This study determined, for the first time, the different subpopulations of germ cells and stereological changes within the cortex of the functional left ovary during germ cell nest breakdown, and ...formation of the primordial follicle pool in the domestic turkey. This was accomplished by measuring the size, density, and count of prefollicular germ cells and primordial follicles in turkey poults between 1 and 35 days posthatch (dph). The percent volume (PV) of germ cells and follicles within the cortex was also calculated as a means of validating the counting technique. The total percent volume of germ cells and primordial follicles within the cortex ranged between 42 and 84%, suggesting that the counting technique was valid. Our findings show that before germ cell nest breakdown (5 dph), there were roughly 1,000,000 prefollicular germ cells within the cortex of the left ovary and that germ cell nest breakdown initiated between 5 and 7 dph, characterized by a decrease (P ≤ 0.001) in prefollicular germ cell density and the subsequent appearance of primordial follicles. Nest breakdown is followed on day 9 by the first increase (P ≤ 0.05) in size of prefollicular germ cells. These cells continue to grow throughout nest breakdown. The majority (>90%) of germ cell nest breakdowns concluded by 15 dph; although, the primordial follicle pool was not fully established until 35 dph, as determined by a total lack of prefollicular germ cells. At this point, the pool was comprised of an estimated 60,000 primordial follicles and shows that during nest breakdown and follicle pool formation, ∼94% of germ cells were lost. This 94% decrease in the number of germ cells during nest breakdown in the turkey is comparable to the domestic chicken but is greater than the average two-thirds which are lost in mammalian species.
Biobanking of turkey ovarian tissue appears to be the most cost-effective method for the long-term preservation of female genetics. However, to ensure the successful transplantation of biobanked ...ovarian tissue for breed or line revival, the transplantation and development of fresh ovarian tissue must be evaluated. To assess transplantability, ovaries from poults 1 to 15 days posthatch (dph) were cultured in ovo in chicken eggs for 6 d and compared with the equivalent fresh tissue. The viability of cultured ovarian tissue was evaluated visually, whereas the level of late-stage apoptosis was measured via the TUNEL assay. In addition, the diameter and density of prefollicular germ cells and follicles (primordial and primary) were measured to assess maturation. Results showed that all cultured grafts (74/74), on surviving chicken chorioallantoic membrane, were viable with low levels (0.8 ± 0.1%) of late-stage apoptosis. The diameter of prefollicular germ cells in cultured ovaries from poults at 5 and 7 dph were larger (P < 0.002) than that of their preculture counterparts but were not able to reach their in vivo size. No significant follicular growth was observed in ovaries cultured in ovo; however, prefollicular germ cell density was over 4-fold greater in ovaries cultured from 7 dph poults (81,030 ± 17,611/mm3) than in their in vivo counterpart (16,463 ± 6,805/mm3). Interestingly, cultured ovaries from all other ages displayed equal or lower (P ≤ 0.05) prefollicular germ cell densities than their in vivo counterparts. Cultured ovaries from poults at 5 and 7 dph also exhibited an increase (P ≤ 0.05) in follicle density compared with their preculture counterparts; whereas, cultured ovaries from 15 dph poults had decreased densities (P < 0.001) compared with their preculture counterparts. This study demonstrated that, although age of ovarian tissue cultured in ovo did not affect the overall viability, 7 dph ovaries appeared to have a better cellular morphology after culturing in ovo than other ages. In addition, we also demonstrated for the first time that avian follicles can form during tissue culturing in ovo.
Biobanked poultry ovaries can be revived via transplantation into a recipient female, which upon maturity will produce donor-derived progeny. Previously, a large portion of these recipients also ...produced recipient-derived progeny, making them gonadal chimeras. These were potentially created when portions of the recipient's ovary were inadvertently left behind. Completely removing the recipient ovary would solve this problem; however, leaving a portion of the recipient's ovary may have inadvertently increased the transplant attachment rate by providing a damaged area for attachment. To test this hypothesis in the turkey, we removed various portions (33–100%) of recipient ovarian tissue and determined the transplant attachment rate. Furthermore, the use of the abdominal air sac membrane as an additional anchoring point was tested. The overall attachment rate of transplants was 91% (27/30), while the average size of the transplants was 4.2 ± 0.6 mm2, 6 d postsurgery. There was no difference (P > 0.05) in the attachment rates, or transplant size between groups with varying amounts of recipent tissue removed, or by using the abdominal air sac membrane as an anchor. Finally, the immunological status of the grafts were evaluated by analyzing the presences of CD3 and MUM-1 (T and B cell markers). This showed that all transplants were infiltrated by large numbers of T and B cells. Shown by a high (P ≤ 0.001) percentage of CD3-positive immunostained cytoplasmic area (49.78 ± 3.90%) in transplants compared to remnant recipient tissue (0.30 ± 0.10%), as well as a high (P ≤ 0.001) percentage of MUM-1-positive immunostained nuclear area (9.85 ± 1.95%) in transplants over remnant recipient tissues (0.39 ± 0.12%). From this study we would recommend removing the entire recipient ovary, and not covering the transplants with the abdominal air sac membrane, to prevent gonadal chimeras. The high levels of lymphocytes within the grafts indicate possible tissue rejection, which could be overcome via immunosuppression with or without histocompatibility matching between donors and recipients.
Photoperiod is essential in manipulating sexual maturity and reproductive performance in avian species. Light can be perceived by photoreceptors in the retina of the eye, pineal gland, and ...hypothalamus. However, the relative sensitivity and specificity of each organ to wavelength, and consequently the physiological effects, may differ. The purpose of this experiment was to test the impacts of light wavelengths on reproduction, growth, and stress in laying hens maintained in cages and to determine whether the retina of the eye is necessary. Individual cages in 3 optically isolated sections of a single room were equipped with LED strips providing either pure green, pure red or white light (red, green, and blue) set to 10 lx (hens levels). The involvement of the retina on mediating the effects of light wavelength was assessed by using a naturally blind line (Smoky Joe) of chickens. Red and white lights resulted in higher estradiol concentrations after photostimulation, indicating stronger ovarian activation, which translated into a significantly lower age at first egg when compared with the green light. Similarly, hens maintained under red and white lights had a longer and higher peak production and higher cumulative egg number than hens under green light. No significant difference in BW gain was observed until sexual maturation. However, from 23 wk of age onward, birds exposed to green light showed higher body growth, which may be the result of their lower egg production. Although corticosterone levels were higher at 20 wk of age in hens under red light, concentrations were below levels that can be considered indicative of stress. Because no significant differences were observed between blind and sighted birds maintained under red and white light, the retina of the eye did not participate in the activation of reproduction. In summary, red light was required to stimulate the reproductive axis whereas green light was ineffective, and the effects of stimulatory wavelengths do not appear to require a functional retina of the eye.
The effect of ME intake (MEI) on the reproductive system was evaluated. Ross 308 broiler breeder pullets (n = 140) were assigned to 2 treatments from 22 to 26 wk of age: (1) Low-energy diet fed ...restricted (2,807 kcal/kg, low MEI) and (2) high-energy diet fed unrestricted (3,109 kcal/kg, high MEI). Daylength was increased from 8 to 14 h at 22 wk of age with a light intensity of 30 lux. Daily palpation was used to detect sexual maturity via the presence of a hard-shelled egg in the shell gland. Expression of gonadotropin releasing hormone-I (GnRH) and gonadotropin inhibitory hormone (GnIH) genes in the hypothalamus and GnRH receptor (GnRH-RI) and GnIH receptor (GnIH-R) genes in the anterior pituitary gland of each pullet was evaluated from 22 to 26 wk of age using quantitative real time-PCR. Blood samples were taken weekly and luteinizing hormone (LH), follicle stimulating-hormone (FSH), and 17-beta-estradiol (E2) determined using commercial ELISA kits. Carcass samples were used for determination of CP and fat content. Data were analyzed using the MIXED procedure in SAS, and differences were reported where P ≤ 0.05. High MEI treatment pullets had 2.3-fold higher GnRH and 1.8-fold higher GnRH-RI mRNA levels than low MEI pullets. MEI affected neither expression of GnIH and GnIH-R nor carcass protein content. For high MEI (489 kcal/D) and low MEI treatments (258 kcal/D), respectively, from 22 to 26 wk of age (P ≤ 0.05), LH concentration was 3.05 and 1.60 ng/mL; FSH concentration was 145 and 89.3 pg/mL; E2 concentration was 429 and 266 pg/mL, and carcass lipid was 13.9 and 10.3%. The onset of lay for pullets in the high MEI treatment advanced such that 100% had laid by 26 wk of age compared with 30% in the low MEI treatment. We concluded that higher MEI advanced the activation of the hypothalamic–pituitary–gonadal axis and also increased body lipid deposition, and moreover, stimulated reproductive hormone levels which overall accelerated puberty in broiler breeder pullets.
ABSTRACT
Body weight (BW) and rearing photoperiod are important factors affecting sexual maturation rate and reproductive performance in broiler breeders. The current experiment used a 2 × 3 ...factorial arrangement of treatments to study the interaction between BW and rearing photoperiod on reproductive performance in group housed broiler breeder hens, while minimizing variation in BW. Hens (n = 180) were fed with a precision feeding system to allocate feed individually to achieve the breeder-recommended target curve (Standard) or to a target curve that reached the 21 wk BW at 18 wk (High). Hens were on 8L:16D, 10L:14D, or 12L:12D photoschedules during rearing and were photostimulated at 21 wk with a 16L:8D photoschedule. Sexual maturity (defined as age at first egg) and individual egg production to 55 wk were recorded. At 55 wk, proportional weights of individual body components were determined by dissection. Differences were reported as significant at P ≤ 0.05. A significant interaction between BW and rearing photoschedule affected age at sexual maturity and egg production. In the High BW treatment, age at sexual maturity did not differ between hens under the 8L:16D and 10L:14D photoschedules (173 vs. 172 d, respectively). In the Standard BW treatment, the 12L:12D rearing photoperiod delayed sexual maturity compared with the 8L:16D rearing photoperiod (266 vs. 180 d, respectively). All hens on the High BW treatment laid at least 1 egg before the end of the experiment. Conversely, 3.3, 18.1, and 37.6% of Standard BW hens on the 8L:16D, 10L:14D, and 12L:12D photoschedules, respectively, never commenced egg production. At the end of the experiment, proportional breast weight was higher and proportional fatpad weight was lower in Standard compared to High BW hens (25.8 vs. 27.5% and 2.4 vs. 1.5% of BW, respectively). We conclude that increased BW partially counters the effect of longer photoschedules on sexual maturity in broiler breeders and that dissipation of the photorefractory state depends on BW.
Abstract
To synchronize the onset of sexual maturity in the face of high BW variation, the age at photostimulation has been increasing in the broiler breeder industry. This experiment studied the ...effects of increased BW and earlier photostimulation on broiler breeder reproductive performance where within-treatment BW uniformity was very high. The experiment tested BW and age at photostimulation treatments in a 2 × 2 factorial arrangement. Hens (n = 120) were fed with a precision feeding system to allocate feed individually following the breeder-recommended target BW (Standard) or to a 22% heavier target BW curve reaching the Standard 21 wk BW at 18 wk (High). Hens were photostimulated at either 18 wk (18WK) or 21 wk (21WK) with a 16L:8D photoschedule. Age at first egg (AFE) and individual egg production to 55 wk were recorded. Differences were reported as significant if P ≤ 0.05. The AFE was decreased and maturation interval between photostimulation and AFE was shorter for hens on the High BW treatment compared to the Standard BW treatment (178.1 vs. 194.7 d and 41.8 vs. 58.2 d, respectively). Hens on the 21WK treatment had a decreased AFE compared to the 18WK treatment (177.0 d vs. 195.9 d) and their maturation interval was shorter (30.0 d vs. 69.9 d). The CV for AFE was higher in the 18WK treatment compared to the 21WK treatment (28.2% vs. 11.2%). Total egg production was higher for hens on the High BW treatment compared to the Standard BW treatment (129.4 vs. 92.8, respectively). Total egg production was higher for hens on the 21WK treatment compared to the 18WK treatment (138.4 vs. 83.8, respectively). Egg weight of Standard BW × 18WK hens was lower compared to High BW × 18WK hens. Current recommended breeder BW may be too low for optimal sexual maturation after photostimulation. It is concluded that even when BW variation is minimized, photostimulation at 18 wk of age is not recommended.
Understanding energy partitioning in broiler breeders is needed to provide efficiency indicators for breeding purposes. This study compared 4 nonlinear models partitioning metabolizable energy (ME) ...intake to BW, average daily gain (ADG), and egg mass (EM) and described the effect of BW and rearing photoperiod on energy partitioning. Ross 708 broiler breeders (n = 180) were kept in 6 pens, controlling individual BW of free run birds with precision feeding stations. Half of the birds in each chamber were assigned to the breeder-recommended target BW curve (Standard) or to an accelerated target BW curve reaching the 21-week BW at week 18 (High). Pairs of chambers were randomly assigned to 8L:16D, 10L:14D, or 12L:12D rearing photoschedules and photostimulated with 16L:8D at week 21. Model I was: MEId = a × BWb + c × ADG × BWd + e × EM + ε, where MEId = daily ME intake (kcal/day); BW in kg; ADG in g/day; EM in g/day. Models II–IV were nonlinear mixed versions of model I and included individual II, age-related III, or both individual and age-related IV random terms to explain these sources of variation in maintenance requirement (a). Differences were reported as significant at P ≤ 0.05. The mean square error was 2,111, 1,532, 1,668, and 46 for models I–IV respectively, inferring extra random variation was explained by incorporating 1 or 2 random terms. Estimated ME partitioned to maintenance IV was 130.6 ± 1.15 kcal/kg0.58, and the ME requirement for ADG and EM were 0.63 ± 0.03 kcal/g/kg0.54 and 2.42 ± 0.04 kcal/g, respectively. During the laying period, maintenance estimates were 124.2 and 137.4 kcal/kg0.58 for standard and high BW treatment, and 130.7, 132.2, and 129.5 kcal/kg0.58 for the 8L:16D, 10L:14D, or 12L:12D treatments, respectively. Although hens on the standard BW treatment with a 12L:12D rearing photoschedule were most energetically conservative, their reproductive performance was the poorest. Model IV provided a new biologically sound method for estimation of life-time energy partitioning in broiler breeders including an age-related random term.
Artificial lighting is used to control growth and reproduction. Lighting protocols are defined by the quantity (photoperiod, intensity) and the quality (wavelength) of light. Recently, with the ...introduction of light emitting diode (LED) bulbs, interest has grown in investigating the effect of spectrum lighting. Thus, the aim of this study was to examine the effect of red and green light on growth during pullet rearing, and on possible carry-over effects during the subsequent adult phase. Lohmann Brown-Lite chicks were raised in a 2-story free run barn divided into 4 sections and exposed to either 60% red LED (RL) or 60% green LED (GL) light treatments. At 19 wk, all birds were moved to an adult free-run barn with RL and GL pullets placed on separate halves of the barn. In the adult barn, all birds were exposed to RL. Body weight, egg production, ovarian morphology, estradiol and calcium levels, as well as bone structure were recorded until 70 wk. Although no consistent significant difference was observed in body weight or general reproductive parameters, RL pullets tended to sexually mature earlier. As well, no carry-over effect was detected. Regardless of pullet treatment, egg production remained high throughout, especially towards the end of lay. In conclusion, spectrum lighting during the rearing of layer pullets did not impact growth or subsequent production performance; however, exposing adult hens to RL may be beneficial to maintain high egg production.
As broiler breeders face increased reproductive challenges specifically related to overfeeding, a clear understanding of the physiological effects of BW and rearing photoperiod on reproductive ...development is needed. The objective was to use mathematical models to compare plasma estradiol-17β (E2) concentration to characterize the effect of BW and rearing photoperiod on E2 levels. A 2 × 3 factorial arrangement of treatments was used. Hens (n = 180) were fed with a precision feeding system to allocate feed individually to achieve the breeder-recommended BW curve (Standard) or to a BW curve reaching the 21 wk target at 18 wk (High). Hens were on 8L:16D, 10L:14D, or 12L:12D photoschedules during rearing and were photostimulated at 21 wk. Age at first egg (AFE) was recorded. Plasma E2 levels were determined weekly between week 20 and 28. Two modified Gompertz models described E2 level as a function of (a) chronological or (b) physiological (relative to AFE) age. Timing of E2-inflection point was compared between models and treatments. Differences were reported as significant at P ≤ 0.05. The chronological age model inferred that High BW reduced the duration between the E2-inflection point and AFE, whereas the physiological age model inferred that High BW only reduced the duration between photostimulation and the E2-inflection point. Hens on the Standard BW treatment had a longer period between photostimulation and the E2-inflection point compared to hens on the High-BW treatment (11.03 vs. 1.50 wk, respectively, based on physiological age). Hens on the 12L:12D photoschedule had a longer period between photostimulation and the E2-inflection point compared to hens on the 8L:16D or 10L:14D photoschedule, both in the Standard and High BW (28.91 vs. 1.78 and 2.40 wk, 2.65 vs. 0.93 and 0.94 wk, respectively, based on physiological age). The described methodology and results provide quantitative insight into E2 dynamics and serves as a model for future endocrinological studies in poultry reproduction.
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