The adoption of genetically engineered (GE) crops in agriculture has increased dramatically over the last few decades. Among the transgenic plants, those tolerant to the herbicide glyphosate are among the most common. Weed resistance to glyphosate-based herbicides (GBHs) has been on the rise, leading to increased herbicide applications. This, in turn, has led to increased glyphosate residues in feed. Although glyphosate has been considered to be generally safe to animal health, recent studies have shown that GBHs have potential to cause adverse effects in animal reproduction, including disruption of key regulatory enzymes in androgen synthesis, alteration of serum levels of estrogen and testosterone, damage to reproductive tissues and impairment of gametogenesis. This review emphasizes known effects of GBHs on reproductive health as well as the potential risk GBH residues pose to animal agriculture.

Sperm mobility is a major determinant of sperm quality in the domesticated chicken (Gallus domesticus) and is therefore an area of interest for improving fertility. Sperm-associated antigen 6 (SPAG6) is an important flagellar protein implicated to be necessary for flagellar function but negatively associated with rooster fertility. This study was aimed to characterize the expression of SPAG6 and investigate its utility as a protein biomarker of sperm mobility. By western analysis, relative SPAG6 abundances were compared between the testicular, epididymal, and vasal tissues and in sequentially maturing sperm. Immunocytochemistry techniques were used to detect localization of SPAG6 in chicken sperm. Last, western analysis was used to compare relative SPAG6 abundances in sperm of differing mobility. SPAG6 was found in higher abundance in epididymal tissues and in highest abundance in vasal tissues, relative to that of the testis. SPAG6 was also found to sequentially increase in abundance in maturing sperm. SPAG6 localizes between the axonemal central pair of microtubules in the sperm flagella, but it is also found in lower concentration in the acrosomal region. SPAG6 was not a significant predictor of sperm mobility. SPAG6 abundance, alone, is not a strong predictor of sperm mobility. Its impact on rooster fertility is likely unrelated to its impact on sperm mobility.

Glyphosate-based herbicides (GBHs) such as RoundUp® are a staple of modern crop production, and as a result, residues of their ingredients are typically found in animal feeds. GBH ingredients have repeatedly been shown to impact the male reproductive health of various animals, but at present, the impact of GBH exposures on reproductive health have not been investigated in broiler breeder roosters. This study sought to determine the effect of these exposures on roosters as well as the ability of humic acids (HA) to counteract the effect of GBH exposure. Through 18 weeks of treatment with Roundup®-added or HA-added feeds compared against a common broiler breeder mash, negative effects of Roundup® exposure were seen on testis morphology as well as sperm quality. Increased exposure to Roundup® ingredients resulted in increased vacuolation of seminiferous tubule epithelium. Exposure to Roundup® impacted assessments of sperm quality including sperm mobility, viability and count during the experimental trail. HA supplementation served as a promising adsorptive additive by improving both morphology and sperm quality during the trial. Roundup® exposure was shown to have a negative influence on broiler breeder rooster reproductive health while HA improved reproductive health. The use of HA as an absorbent additive in broiler breeder feeds shows promise in improving reproductive efficiency in broiler breeders.

To prevent broiler breeders from growing too quickly and becoming too large for optimum reproduction, their dietary intake is restricted. While current restricted feeding programs, such as skip-a-day feeding (SAD), improve the economic efficiency of broiler breeder operations, this management practice impacts bird welfare. There is an interest in finding strategies that could reduce the impact of feed restriction during broiler breeder rearing. This research investigated the effects of feeding pullets on an advanced growth curve for early photostimulation at 15 wk (15P) or standard growth curve for photostimulation at 21 wk (21P), using either an every-day-spin feeding program (EDS) or SAD feeding, on the reproductive parameters of broiler breeder hens in a 2 × 2 factorial arrangement. Overall, advancing the growth curve (15P) decreased blood corticosterone levels compared to 21P, but EDS resulted in higher blood corticosterone levels compared to SAD. At the end of rearing in both 15P and 21P, EDS pullets weighed less than SAD pullets. The onset of egg production was 20 and 24 wk of age for the 15P and 21P hens, respectively. Despite an earlier onset, 15P hens did not produce more eggs than 21P hens through 65 wk of age. Egg weight was reduced for 15P compared to 21P until 30 wk of age. The 15P hens had a greater number of double yolk eggs than the 21P hens. Fertility and hatch were not impacted by the advanced growth curve and early photostimulation. Although the current research indicates the potential to reduce feed restriction associated welfare issues by rearing broiler breeder pullets for an earlier photostimulation onset, further research in needed to determine if this management technique can be improved to optimize hen reproductive efficiency.

In sexual reproduction, a male gamete or sperm cell fuses with a female gamete to bring about fertilization. However, a large number of sperm cells with fertilizing ability are required to interact with a female gamete to ensure fertilization. As such, the fertilizing ability of individual sperm cells is critical for successful reproduction. Density gradient centrifugation has been utilized for several decades as a reproducible, fast, efficient, effective and extremely adaptable method to collect only high-quality sperm to be used in assisted reproductive technology. The protocols we described herein focus on the utilization of the discontinuous Percoll density gradient centrifugation (PDGC) technique to isolate three distinct populations of rooster sperm by their quality. We were able to collect low-, medium- and high-quality sperm. We also describe reproducible protocols that entail determining fertility potential of sperm by assessing their viability, mobility and penetrability. Collection of sperm by their quality using PDGC technique would be useful to accurately and thoroughly characterize sperm with differential fertility potential.

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.