Soybean Bioactive Peptide Supplementation Affects the Intestinal Immune Antioxidant Function, Microbial Diversity, and Reproductive Organ Development in Roosters.

Soybean is an important source of high-quality vegetable protein with various health-improving properties, and its main bioactive substances are small peptides produced by in vitro enzymatic hydrolytic processes. In traditional layer breeding, the nutritional health of roosters is frequently neglected, ultimately affecting the quality and quantity of offspring. This study investigated the effects of various quantities (0%, 0.15%, 0.30%, 0.45%, and 0.60%) of soybean bioactive peptide (SBP) feed additives on immunological and antioxidant functions, gut health, and reproductive performance of roosters. SBP supplementation significantly improved male growth and reproductive performance, including growth rate, feed conversion ratio, reproductive organ development, and semen quality. SBP also increased immune and antioxidant levels, boosted the integrity of the small intestinal physiological structure and barrier function, and diversity of cecal microbes, and decreased the apoptotic ratio of small intestinal epithelial cells. The effects of SBP on various functions of males showed a quadratic trend, with the optimal concentration determined to be 0.45%.

Effects of hydroxytyrosol on post-thaw quality of rooster sperm.

Semen cryopreservation is one of the most important reproduction techniques in the livestock and poultry industry. Cryopreservation induces cold stress, generating reactive oxygen species (ROS) and oxidative stress causing structural and biochemical damages in sperm. In this study, we evaluated the effects of the hydroxytyrosol (HT), as an antioxidant, at the concentrations of 0, 25, 50, and 100 µg/mL on post-thaw semen quality metrics in rooster. Semen samples were collected twice a week from 10 roosters (29 weeks), processed and frozen according to experimental groups. Different quality parameters, including total motility, progressive motility, viability, morphology, membrane integrity, and malondialdehyde were measured after thawing. Results showed that 25 and 50 µg/mL of HT produced the highest percentage of total motility (51.01 ± 2.19 and 50.15 ± 2.19, respectively) and progressive motility (35.74 ± 1.34 and 35.15 ± 1.34, respectively), membrane integrity (48.00 ± 2.18 and 46.75 ± 2.18, respectively) as well as viability (53.00 ± 2.17 and 52.50 ± 2.17, respectively) compared with the other groups (p < .05). The group with 25 µg/mL of HT showed the lowest significant (p < .05) MDA concentration (1.81 ± 0.25). Our results showed that the effect of HT was not dose-dependent and optimum concentration of HT could improve functional parameters of rooster sperm after freezing-thawing. These findings suggest that HT may have protective effects on the rooster sperm during the freezing-thawing process.

Gallic acid supplementation partially ameliorates reproductive aging in rooster breeders by improving semen quality, sperm kinetics, hormones, and antioxidant status.

Aging leads to decreased fertility in roosters, which is likely due to increased oxidative stress. This study evaluated the antioxidant effects of gallic acid (GA) supplementation on sperm quality and fertility of aged roosters. This study evaluated whether GA supplementation can mitigate age-related fertility decline. Roosters were randomly assigned to: control, 100 mg/kg GA, or 200 mg/kg GA. Semen parameters, sperm kinetics, hormone levels, fertility rate, and hatchability were assessed. GA increased semen concentration, membrane integrity and viability while decreasing defects versus control (P < 0.01). Testosterone was higher in GA groups (P<0.01) without affecting gonadotropins. Furthermore, 200 mg/kg GA optimized motility, velocity, linearity, and beat cross frequency versus control and 100 mg/kg GA (P < 0.01). Fertility and hatchability were higher in both GA groups. In conclusion, GA supplementation in aged roosters improves sperm quality, antioxidant status, testosterone, and fertility outcomes, likely by mitigating oxidative stress. The 200 mg/kg dose elicited optimal effects on motion parameters.

Barley sprouts and D-Aspartic acid supplementation improves fertility, hatchability, and semen quality in aging male broiler breeders by up-regulating StAR and P450SCC gene expressions.

At 50 wk of age, broiler breeder roosters exhibit a significant decline of fertility. Therefore, the aim of this study was to assess the impact of incorporating barley sprout (BS) powder, D-aspartic acid (DA), or their combination into the diet on fertility, hatchability, semen quality, and the relative expression of StAR and P450SCC genes in aging broiler roosters. Aging (50 wk) male broiler breeders (n=32) were randomly assigned to one of four dietary treatments (2 × 2 factorial) with 2 levels of BS (0 or 2% basal diet) and DA (0 or 200 mg/kg/BW) for 12 wk. Roosters were individually housed under a 14-h light and 10-h dark cycle, with 150 g/d feed allocation and free access to fresh water, then euthanized. Throughout the study, the body weight of the broiler breeders was measured, along with various parameters related to semen quality, on a weekly basis. Additionally, artificial insemination was performed during the last 2 wk to evaluate reproductive endpoints. The results revealed that both BS and DA decreased (P < 0.01) body weight. Interestingly, the inclusion of BS, either alone or in combination with DA, resulted in a significant increase in total and forward sperm motility. Furthermore, it was demonstrated that the seminal concentration of malondialdehyde, a marker of oxidative stress, was significantly decreased by more than 20% in all groups compared to the control. The combination of both BS and DA led to the highest levels of circulating testosterone, as well as the functionality and membrane integrity of sperms. Additionally, it resulted in increased sperm concentrations, production, and penetration, ultimately leading to improved fertility rate and hatchability percentage. Moreover, a positive association between total motility and fertility was observed (P < 0.01). Furthermore, the combined supplementation of BS and DA up-regulated the relative mRNA expression of P450scc and StAR (P < 0.01). To summarize, dietary inclusion of BS, DA, or their combination have a potential to improve various aspects of reproductive performance in aging roosters.

Protective Effect of N-Acetylcysteine on Rooster Semen Cryopreservation.

Cryopreservation of avian semen is a useful reproductive technique in the poultry industry. However, during cooling, elevated reactive oxygen species (ROS) levels have destructive effects on both quality and function of thawed sperm. The aim of the current study is to investigate the antioxidant effects of N-acetylcysteine (NAC) during rooster semen cryopreservation. Semen samples were collected from ten Ross 308 broiler breeder roosters (32 weeks) and mixed. The mixed samples were divided into five equal parts and cryopreserved in Lake Buffer extender that contained different concentrations (0, 0.01, 0.1, 1, and 10 mM) of NAC. The optimum concentration of NAC was determined based on quality parameters of mobility, viability, membrane integrity, acrosome integrity, lipid peroxidation, and mitochondrial membrane potential after the freeze-thaw process. There was a higher percentage (p < 0.05) of total motility (TM) (60.9 ± 2.4%) and progressive motility (PM) (35.6 ± 1.9%) observed with the NAC-0.1 group compared to the other groups. Significantly higher percentages of viability (74.4 ± 2.3% and 71 ± 2.3%), membrane integrity (76.4 ± 1.5% and 74.7 ± 1.5%) and mitochondrial membrane potential (67.1 ± 1.6% and 66.3 ± 1.6%) were observed in the NAC-0.1 and NAC-1 groups compared to the other frozen groups (p < 0.05). The lowest percentage of lipid peroxidation and nonviable sperm was found in the NAC-0.1 and NAC-1 groups compared to the other groups (p < 0.05). The average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), and acrosome integrity, were not affected by different concentrations of NAC in the thawed sperm (p > 0.05). Both NAC-0.1 and NAC-1 appear to be beneficial for maintaining the quality of rooster sperm after thawing.

Zinc oxide nanoparticles preserve the quality and fertility potential of rooster sperm during the cryopreservation process.

Sperm cryopreservation is one of the main methods for preserving rooster sperm for artificial insemination (AI) in commercial flocks. Yet, rooster sperm is extremely susceptible to reactive oxygen species (ROS) produced during the freezing process. Oxidative stress could be prevented by using nanoparticles containing antioxidants. The present study was conducted to investigate the effect of zinc oxide nanoparticles (ZnONP) in rooster semen freezing extender on quality parameters and fertility potential. For this aim, semen samples were collected and diluted in Lake extenders as follows: control: Lake without ZnONP, ZnO100: Lake with 100-µg zinc oxide (ZnO), ZnONP50: Lake with 50-µg ZnONP, ZnONP100: Lake with 100-µg ZnONP and ZnONP200: Lake with 200-µg ZnONP. After freezing and thawing, sperm motility, viability, membrane integrity, morphology, mitochondrial activity, acrosome integrity, DNA fragmentation, lipid peroxidation and ROS, as well as fertility and hatchability were assessed. According to the current results, higher rates of motility, membrane integrity, mitochondrial activity, acrosome integrity and live cells were detected in the ZnO100, ZnONP50 and ZnONP100 groups compared to other groups (p ≤ .05). Yet, the percentage of dead cells, DNA fragmentation, lipid peroxidation and ROS levels were lower in the mentioned groups (p ≤ .05). Furthermore, a higher percentage of fertility was observed in the ZnO100 and ZnONP100 groups than in the control group (p ≤ .05). In conclusion, the use of 100-µg ZnO and 50- to 100-µg ZnONP represents a valuable and safe additive material that could be used to improve the quality and fertility potential of rooster sperm under cryopreservation conditions.

Metabolites assay offers potential solution to improve the rooster semen cryopreservation.

Semen cryopreservation represents a promising technology utilized for preserving high-quality chicken varieties in husbandry practices. However, the efficacy of this methodology is significantly impeded by the diminished quality of sperm. Metabolites, as the end products of metabolic reactions, serve as indicators of biological processes and offer insights into physiological conditions. In this study, we investigaged the sperm quality and alteration in metabolic profiles during the cryopreservation of Longyou Partridge Chicken semen. Following artificial semen collection, four groups of semen samples were established based on four points of the cryopreservation process (Ⅰ, fresh semen; Ⅱ, semen added extender and chilled at 4 °C for 30 min; Ⅲ, semen added cryoprotectants; Ⅳ, semen gradient freezed and stored in liquid nitrogen). Semen cryopreservation has a negative effect on the percentage of sperm in a straight-line trajectory (LIN), has no significant effect on total motile sperms (TM) or the proportion of sperm with typical morphology (NM). Metabolites were identified using LC-MS technique and analyses including Principal Component Analysis (PCA), Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA), Univariate statistical analysis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database were employed to identify metabolites. A total of 2471 metabolites had been identified, with the majority of the list being made up of amino acids and their metabolites as well as benzene and substituted derivatives. Group II exhibits 882 metabolites with significantly elevated abundance relative to Group I, alongside 37 metabolites displaying decreased abundance. In Group III, 836 metabolites demonstrate notably augmented abundance compared to Group II, while 87 metabolites exhibit reduced abundance. Furthermore, Group IV showcases 513 metabolites with markedly heightened abundance in comparison to Group III, and 396 metabolites with decreased abundance. Specific metabolites such as 5-Hydroxylysine, Phosphocholine, and alpha-d-glucose-6-phosphate exhibited a progressive decline during the cryopreservation process, correlating with either dilution and chilling, cryoprotectant addition, or freezing. In conclusion, our investigation systematically examined the changes of seminal metabolome and sperm quality throughout the cryopreservation process of rooster semen.

Melatonin alleviates glyphosate-induced testosterone synthesis inhibition via targeting mitochondrial function in roosters.

Glyphosate (GLY) is a widely used herbicide that has been revealed to inhibit testosterone synthesis in humans and animals. Melatonin (MET) is an endogenous hormone that has been demonstrated to promote mammalian testosterone synthesis via protecting mitochondrial function. However, it remains unclear whether MET targets mitochondria to alleviate GLY-inhibited testosterone synthesis in avian. In this study, an avian model using 7-day-old rooster upon chronic exposure to GLY with the treatment of MET was designed to clarify this issue. Data first showed that GLY-induced testicular Leydig cell damage, structural damage of the seminiferous tubule, and sperm quality decrease were mitigated by MET. Transcriptomic analyses of the testicular tissues revealed the potentially critical role of mitophagy and steroid hormone biosynthesis in the process of MET counteracting GLY-induced testicular damage. Also, validation data demonstrated that the inhibition of testosterone synthesis due to GLY-induced mitochondrial dynamic imbalance and concomitant Parkin-dependent mitophagy activation is alleviated by MET. Moreover, GLY-induced oxidative stress in serum and testicular tissue were significantly reversed by MET. In summary, these findings demonstrate that MET effectively ameliorates GLY-inhibited testosterone synthesis by inhibiting mitophagy activation, which provides a promising remedy for the application of MET as a potential therapeutic agent to antagonize reproductive toxicity induced by GLY and similar contaminants.

Evaluation of the effect of the addition of an olive oil-derived antioxidant (Pectoliv-80A) in the extender for cryopreservation of rooster sperm through the use of a discriminant statistical tool.

During the poultry sperm cryopreservation process, an excess of reactive oxygen species is generated resulting in oxidative stress which harms the quality of avian spermatozoa. To counteract this effect, the addition of exogenous antioxidants, such as Pectoliv-80A (a by-product of olive oil), to the cryopreservation diluent is interesting. For this purpose, 16 roosters belonging to the Utrerana avian breed were used. Six semen pools (from the 6 different replicates) were divided into 4 aliquots corresponding to different concentrations of Pectoliv-80A that were tested (0, 300, 400, and 500 µg/mL), and the cryopreservation process was carried out. To evaluate post-thawing semen quality, different parameters such as motility, membrane functionality, reactive oxygen species production, lipid peroxidation, and acrosome integrity were studied. A discriminant canonical analysis was used to determine both the differences between the Pectoliv-80A concentration groups and the discriminant power of the aforementioned parameter used for semen evaluation. Total motility and membrane functionality were reported to be the most discriminant variables for differentiating the different antioxidant enrichment groups and concluded that concentrations of 300 µg/mL showed the most desirable quality of post-thawing semen. The present study could lead to the optimization of both cryopreservation and quality evaluation techniques of the sperm of rooster species, that support the conservation program of endangered local breeds.

Alpha-lipoic acid improves cryopreservation of rooster semen by reducing oxidative stress.

Inhibiting oxidative stress is key for ensuring sperm motility during semen cryopreservation. The aim of this study was to investigate the effect of adding alpha-lipoic acid (ALA) as an extender in rooster semen cryopreservation. Different concentrations of ALA were added to the frozen diluent of rooster semen; subsequently, computer-aided semen analysis was used to determine membrane functional integrity, acrosome integrity, antioxidant capacity (based on T-AOC, GSH-Px, SOD, CAT, and MDA contents), and mitochondrial integrity. The frozen sperm ultrastructure was observed using transmission electron microscopy. The results showed that the addition of different concentrations of ALA partially to greatly improved the quality of frozen sperm; in particular, 8 µg/mL ALA significantly improved multiple parameters of sperm quality, including sperm motility and antioxidant enzyme activity, after freeze-thaw. The results of this study provide empirical and theoretical support for effective rooster semen cryopreservation and can inform the development of new protective agents in the field of livestock reproduction.

Effects of dietary Moringa oleifera leaf extract on semen characteristics, fertility, and hatchability in aged broiler breeder roosters.

Declining semen quality will have a negative impact on the fertility of aged roosters. Various factors influence this decrease in quality. This study was conducted to investigate the effects of different levels of Moringa plant extract on semen characteristics, fertility, and hatchability in aged broiler breeder roosters. A total of 24 roosters were fed 1 of 4 dietary supplements for 10 wk: Control, 100 µL/kg (Moringa oleifera leaf extract [MOLE]-100), 200 µL/kg (MOLE-200), or 400 µL/kg body weight (MOLE-400) of Moringa oleifera extract. Results showed supplementation with MOLE-200 significantly improved (P < 0.05) semen concentration, total motility, progressive motility, sperm membrane integrity compared to other treatments. However, semen volume and body weight were unaffected (P > 0.05). Sperm lipid peroxidation, as indicated by malondialdehyde concentration, was lowest in MOLE-200. There was a significant difference observed among the treatments in terms of total antioxidant capacity (TAC) results. The testosterone concentration in the MOLE-200 treatment was significantly higher than the other treatments (P < 0.05). However, no significant differences were observed in the levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) hormones among the experimental treatments. Fertility and hatchability rates were measured at the end of the trial. Fertility, defined as the number of fertilized eggs, was greatest in the MOLE-200 treatment compared to the other treatments. Similarly, hatchability (hatched chicks/fertilized eggs %) was highest at 88.02% for MOLE-200. In conclusion, dietary supplementation with M. oleifera extract improved semen quality, fertility, and hatchability in aged broiler breeder roosters.

Research Note: Evaluation of phytic acid disappearance, ileal P digestibility, and total tract P retention in canola meal supplemented with increasing levels of exogenous phytase using conventional and cecectomized precision-fed roosters and growing chicks.

Apparent ileal digestibility (AID) of P, apparent total tract retention (ATTR) of P, and phytic acid disappearance in canola meal were evaluated in the presence of increasing levels of exogenous phytase. In Experiment 1, a precision-fed rooster assay was used to determine phytic acid (myo-inositol 1,2,3,4,5,6-hexakis; InsP6) and inositol phosphate (InsP6-3; InsP-P) disappearance in conventional and cecectomized Leghorn roosters. Roosters were crop intubated with 25 g of canola meal mixed with 0, 500, 1,000, or 2,000 FTU/kg of exogenous phytase. In Experiment 2, InsP6 and InsP-P disappearance and AID and ATTR of P were determined using ad libitum-fed broiler chickens. Treatments consisted of semi-purified diets containing 45% canola meal as the sole source of P. Phytase was added to increase phytase activity by 0, 500, 1,000, or 2,000 FTU/kg. Experiments contained 6 replicates per treatment. Canola meal contained a high phytase activity (1,630 FTU/kg as-fed) due to contamination with a commercially available phytase at the feed mill from which the canola meal was sourced. In Experiment 1 with precision-fed roosters, there was no effect (P > 0.05) of phytase or bird type on InsP6 and InsP-P disappearance; however, phytase linearly reduced (P < 0.05) InsP3 concentrations in excreta. In Experiment 2 with ad libitum-fed chickens, phytase linearly increased (P < 0.05) ileal InsP6 and InsP-P disappearance, and phytase had a quadratic effect (P < 0.05) on excreta InsP6 and InsP-P disappearance. Increasing dietary phytase activity resulted in a linear increase (P < 0.05) in AID of P and phytase had a quadratic effect (P < 0.05) on ATTR of P. In conclusion, titration of high levels of phytase (1,600 to 3,600 FTU/kg as-fed) reduced InsP3 concentrations in precision-fed roosters but did not affect overall phytic acid hydrolysis, which was 78% or greater for all treatments; however, increasing the total phytase activity from 700 to 2,700 FTU in ad libitum-fed broiler chickens increased phytic acid disappearance and P digestibility.

Apigenin supplementation substantially improves rooster sperm freezability and post-thaw function.

This pioneering research investigated apigenin potential to augment rooster sperm cryosurvival in an extender model. Apigenin is a natural antioxidant flavonoid showing promise for improved post-thaw sperm function. However, its effects on avian semen cryopreservation remain unexplored. This first study supplemented rooster sperm Lake extender with 0, 50, 100, 200, 400 µmol/L apigenin to determine the optimal concentrations for post-thaw quality. Supplementation with 100 µmol/L apigenin resulted in significant enhancements in total motility (from 41.5% up to 71.5%), progressive motility (18.1% to 29.1%) (p < 0.05), membrane integrity (40% to 68%), mitochondrial function (p < 0.001), viability (37% to 62%) and total antioxidant capacity (p < 0.001) compared to the control. It also substantially reduced percentages of abnormal morphology, reactive oxygen species and apoptosis (p < 0.001). Although 200 µmol/L apigenin significantly enhanced some attributes, effects were markedly lower than 100 µmol/L. Higher doses did not improve cryoprotective parameters. This indicates 100 µmol/L as the optimal apigenin concentration. This represents the first report of apigenin protecting rooster sperm from cryodamage. The natural antioxidant improved post-thaw sperm quality, likely by suppressing oxidative stress and apoptosis. Apigenin shows promise for enhancing rooster sperm cryosurvival.

Improving the quality of chilled semen from Thai native chicken using phosphorus and vitamin B12 supplementation in semen extender.

This study aimed to determine phosphorus and vitamin B12 supplementation effect in semen extender on the quality and fertility ability of chilled Thai native rooster semen. Eighty-four ejaculates of semen from 26 Thai native roosters (Burmese × Vietnam crossbreed) were included. Semen was collected by applying dorsal-abdominal massage once a week, pooled, diluted to 500 million sperms per dose, and divided into 6 groups. The semen samples used for control group were diluted with modified Beltsville poultry semen extender (BPSE). For the treatment groups 2 to 6: semen samples were diluted with modified BPSE and enriched with phosphorus and vitamin B12 (Octafos Octa Memorial Co., Ltd., Bangkok, Thailand) at concentrations 0.02, 0.04, 0.06, 0.08, and 0.10%. Semen fertility ability was tested in 6 replications by inseminating layer hens. Thirty-six Thai native hens were randomly assigned to 3 groups (control, 0.04, and 0.08%) of 12 hens and were inseminated with a dose of 0.2 mL on collecting day. Sperm motion characteristics (i.e., sperm motility, sperm progressive motility, and sperm kinetic parameters) were measured using a computer-assisted sperm analysis system (SCA, Proiser S.L., Valencia, Spain). Sperm viability, mitochondrial activity, acrosome integrity, plasma membrane integrity, and malondialdehyde (MDA) concentration were also evaluated. The sperm motion characteristics were the highest in the 0.04% supplementation group on all days of collection, especially the VCL and VAP (P < 0.05). The viability, mitochondrial activity, plasma membrane and acrosome integrity of spermatozoa were greater in the 0.04% supplementation group than in the control groups (P < 0.05). The 0.04% supplementation group had the lowest MDA concentration in all days of collection. The 0.04% supplementation group were higher both fertility (66.59 vs. 48.50%: P < 0.05) and hatching rates (58.80 vs. 43.18%: P < 0.05) than in the control group. In conclusion, 0.04% phosphorus and vitamin B12 concentrations supplementation in semen extender improved rooster semen quality and fertility in chilled rooster semen.

Preservation of rooster post-thawed sperm epigenetic modifications, fertility potential and other quality parameters in different extenders using reduced glutathione.

Although rooster semen cryopreservation is an efficient procedure to spread qualified semen samples for reproductive goals, some post-thawed qualified semen samples resulted in poor fertility rate that could be related to epigenetic modifications during the cryopreservation process. This research was conducted to investigate the effect of reduced glutathione (GSH) in different cryopreservation extenders (Lake and Beltsville) on preservation of epigenetic modifications, fertility potential and other quality parameters of rooster sperm after thawing. Semen samples were collected and diluted in Lake and Beltsville extenders as follows: L-0: Lake without GSH, L-G: Lake with GSH, B-0: Beltsville without GSH, and B-G: Beltsville with GSH. After freeze-thawing process, sperm motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration, epigenetic modifications and fertility potential were evaluated. In results, the type of extender had no effect (P > 0.05) of post-thawed sperm quality. The treatments containing GSH presented higher (P ≤ 0.05) total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, DNA methylation, fertility as well as lower (P ≤ 0.05) lipid peroxidation, apoptosis, DNA fragmentation and ROS concentration than other treatments. Extender supplementation with GSH had no effect (P > 0.05) on histone methylation, histone acetylation and hatching rate. In conclusion, supplementation of rooster sperm cryopreservation extender with GSH could be an effective strategy to preserve post-thawed sperm DNA methylation, fertility and other quality parameters during reproductive programs.

Prediction equations of the metabolizable energy in corn developed by chemical composition and enzymatic hydrolysate gross energy for roosters.

Two experiments were conducted to establish the prediction equations for AME and TME of corn based on chemical composition and enzymatic hydrolysate gross energy (EHGE) in roosters. In experiment 1, eighty 32-wk-old Hy-line Brown roosters with an average body weight of 2.55 ± 0.21 kg were randomly assigned to 10 diet treatments in a completely randomized design to determine AME and TME by the force-feeding method. Each treatment had 8 replicates with 1 bird per replicate. The 10 test diets used in the experiment were formulated with corn (including 96.10%) as the sole source of energy. In experiment 2, the EHGE of 14 corn samples was measured by the computer-controlled simulated digestion system (CCSDS) with 5 replicates of each sample. The average AME and TME values of corn were 14.58 and 16.46 MJ/kg DM, respectively. The EHGE of 14 corn samples ranged from 14.66 to 15.89 (the mean was 15.24) MJ/kg DM. The best-fit equations for corn based on chemical composition were AME (MJ/kg DM) = 14.5504 + 0.1166 × ether extract (EE) + 0.5058 × Ash - 0.0957 × neutral detergent fiber (NDF) (R2 = 0.8194, residual standard deviation (RSD) = 0.0860, P < 0.01) and TME (MJ/kg DM) = 16.0625 + 0.1314 × EE + 0.4725 × Ash - 0.0872 × NDF (R2 = 0.7867, RSD = 0.0860, P < 0.01). The best-fit equations for corn based on EHGE were AME (MJ/kg DM) = 7.8883 + 0.4568 × EHGE (R2 = 0.8587, RSD = 0.0693, P < 0.01) and TME (MJ/kg DM) = 10.0099 + 0.4228 × EHGE (R2 = 0.8720, RSD = 0.0608, P < 0.01). The differences between determined and predicted values from equations established based on EHGE were lower than those observed from chemical composition equations. These results indicated that EHGE measured with CCSDS could predict the AME and TME of corn for roosters with high accuracy.

Melatonin Inhibits Testosterone Synthesis in Rooster Leydig Cells by Targeting CXCL14 through miR-7481-3p.

Melatonin has been proved to be involved in testosterone synthesis, but whether melatonin participates in testosterone synthesis by regulating miRNA in Leydig cells is still unclear. The purpose of this study is to clarify the mechanism of melatonin on Leydig cells testosterone synthesis from the perspective of miRNA. Our results showed that melatonin could significantly inhibit testosterone synthesis in rooster Leydig cells. miR-7481-3p and CXCL14 were selected as the target of melatonin based on RNA-seq and miRNA sequencing. The results of dual-luciferase reporter assays showed that miR-7481-3p targeted the 3'-UTR of CXCL14. The overexpression of miR-7481-3p significantly inhibited the expression of CXCL14 and restored the inhibitory role of melatonin testosterone synthesis and the expression of StAR, CYP11A1, and 3ß-HSD in rooster Leydig cells. Similarly, interference with CXCL14 could reverse the inhibitory effect of melatonin on the level of testosterone synthesis and the expression of StAR, CYP11A1, and 3ß-HSD in rooster Leydig cells. The RNA-seq results showed that melatonin could activate the PI3K/AKT signal pathway. Interference with CXCL14 significantly inhibited the phosphorylation level of PI3K and AKT, and the inhibited PI3K/AKT signal pathway could reverse the inhibitory effect of CXCL14 on testosterone synthesis and the expression of StAR, CYP11A1 and 3ß-HSD in rooster Leydig cells. Our results indicated that melatonin inhibits testosterone synthesis by targeting miR-7481-3p/CXCL14 and inhibiting the PI3K/AKT pathway.

Effects of the Calls and Presence of Roosters on Egg Incubation Behavior of Nagoya Laying Hens.

The incubation behavior of the Japanese Nagoya chicken breed is a commercial issue because it often causes a sudden and sharp drop in egg production. In this study, whether the incidence of incubation behavior in Nagoya laying hens was associated with calls and the presence of roosters in the same laying house was investigated. Four experiments were conducted using commercial layer-type Nagoya hens where the hatching time of the experimental birds and the treatment order in the presence of males were changed . In Experiment 1, the proportion of incubation behavior in the presence of roosters kept in another pen located between pen-rearing hens (51.3%) was higher than that in their absence (15.9%) or with only rooster calls (23.8%). In Experiments 2, 3, and 4, the proportion of incubation behavior in the presence of roosters (47.3%, 33.3%, and 37.9%, respectively) was higher than that in their absence (33.3%, 17.4%, and 25.6%, respectively). In all experiments, approximately 70% of the incubating hens observed in the absence of roosters exhibited incubation behavior, even in the presence of roosters. Therefore, the presence of roosters may enhance egg incubation behavior in Nagoya laying hens.

Estimation of the seminal parameters of rooster and its association with fertility traits in synthetic dam line.

The objective of the present study was to assess the seminal parameters of rooster and its association with fertility traits (%), viz., hatchability of the fertile egg set (HFES), hatchability of the total egg set (HTES), and fertility (FERT). The data records pertained to traits of interest were obtained from various registers maintained at Poultry farm, of the Department of Animal Genetics and Breeding, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar (India). The relationship between seminal and fertility characteristics was investigated using regression analysis and correlation. Moreover, the efficacy of seminal characteristics to distinguish between roosters with low and high fertility traits was evaluated using linear discriminant analysis (LDA). The findings showed that reproductive traits and seminal characteristics were significantly (P < 0.05) correlated. The LDA showed that the seminal parameters can effectively separate the roosters into those with high and poor reproductive features. It was revealed from LDA that seminal features showed higher classification accuracy for FERT (80.77%). Hatchability is dependent on eggs that have been artificially incubated; hence, these crucial traits are comparatively weaker for HTES (65.38%) and HFES (67.31%). Cross-validation of the seminal parameter LDA corroborated the aforementioned and related conclusions. It is suggested that the studied LDA function may be utilised to choose genotypes with improved reproductive traits based on seminal variables.

Standardized amino acid digestibility and nitrogen-corrected true metabolizable energy of frozen raw, freeze-dried raw, fresh, and extruded dog foods using precision-fed cecectomized and conventional rooster assays.

Processing conditions, particularly temperature and duration of heating, impact pet food digestibility. Various commercial pet food formats are now available, but few have been tested thoroughly. The objective of this study was to determine the amino acid (AA) digestibilities and nitrogen-corrected true metabolizable energy (TMEn) values of frozen raw, freeze-dried raw, fresh (mildly cooked), and extruded dog foods using the precision-fed cecectomized and conventional rooster assays. The diets tested were Chicken and Barley Recipe [Hill's Science Diet, extruded diet (EXT)], Chicken and White Rice Recipe [Just Food for Dogs, fresh diet (FRSH)], Chicken Formula [Primal Pet Foods, frozen raw diet (FRZN)], Chicken and Sorghum Hybrid Freeze-dried Formula [Primal Pet Foods, hybrid freeze-dried raw diet (HFD)], and Chicken Dinner Patties [Stella & Chewy's, freeze-dried raw diet (FD)]. Two precision-fed rooster assays utilizing Single Comb White Leghorn roosters were conducted. Cecectomized roosters (n = 4/treatment) and conventional roosters (n = 4/treatment) were used to determine standardized AA digestibilities and TMEn, respectively. All roosters were crop intubated with 12 g of test diet and 12 g of corn, with excreta collected for 48 h. In general, FD had the highest, while EXT had the lowest AA digestibilities; however, all diets performed relatively well and few differences in AA digestibility were detected among the diets. Lysine digestibility was higher (P < 0.05) in FD and FRZN than EXT, with other diets being intermediate. Threonine digestibility was higher (P < 0.05) in FD than EXT, with other diets being intermediate. Digestibilities of the other indispensable AA were not different among diets. The reactive lysine:total lysine ratios were 0.94, 0.96, 0.93, 0.93, and 0.95 for EXT, FRSH, FRZN, HFD, and FD, respectively. TMEn was higher (P < 0.05) in FRZN than FD, FRSH, and EXT, higher (P < 0.05) in HFD than FRSH and EXT, and higher (P < 0.05) in FD than EXT. In conclusion, our results support the notion that AA digestibilities are affected by diet processing, with FD, HFD, FRZN, and FRSH diets having higher AA digestibility coefficients and greater TMEn values, than the EXT diet; however, other factors such as ingredient inclusion and macronutrient composition may also have affected these results. More research in dogs is necessary to test the effects of format on diet palatability, digestibility, stool quality, and other physiologically relevant outcomes.

Processing conditions, particularly temperature and duration of heating, impact pet food digestibility. This study tested the standardized amino acid (AA) digestibilities and nitrogen-corrected true metabolizable energy (TMEn) values of five commercial dog diets: extruded diet (EXT), fresh (mildly cooked) diet (FRSH), frozen raw diet (FRZN), hybrid freeze-dried raw diet (HFD), and freeze-dried raw diet (FD). The first study, to determine AA digestibility, used 20 roosters who had their ceca (the main site of microbial fermentation in chickens) surgically removed. The second study used 20 conventional roosters to determine the TMEn of the diets. In general, FD had the highest AA digestibilities, while EXT had the lowest AA digestibilities. True metabolizable energy concentration was higher in the FRZN diet than the FD, FRSH, and EXT diets, higher in the HFD diet than the FRSH and EXT diets, and higher in the FD diet than the EXT diet. Our results support the notion that differences in diet processing, as well as factors such as macronutrient composition, and ingredient source, characteristics, and inclusion may impact AA digestibility and TMEn of dog diets. More research should be conducted to determine exactly how, and to what extent, these different factors impact digestibility in dogs.