Fertilization Rate Research Articles

Agronomic and environmental effects of forage-cutting schedule and nitrogen fertilization for bermudagrass (Cynodon dactylon, L.)

Bermudagrass (Cynodon dactylon, L.) is widely used as a forage in ruminant diets owing to its nutritional value and its capacity to grow under various agroecological conditions. For the southeastern United States climate conditions, previous research on bermudagrass recommended the cultivar Tifton 85 for forage production and a monthly frequency for forage-cutting. Even though bermudagrass cultivars are known to positively respond to nitrogen (N) fertilization, the interplay of the forage-cutting and N fertilization rates is not well understood. As a result, there is not sufficient guidance for adequate harvesting schedules and fertilization management to optimize bermudagrass forage production. Hence, this study aims to clarify the interplay of biomass-cutting events and N fertilization rates on forage quality and quantity, and N footprint. The study used experimental data of bermudagrass Tifton 85 forage production under three N fertilization rates (i.e., high = 504 kg N ha−1, medium = 336 kg N ha−1, and low = 168 kg N ha−1), to calibrate and validate Root Zone Water Quality Model 2 (RZWQM2) for biomass weight and biomass N content. For each N fertilization rate, multiyear simulations of four biomass-cutting scenarios were used to investigate the joint effects of harvesting schedules and N fertilization rates on biomass weight, biomass N content, N use efficiency (NUE), and N leaching. Results show statistically significant effects of the biomass-cutting scenarios on biomass weight and biomass N content for both high and medium N fertilization rates. The interplay of biomass-cutting and N fertilization reflected differently on forage quality and quantity, and N footprint. The low N fertilization did not show any statistically significant effect except for the biomass weight. NUE values were higher with both medium and low N fertilization rates compared to the high N fertilization which showed a relatively high N leaching. The outcomes of this study can be used to inform bermudagrass cutting and fertilization options to achieve forage yield goals with an understanding of the potential environmental consequence of N leaching and low NUE.

Effects of ovarian stimulation on embryo euploidy: an analysis of 12874 oocytes and 3106 blastocysts in cycles with preimplantation genetic testing for monogenic disorders.

Does ovarian stimulation and the ovarian response affect embryo euploidy? Ovarian stimulation and the ovarian response in women undergoing preimplantation genetic testing for monogenic disorders (PGT-M) cycles did not affect the rates of blastocyst euploidy. Whether or not ovarian stimulation in IVF-embryo transfer has potential effects on embryo euploidy is controversial among studies for several reasons: (i) heterogeneity of the study populations, (ii) biopsies being performed at different stages of embryo development and (iii) evolution of the platforms utilized for ploidy assessment. Patients who undergo PGT-M cycles typically have no additional risks of aneuploidy, providing an ideal study population for exploring this issue. A retrospective cohort study including embryos undergoing PGT-M was conducted at a single academically affiliated fertility clinic between June 2014 and July 2021. A total of 617 women with 867 PGT-M cycles involving 12874 retrieved oocytes and 3106 trophectoderm biopsies of blastocysts were included. The primary outcome of the study was median euploidy rate, which was calculated by dividing the number of euploid blastocysts by the total number of biopsied blastocysts for each cycle. Secondary outcomes included the median normal fertilization rate (two-pronuclear (2PN) embryos/metaphase II oocytes) and median blastulation rate (blastocyst numbers/2PN embryos). Comparable euploidy rates and fertilization rates were observed across all age groups, regardless of variations in ovarian stimulation protocols, gonadotropin dosages (both the starting and total dosages), stimulation durations, the inclusion of human menopausal gonadotrophin supplementation, or the number of oocytes retrieved (all P > 0.05). Blastulation rates declined with increasing starting doses of gonadotropins in women aged 31-34 years old (P = 0.005) but increased with increasing gonadotrophin starting doses in women aged 35-37 years old (P = 0.017). In women aged 31-34, 35-37, and 38-40 years old, blastulation rates were significantly reduced with increases in the number of oocytes retrieved (P = 0.001, <0.001, and 0.012, respectively). Limitations include the study's retrospective nature and the relatively small number of patients of advanced age, especially patients older than 40 years old, leading to quite low statistical power. Second, as we considered euploidy rates as outcome measures, we did not analyze the effects of ovarian stimulation on uniform aneuploidy and mosaicism, respectively. Finally, we did not consider the effects of paternal characteristics on embryo euploidy status due to the fact that blastocyst aneuploidy primarily originates from maternal meiosis. However, sperm factors might have an effect on embryo development and the blastulation rate, and therefore also the number of blastocysts analyzed. The exclusion of patients with severe teratozoospermia and the fact that only ICSI was used as the insemination technique for women undergoing PGT-M contributed to minimize the effect of paternal factors. Ovarian stimulation and response to stimulation did not affect blastocyst euploidy rates in women undergoing PGT-M cycles. However, in women aged 31-40 years old, there was a significant decline in blastulation rates as the number of retrieved oocytes increased. This study was supported by the National Natural Science Foundation of China (Grant No. 81701407, 82301826); the National Key Research and Development Program of China (2022YFC2702901, 2022YFC2703004); China Postdoctoral Science Foundation (2022M710261), and China Postdoctoral Innovation Talent Support Program (BX20220020). There is no conflict of interest. N/A.

A Mixture of Summer Legume and Nonlegume Cover Crops Enhances Winter Wheat Yield, Nitrogen Uptake, and Nitrogen Balance

Cover crops protecting soil erosion during the summer fallow in the monsoon weather may enhance dryland winter wheat yield and N relations. We examined the effects of four summer cover crops (soybean (Glycine max L., SB), sudangrass (Sorghum sudanense {Piper} Stapf, SG), soybean and sudangrass mixture (SS), and no cover crop (CK)) and three N fertilization rates (0, 60, and 120 kg N ha−1) on winter wheat yield, quality, and N relations from 2017–2018 to 2020–2021 in the Loess Plateau of China. Cover crop biomass and N accumulation, soil mineral N, and winter wheat yield, protein concentration, and N uptake were greater for SB and SS than other cover crops at most N fertilization rates and years. The N fertilization rate had variable effects on these parameters. Winter wheat aboveground biomass and grain N productivities were greater for CK than other cover crops at all N fertilization rates and years. Nitrogen balance was greater for SS than other cover crops at 60 and 120 kg N ha−1 in all years. The SS with 120 kg N ha−1 can enhance soil mineral N, winter wheat yield and quality, and N balance compared to CK and SG with or without N fertilization rates.

Comparison of highly purified human menopausal gonadotropin and recombinant follicle stimulating hormone use in patients undergoing in vitro fertilization with progestin-primed ovarian stimulation protocol: a single center retrospective analysis.

Recently, progesterone has been used to prevent LH surge instead of GnRH analogues during ART treatments, which is known as progesterone-primed ovarian stimulation (PPOS) protocol. During ART treatment, highly purified human menopausal gonadotropin (HP-hMG) and recombinant follicle stimulating hormone (rFSH) are two of the agents used for stimulation of antral follicles. The aim of this study is to compare the efficacy and success of HP-hMG and rFSH agents in the ovarian stimulation step of the PPOS protocol, which has not been previously reported in the literature. This retrospective study was conducted at a university hospital with patients who underwent IVF treatment using PPOS protocols in between January 2019 and July 2021. For ovarian stimulation, rFSH was used in group I and HP-hMG was used in group II. Mature oocyte ratio was the primary outcome, and live birth rate was the secondary outcome. Mann-Whitney and Chi-square tests were used for statistical analysis. All p values below 0.05 were considered significant. Total numbers of follicles, oocytes, MII, and 2PN numbers obtained were similar between the two groups. The fertilization rates were 66.7% in the rFSH group and 64.3% in the HP-hMG group (p > 0.05). The pregnancy rates were 53.5% and 46.7% in the rFSH and HP-hMG groups, respectively. There was no statistically significant difference between pregnancy, abortus, and live birth rates. In this study, it is demonstrated that stimulation of oocytes with either rFSH or hMG in the PPOS protocol, which has been added to IVF treatment protocols in recent years, had no statistical difference regarding mature oocyte numbers and live birth rates between the two groups. These results are consistent with the previous literature which compared rFSH and hMG in GnRH agonist and antagonist protocols.

Effects of Planting Density and Nitrogen Fertilization on Growth Traits and Leaf and Wood Characteristics of Three Poplar Clones

A comprehension of the effects planting density and nitrogen (N) fertilization have on the physiological and morphological characteristics of trees is critical for optimizing the require size and characteristics of wood products. We evaluated the growth traits and the leaf and wood characteristics of three clone poplars including Populus simonii × P. nigra ‘Xiaohei’, ‘Xiaohei-14’ and ‘Bailin-3’ under five planting densities (1666, 1111, 833, 666, and 555 tree ha−1) and four N fertilization rates (0, 100, 160, and 220 g tree−1 year−1). The results show that the clone type significantly affected all observed indicators, while planting density and N fertilization treatments had a significant effect on growth traits and leaf characteristics, but not on wood characteristics. Specifically, the clone ‘Bailin-3’ exhibited the largest annual increments in tree height and diameter at breast height (DBH), leaf width, N content, and soluble protein content. A decrease in initial planting density (from 1666 to 555 tree ha−1) led to an increased annual incremental tree height and DBH, regardless of clone type and N fertilization treatment. N fertilization treatment significantly impacted the annual increment in DBH, but not that of tree height. Further, the annual increments in tree height and DBH were positively correlated with leaf width, N content, chlorophyll content, and soluble protein content, and negatively correlated with hemicellulose content. In addition, the chlorophyll and soluble protein contents were identified as the most reliable predictors of the annual increments in tree height and DBH. Our results demonstrate the clone ‘Bailin-3’ with 555 tree ha−1 under 160 g N tree−1 yr−1 showed superior growth traits and leaf characteristics. Thus, it is recommended for future poplar silviculture of larger diameter timber production at similar sites. The results contribute to understanding of the effects of planting density and fertilization on the growth traits and the leaf and wood characteristics of three poplar clones, offering valuable guidance for the sustainable development and long-term productivity of poplar plantations.

Maize–peanut intercropping and N fertilization changed the potential nitrification rate by regulating the ratio of AOB to AOA in soils

Maize–peanut intercropping could potentially mitigate nitrogen (N) loss from the soil, a process primarily governed by the net nitrification rate. However, the impact of maize–peanut intercropping on the potential nitrification rate (PNR) and its relationships with key players, such as ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), are not well understood. Herein, we conducted a field experiment involving two management systems and two crops, namely, maize (MPm) and peanut (MPp) intercropping, maize monoculture (MM), and peanut monoculture (PM), under three N fertilization rates (no N fertilization, 150 kg N ha−1, and 300 kg N ha−1). Under intercropping (MPm and MPp), the abundance of AOA amoA gene increased by 64.8% and 60.3% and the abundance of AOB amoA gene increased by 63.2% and 68.2% compared to the MM and PM monoculture systems, respectively. Furthermore, the abundances of AOA and AOB decreased in MPp and MM, while AOB increased in MPm and PM across the N fertilization gradient. The PNR increased corresponding to the N fertilization rates, with intercropping enhancing the PNR in peanut-planted soil but reducing the PNR in maize-planted soil compared to monocropping. Notably, no significant positive relationship between the abundances of AOA or AOB and the PNR. Random forest analysis indicated that the AOB/AOA ratio was an important predictor of the PNR. N fertilization and intercropping regulated the AOB/AOA ratio mainly through controlling the ammonia content and the soil C/N, respectively. These findings highlight the substantial impacts of N fertilization and intercropping on PNR, with the AOB/AOA ratio emerging as a valuable predictive indicator for the PNR.

GnRH antagonist protocol with hCG triggering ameliorates fertilization defect caused by failure of cumulus cell pentraxin-3 expression in unilateral endometriomas.

The aim of the study was to determine the expression pattern of long pentraxin 3 (PTX3) mRNA in cumulus cells (CCs) isolated from metaphase II oocytes of women with unilateral endometrioma undergoing controlled ovarian stimulation using a gonadotropin-releasing hormone antagonist (GnRHa) protocol. A total of 60 CC samples, 30 from the affected ovary and 30 from the contralateral ovary, were collected from 12 patients with unilateral endometrioma who underwent flexible GnRHa protocol with recombinant human chorionic gonadotropin (rhCG) trigger. Thirty CC samples collected from the left ovary of 12 women with male factor infertility were used as external controls. Long PTX3 mRNA expression in each group was analyzed by real-time polymerase chain reaction (RT-PCR). Relative gene expression (fold-change) was calculated according to the 2-ΔΔCt equation. Fertilization rates after intracytoplasmic sperm injection (ICSI) were recorded for each group. CC-PTX3 mRNA expression in the unilateral endometrioma group was significantly lower than the mRNA expression of the disease-free ovary (0.90±0.01 vs. 0.25±0.02, p<0.01). CC-PTX3 mRNA expression of MII oocytes of the disease-free ovary was found to be similar to the control group (1.02±0.03 vs. 0.90±0.01, p=0.107). A significant 3.6-fold downregulation was observed in the CC-PTX3 mRNA expression of the endometrioma group compared to the CC-PTX3 mRNA expression in the contralateral ovary. CC-PTX3 mRNA expression in the endometrioma group was downregulated 4.08-fold compared to the CC-PTX3 mRNA expression of the control group (1.02±0.03 vs. 0.25±0.02, p<0.001). The cumulus morphologies of the endometrioma group with low CC-PTX3 expression and the groups with normal CC-PTX3 levels were similar. Fertilization rates of the endometrioma group were similar to the contralateral ovary and control groups. Unilateral endometrioma reduces CC-PTX3 expression but does not affect disease-free ovaries. The GnRHa protocol improved the fertilization rates, suggesting that failed CC-PTX3 expression is an in vivo pathology.

Water dispersible carbon nanomaterials reduced N, P, and K leaching potential in sandy soils: A column leaching study

Carbon nanomaterials (CNMs) — amendments with carbon in nanoscale form —could potentially enhance fertilizer delivery efficiency in agriculture, but their interaction with soil properties and nutrient co-mobility, especially in coarse-textured soils, remain poorly understood. We conducted a column leaching study in repacked soil columns to compare the co-leaching of novel water-dispersible CNMs and soil nutrients across two levels of CNMs applications (200 & 400 mg kg−1), two fertilization rates (low:80 mg kg−1 of N, P and K and high: 200 mg N kg−1, 100 mg P kg−1, 200 mg K kg−1, applied as ammonium nitrate, potassium phosphate, and potassium nitrate) and two soils (Spodosol with pH = 5.1, Alfisol with pH = 6.5). We imposed 12 leaching events to each column, with each leaching event adding water equivalent to the soil-pore volume (250 mL), resulting in cumulative leaching of 3000 mL of water through each column. CNMs applications reduced cumulative leaching losses of NO3-N (Spodosol: 8–12 %, Alfisol: 9–19 %), NH4-N (Spodosol: 2–14 %, Alfisol: 9–14 %), P (Spodosol: 23–27 %, Alfisol: 23–36 %) and K (Spodosol: 17–23 %, Alfisol: 24–26 %) compared to fertilized columns without CNMs. CNMs increased soil pH by up to 0.3 units (Spodosol) or 0.5 units (Alfisol), while lowering electrical conductivity by 15–20 % at the high fertilization rate in both soils. Columns with water-dispersible CNMs accumulated 25–30 % more total C in the base sections of the Alfisol compared to the Spodosol, indicating faster downward movement through the soil profile. Overall, we demonstrated that CNMs have the potential to reduce nutrient leaching in coarse-textured soils, which could be particularly beneficial in high-input intensive agricultural systems.

In Vitro Fertilization (IVF) Outcomes in Patients With Endometriosis Compared to Patients With Tubal Factor Infertility in Indonesia: A Retrospective Study.

Endometriosis is a disease characterized by dysmenorrhea, chronic pelvic pain, and infertility. The pathogenesis of endometriosis and its relationship to infertility are still not fully understood. In vitro fertilization (IVF) is considered an effective treatment for patients with endometriosis-associated infertility. This study compared the pregnancy rates of endometriosis patients who underwent IVF with those of patients with tubal obstruction. This was a retrospective cross-sectional study of 225 patients with endometriosis and tubal factor infertility who underwent IVF at Yasmin Clinic, Dr. Cipto Mangunkusumo National General Hospital, Jakarta, Indonesia, between January 2013 and August 2021. Demographic data, anti-Mullerian hormone (AMH) levels, the initial and total dose of recombinant follicle stimulating hormone (rFSH), the total dose of recombinant luteinizing hormone (rLH), stimulation duration, oocyte maturation rates, fertilization rates, embryo cleavage rates, and pregnancy rates (biochemical, clinical and ongoing) were obtained. AMH level, number of oocytes retrieved, and embryo cleavage rates were significantly lower in the endometriosis group. Initial and total doses of rFSH and total dose of rLH were higher, and the duration of stimulation was longer in the endometriosis group compared to the tubal factor group. In the endometriosis group, the biochemical (47.3% vs. 52.7%, p=0.375), clinical (43.1% vs. 56.9%, p=0.215), and ongoing (45.5% vs. 54.5%, p=0.511) pregnancy rates were lower than those in the tubal factor group. However, there were no statistically significant differences between the two groups. There was no significant difference in pregnancy rates between the short antagonist and ultra-long protocols (three months of downregulation). From multivariate analysis, only rLH supplementation was found to be significantly related to pregnancy outcomes in patients with endometriosis. AMH levels, number of oocytes retrieved, and embryo cleavage rates were lower in patients with endometriosis. However, pregnancy rates were not significantly different from patients with tubal factors. Supplementation with rLH may improve pregnancy rates in patients with endometriosis who have undergone IVF programs.

Comparative analysis of short versus long co-incubation of gametes on post-insemination outcomes and embryo morphokinetics: a sibling oocyte randomized study

Research questionDoes a short co-incubation of gametes in conventional IVF impact post-insemination outcomes and embryo morphokinetics? DesignSibling oocyte randomized pilot study performed between December 2020 and March 2023. Eligible couples (n=55) were female aged 18-43 years with body mass index (BMI) ≤35 and male normal semen parameters. Cumulus oocyte complexes (COCs) (min 6 - max 12) were randomized in a 1:1:2 proportion in long (16-18 hrs), or short (2 hrs) co-incubation IVF exposure, and ICSI, respectively. All oocytes inseminated via IVF were monitored by time-lapse (TL). Blastocysts of ≥BL3CC were subjected to trophectoderm (TE) biopsy on day (D)5, D6 or D7 by next generation sequencing (NGS). ResultsA total of 954 COCs were distributed to long co-incubation (n=235, 24.6%), short co-incubation (n=235, 24.6%), and ICSI (n=484, 50.3%). In total, 202 were MII after long co-incubation, 209 after short co-incubation and 394 at ICSI. Regression analyses showed both short co-incubation groups (OR: 0.72, 95% CI: 0.46–1.11, P=0.139) and ICSI (OR: 1.43, 05% CI: 0.95–2.15, P=0.090) did not significantly impact normal fertilization rates (2PN) when compared to long co-incubation. Usable blastocysts per MII, per 2PN, and euploid blastocysts per MII, per 2PN and per tested blastocysts were similar among the groups. The rate of embryo arrest (HR: HR: 0.79, 95% CI: 0.57-1.11, P=0.183) and timings to reach morphokinetic milestones among embryos reaching blastulation were similar between IVF groups (P>0.05, for all). ConclusionA short 2-hour co-incubation of gametes in IVF yields comparable fertilization, blastocyst, and euploidy rates without adverse embryo morphokinetic events, compared to standard overnight co-incubation.

Rat copper transport protein 2 (CTR2) is involved in fertilization through interaction with IZUMO1 and JUNO

In mammalian reproduction, testis-specific protein IZUMO1 and its receptor JUNO on the oocyte surface are essential for sperm-oocyte recognition, binding, and membrane fusion. However, these factors alone are insufficient to accomplish cytoplasmic membrane fusion. It is believed that other gametic proteins interact with them to facilitate sperm-oocyte interaction on the head and mid-tail of rat spermatozoa as well as on the surface of oocytes. In this study, Copper Transport Protein 2 (CTR2) has been identified on the head and mid-tail of rat spermatozoa as well as on the surface of oocytes. CTR2 directly interacts with both IZUMO1 and JUNO, colocalizing with IZUMO1 on the sperm head and with JUNO on the oocyte membrane. Treatment of the capacitated sperm and zona pellucida-free oocytes with anti-CTR2 antibody resulted in a significant decrease in fertilization rates in IVF experiments. These findings suggest that CTR2 plays an important role in mammalian fertilization by interacting with IZUMO1 and JUNO, providing new insights into the molecular mechanisms of mammalian sperm-oocyte adhesion and fusion.

Exposure of antral follicles to medroxyprogesterone acetate during stimulation does not cause molecular perturbations in gonadotropin-responsiveness and steroidogenic function of granulosa cells in progestin-primed cycles.

Does medroxyprogesterone acetate (MPA) exposure in progestin-primed ovarian stimulation (PPOS) cycles cause molecular perturbations in the steroidogenic function and gonadotropin responsiveness of the granulosa cells? PPOS cycles are identical to traditional GnRH antagonist cycles not only for clinical IVF characteristics but also for gonadotropin receptor expression, response to gonadotropins, and steroidogenic function at the molecular level. PPOS is increasingly used as an alternative to GnRH antagonists due to the inhibitory effect of progesterone on LH release by reducing GnRH pulsatility at the hypothalamic level. Although a growing body of evidence from clinical studies did not indicate significant differences between PPOS and antagonist protocols for IVF cycle characteristics and obstetrical outcomes, it is still unknown whether exposure of the antral follicle cohort to progesterone or its synthetic derivatives during ovarian stimulation causes any subtle molecular aberrations in terms of steroidogenesis and gonadotropin responsiveness. To address this issue, detailed comparative molecular analyses were conducted in the luteinized mural granulosa cells (GCs) obtained from normal responding IVF patients undergoing PPOS and antagonist cycles. A clinical translational research study was conducted with IVF patients. This study included 55 normal responding IVF patients who underwent ovarian stimulation with either PPOS using MPA (5 mg twice daily) or GnRH antagonist cetrorelix acetate. Recombinant forms of FSH and hCG were used for ovarian stimulation and ovulation triggering, respectively. Luteinized mural GCs obtained during the oocyte retrieval procedure were used for the experiments. Cell culture, quantitative real-time PCR, immunoblotting, confocal time-lapse live cell imaging, and hormone assays were used. Demographic and IVF cycle characteristics of the patients undergoing ovarian stimulation with PPOS and GnRH antagonist were similar, including ovarian response, mature oocyte yield, and fertilization rates. Molecular analyses revealed that the expression of the enzymes involved in sex-steroid synthesis (StAR, SCC, 3β-HSD, 17β-HSD, aromatase) and the uptake/storage/utilization of cholesterol (LDL receptor, Hormone-sensitive lipase, hydroxy-methyl glutaryl Co-enzyme-A reductase, and Sterol O-acyltransferase1) in the GCs of the PPOS cycles were comparable to those of the antagonist cycles. The expression of the receptors for gonadotropins, estrogen, and progesterone hormones was also similar. Basal and hCG-induced increases in 3β-HSD expression and progesterone production and basal and FSH-induced increases in aromatase expression and E2 output of the GCs from PPOS patients did not exhibit any meaningful differences when compared with GCs from antagonist cycles. Furthermore, basal and hCG-induced up-regulation in the LDL receptor expression and cholesterol uptake did not differ between the groups. Confocal imaging also revealed similar patterns of expression for the steroidogenic enzymes and their co-localization with mitochondria. Lastly, the expression of the other important genes regulating cumulus expansion, ovulation, and luteal function [Relaxin, ADAMTS-1, and epidermal growth factor (EGF)-like growth factor amphiregulin] in the GCs of the PPOS and antagonist cycles were similar. N/A. Caution should be exercised when interpreting our data which was derived from normally responding patients whose ovulation was triggered with hCG. It is unclear whether the molecular parameters assessed vary according to infertility etiologies, magnitude of ovarian response, mode of trigger, and any other underlying ovarian pathologies or systemic diseases. MPA was the progestin used for PPOS and whether these findings can be generalized to other progestins is unknown. This study provides reassuring molecular evidence that exposure of antral follicle cohorts to MPA during the follicular growth phase does not have any detrimental effects on steroidogenic, ovulatory, and luteal functions when compared with GnRH antagonist cycles. This study was funded by the School of Medicine, the Graduate School of Health Sciences of Koc University and Koç University Research Center for Translational Medicine (KUTTAM), and equally funded by the Republic of Turkey Ministry of Development Research Infrastructure Support Program. All authors declare no conflict of interest. N/A.

Nitrogen fertilization management and seeding density differently affect net blotch incidence and grain yield in one two-row and one six-row cultivar of barley

A two-year field experiment was carried out on barley (Hordeum vulgare L.) to test the effect of cultivar, N fertilization management and seeding density on crop growth, net blotch (NB) disease occurrence and yield. The three variables were factorially combined in a split-split-plot design as follows: 1) two barley cultivars, one two-row (H. v. distichum, cv Callas) and one six-row (H. v. hexastichum, cv Azzurro); 2) four N application schedules differing for rates and/or timing of N supply: unfertilized control; fertilized with 35kgha-1 of N at BBCH29 plus 30kgha-1 of N at BBCH31; fertilized with 70kgha-1 of N at BBCH29 plus 60kgha-1 of N at BBCH31; fertilized with 130kgha-1 of N simultaneously at BBCH31; 3) four seeding densities: 100, 200, 400 and 800 viable seeds m2.The three variables affected crop growth and NB occurrence resulting in different grain yield and quality. Results varied with the seasonal trend, in particular with rainfall regime and air temperature, for their possible effect on soil N dynamics and the presence of NB, which resulted in different source capacity, and for their effect on grain setting and filling, i.e., the sink capacity. Both N fertilization and seeding density increased NB pressure, with seeding density affecting the disease even under unfavourable weather conditions for NB development. At the maximum N fertilization rate, delaying N application helped limit plant height and the related risk of lodging, as well as NB symptoms, resulting in greater hectolitre weight and protein content together with no appreciable decrease of grain yield, provided seeding density was close to optimal (i.e., 400 seeds m-2) or higher.

Clinical Applications of Assisted Oocyte Activation in Couples with Various Reproductive Problems: A Systematic Review.

ICSI may face fertilization failure, prompting the use of assisted oocyte activation (AOA) techniques. While AOA is implemented in infertility clinics, its target patients and definitive application remain uncertain. This systematic review adheres to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines to assess reproductive outcomes in ICSI-AOA cycles compared to conventional ICSI and evaluate AOA effectiveness in various infertility disorders. A literature search encompassed PubMed, Web of Science, EMBASE, and Scopus databases until December 2023 for relevant English studies. Included studies compared ICSI-AOA with conventional ICSI in couples with prior fertilization failure, utilizing diverse AOA methods. Control groups consisted of sibling oocytes, previous cycles of the same couples, or couples undergoing conventional ICSI. Evaluated outcomes included fertilization, cleavage, embryo quality, implantation, pregnancy, and live birth rates. Article screening and data extraction were performed by two authors, with risk of bias assessed by another investigator. Out of 3088 initially identified articles, 30 studies were included, focusing on fertilization failure (n = 10), female infertility (n = 3), PLCζ defects (n = 4), poor sperm quality (n = 4), Globozoospermia (n = 4), and surgically retrieved sperm (n = 8). Most studies concluded that AOA could overcome fertilization failure, but success rates varied based on sperm-related or oocyte-related factors in ICSI-AOA cycles. Due to differences in patient inclusion criteria and sample sizes, most studies were not sufficiently similar for pooled analysis, limiting robust conclusions. There is insufficient evidence, particularly from randomized controlled trials (RCTs), to determine the efficacy or safety of ICSI-AOA as a treatment strategy. Registration number is PROSPERO, CRD42024551221.

A global synthesis of nitrous oxide emissions across cotton-planted soils

Globally, acquiring information on region- and crop-specific nitrous oxide (N2O) emissions is vital for establishing effective N2O mitigation strategies. Soil cultivated with cotton (Gossypium hirsutum L.) is an important source of N2O in agricultural production. However, little is known about the magnitudes and main drivers of soil N2O emissions from cotton fields worldwide. In this meta-analysis, we were the first to synthesize 34 peer-reviewed papers (298 observational datasets) to quantify the magnitudes and controlling factors of area-scaled N2O emissions (N2Oarea), direct N2O emission factors (EFd), and yield-scaled N2O emissions (N2Oyield) from the soils of cotton fields and to explore associated potential mitigation strategies. On average, the N2Oarea from global cotton-planted soils was 2.10 kg N ha−1, with a mean EFd of 0.92 %, which is comparable to those reported for cereal crops (e.g., maize, 1.02 %) and the Intergovernmental Panel on Climate Change default value of 1 % for global croplands. The global mean N2Oyield estimated here was 622 g N Mg−1. At the global scale, the variations in all N2O-related indices in the soils of cotton fields were demonstrated to be primarily controlled by climatic conditions (e.g. climate type) and soil properties (e.g., bulk density, pH, C/N or soil texture) rather than by well-recognized management practices (e.g., N fertilization rate). Furthermore, our analysis showed that the application of urease and/or nitrification inhibitors significantly reduced soil N2O emissions while maintaining seed cotton yields. These findings emphasize that cotton production has an obvious climate footprint and provide potential N2O mitigation options for the sustainable intensification of cotton production.

Blastocysts originated from oocytes with smooth endoplasmic reticulum aggregates have a reduced euploidy rate: a retrospective cohort study.

Does the presence of smooth endoplasmic reticulum aggregates (SERa) in oocytes adversely impact the euploidy rate of subsequent blastocysts? We performed a retrospective cohort study with 671 young patients (< 38 years) undergoing their first preimplantation genetic testing for aneuploidy (PGT-A) between January 2019 and October 2022 at a reproductive medical center of university affiliated teaching hospitals in China. Cycles were categorized as either SERa(+) cycles (containing at least one SERa(+) oocyte) or SERa(-) cycles (all oocytes without SERa). In SERa(+) cycles, oocytes were further subdivided into the SERa(+) oocyte group and the sibling SERa(-) oocyte group, comprising oocytes with normal morphology. No significant differences were observed in the normal fertilization rate (72.9% vs. 75.4% vs. 72.6%, P=0.343), and cleavage rate (96.8% vs. 97.1% vs. 96.4%, P=0.839) among the SERa(-) cycle group, the SERa(-) oocyte group, and the SERa(+) oocyte group. Additionally, there were no statistically significant differences in the rates of good quality embryos (44.7% vs. 48.8% vs. 46.2%, P=0.177) or blastocyst formation (60.1% vs. 60.9% vs. 60.5%, P=0.893) among the groups. However, the euploidy rate of blastocysts derived from SERa(+) oocytes was significantly lower compared to those from SERa(-) oocytes in SERa(+) cycles and normal oocytes in SERa(-) cycles (39.3% vs. 51.2% vs. 54.5%, P=0.005). Despite this, there were no significant differences in pregnancy and neonatal outcomes after euploid embryo transfer among the three groups. Blastocysts derived from SERa(+) oocytes have a lower euploidy rate than those derived from SERa(-) oocytes. Nevertheless, comparable reproductive outcomes were achieved following euploid embryo transfer from both SERa(+) and SERa(-) oocytes.

Genome-wide re-sequencing reveals regulatory genes and variants involved in the regulation of intermittent fertilization intensity in Wenchang chickens.

Intermittent fertilization intensity (IFI) is closely related to higher fertilization in chickens, but the genetic basis of IFI is not clearly understood. Here, we sampled a total of 939 Wenchang chickens with IFI. The IFI traits had negative correlation with the fertilization rate and exhibited huge phenotypic variations among individuals of the same strain. Based on SNPs derived from a subset of 499 whole genome data, a genome-wide association study with mixed linear model and further linkage disequilibrium analysis were performed to test potential associations between IFI traits and genomic variants. We identified 35 SNP variants and a 19.82 kb linkage disequilibrium block on chr8 significantly associated with IFI. This block is in the intron of LOC101750715, which shows significant homology with the human LMO4. Therefore, LOC101750715 and LMO4 may regulate IFI. The oviduct's immune regulation is crucial for fertilization. LMO4, activated by IL-6 and IL-23, promotes inflammation in epithelial cells. Thus, LOC101750715 and LMO4 may affect fertilization by regulating oviductal inflammation, impacting IFI. Our findings will provide targets for molecular-marker selection and genetic manipulation for lines of chickens with lower IFI.

Effect of Starvation and Refeeding on Gamete Quality and Fertilization in Rainbow Trout (Oncorhynchus mykiss) Broodstock

In this study, the impact of starvation and refeeding on broodstocks was observed by examining quality parameters of gametes and fertilization. While the control group fish were fed every day, the male and female members of the group were fed one week apart and placed in starvation after a week of feeding. Weight gain in the control group was the highest. The feed conversion ratio was normal in all groups (0.9-1.3%). Relative fecundity was (935±62 eggs/kg) in control group females; it was found to be (1317±241 eggs/kg) in starving females and statistically different in the groups (p &amp;lt; .05). The egg diameter of the group receiving intermittent feeding for a week (3.36±0.2 mm) was found to be the lowest. All spermatological parameters were similar between groups, except for sperm volume. The most sperm count was seen in the group that received one-week intermittent feeding (46.9± 20 ml). The results of fertilization with the control female in the fertilization study based on male individuals showed similarity for the male individuals of the trial group, the highest fertilization rate was seen in fertilization using the control female and control male.

High rate of detected variants in male PLCZ1 and ACTL7A genes causing failed fertilization after ICSI.

What is the frequency of PLCZ1, ACTL7A, and ACTL9 variants in male patients showing fertilization failure after ICSI, and how effective is assisted oocyte activation (AOA) for them? Male patients with fertilization failure after ICSI manifest variants in PLCZ1 (29.09%), ACTL7A (14.81%), and ACTL9 (3.70%), which can be efficiently overcome by AOA treatment with ionomycin. Genetic variants in PLCZ1, and more recently, in ACTL7A, and ACTL9 male genes, have been associated with total fertilization failure or low fertilization after ICSI. A larger patient cohort is required to understand the frequency at which these variants occur, and to assess their effect on the calcium ion (Ca2+) release during oocyte activation. AOA, using ionomycin, can restore fertilization and pregnancy rates in patients with PLCZ1 variants, but it remains unknown how efficient this is for patients with ACTL7A and ACTL9 variants. This prospective study involved two patient cohorts. In the first setting, group 1 (N = 28, 2006-2020) underwent only PLCZ1 genetic screening, while group 2 (N = 27, 2020-2023) underwent PLCZ1, ACTL7A, and ACTL9 genetic screening. Patients were only recruited when they had a mean fertilization rate of ≤33.33% in at least one ICSI cycle with at least four MII oocytes. Patients underwent a mouse oocyte activation test (MOAT) and at least one ICSI-AOA cycle using calcium chloride (CaCl2) injection and double ionomycin exposure at our centre. All patients donated a saliva sample for genetic screening and a sperm sample for further diagnostic tests, including Ca2+ imaging. Genetic screening was performed via targeted next-generation sequencing. Identified variants were classified by applying the revised ACMG guidelines into a Bayesian framework and were confirmed by bidirectional Sanger sequencing. If variants of uncertain significance or likely pathogenic or pathogenic variants were found, patients underwent additional determination of the sperm Ca2+-releasing pattern in mouse (MOCA) and in IVM human (HOCA) oocytes. Additionally, ACTL7A immunofluorescence and acrosome ultrastructure analyses by transmission electron microscopy (TEM) were performed for patients with ACTL7A and/or ACTL9 variants. Overall, the frequency rate of PLCZ1 variants was 29.09%. Moreover, 14.81% of patients carried ACTL7A variants and 3.70% carried ACTL9 variants. Seven different PLCZ1 variants were identified (p.Ile74Thr, p.Gln94*, p.Arg141His, p.His233Leu, p.Lys322*, p.Ile379Thr, and p.Ser500Leu), five of which are novel. Interestingly, PLCZ1 variants p.Ser500Leu and p.His233Leu occurred in 14.55% and 9.09% of cases. Five different variants were found in ACTL7A (p.Tyr183His, p.Gly214Ser, p.Val340Met, p.Ser364Glnfs*9, p.Arg373Cys), four of them being identified for the first time. A novel variant in ACTL9 (p.Arg271Pro) was also described. Notably, both heterozygous and homozygous variants were identified.The MOCA and HOCA tests revealed abnormal or absent Ca2+ release during fertilization in all except one patient, including patients with PLCZ1 heterozygous variants. TEM analysis revealed abnormal acrosome ultrastructure in three patients with ACTL7A variants, but only patients with homozygous ACTL7A variants showed reduced fluorescence intensity in comparison to the control.AOA treatment significantly increased the fertilization rate in the 19 patients with detected variants (from 11.24% after conventional ICSI to 61.80% after ICSI-AOA), as well as positive hCG rate (from 10.64% to 60.00%) and live birth rate (from 6.38% to 37.14%), resulting in 13 healthy newborns. In particular, four live births and two ongoing pregnancies were produced using sperm from patients with ACTL7A variants. Genetic screening included exonic and outflanking intronic regions, which implies that deep intronic variants were missed. In addition, other male genes or possible female-related factors affecting the fertilization process remain to be investigated. Genetic screening of PLCZ1, ACTL7A, and ACTL9 offers a fast, cost-efficient, and easily implementable diagnostic test for total fertilization failure or low fertilization after ICSI, eliminating the need for complex diagnostic tests like MOAT or Ca2+ analysis. Nonetheless, HOCA remains the most sensitive functional test to reveal causality of uncertain significance variants. Interestingly, heterozygous PLCZ1 variants are sufficient to cause inadequate Ca2+ release during ICSI. Most importantly, AOA treatment using CaCl2 injection followed by double ionomycin exposure is highly effective for this patient group, including those with ACTL7A variants, who also display a Ca2+-release deficiency. This study was supported by the Flemish Fund for Scientific Research (FWO) (TBM-project grant T002223N awarded to B.H.) and by the Special Research Fund (BOF) (starting grant BOF.STG.2021.0042.01 awarded to B.H.). A.C.B., R.R.G., C.C., E.V.D.V., A.R., D.S., L.L., P.C., S.S., A.B., and F.V.M. have nothing to disclose. B.H. reports a research grant from FWO and BOF, and reports being a board member of the Belgian Ethical Committee on embryo research. N/A.

Ascorbic acid 2-glucoside improves survival, quality, and fertility of frozen-thawed C57Bl/6J and C57Bl/6N mouse spermatozoa.

Ascorbic acid 2-glucoside (AA2G) is a stabilized form of ascorbic acid and a potent antioxidant. Ascorbic acid is present in the testes and epididymis and helps maintain the physiological integrity of reproductive organs. Its properties have been utilized to protect spermatozoa of different species from oxidative stress. Spermatozoa of C57Bl/6J and C57Bl/6N strains were frozen and analyzed, after thawing, by supplementing the capacitation medium with AA2G, both in the presence and absence of methyl-β-cyclodextrin (MBCD). The effect of treatment was evaluated by SCA System (Microptic) analyzing the velocity, vitality, morphology, and the DNA fragmentation. We also examined sperm capacitation (CTC), acrosome reaction (Coomassie Brillant Blue), and fertility (in vitro fertilization) of treated spermatozoa. AA2G improved sperm quality and fertility particularly in association with MBCD. We observed a significant increase of sperm motility, velocity, and vitality associated with an enhanced capacitation and acrosome reaction. These improvements resulted in a marked increase in in vitro fertilization success. Embryos obtained were cultured and reached normally the blastocyst stage. This study aimed to determine if AA2G could safeguard mouse spermatozoa during cryopreservation. We found a protective effect of AA2G that increased sperm survivability resulting in higher fertilization rate. This newly improved protocol shows potential for reanimating cryopreserved GEMMs stored in mouse biobanks and international repositories, such as the European Mouse Mutant Archive (EMMA). This can serve as a pivotal tool in fulfilling the 3Rs mission (replacement, reduction, and refinement), promoting ethical and humane research practices.