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Semen quality is important for roosters' fertility. The correlations between semen quality traits and fertility has less been analyzed, impeding the selection of effective parameters for roosters selection. This research aimed to investigate the variation in semen quality traits in relation to fertilization capacity between 2 chicken breeds. A total of 156 Rhode Island Red (n = 78) and White Leghorn (n = 78) roosters of 53 wk were selected for semen quality estimation including semen volume, pH, sperm concentration, motility, viability, abnormal sperm percentage, and sperm kinematic parameters. Individual fertility was measured by artificial insemination using each 30 birds from 2 breeds. Significant variations were observed between breeds in semen volume, pH, sperm motility (MOT), viability, and abnormal sperm percentage (P < 0.05). The volume, MOT, and viability in Rhode Island Red were higher than those of White Leghorn roosters (P < 0.001). In addition, sperm kinematic parameters such as curvilinear velocity (VCL), straight line velocity (VSL), amplitude lateral head displacement (ALH), and average path velocity (VAP) in Rhode Island Red were higher than those of White Leghorn (P < 0.001). Fertility rate was positively correlated with MOT (r = 0.57), concentration (r = 0.43), viability (r = 0.39), VSL (r = 0.36), ALH (r = 0.43), and ALH (r = 0.38) for Rhode Island Red roosters (P < 0.05). Fertility rate of White Leghorn roosters was positively correlated with MOT (r = 0.71), concentration (r = 0.39), VCL (r = 0.52), ALH (r = 0.50), and VAP (r = 0.39) (P < 0.05). Principal component analysis of sperm kinematic descriptors revealed 2 principal components explaining more than 65% of total variance. In addition, for both genetic lines, the whole population was divided into 3 independent clusters. These results indicated that selection of roosters based on semen quality traits for may improve the fertility, and multivariate analysis may help to precise selection by comprehensive usage of different measures of sperm quality., (Copyright © 2020. Published by Elsevier Inc.)

The objective of this study was to determine the effect of age at photostimulation on sexual maturity and performance of layer breeders. A total of 192 fourteen-wk-old White Leghorn (WL) breeder hens were randomly allocated to 4 treatments of 48 birds each, with 2 replicates per treatment. The birds were photostimulated at 16 (PS16), 18 (PS18), 20 (PS20), and 22 (PS22) wk of age. Four birds per treatment were randomly selected to evaluate sexual organ development at 1 D before photostimulation and 2, 4, and 6 wk after photostimulation. The ovary weight, large yellow follicles number (LYF), oviduct weight, and oviduct length of PS18 increased sharply after photostimulation. Conversely, the increase in PS16 was not observed until 2 wk after photostimulation. There was no difference in age at sexual maturity between treatments (P > 0.05). The PS16 had the longest interval (28 D) from photostimulation to 5% egg production, while PS22 reached 5% egg production 7 D before photostimulation. The PS22 had lower peak production (P = 0.02) and less egg production (P = 0.02) than other treatments. The PS16 had more broken and abnormal eggs (P = 0.01) and lower hatchability (P = 0.04) than other treatments. In conclusion, photostimulation at 16 and 22 wk of age decreases hatchability and egg production, respectively, and photostimulation at 18 wk is appreciated for the WL breeder hens., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

This work studied the effect of age at photostimulation on reproductive performance of Beijing-You Chicken (BYC) breeders. A total of 384 fourteen-week-old BYC breeder hens were randomly allocated to 4 treatments of 96 birds each, with 2 replicates per treatment. The treatments represent photostimulation at 16, 18, 20, and 22 wk of age, respectively (PS16, PS18, PS20, and PS22) by incrementally increasing day length from 8L:16D to 14L:10D and by increasing lighting intensity from 10 to 80 lx. Egg production was recorded for each replicate until 51 wk. Four birds randomly selected from each treatment were sacrificed to characterize sexual organ development at 4 time points: 1 D before photostimulation and 2, 4, and 6 wk after photostimulation. Eggshell quality at peak laying was measured. The results showed that the changes in ovary and oviduct weight in PS22 were 8.68- and 4.27-fold higher than in PS16 at 6 wk after photostimulation. PS16 had an earlier age at 5% egg production than PS20 and PS22 (P = 0.003). The interval from photostimulation to age at 5% egg production in PS20 and PS22 was shorter than in other treatments (P = 0.005). Later peak egg production was seen in PS20 (211.0 D) and PS22 (218.0 D) than in PS16 (183.0 D) and PS18 (190.0 D, P = 0.020), but the laying rate of PS20 decreased slowly after peak laying. PS20 and PS22 had higher egg weights than PS16 and PS18 at peak laying (P = 0.018). Age at photostimulation had no effect on egg number, defective egg number, or hatchability (P > 0.05). In summary, the onset of lay can be advanced by photostimulation at early ages, but there is no difference on egg production until 51 wk. Photostimulation at 20 wk improves peak of laying and laying uniformity., (© 2019 Poultry Science Association Inc.)

Sperm motility is used to predict the semen quality and fertilization ability of breeder roosters. Indigenous chicken breeder roosters show variation in sperm motility, but less has been known about its relationship with other reproductive attributes. In this study, 40- to 44-wk-old Beijing-You breeder roosters with the high and low sperm motility phenotypes (n = 15 in each group) were identified and compared, with regard to their semen characteristics, reproductive hormone concentrations, testicular histomorphology, fertility capacity, and genes expression of hematopoietic prostaglandin D synthase (H-PGDS) and wingless-type MMTV integration site family, member 2 (WNT2). The low sperm motility roosters showed lower semen volume (P = 0.003) and sperm viability (P = 0.008). Like semen donors in the artificial insemination test, the low sperm motility roosters showed lower fertility than the high sperm motility roosters (P < 0.001). No difference in the serum concentration of follicle stimulating hormone (FSH, P = 0.347), luteinizing hormone (LH, P = 0.376), or testosterone (T, P = 0.068) was found between the high and low sperm motility roosters. Furthermore, among roosters with similar BW, the low sperm motility roosters had smaller testis mass (P = 0.037), seminiferous tubules diameter (P = 0.031), spermatogenic epithelium height (P = 0.001), and Johnsen score (P = 0.005) than the high sperm motility ones. Finally, the expression of both H-PGDS and WNT2, which are involved in the prostaglandin D2 and WNT signaling pathways, respectively, was low in the testis of the low sperm motility phenotype roosters (P < 0.05). Thus, the findings of the present study indicated that the testis of low sperm motility showed impaired testicular histomorphology as compared to normal; the BW, testis weight, and serum FSH, LH, and T levels are associated with sperm production, but not with sperm motility, and that low expression of H-PGDS and WNT2 may be related with the abnormal testicular morphology of low sperm motility birds which deserves further functional validation studies., (© 2019 Poultry Science Association Inc.)

Sperm motility is one of the most important indicators for evaluating roosters' fecundity. However, molecular regulation underlying chickens' sperm motility remains to be understood. MicroRNA (miRNA) plays an epigenetic role in reproduction. In this study, we compared the testicular miRNAs of Beijing-you roosters with high (HS) and low (LS) sperm motility using Illumina sequencing to try to understand the associated molecular regulation. Length distribution analysis showed that the dominant size of small RNAs detected in the testes of the chicken samples was 24-28 nt. In total, 518 known and 106 novel miRNAs were identified. A total of 23 miRNAs were found differentially expressed (P < 0.05, |log 2 fold change| ≥ 1) between the HS and LS groups, including 18 known and 5 novel miRNAs. Functional enrichment of the predicted target genes of the differentially expressed miRNAs indicated that these miRNAs were involved in the pathways of GnRH, MAPK and Wnt signaling. The miRNA-gene interaction network revealed two key candidate miRNA-gene pairs that might affect chicken sperm motility, viz, gga-miR-155/KCNA1 and gga-miR-7480-5p/AHI1. qPCR was then used to further validate their expressions. The results here provided a deep insight into the expressions of the miRNAs in the testes of chickens and suggested their roles in sperm motility regulation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The freezability of chicken spermatozoa is low, therefore, effective cryoprotectants is desiderated. Antifreeze proteins (AFPs) are widely found in cold-tolerant species and help them to survive in freezing environments. This study was the first to evaluate the effects of different concentrations of plant-originated antifreeze glycoprotein (AFGP) (0, 0.1, 1, and 5 μg/mL) on post-thawed sperm motion characteristics, morphology, mitochondrial function, antioxidant activity, and fertilizing potential in chickens. Results showed that the total motility of 0.1 to 1 μg/mL AFGP groups were significantly higher than those of the 5 μg/mL AFGP group (P < 0.05). The post-thawed sperm viability of 0.1 μg/mL AFGP group was significantly higher than any of test groups (P < 0.05). Higher abnormal morphology rate of post-thawed sperm was observed in the control group (0 μg/mL AFGP) than in the 0.1, 1, and 5 μg/mL AFGP groups (P < 0.05). The concentrations of malondialdehyde (MDA) decreased gradually with the increase of AFGP concentration. ATP was significantly higher in the 0.1 and 1 μg/mL AFGP groups than those of control and any of test groups (P < 0.05). The 0.1 to 1 μg/mL AFGP groups had increased mitochondrial membrane potential (MMP) level (P > 0.05). The 0.1 μg/mL AFGP group had the highest average fertility (61.36%) compared with control group (57.02%) and any of test groups of chickens at 31 wk of age, and the 1 μg/mL AFGP group had the highest average fertility (37.72%) compared with control group (21.73%) and any of test groups of chickens at 65 wk of age. In conclusion, the results from this study suggest lower concentration of AFGP (0.1–1 μg/mL) showed positive effect for sperm function. This study inspires the continuous evaluation and seeking right way of adopting different kinds of AFPs in rooster semen cryopreservation. [ABSTRACT FROM AUTHOR]

Simple Summary: Although sperm cryopreservation has a great potential for application in the poultry industry, it is still very limited at the economical scale due to the low fertility of frozen/thawed avian spermatozoa. Dimethylacetamide (DMA) has been widely employed as a cryoprotectant for the cryopreservation of chicken spermatozoa, but with variable results in motility and fertility. To determine the appropriate concentration of DMA in the chicken semen freezing extender, in vitro sperm quality and motility analyses and in vivo field trials based on the fertility of the frozen/thawed rooster sperm were conducted in the present study. The antioxidant biomarkers and novel anti-freeze-associated genes were also analyzed in the post-thawed sperm. Results conclude that DMA is preferably used at a low concentration to obtain applicable results in sperm motility and fertility after thawing without harming the quality and antioxidant system defense. The current study could highlight the possible molecular mechanisms behind the obtained fertility results of DMA-frozen sperm, especially when artificial insemination is implemented in the poultry industry. Sperm cryopreservation is of great importance for the poultry industry but still needs to be optimized. The high susceptibility of poultry sperm to cryodamage leads to low fertility rates after cryopreservation. Therefore, the present study aimed at evaluating the effect of including a cryoprotectant, dimethylacetamide (DMA), in the chicken semen freezing extenders at a final concentration of 3%, 6%, or 9% on the post-thawed sperm motility, quality, antioxidant biomarkers, anti-freeze gene expression, and fertilizing ability. Results showed that the total motile sperm, progressivity, and viability were quadratically increased (p < 0.05) in the 6% DMA group. The antioxidant enzyme activity and lipid peroxidation were negatively (p < 0.05) affected by the increase in DMA concentration. Furthermore, some anti-freeze-associated genes such as heat shock protein 70 (HSP70) and ras homolog family member A (RHOA) were linearly and quadratically down-regulated (p < 0.05) with the high concentration of DMA. Finally, the fertility and hatchability rates did not indicate statistical differences between DMA groups. It can be concluded that using the low concentration of 3–6% DMA in the freezing semen extender is preferable to obtain acceptable results in the post-thawed sperm quality and fertility. [ABSTRACT FROM AUTHOR]

The cytoplasmic male sterility/restorer-of-fertility (CMS/Rf) system plays a vital role in high-efficiency hybrid seed production in crops, including soybean (Glycine max (L.) Merr.). The markers linked to fertility restoration and the restorer-of-fertility (Rf) genes are essential because they can facilitate the breeding of new CMS lines and production of commercial hybrid soybean seeds. To date, several soybean Rf genes have been mapped to various genetic loci in diverse genetic populations. However, the mapping range of restorer genes remains narrow, with relatively limited practical applicability. Therefore, in the present study, F2 and F3 segregating populations derived from the CMS line JLCMS5A crossed with the restorer line JLR2 were developed and used for Rf3 gene fine mapping. Genetic investigation indicated that the restorer line JLR2 was controlled by a single dominant gene, Rf3. By integrating bulk-segregant analysis and next-generation sequencing, a 4 Mb region on chromosome 9 was identified, which was most likely the target region harboring the candidate gene responsible for fertility restoration. This region was further narrowed down to 86.44 Kb via fine mapping in F2 and F3 populations using SSR, InDel, and dCAPS markers. This region contained 10 putative genes (Glyma.09G171100–Glyma.09G172000). Finally, Glyma.09G171200, which encodes a mitochondria-targeted pentatricopeptide repeat protein, was proposed as the potential candidate for Rf3 using sequence alignment and expression analysis in restorer and CMS lines. Based on single-nucleotide polymorphisms in Glyma.09G171200, a CAPS marker co-segregated with Rf3 named CAPS1712 was developed. Our results will be fundamental in the assisted selection and creation of potent lines for the production and rapid selection of novel restorer lines. [ABSTRACT FROM AUTHOR]

Background: Photoperiod and/or thermo-sensitive male sterility is an effective pollination control system in crop two-line hybrid breeding. We previously discovered the spontaneous mutation of a partially male sterile plant and developed a thermo-sensitive genic male sterile (TGMS) line 373S in Brassica napus L. The present study characterized this TGMS line through cytological observation, photoperiod/ temperature treatments, and genetic investigation. Results: Microscopic observation revealed that the condensed cytoplasm and irregular exine of microspores and the abnormal degradation of tapetum are related to pollen abortion. Different temperature and photoperiod treatments in field and growth cabinet conditions indicated that the fertility alteration of 373S was mainly caused by temperature changes. The effects of photoperiod and interaction between temperature and photoperiod were insignificant. The critical temperature leading to fertility alteration ranged from 10 °C (15 °C/5 °C) to 12 °C (17 °C/7 °C), and the temperature-responding stage was coincident with anther development from pollen mother cell formation to meiosis stages. Genetic analysis indicated that the TGMS trait in 373S was controlled by one pair of genes, with male sterility as the recessive. Multiplex PCR analysis revealed that the cytoplasm of 373S is pol type. Conclusions: Our study suggested that the 373S line in B. napus has a novel thermo-sensitive gene Bnmst1 in Pol CMS cytoplasm background, and its fertility alteration is mainly caused by temperature changes. Our results will broaden the TGMS resources and lay the foundation for two-line hybrid breeding in B. napus. [ABSTRACT FROM AUTHOR]

The long-term semen cryopreservation is increasingly crucial for conservation of endangered livestock and poultry species. Glycerol is the most widely used cryoprotectant for freezing chicken semen. Continuous improvement in details with glycerol may help increase the fertility of post-thawed semen. Two experiments were performed in the present study to investigate the effects of glycerol concentration, removal method, and straw type on the quality of post-thawed sperm. In experiment 1, glycerol concentration (3%, 5%, 7%, 9%, 11%, and 13%) and glycerol removal method (final dilution ratio 1:1, 1:2, 1:4, 1:8, 1:16, and 1:20) combination groups were investigated for post-thawed sperm quality, residual glycerol concentration, and fertility to find the best combinations. Experiment 2 was performed to evaluate the effects of straw type (0.25 and 0.5 mL) and glycerol concentration (3%, 5%, 7%, 9%, 11%, and 13%) on the post-thawed sperm quality. Results showed that post-thawed sperm motility of 6 glycerol concentration groups were different (P < 0.01). Sperm motility of 5%, 7%, 9%, 11% and 13% was higher than that of 3% (P < 0.01). There was no difference among different concentrations of glycerol in VSL, VCL, VAP, ALH, WOB, BCF, LIN, or STR (P > 0.05). As for the glycerol removal method, sperm motility of 1:8 dilution was the highest, followed by 1:1 and 1:2, while the difference among groups was not statistically significant (P = 0.11). Glycerol concentration and removal method had no interaction effect on sperm motion parameters (P > 0.05). The highest fertility (48.70%) was found for the 5% and 1:2 combination. There was no difference for sperm motility between 0.25 and 0.5 mL straws (P > 0.05). Glycerol concentration and straw type had no interaction effect on the sperm motion parameters (P > 0.05). It can be concluded from these observations that the combination of 5% glycerol and 1:2 dilution rendered higher fertility should be suggested in practice, and that both 0.25 and 0.50 mL straws fit the present procedure. [ABSTRACT FROM AUTHOR]

The 2 × 4 factorial experiment was designed to determine the effect of strain and photostimulation age on sexual maturity and reproductive performance of rooster breeders. A total of 96 White Leghorn (WL) and 120 Beijing You Chicken (BYC) roosters were randomly allocated to 4 treatments at 14 wk of age. The treatments represent photostimulation at 16, 18, 20, and 22 wk of age, respectively (PS16 , PS18 , PS20 , and PS22), in both strains. Photostimulation was achieved by increasing the day length from 8L:16D to 14L:10D and by increasing lighting intensity from 10 lx to 80 lx. Three birds from each interaction were sacrificed to characterize the comb and testis weights at 4 time points: 1 d before photostimulation and 2, 4, and 6 wk after photostimulation. Semen quality and hatching performance with the semen of the experimental roosters were measured at 30 and 45 wk of age, respectively. Results showed that the testis weight of PS20 and PS22 in WL and BYC was 6.4- and 2.9-fold higher than that of PS18 before photostimulation, while testis weight of PS18 in both strains increased sharply after photostimulation. The diameter of seminiferous tubules increased in the photostimulated roosters as compared with the nonphotostimulated ones, and mature spermatozoa were produced 4 wk after photostimulation and at 20 wk of age for PS16. The WL had lower semen volume and total sperm count than BYC (P < 0.01), but there was no difference on effective sperm count (P > 0.05). In addition, semen quality traits were not affected by age at photostimulation (P > 0.05) in both strains. The fertility and hatching performance were not affected by strain or photostimulation age (P > 0.05). In summary, the sexual maturation of rooster breeders can be advanced by photostimulation at an early age, which does not lead to a difference in semen quality or hatching performance at adult stage. [ABSTRACT FROM AUTHOR]