia coli challenge and transport stress on hematology and serum chemistry values of three genetic lines of turkeys.

Three lines of turkeys were compared for response to an Escherichia coli challenge followed by transport stress (transport). The turkey lines were a slow-growing line selected for increased egg production (egg line), a fast-growing line selected for increased 16-wk BW (F line), and a commercial line (Comm line). Birds were challenged at 14 wk of age with an air sac injection of 5,000 to 10,000 cfu of E. coli. At 8 d postchallenge, birds were subjected to a transport stress procedure that included 12 h of holding time in a transport vehicle. The following morning all birds (n = 10 to 19 birds/line) were bled. Whole blood was analyzed using the Cell-Dyn 3500 blood analysis system (Abbott Diagnostics), and serum chemistry was measured using the Express Plus analyzer (Ciba-Corning Diagnostics Corp.). Transport significantly decreased the levels of hematocrit, hemoglobin, mean cell volume, mean corpuscular hemoglobin, glucose, triglycerides, cholesterol, phosphorus, iron, albumin, and alkaline phosphatase (AP) and increased the levels of uric acid, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, and creatine kinase. Line differences were variable, but the levels of both iron and AP were least in the fastest-growing Comm line birds and greatest in the slowest-growing egg-line birds with intermediate values in the F line. Iron and AP were also the only parameters influenced by sex, with males having greater levels of both compared with females. The creatine kinase levels were more than 6-fold greater in transported Comm line birds, and iron levels of transported Comm males were 3-fold less than controls. Previously, the growth rate of these lines was positively correlated with increased heterophil to lymphocyte ratios and susceptibility to colibacillosis. The differences seen in the Comm line for these commonly measured blood parameters suggest that they may be useful for profiling flocks to determine their response to transport stress and feed withdrawal.

Development of a real-time (Q) PCR assay to measure variation in expression of prolactin receptor mRNA in the hypothalamus and pituitary gland during late embryogenesis in turkeys and chickens.

Changes in levels of PRLR mRNA in the pituitary gland and hypothalamus of chickens and turkeys from embryonic day (ED) 15 and ED21 to 1 day post-hatch, respectively, were measured by real-time PCR. In both species, PRLR mRNA increased from low levels during the last week of ED to reach maxima at the peri-hatch period. Similarly, circulating levels of PRL also increased during this interval and were highly correlated with levels of the PRLR mRNA in both the pituitary gland and hypothalamus. This suggests that PRL was up-regulating its receptor. In support of this, stimulation of the turkey pituitary gland with VIP on ED24 resulted in a 4- and 3-fold increase in PRL and PRLR, respectively. Since VIP had no direct effect on the levels of PRLR transcript in the hypothalamus, it is likely that VIP is acting indirectly through increased PRL to up-regulate the number of receptors.

Poult performance as influenced by age of dam, genetic line, and dietary vitamin E.

An experiment was conducted to measure the effects of age of dam, genetic line, and dietary levels of vitamin E on growth and immunocompetence of poults. Age of dam was defined as younger (in early egg production) and older (past peak production); line consisted of a commercial sire and dam line; and dietary vitamin E was supplemented into the diet at 10 and 300 IU/kg of feed. Traits measured included body, liver, gizzard, and yolk sac weights at hatch; BW and feed conversion to 9, 28, and 42 d; response to SRBC, Phaseolus vulgaris agglutinin-P, and Escherichia coli administered at 28 d of age; and response to a cold stress on d 5 posthatch. Differences among genetic lines were evident with growth greater for poults from the sire than from the dam line. Performance of poults from older dams was generally superior to that of poults from younger dams. The higher level of vitamin E resulted in a greater than 7-fold increase in blood plasma vitamin E and reduced mortality. There were interactions among the main effects in which the fitness of poults from younger dams was enhanced by the higher level of vitamin E and the effect of breeder age differed among genetic lines.

Isolation, cloning and sequencing of transferrins from red-eared turtle, African ostrich, and turkey.

Transferrins form an important class of iron-binding proteins widely distributed in the physiological fluids of vertebrates and invertebrates. In vertebrates they are present mostly in serum as serotransferrins. In birds and reptiles transferrins are also found in eggs as ovotransferrins. However, until now only chicken and duck ovotransferrin sequences have been published. This paper presents data on the purification, biochemical analysis, cloning and sequencing of ovotransferrins from red-eared turtle, African ostrich and turkey, revealing their significant homology with other known ovotransferrin sequences. The proteins were purified by size-exclusion and anion-exchange chromatography. Isoelectric points, iron-saturated and iron-free spectra, as well as the mRNA nucleotide sequences of 2,409 nt (ORF: 2,106 nt encoding a 701-amino-acid polypeptide; ), 2,418 nt (ORF 2,118 nt encoding a 705-amino-acid polypeptide; ), and 2,397 nt (ORF: 2,118 nt encoding a 705-amino-acid polypeptide; ) were determined for ostrich (OtrF), red-eared turtle (TtrF), and turkey (MtrF) ovotransferrin, respectively.

Identification of two alphaRYR alleles and characterization of alphaRYR transcript variants in turkey skeletal muscle.

We hypothesized that a mutation in the turkey skeletal muscle ryanodine receptor alpha isoform (alphaRYR) underlies turkey meat quality problems which are strikingly similar to pale, soft, exudative (PSE) pork. RT-PCR analysis of turkey alphaRYR mRNA covering amino acids 376 to 615 (numbered according to the human sequence) revealed at least three transcript variants. One transcript was homologous to the mammalian skeletal muscle RYR1 sequence in this region. The second transcript variant (AS-81) was characterized by the absence of 81 bases located at the beginning of exon 13, while the third transcript variant (AS-193) carried a deletion of 193 bases, corresponding to the entire exon 13. Two alphaRYR genomic DNA alleles (alphaRYR-I and alphaRYR-II) carrying the region of deletions in the turkey cDNA sequences were identified. Nucleotide sequence analysis demonstrated that the two alleles are identical in exon sequences but different in intron sequences. Comparison of genomic and cDNA sequences indicated that both AS-81 and AS-193 transcript variants probably arose via alternative splicing. Consistent with this mechanism, the last eight nucleotides of the 81 bases form a consensus sequence for a splice acceptor site. Both alleles could give rise to the AS-81 and AS-193 transcript variants via alternative splicing. Birds homozygous for alphaRYR-II tended to have superior meat quality indicators including significantly higher muscle pH at 15-min post mortem and lower muscle exudate at 24-h post mortem, compared to birds homozygous for alphaRYR-I.

An R111C polymorphism in wild turkey cardiac troponin I accompanying the dilated cardiomyopathy-related abnormal splicing variant of cardiac troponin T with potentially compensatory effects.

Cardiac muscle contraction is regulated by Ca(2+) through the troponin complex consisting of three subunits: troponin C (TnC), troponin T (TnT), and troponin I (TnI). We reported previously that the abnormal splicing of cardiac TnT in turkeys with dilated cardiomyopathy resulted in a greater binding affinity to TnI. In the present study, we characterized a polymorphism of cardiac TnI in the heart of wild turkeys. cDNA cloning and sequencing of the novel turkey cardiac TnI revealed a single amino acid substitution, R111C. Arg(111) in avian cardiac TnI corresponds to a Lys in mammals. This residue is conserved in cardiac and skeletal muscle TnIs across the vertebrate phylum, implying a functional importance. In the partial crystal structure of cardiac troponin, this amino acid resides in an alpha-helix that directly contacts with TnT. Structural modeling indicates that the substitution of Cys for Arg or Lys at this position would not disrupt the global structure of troponin. To evaluate the functional significance of the different size and charge between the Arg and Cys side chains, protein-binding assays using purified turkey cardiac TnI expressed in Escherichia coli were performed. The results show that the R111C substitution lowered binding affinity to TnT, which is potentially compensatory to the increased TnI-binding affinity of the cardiomyopathy-related cardiac TnT splicing variant. Therefore, the fixation of the cardiac TnI Cys(111) allele in the wild turkey population and the corresponding functional effect reflect an increased fitness value, suggesting a novel target for the treatment of TnT myopathies.

Molecular cloning and characterization of alternatively spliced transcripts of the turkey pituitary adenylate cyclase-activating polypeptide.

Pituitary adenylate cyclase-activating polypeptide (PACAP) increases the release of growth hormone (GH) and prolactin (PRL) in mammals. However, the evolutionary and functional relationships of PACAP, GH, and PRL are not clear. To understand how PACAP is regulated in the turkey, a turkey PACAP (tPACAP) cDNA has been cloned by the combination of reverse transcription-polymerase chain reaction and the rapid amplification of cDNA 5'- and 3'-ends. The deduced amino acid sequence of tPACAP-38 and turkey PACAP-related peptide (tPRP) displayed 87-97 and 52-63% similarity when compared to a variety of known PACAP-38 and PRP sequences, respectively. Two major transcripts (1.3 and 3.0 kb) of tPACAP were detected by Northern blot analysis. The highest levels of tPACAP mRNA were shown to be expressed in the hypothalamus, the cerebellum, and the cerebrum. In contrast, most of the other tissues tested expressed relatively low steady-state levels of tPACAP mRNA. Alternative splicing of tPACAP resulted in the expression of two different isoforms. The smaller form of tPACAP was expressed in the hypothalamus during early embryonic development and decreased significantly in later stages.

Turkey osteomyelitis complex.

Turkey osteomyelitis complex (TOC) is defined by the US Food Safety Inspection Service (FSIS) to include normal-appearing processed turkey carcasses that contain lesions including green discoloration of the liver, arthritis/synovitis, soft-tissue abscesses, and osteomyelitis of the proximal tibia. The lesions are associated with many different opportunistic organisms, mainly Staphylococcus aureus and Escherichia coli, suggesting that TOC incidence may be influenced more by deficiencies in the host immune response rather than by the virulence of any one organism. This syndrome is primarily a disease of adolescent male turkeys, and birds with TOC lesions have decreased indices of cell-mediated immunity, leading to the hypothesis that defects in the immune response of individuals within flocks of male turkeys may be responsible for the occurrence of these opportunistic infections. We have developed an experimental model for this disease in which treatment with dexamethasone (DEX), either with or without air sac inoculation with Escherichia coli, produces all of the lesions associated with TOC. These studies suggest that TOC is a result of stress-induced immunosuppression in a subpopulation of male turkeys that respond to the stressors in modern poultry production in a detrimental manner. Supplemental vitamin D3 treatment protected male turkeys from the immunosuppression induced by multiple treatments with DEX and resulted in decreased incidence of mortality, TOC, green liver, and isolation of bacteria from tissues, lower air sacculitis scores, and lower heterophil to lymphocyte ratios than nonsupplemented controls. Vitamin D3 also protected BW; relative weights of the liver, heart, spleen, and bursa; and clinical chemistry values from the effects of DEX treatment. The ability of vitamin D3 supplementation to protect turkeys from the immunosuppressive effects of severe stress emphasizes its role as a prohormone that affects health and disease resistance in turkeys and suggests that variation in the vitamin D receptor genotype may be involved in this disease process. This model has potential value in the identification of other nutritional and physiological immunomodulators that can decrease TOC incidence and will provide a means for the divergent selection of birds more resistant to the stressors of turkey production. In addition, this model will provide justification for management options designed to minimize stress.

Three different turkey luteinizing hormone receptor (tLH-R) isoforms I: characterization of alternatively spliced tLH-R isoforms and their regulated expression in diverse tissues.

Using combinations of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends, three different, alternatively spliced, partial turkey LH receptor (tLH-R) cDNA isoforms were characterized from ovarian mRNA. The first cDNA (tLH-R(intact)) showed 98% and 72-75% similarity with chicken and mammalian LH-R sequences, respectively. The second cloned cDNA isoform (tLH-R(insert)) contained an in-frame TGA stop codon within an 86-base pair insertion that was located in the extracellular domain of the seven-transmembrane region. The tLH-R(insert) isoform could encode a truncated soluble protein isoform that lacked the transmembrane region. The third cDNA isoform truncated the transmembrane region (tLH-R(trunc)) and was derived by the deletion of the last exon by incomplete splicing. Generation of multiple transcripts by alternative splicing was elucidated by partial characterization of tLH-R genomic sequences. The differentially regulated expression of the tLH-R mRNA isoforms in nongonadal tissues and ovarian stromal tissues during various reproductive stages was quantified and analyzed by Northern blot and/or RT-PCR. Alternatively spliced tLH-R isoforms were differentially expressed in a tissue-specific manner in most of the tissues examined. The steady-state levels of tLH-R mRNA isoforms were relatively high in the hypothalamus and optic nerve and relatively low in the cortex, pituitary, and cerebellum when compared to levels in ovarian follicles. In nongonadal reproductive tissues, the steady-state levels of tLH-R mRNA isoforms were relatively high in the uterus and infundibulum and relatively low in the isthmus, oviduct, and magnum. In addition, in the nongonadal peripheral tissues, the steady-state levels of tLH-R isoforms were relatively high in the thyroid gland and relatively low in the spleen, adrenal gland, kidney, skin, bursa, and muscle. The present study suggests that the alternative splicing of LH-R transcripts occurs in a tissue-specific manner and has been evolutionarily conserved (similar results were obtained in chicken and swine). These results raise fundamental questions as to the function of LH-R isoforms in nongonadal tissues.

Effect of supplemental oxygen on blood plasma organic acids within embryos from selected lines of turkeys.

The hypothesis was tested that increased oxygen tensions during the plateau stage of oxygen consumption (25 and 26 d of incubation) would cause different metabolic responses from embryos selected for increased egg production or growth. Embryos were exposed to 171 or 152 mm Hg partial pressure of oxygen from 25 to 28 d of incubation, a time when the oxygen conductance properties of the eggshell are exceeded by the embryonic tissue demands for oxygen. Carbohydrate and lipid metabolism were observed by measuring plasma organic acids in embryos from selected lines and randombred controls. (E was selected for increased egg production; RBC1 is the randombred line from which it was selected. F was selected for increased BW; RBC2 is the randombred line from which it was selected.) During the plateau stage in oxygen consumption, RBC2 embryos responded to added oxygen by utilizing fat rather than carbohydrate, whereas F embryos responded by using less fat as well as less carbohydrate from the liver and kidney. The response of F embryos to added oxygen is the opposite that might be expected for aerobic metabolism. The reason that selection for growth has resulted in such a metabolism is unknown. The E embryos displayed depressed lactate and beta-hydroxybutyrate levels, but plasma urates were elevated compared with RBC1, suggesting that the selection for egg production has also resulted in a unique metabolism. The embryonic processes described in the current study suggest that selected embryos are unable to respond to elevated partial pressure of oxygen by adjusting energy metabolism, which may result in increased embryonic mortality during this stage.

Effect of genetics and maternal dietary iodide supplementation on turkey embryonic growth.

Embryonic growth of a turkey lines selected for 16-wk BW (F) or 180-d egg production (E) was measured and compared to randombred controls (RBC2 or RBC1). Egg weight at setting relative to poult weight at hatching indicated increased growth in F as well as E embryos compared to randombred controls. Eggs from F weighed 10 g more than those of RBC2 (P < or = 0.0001) but the poults at hatching were only 8 g heavier (P < or = 0.0001). Water vapor loss during incubation indicated that only 0.9% of the difference could be accounted for by water vapor loss. Selection for increased 16-wk BW resulted in decreased embryo growth rates relative to hatchling mass (P < or = 0.0001) beginning at Day 16 of incubation compared to that of RBC2. Eggs from E weighed 15 g less than RBC1 (P < or = 0.0001) but produced poults weighing only 7 g less (P < or = 0.0001). Incubation water vapor loss was depressed in E compared to RBC1 (P < or = 0.0001) but accounted for only 1.4% of the difference between hatchling weights. Selection for egg production increased embryo growth rates (relative to hatchling mass) measured at 4-d intervals compared to those of the RBC1 line (P < or = 0.05). Iodide supplementation of the maternal diet depressed (P < or = 0.05) glycogen in F embryos but interacted with line to generally increase glycogen in E embryos. Increased glycogen was related to increased growth rates in E but not F line embryos. It may be concluded that iodide supplementation of the material diet and genetics are determinants of embryonic growth in turkeys.

Effects of genetics and maternal dietary iodide supplementation on glycogen content of organs within embryonic turkeys.

In prior studies it was shown that the growth of turkey embryos was dependent upon maternal dietary iodide as well as genetic selection. The current study posed the question of which organ systems respond to these variables. Embryos from lines selected for 16-wk BW grew at the same rate as unselected embryos from the randombred population serving as the initial source of the selected line until approximately 21 d of incubation (selected = F; randombred control = RBC2). Line differences in growth of F embryos could be accounted for increased liver and heart growth at the expense of muscle growth. Muscle growth increased in the growth-selected line prior to pipping. Muscle growth was affected less when dams were selected for egg production (selected = E; randombred control = RBC1). Muscle growth was slowed in E line embryos compared to that of RBC1, and liver and heart growth were slowed at internal and external pipping stages in E embryos compared to RBC1. Early muscle growth was augmented when F dams were fed supplemental iodide. A similar response was observed in E line embryos but occurred at a later stage of development. Measurements indicated decreased tissue glycogen in liver, heart, and muscle of selected lines may be one possible mechanism by which growth or organ function may come in conflict.

Bobber: a cervical ataxic mutant in the domestic turkey responsive to sunlight.

Bobber is a genetic disorder in the domestic turkey that is usually expressed between two and four weeks of age. The condition is permanent and is characterized by ventrocaudad bending of the neck accompanied by a lateral pendulumlike motion of the head between the legs. Expressivity of the defect is variable and may be exhibited in some turkeys as a stargazing posture or a rapid clockwise twirling motion. When suspended by the legs in a head-down orientation, afflicted turkeys exhibit an inward turning of the neck and head toward the breast as opposed to an outward turning in normal turkeys. The disorder is inherited as a sex-linked recessive trait. The symbol bo is used for the gene. The defect can be corrected by exposure to intense light in the visible spectrum.

Influence of genetic increases in egg production on traits associated with broodiness in turkeys.

Egg production, nesting frequency, and serum levels of prolactin, estradiol, and total phosphorus were monitored in relatively nonbroody (egg) and relatively broody (RBC1) strains of turkey hens during a reproductive period. In the egg strain, prolactin levels were increased in a group with a relatively high frequency of nesting in comparison to a group with a relatively low frequency of nesting. No differences between these two groups were detected for serum estradiol, total phosphorus, or egg production. In the RBC1 strain, prolactin levels did not differ between a group of hens that did not exhibit broodiness and a group that exhibited one or more bouts of broodiness. A broodiness treatment was used for the latter group. The broody group exhibited extremely variable levels of prolactin. In individual broody hens, the levels of prolactin were relatively high. After broodiness in 12 of 13 hens, the level of prolactin fell to relatively low levels. In the one hen not responding to broodiness treatment, the level of prolactin became low levels. In the one hen not responding to broodiness treatment, the level of prolactin became extremely high. Nesting frequency, total serum phosphorus, and egg production were generally not different between the two groups. The level of prolactin showed seasonal changes in both strains of hens, starting low, increasing to maximal levels between 40 and 80 days of production, and then declining to low levels late in the reproductive period. Laying hens always had higher levels of prolactin than nonlaying, nonbroody hens.