Genetic control of embryonic cardiac growth and functional maturation in turkeys.

Turkey experimental lines E (selected 44 yr for increased total egg production) and F (selected 38 yr for increased 16-wk BW) were mated reciprocally with the randombred control lines from which they were derived (RBC1 and RBC2, respectively), and the pureline and reciprocal cross poults were compared for their BW, heart weight, heart rates, myocardial glycogen and lactate concentrations, and plasma creatine kinase (CK) and lactate dehydrogenase (LDH) activities. The CK and LDH were used as indicators of cardiac insufficiency. Orthogonal contrasts of the data from the pureline and reciprocal cross data were used to estimate additive genetic effects, reciprocal effects (confounded maternal and sex-linked effects), and heterosis for each of the traits measured. Long-term selection for increased egg production in the E line has reduced embryo heart weight and has altered the energy metabolism of the myocardium. The differences in energy metabolism may be due to the more rapid heart rates. Conversely, long-term selection for increased 16-wk BW has significantly decreased the heart rate of F line embryos and has not changed the weight of the heart relative to the BW until the embryo has passed through the plateau stage. The F line embryos show a different energy metabolism that relies much more on gluconeogenesis. Embryo deaths occur more frequently in turkey embryos when the energy metabolism of the myocardium shows elevated glycogen to lactate ratios as it did in the pure E and F lines.

Effects of incubator temperature and oxygen concentration during the plateau stage of oxygen consumption on Turkey embryo long bone development.

Temperature (TEM) and O(2) concentrations during the plateau stage of oxygen consumption are known to affect yolk utilization, tissue development, and thyroid metabolism in turkey embryos. Three experiments were conducted to evaluate these incubation effects on long bone development. Fertile eggs of Nicholas turkeys were used. In each trial, standard incubation conditions were used to 24 d, when the eggs containing viable embryos were randomly divided into 4 groups. Four experimental cabinets provided 4 TEM (36, 37, 38, or 39 degrees C) or 4 O(2) concentrations (17, 19, 21, or 23% O(2)). In the third experiment, 2 temperatures (36 and 39 degrees C) and 2 O(2) concentrations (17 and 23%) were evaluated in a 2 x 2 factorial design. Body and residual yolk weights were obtained. Both legs were dissected, and shanks, femur, and tibia weights, length, and thickness were recorded. Relative asymmetry of each leg section was calculated. Chondrocyte density was evaluated in slides stained with hematoxylin and eosin. Immunofluorescence was used to evaluate the presence of collagen type X and transforming growth factor beta. Hot TEM caused reduction of tibia weights and increase of shank weight when compared with cool TEM. The lengths of femur, tibia, and shanks were reduced by 39 degrees C. The relative asymmetry of leg weights were increased at 38 and 39 degrees C. Poult body and part weights were not affected by O(2) concentrations, but poults on 23% O(2) had bigger shanks and heavier tibias than the ones on 17% O(2). High TEM depressed the fluorescence of collagen type X and transforming growth factor beta. The O(2) concentrations did not consistently affect the immunofluorescence of these proteins. The chondrocyte density was affected by TEM and O(2) in resting and hypertrophic zones. In the third experiment, high TEM depressed BW, leg muscle weights, and shank length. Low O(2) reduced tibia and shanks as a proportion of the whole body. We concluded that incubation conditions affect long bone development in turkeys.

Broiler embryo bone development is influenced by incubator temperature, oxygen concentration and eggshell conductance at the plateau stage in oxygen consumption.

1. Four experiments were conducted to evaluate the effects of temperature (TEM) and oxygen (O(2)) concentrations during the last 4 d of incubation on bone development. Fertile eggs from two strains were obtained that either exhibited Low or High eggshell conductance (G). 2. Four experimental cabinets provided either four TEM (36, 37, 38 or 39 degrees C) or four O(2) concentrations (17, 19, 21 or 23% O(2)). Data were analysed as a 2 x 2 factorial design. In the fourth experiment, two temperatures (36 and 39 degrees C), two O(2) concentrations (17 and 23%) and the same Low and High G strains were evaluated in a 2 x 2 x 2 factorial design. 3. Body weights (BW) and residual yolks were obtained, both legs were dissected. Femur, tibia and shank weights, length and thickness were recorded. Relative asymmetry (RA) of each leg section was calculated. 4. The results indicated that elevated TEM during incubation increased RA between the two legs, mainly in the Low G strain. Chickens at the lowest O(2) concentrations had lighter and shorter tibias, lighter shanks, and increased RA of femur length compared to chickens in the 23% O(2). In the fourth experiment no interactions were observed between O(2) and TEM. High TEM depressed BW of Low G broilers, but no significant effect of treatments was observed on BW of High G broilers. Nevertheless, the high TEM or low O(2) independently caused reduced femur and tibia weights and length, shank length and thickness, and both low O(2) and high TEM together increased RA in shank weight. 5. These results suggest that late incubation conditions affect long bone development in broilers.

Dam line and sire line effects on turkey embryo survival and thyroid hormone concentrations at the plateau stage in oxygen consumption.

Inheritance of embryo thyroid function was measured in lines of turkeys. Two lines that had been selected for either increased egg production (E) or increased 16-wk BW (F) and their respective randombred controls (i.e., RBC1 and RBC2) were examined. Reciprocal crosses of dams and sires from each selected line and its randombred control were made to estimate sire line and dam line effects. Orthogonal contrasts were used to determine if the differences found were due to the presence of additive, nonadditive, or maternal, sex-linked, or both, gene effects. With the data involved, sex-linkage and maternal effects could not be separated. Embryo survival was measured for all lines and their reciprocal crosses. Crossing the RBC1 sire and E dam also resulted in better embryo survival and lower death losses at pipping than for the other cross- or purelines. Reciprocal crosses of the F and RBC2 lines showed better total embryo survival, and they survived pipping better than the F or RBC2 purelines. Thyroxine (T(4)) and triiodothyronine (T(3)) concentrations differed between the reciprocal crosses at external pipping, but the effects were inconsistent for the 2 data sets. Reciprocal tests indicated that maternal, sex-linked, or both, effects were present for T(3) concentrations at internal pipping in the E and RBC1 lines and at external pipping for the F and RBC2 lines. Reciprocal effects were significant for T(4) at internal pipping for both data sets. The RBC1 sire embryos had significantly higher T(3):T(4) ratios than the E line sire embryos at internal and external pipping, and the pureline RBC1 embryos had consistently higher ratios than the pureline E embryos. The differences for the T(3):T(4) ratios between these 2 lines at internal pipping, external pipping, and hatch appeared to be consistently additive in nature, although significant nonadditive or heterotic effects were present for the ratio at external pipping. Similar effects on the T(3):T(4) ratio were observed for the F and RBC2 lines at external pipping.

Follicular development and expression of the messenger ribonucleic acid for the inhibin/activin subunits in two genetic lines of turkey hens that differ in total egg production.

The characterization of the follicular hierarchy and the expression of the mRNA for the inhibin/activin subunits was investigated in the follicles of 2 lines of turkey hens selected for over 40 generations for increased egg production (Egg line) or increased body weight (Growth line). The follicular hierarchies of 6 hens from the Egg and Growth lines were characterized in middle (45 wk of age) and late production (58 wk of age). Relative follicular weights for individual hierarchical follicles (>12 mm), pooled small yellow follicles (5 to 12 mm), and large white follicles (2 to 5 mm) were calculated. Total RNA was extracted for Northern blot analysis from individual granulosa cell layers of the F1 through F4 follicles, and from the combined granulosa and theca layers of small yellow follicles and large white follicles from an additional 6 hens from each genetic line. Egg line hens displayed a more distinct follicular size hierarchy than Growth line hens at 45 and 58 wk. Although total follicular weight relative to body size was greater at 45 and 58 wk of age for the Egg line hens than the Growth line hens, the total number of hierarchical follicles was greater in the Growth line hens at 45 and 58 wk of age. Expression of follistatin and the inhibin beta(B)-subunit was highest in nonhierarchical follicles, whereas the expression of the inhibin alpha- and beta(A)-subunits was highest in the hierarchical follicles. The inhibin alpha- and beta(A)-subunit mRNA expression pattern in the 4 largest follicles of the Growth line hens was not similar to the Egg line hens or characteristic of laying hens that have a high rate of egg production. The unusual inhibin subunit mRNA expression in the largest hierarchical follicles of the Growth line hens may account for their development of an abnormal follicular size hierarchy and for their poor egg production.

Genetic control of neonatal growth and intestinal maturation in turkeys.

Turkey experimental lines E (selected 44 yr for increased egg production) and F (selected 38 yr for increased 16-wk BW) were mated reciprocally with the randombred control lines from which they were derived (RBC1 and RBC2, respectively), and the pure line and reciprocal cross poults were compared according to their hatch, 3- and 7-d BW, jejunum weight, jejunum length, and jejunal maltase and alkaline phosphatase activities. Orthogonal contrasts of the data from the pure line and reciprocal cross-poult data were used to estimate additive genetic effects, reciprocal effects (confounded maternal and sex-linked effects), and heterosis for each of the traits measured. Body weights at hatch and at 3 and 7 d of age were increased in the F line relative to the RBC2 line and were decreased in the E line relative to the RBC1 line. The genetic changes from long-term selection in the E and F lines have had concomitant effects on jejunum growth and function that parallel the changes in growth rate. The increased BW of the F line poults and the decreased BW of the E line poults relative to their randombred controls may be due to increases in the absorption of nutrients because of greater intestinal mass rather than to differences in glucose digestion. Concomitant changes in egg weight in the 2 selected lines appear to have resulted in maternal effects that have significantly affected neonatal BW and digestive system maturation.

The effect of broiler breeder genetic strain and parent flock age on eggshell conductance and embryonic metabolism.

The effect of genetic strain (Ross 308; Cobb 500) and parent flock age [young (29 wk), peak (Ross = 34 wk; Cobb = 36 wk), postpeak (40 wk), mature (45 wk), old (55 wk), and very old (59 wk)] on eggshell conductance and embryonic metabolism were examined. At each flock age, eggs from each strain were incubated for 21.5 d in individual metabolic chambers to measure embryonic O(2) intake and CO(2) output. From these data, the respiratory quotient (RQ) and metabolic heat production were calculated. Data were analyzed by the GLM procedure of SAS at P < or = 0.05. Neither strain nor flock age influenced conductance. Total embryonic O(2) consumption, CO(2) output, RQ, and metabolic heat production over the entire incubation period were not affected by strain. Daily differences existed between strains for embryonic O(2) intake (1, 7, 16, 17, 19, 20 d of incubation), CO(2) output (1 to 4, 16 to 20 d of incubation), and heat production (4, 7, 16 to 19 d of incubation). Embryos from young, mature, old, and very old flocks produced significantly more total embryonic heat over the entire 21 d (1,712, 1,677, 1,808, and 1,832, respectively) than embryos from peak (1,601) and postpeak (1,693) flocks. Average RQ for the entire incubation period was higher in embryos from mature flocks compared with all other flock ages. Daily differences among embryos from different flock ages were shown for O(2) consumption (all but d 8 of incubation), CO(2) production (all but d 7 and 9 of incubation), and heat output. The results showed that genetic strain and parent flock age influence daily embryonic metabolism, especially during the early and latter days of incubation. These daily differences coincide with the days of incubation having a higher incidence of embryonic mortality; these 2 factors may be related. Further investigation into the relationship between embryonic metabolic heat production and mortality during incubation may lead to the development of specific incubation conditions for different genetic strains and flock ages.

Zona pellucida protein B2 messenger ribonucleic acid expression varies with follicular development and granulosa cell location.

The freshly ovulated ovum in avian species is surrounded by a protein layer called the inner perivitelline layer (IPVL). The IPVL contains zona pellucida proteins and 6 distinct zona pellucida genes have been identified (ZPA, ZPB1, ZPB2, ZPC, ZPD and ZPX1) in the chicken. In the present research, the expression of the mRNA for ZPA, ZPB2, and ZPX1 was investigated in 2 lines of turkey hens selected for either increased egg production (E line) or increased body weight (F line). Theca and granulosa cell expression of the mRNA for ZPA and ZPB2 was also investigated in hierarchical and prehierarchical follicles from broiler breeder hens. Granulosa tissue was collected from F1 through F4 and F1 through F10 follicles in E line and F line hens, respectively. A one cm2 section of the granulosa layer around the germinal disc (GD) and an equivalent sized nongerminal disc (NGD) area was also collected from the F1 and F2 follicles from other hens from each genetic line. Granulosa and theca tissue was collected from hierarchical and prehierarchical follicles of broiler breeder hens. Total RNA was extracted from the samples. Minor groove-binding probes and primers for detecting ZPA, ZPB2, and ZPX1, were made for real-time PCR analyses. Expression of ZPA, ZPB2, and ZPX1 was detected in all follicle sizes from both genetic lines of hens. No significant differences in ZPA and ZPX1 mRNA expression were detected between the GD and NGD granulosa cells. However, the expression of the mRNA for ZPB2 was significantly greater in the GD granulosa cells when compared to the NGD granulosa cells in F1 and F2 follicles from E line and F line hens. In broiler breeder hens, the mRNA expression of ZPA and ZPB2 was greatest in the smallest prehierarchical follicles. The results suggest that higher expression of ZPB2 in the germinal disc area may be important for the preferential binding of sperm to this region of the IPVL.

Effect of dietary phosphorus and phytase levels on the reproductive performance of large white turkey breeder hens.

An experiment was conducted to determine the effect of dietary P levels and dietary phytase enzyme (E) inclusion on Large White turkey breeder hen reproductive performance from 31 to 62 wk of age. Hens were placed in a curtain-sided house with 48 pens (10 birds per pen; 8 pens per treatment) at 31 wk and were fed a breeder ration with treatments as follows: HP, dietary available P = 0.55%; HPE, HP + E; MP, dietary available P = 0.35%; MPE, MP + E; LP, dietary available P = 0.17%; and LPE, LP + E. Feed and water were available ad libitum for 28 wk of lay. Diets were fed in mash form, and all other nutrients were formulated to meet or exceed NRC requirements. All hens were photostimulated in January (31 wk) with 15.5 h of light daily. Production data were recorded on a pen basis. Individual bird BW and feed consumption, by pen, were determined at monthly intervals from 31 to 62 wk. Hens were observed for weekly reproductive performance for hen housed egg production, hen-day egg production, settable eggs, cumulative settable eggs, hens out of lay, and hen mortality and for biweekly performance for egg fertility, hatchability of all eggs, hatchability of fertile eggs, egg weight loss, conductance, conductance constant (k), and embryonic mortality. Egg weight, eggshell thickness, egg components, and albumen and yolk P were measured monthly. At 62 wk of age, hen tibia P, plasma P, total fecal P, and water-soluble fecal P were determined. Decreasing dietary P resulted in no decreases in reproductive performance for turkey breeder hens to 62 wk. Additionally, decreased dietary P resulted in decreased total fecal P and water-soluble fecal P. Feeding turkey breeder hens dietary phytase enzyme resulted in significantly fewer hens going out of lay; however, this was not reflected in hen housed egg production. It was concluded that phosphorus could be lowered in turkey breeder hen diets, compared with current surveyed industry levels, without impairing reproductive performance.

Anti-sperm antibodies and fertility of turkey hens.

Anti-sperm antibody titers increase with time in serum of turkey hens following a standard production schedule of artificial insemination (AI). In hens receiving intravenous (IV) or intraperitoneal (IP) additional AI, serum anti-sperm antibody levels increase more rapidly after a lag phase. A single injury to the oviduct also resulted in increased anti-sperm antibodies similar to IV and IP groups. This is a new observation that a single injury increased antibody titers to spermatozoa equal in IV and/or IP injections. A negative correlation between serum anti-sperm antibody titers for IV, IP and injury to oviduct and fertility of these groups was observed. Hens of IV and injury to oviduct groups with high levels of anti-sperm antibodies in the last 2 weeks of production had significantly lower fertility than hens with low levels of antibodies and control hens.

A novel delivery of oxytetracycline in turkey breeder hens.

A novel product (SQ12) for subcutaneous (SQ) injectable delivery of oxytetracycline (OTC) has been developed for use in livestock. SQ12 employs microfluidic spheres encasing OTC crystals, which allows for longer release of the OTC compared with other injectable antibiotics. The objectives of the study were to determine serum and tissue levels of SQ12 in turkey breeder hens to 14 days postinjection and to evaluate effects of SQ12 on reproductive status. Thirty photostimulated hens were housed in litter floor pens and provided with 14.5 hr of light per day in a curtain-sided facility. Six hens served as untreated controls. Twelve hens per treatment group received SQ injections in the neck with SQ12 at 11.4 (L dose group) or 22.7 mg/kg (H dose group) to assess low and high doses, respectively. Serum samples were obtained from each hen at predose and 6, 12, 24, 48, 72, 96, 168, 240, and 336 hr postinjection. All hens were euthanatized at 14 and 15 days postinjection. One-half of the hens in each treatment group were sampled (liver, lung, kidneys, and breast muscle) for tissue residue levels of OTC. The control group had no detectable OTC in serum or tissues at any sample collection time. There were no detectable serum levels of OTC in either treatment group prior to injection. The average serum concentrations of the L and H dose groups showed similar depletion curves although the H dose group was 42% higher at maximum concentration than the L group. Average tissue concentration of OTC for all tissues sampled from the H dose group was twice that of the L dose group. All tissue levels were below the OTC residue tolerance limit. SQ12 provided an extended source of OTC in serum of turkey breeder hens with no effect on reproductive status. SQ12 may provide for a novel treatment of bacterial infection in turkey breeder hens with longer lasting serum levels compared with other single injectable OTC products.

Effect of insemination intervals on oviducal sperm storage in turkeys.

Filling and emptying of sperm storage sites of the turkey oviduct were studied using tandem inseminations with semen from bronze and white males. Fertility was better among hens inseminated at weekly intervals than those inseminated at biweekly intervals. The rate of gland filling and emptying was unaffected by weekly and biweekly insemination intervals. However, gland filling and emptying were affected when the birds were inseminated early in a reproductive cycle, and stratification of spermatozoa occurred within the sperm-host glands. Stratification may have occurred to a lesser degree late in the reproductive cycle. It is suggested that sperm storage glands fill and empty at the same rates early and late in a breeding cycle, but fewer glands contain spermatozoa in the latter part of the season, resulting in a decreased fertility.

Supplemental thyroid hormones and hatchability of turkey eggs.

Fertile turkey eggs were injected with thyroid hormones to test the effects of exogenous hormones on hatchability. Physiological doses of thyroxine (T4) and triiodothyronine (T3) of 50 and 25 ng, respectively, injected at setting, depressed hatchability. Conversely, injections of T4 and T3 at 25 days of incubation significantly (P less than .05) improved hatchability. Injections with thyroid-stimulating hormone (TSH) or tyrosine had no significant effect on hatchability when injected at setting or at 25 days of incubation. Injections of thyroid-releasing hormone (TRH) had no effect on hatchability when injected at setting but significantly (P less than .05) improved hatchability when injected at 25 days of incubation. Because T4 and T3 significantly (P less than .05) depressed hatchability when injected whereas TSH and TRH had no effect on hatchability when injected at setting, these data suggest classic negative feedback mechanisms function very early in the embryonic turkey. The improvement in hatchability observed when thyroid hormones were injected at 25 days of incubation offers further evidence that hypothyroidism may be a cause of poor hatchability among turkey eggs.

Water balance in incubating turkey eggs.

The eggshell water vapor conductance capable of maintaining constant water content in incubating turkey eggs was determined for three different egg weight classifications. Metabolism of the developing turkey embryos was altered by increasing or decreasing eggshell permeability. Eggshell permeability accommodated metabolic water in large eggs very well. Both relative poult weights and relative water content of embryos in large eggs at the plateau stage in oxygen consumption decreased or increased allometrically to the permeability treatment. However, among small and medium eggs, if eggshell permeability was increased, relative poult weights at hatching were much less than expected. This occurred despite the fact that relative water content of the eggs at 25 days of incubation was not similarly affected. The data suggest a biological asynchrony for egg size, metabolism, and water content of turkey eggs.

Improved hatchability of turkey eggs at high altitudes due to added oxygen and increased incubation temperature.

Turkey egg hatchability at high altitudes was examined at two different oxygen tensions and two incubation temperatures. Incubating turkey eggs in a 149-torr oxygen environment at 37.7 C gave significantly better hatchability than a 109-torr oxygen environment at 37.5 C. Embryonic mortality data indicated that the higher incubation temperature in combination with increased oxygen tension decreased embryonic mortality during the 3rd and 4th wk of incubation and resulted in higher hatchability. The data suggest that hatchery managers at high altitudes should supplement with oxygen and incubate turkey eggs at higher temperatures than employed at lower altitudes.

Vital gas exchange and hatchability of turkey eggs at high altitude.

Conductance of turkey eggshells was observed to be significantly (P less than .01) greater at 2000 than at 200 m elevation. It was concluded that the increased conductance may have been due to the Chapman-Enskog relation. Eggshells of nonhatching eggs from the high altitude were examined, and it was determined that despite the increased conductance rate, eggshells with significantly (P less than .05) less functional pore area hatched poorly in both oxygenated and nonoxygenated environments. When oxygen was not supplemented to the incubators at high altitudes, eggshells required significantly (P less than .01) fewer pore concentrations to allow embryos to survive to late stages of embryonic development than eggs in oxygen supplemented environments. However, greater pore concentrations on the air space were required in both environments to complete hatching. Cuticle removal from eggshells incubated in oxygen supplemented incubators at high altitudes significantly (P less than .05) reduced late embryonic mortality. It was concluded that eggshell cuticle removal may be more advantageous to hatchability at high altitude than oxygen supplementation.

Influence of diet-mediated maternal thyroid alterations on functional properties of turkey eggs.

Maternal thyroid status was altered by means of diet to determine its effect on functional properties of turkey eggshells. Hens were fed a control diet (CON), the CON diet containing .5 ppm triiodothyronine (T3), the CON diet containing 2.1 ppm iodine as potassium iodide (KI) or the CON diet containing .1% thiouracil (THIO). Feeding T3 decreased plasma thyroxine but elevated plasma T3 concentrations compared to CON. The KI diet had no effect on plasma thyroid hormone concentrations, but feeding THIO depressed plasma thyroxine with no effect on T3, resulting in an elevated ratio of the two hormones compared to the CON ratio. Feeding KI decreased egg volume and T3 increased egg density compared to CON, but no effects on egg weight, surface area, width, or length were noted. Dietary T3 depressed eggshell water vapor conductance compared to CON. Dietary iodine resulted in thinner eggshells with fewer pores than the CON, whereas THIO caused significantly more pores in eggshells than CON but had no effects on shell thickness. Dietary KI had no effects on maternal plasma thyroid hormone concentrations, suggesting that the effects were due to iodine availability rather than to thyroid hormones. It is concluded that the availability of iodine to turkey breeder hens may influence eggshell characteristics.

Effect of three routes of spectinomycin injection in neonatal turkeys with mixed bacterial contamination.

Neonatal poults diagnosed to have mixed bacterial contamination were injected with spectinomycin in one of three different routes: yolk sac, intramuscularly, or subcutaneously. An additional group was uninjected controls. Yolk sac injections, but not other routes, resulted in significantly heavier birds.

Effect of oxygen and maternal dietary iodine on embryonic carbohydrate metabolism and hatchability of turkey eggs.

Fertile turkey eggs were produced by hens fed supplemental iodine to decrease eggshell conductance. The eggs were then incubated in an oxygen-enriched environment during Days 25 and 26 of incubation to test the hypothesis that maternal dietary iodine has separate effects on the eggshell permeability and the developing embryo. Oxygen supplementation improved hatchability regardless of dietary iodine treatment, suggesting that oxygen availability may limit hatching of eggs with normal gas conductance. Oxygen and dietary iodine did not affect body weight but interacted prior to pipping to affect embryonic liver and heart glycogen. Dietary iodine increased liver glycogen at internal pipping. The effects of oxygen on embryos, which were correlated directly with hatchability, were increased liver growth prior to pipping and increased heart weight at external pipping and posthatching. It was concluded that supplementing oxygen to incubating turkey eggs may improve hatchability by increasing liver and heart growth. Dietary iodine played only a minor or modulating role in assisting embryos to survive.

Increasing hatchability of turkey eggs with biotin egg injections.

At 25 days of incubation, eggs from Large White turkey hens in two field trials were injected with 87 micrograms of exogenous d-biotin per egg. Hatchability of fertile biotin-injected eggs was significantly higher than that of control (noninjected) eggs. Hatchability values of fertile eggs were 4 and 5% higher for exogenous d-biotin-injected eggs than for control eggs for December 1985 and May 1986 trials, respectively.