Fertilization Rate Research Articles

The association of sperm BAX and BCL-2 gene expression with reproductive outcome in Oligoasthenoteratozoospermia cases undergoing intracytoplasmic sperm injection: A case-control study.

The B-cell lymphoma 2 (BCL-2) protein is one of the members of the BCL-2 associated X (BAX) protein family that acts as an inducer of apoptosis. The present study aims to investigate the association between BAX and BCL-2 gene expression with reproductive outcome, in cases undergoing intracytoplasmic sperm injection. In this case-control study, 50 men were divided into healthy fertile and oligoasthenoteratozoospermic infertile men (n = 25/each). They were subjected to history taking, clinical examination, and semen analysis. Expression of BAX and BCL-2 genes were measured using real-time polymerase chain reaction. The DNA fragmentation index was measured using the sperm chromatin dispersion assay technique. Using World Health Organization criteria, sperm parameters were evaluated. Evaluation of apoptosis-related genes showed that oligoasthenoteratozoospermic significantly increased mRNA expression of BAX, and significantly decreased mRNA expression of BCL-2, when compared with control. Moreover, the BAX/BCL-2 ratio was significantly higher in oligoasthenoteratozoospermic compared to the normozoospermic group (p = 0.01). Also, this study showed that the BAX and BCL-2 genes expression had a significant correlation with sperm quality, and DNA fragmentation in the oligoasthenoteratozoospermic group (p = 0.01). The oligoasthenoteratozoospermic men, had a considerably lower proportion of fertilization rate and good-quality embryos at the cleavage stage than the normozoospermic subjects (p = 0.01). A significant correlation was observed between the expression of BAX and BCL-2 genes, fertilization, and embryo quality (p = 0.01). We concluded that the sperm BAX/BCL-2 ratio demonstrates a significant correlation with fertilization rate and embryo quality.

Methionine Improves Boar Sperm Quality by Promoting Mitochondrial Translation during Liquid Storage.

Boar sperm quality serves as an important indicator of reproductive efficiency, playing a direct role in enhancing the output of livestock production. It has been demonstrated that mitochondrial protein translation is present in sperm and plays a crucial role in regulating sperm motility, capacitation and in vitro fertilization rate. The present study aimed to determine whether methionine supplementation enhances mitochondrial translation in boar sperm, thereby improving sperm quality. The results showed a significant elevation in the abundance of mitochondrial methionyl-tRNA formyltransferase (MTFMT), a crucial enzyme for mitochondrial protein translation, and mitochondrial DNA-encoded cytochrome c oxidase subunit 1 (COX1) in boar sperm exhibiting high motility. Both amino acids and methionine supplementation significantly enhanced boar sperm motility during storage. Moreover, methionine supplementation mitigates the loss of acrosomal integrity, enhances the expression of COX1, and boosts mitochondrial activity. Furthermore, the positive impact of methionine was negated in the presence of the mitochondrial translation inhibitor chloramphenicol. Together, these findings suggest that boar sperm may utilize methionine as a protein translation substrate to enhance sperm motility by stimulating mitochondrial protein translation. The supplementation of methionine may enhance the quality of boar sperm, thereby providing guidance for the optimization of diluent formulations for liquid storage and the identification of physiological regulators that regulate sperm motility.

CARP HATCHERY MANAGEMENT (Cyprinus carpio L.) NATURALLY AT THE PUNTEN AQUACULTURE INSTALLATION (IPB), BATU CITY, EAST JAVA

The large market demand for consumption fish makes some farming communities interested in farming, especially consumption fish farming. Fish hatchery business is the first step to the success of fish farming, because for each growing season the aquaculture business requires a supply of seeds from the hatchery business. Goldfish hatcheries are relatively easy because carp can spawn naturally or artificially. Goldfish have a relatively high fecundity or number of eggs, which ranges from 84,000-110,000 eggs per kilogram of brood. The implementation of carp hatchery techniques includes several processes, namely pond preparation, brood selection, kakaban installation, brood release, spawning, hatching eggs, larval care, checking water quality, seeding, pest and disease control. calculation of fecundity, fertilization rate (FR), hatching rate (HR), and survival rate (SR). The data collection method used in this street vendor is primary data. Primary data include active participation, observation, interviews, and documentation. The carp hatchery technique (Cyprinus carpio L.) at IPB Punten, Batu City is carried out naturally, the carp mother used comes from Punten using a ratio of male to female parents of 1: 3. The feeding method, namely Blind Feeding, obtained an F value of 40,000 eggs, FR 84%, HR 84.5% and SR 90.2%.

Straw incorporation and nitrogen fertilization regulate soil quality, enzyme activities and maize crop productivity in dual maize cropping system

BackgroundStraw incorporation serves as an effective strategy to enhance soil fertility and soil microbial biomass carbon (SMBC), which in turn improves maize yield and agricultural sustainability. However, our understanding of nitrogen (N) fertilization and straw incorporation into soil microenvironment is still evolving. This study explored the impact of six N fertilization rates (N0, N100, N150, N200, N250, and N300) with and without straw incorporation on soil fertility, SMBC, enzyme activities, and maize yield.ResultsResults showed that both straw management and N fertilization significantly affected soil organic carbon (SOC), total N, SMBC, soil enzyme activities, and maize yield. Specifically, the N250 treatment combined with straw incorporation significantly increased SOC, total N, and SMBC compared to lower fertilization rates. Additionally, enzyme activities such as urease, cellulase, sucrose, catalase, and acid phosphatase reached their peak during the V6 growth stage in the N200 treatment under for both straw management conditions. Compared to N250 and N300 treatments of traditional planting, the N200 treatment with residue incorporation significantly increased yield by 8.30 and 4.22%, respectively. All measured parameters, except for cellulase activity, were significantly higher in spring than in the autumn across both study years, with notable increases observed in 2021.ConclusionsThese findings suggest that optimal levels of SOC, soil total N (STN), and SMBC, along with increased soil enzyme activities, is crucial for sustaining soil fertility and enhancing maize grain yield under straw incorporation and N200 treatments.

Influence of Nitrogen, Potassium and Phosphorus Fertilizers with Foliar Application of Micronutrient of Iron, Zinc and Manganese on Bulb Characteristics of Single Flower Tuberose Plants (Polianthes Tuberosa, L.)

The global market for flowers and ornamental plants is expected to expand at a rate of approximately 6.3% over the next five years, reaching an estimated value of $57.4 billion USD by 2024. Tuberose (Polianthes tuberosa L.), a summer flower cultivated by small-scale growers primarily for export. The study was aimed to cultivate tuberose plants in Al-Jabal Al-Khader region, introducing them as a new cut flower crop, as a source for tuberose, and leveraging their economic potential. A field experiment was conducted over two summer growing seasons to investigate the impact of different levels of compound fertilizer (nitrogen, phosphorus and potassium, NPK 4:4:4) in application rates (0, 500, 1000, and 1500 kg.ha-1) and foliar sprays of micronutrients iron (Fe), zinc (Zn), manganese (Mn), and a )mixture of these three elements) on tuberose bulbs. The experimental design was a completely randomized block design split-plot with three replications. The NPK fertilizer levels were randomly assigned to the main plots, while the micronutrient treatments were allocated to the subplots. The study found that varying rates of NPK fertilization did not significantly affect the number of bulbs per plant over the two growing seasons. In contrast, the application of micronutrient foliar sprays had a notable impact, with the mixture of Fe, Zn, and Mn yielding the highest significant increase in bulb number per plant in both seasons. It is recommended to apply a mixture of the three micronutrients (Fe, Zn, and Mn) as fertilizer to produce high-quality Tuberose plants for various decorative purposes in landscaping.

Impact of Luteinizing Hormone on IVF/ICSI Assisted Reproduction on the Initiation Day of Gonadotropin-releasing Hormone Antagonist Protocol.

The aim of the study was to explore the optimal timing of gonadotropin initiation and the reasonable interval of luteinizing hormone (LH) levels in the gonadotropin-releasing hormone antagonist (GnRH-A) protocol. A retrospective cohort study was conducted to analyze the data concerning the oocyte retrieval cycles from 1,361 cases with the GnRH-A protocol implemented. The ovarian responses (including AMH, AFC) in these patients were divided into the poor ovarian response group (an antral follicle count [AFC] ≤ 6, n = 394), the normal ovarian response group (an AFC > 6 and < 15, n = 570), and the high ovarian response group (an AFC ≥ 15, n = 397), according to the AFC. The patients were sub-grouped according to LH levels on the protocol initiation day, and the clinical outcomes (including dose of Gn initiation, Gn administration days, GnRH-ant administration days, P levels on the HCG day, E2 levels on the HCG day, LH levels on the HCG day, number of embryos transferred, total fertilization rate, embryo implantation rate(%), proportion of 2PN, proportion of good-quality embryos, endometrial thickness on the hCG injection day(mm), moderate to severe OHSS, AFC on the initiation day, proportion of type A endometrium on the hCG injection day, clinical pregnancy rate, biochemical pregnancy rate, early abortion rate, ectopic pregnancy rate) were compared. On the GnRH-A protocol initiation day, among all patients with different ovarian responses, the body mass index (BMI) in those with an LH ≥ 5 IU/L was lower. The differences in pregnancy outcomes between the LH < 5 IU/L group and the LH ≥ 5 IU/L group were not statistically significant across the different ovarian response groups, but the LH < 5 IU/L group had a higher proportion of good-quality embryos (80.3±24.9 vs. 74.8±26.9, P =0.035) than the LH≥5IU/Lgroup in those with poor ovarian response. The total fertilization rate (82.2±18.1 vs 85.4±15.1, P =0.021) and proportion of two pronuclei (2PN) (69.0±20.9 vs 72.7±19.9, P =0.035) were higher in the LH ≥ 5 IU/L group than the LH<5 IU/L group for those with normal ovarian responses. The embryo implantation rate (41.4±41.3 vs 52.6±43.4, P =0.012) was higher in the LH ≥ 5 IU/L group than in the LH<5 IU/L group in those with high ovarian response. The results of the multivariate logistic analysis showed that the age of the female partner, number of embryos transferred, proportion of good-quality embryos, endometrial thickness on the hCG injection day, and moderate- to-severe ovarian hyperstimulation syndrome (OHSS) were independent factors correlated with the outcome of live births (P < 0.05). The LH levels on the gonadotropins (Gn) initiation day in the GnRH-A protocol will not affect pregnancy outcomes.

Prediction of reproductive performance in wintering females of Siberian sturgeon (Acipenser baerii, Brandt) based on in vitro ovulation rate of isolated follicles

Early prediction of the reproductive success of sturgeon females is important both for artificial breeding conditions in sturgeon farms and for monitoring wild fish populations approaching spawning. This study demonstrated that ovarian follicles removed from wintering Acipenser baerii females can undergo in vitro ovulation when exposed to 17α,20β-dihydroxy-4-pregnen-3-one. The research was conducted from 2015 to 2021 and involved the examination of 110 wintering Siberian sturgeon females (A. baerii), with an average weight of 8–11 kg and an age range of 8–10 years. Selection of females for spawning was based on the polarization index (PI) at the time when the river temperature dropped to +8 °C. The selected females were overwintered in concrete flow-through tanks with water from the Volga River for approximately two months. At the end of this period, oocyte samples were collected for in vitro incubation. An increase in water temperature to 15 °C, which precedes spawning, was achieved by mixing with warm effluent from a regional power station. The reproductive success of females was evaluated based on fertilization and embryonic survival rates. High reproductive success was characterized by females with fertilization rates above 70% and hatching rates above 60%. We identified a positive correlation between the percentage of in vitro ovulated follicles and the in vivo fertilization and embryo survival rates, with correlation coefficients of Rsp = 0.60 and Rsp = 0.48, respectively. Statistical analysis showed that females whose in vitro ovulation rate of follicles exceeded 30% displayed high reproductive performance compared to those with an in vitro ovulation rate below 30%. The predictive accuracy of this criterion was determined by calculating the ratio of ranks between females with high and low reproductive performance, averaging 80.9% for fertilization and 77% for embryonic survival. Therefore, we propose a new criterion to select females of A. baerii for spawning based on the in vitro oocyte ovulation rate obtained from wintering individuals.

Integrative phenotyping analyses reveal the relevance of the phyB-PIF4 pathway in Arabidopsis thaliana reproductive organs at high ambient temperature

BackgroundThe increasing ambient temperature significantly impacts plant growth, development, and reproduction. Uncovering the temperature-regulating mechanisms in plants is of high importance, for increasing our fundamental understanding of plant thermomorphogenesis, for its potential in applied science, and for aiding plant breeders in improving plant thermoresilience. Thermomorphogenesis, the developmental response to warm temperatures, has been primarily studied in seedlings and in the regulation of flowering time. PHYTOCHROME B and PHYTOCHROME-INTERACTING FACTORs (PIFs), particularly PIF4, are key components of this response. However, the thermoresponse of other adult vegetative tissues and reproductive structures has not been systematically evaluated, especially concerning the involvement of phyB and PIFs.ResultsWe screened the temperature responses of the wild type and several phyB-PIF4 pathway Arabidopsis mutant lines in combined and integrative phenotyping platforms for root growth in soil, shoot, inflorescence, and seed. Our findings demonstrate that phyB-PIF4 is generally involved in the relay of temperature signals throughout plant development, including the reproductive stage. Furthermore, we identified correlative responses to high ambient temperature between shoot and root tissues. This integrative and automated phenotyping was complemented by monitoring the changes in transcript levels in reproductive organs. Transcriptomic profiling of the pistils from plants grown under high ambient temperature identified key elements that may provide insight into the molecular mechanisms behind temperature-induced reduced fertilization rate. These include a downregulation of auxin metabolism, upregulation of genes involved auxin signalling, miRNA156 and miRNA160 pathways, and pollen tube attractants.ConclusionsOur findings demonstrate that phyB-PIF4 involvement in the interpretation of temperature signals is pervasive throughout plant development, including processes directly linked to reproduction.

Intracytoplasmic sperm injection versus conventional in vitro insemination in couples with non-male infertility factor in the ‘real-world’ setting: analysis of the HFEA registry

BackgroundIn assisted reproductive technology (ART), the choice between intracytoplasmic sperm injection (ICSI) and conventional in vitro insemination (IVF) remains a pivotal decision for couples with female or unexplained infertility. The hypothesis that ICSI may not confer significant improvements in live birth rates in the absence of a male infertility factor was explored in this study.MethodsThis was a retrospective collection of data recorded by the Human Fertilisation and Embryology Authority (HFEA) in the UK from 2005 to 2018 and analysed through regression analysis models on both the entire dataset and a matched-pair subset. First fresh ART cycles were analysed according to the insemination technique in order to compare live birth as the main outcome. Cycles were included if complete information regarding infertility cause, female age, number of oocytes retrieved, allocation to ICSI or IVF, and treatment outcome in terms of live birth was available. Matching was performed at a 1:1 ratio between IVF and ICSI cycles according to the cause of infertility, female age, number of oocytes, and year of treatment.ResultsThis study, based on 275,825 first cycles, revealed that, compared with IVF, ICSI was associated with higher fertilization rates and lower cycle cancellations rates. However, ICSI was associated with a lower chance of implantation and live birth than IVF in cycles with female-only infertility: in the entire dataset, the adjusted odds of having a live birth decreased by a factor of 0.95 (95% CI 0.91–0.99, p = 0.011), while in the matched-pair analyses it decreased by a factor of 0.91 (95% CI 0.86–0.96, p = 0.003) using ICSI compared to IVF. For unexplained infertility cycles, the adjusted odds ratios for live birth in ICSI compared to IVF cycles were 0.98 (95% CI 0.95–1.01) in the entire dataset and 0.97 (95% CI 0.93–1.01) in the matched-pair analysis.ConclusionsCompared with IVF, ICSI was associated with a reduction in live births when ART was indicated due to female-only factors. Additionally, no significant improvements were associated with the use of ICSI in cycles with unexplained infertility. Our findings impose a critical reevaluation regarding the use of ICSI over IVF for cases with female-only factors and unexplained infertility.

The impact of implementing a non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) embryo culture protocol on embryo viability and clinical outcomes.

Are modifications in the embryo culture protocol needed to perform non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) affecting clinical reproductive outcomes, including blastocyst development and pregnancy outcomes? The implementation of an embryo culture protocol to accommodate niPGT-A has no impact on blastocyst viability or pregnancy outcomes. The recent identification of embryo cell-free (cf) DNA in spent blastocyst media has created the possibility of simplifying PGT-A. Concerns, however, have arisen at two levels. First, the representativeness of that cfDNA to the real ploidy status of the embryo. Second, the logistical changes that need to be implemented by the IVF laboratory when performing niPGT-A and their effect on reproductive outcomes. Concordance rates of niPGT-A to invasive PGT-A have gradually improved; however, the impact of culture protocol changes is not as well understood. As part of a trial examining concordance rates of niPGT-A versus invasive PGT-A, the IVF clinics implemented a specific niPGT-A embryo culture protocol. Briefly, this involved initial culture of fertilized oocytes following each laboratory standard routine up to Day 4. On Day 4, embryos were washed and cultured individually in 10 μl of fresh media. On Day 6 or 7, blastocysts were then biopsied, vitrified, and media collected for the niPGT-A analysis. Six IVF clinics from the previously mentioned trial were enrolled in this analysis. In the concordance trial, Clinic A cultured all embryos (97 cycles and 355 embryos) up to Day 6 or 7, whereas in the remaining clinics (B-F) (379 cycles), nearly a quarter of all the blastocysts (231/985: 23.5%) were biopsied on Day 5, with the remaining blastocysts following the niPGT-A protocol (754/985: 76.5%). During the same period (April 2018-December 2020), the IVF clinics also performed standard invasive PGT-A, which involved culture of embryos up to Days 5, 6, or 7 when blastocysts were biopsied and vitrified. In total, 428 (476 cycles) patients were in the niPGT-A study group. Embryos from 1392 patients underwent the standard PGT-A culture protocol and formed the control group. Clinical information was obtained and analyzed from all the patients. Statistical comparisons were performed between the study and the control groups according to the day of biopsy. The mean age, number of oocytes, fertilization rates, and number of blastocysts biopsied were not significantly different for the study and the control group. Regarding the overall pregnancy outcomes, no significant effect was observed on clinical pregnancy rate, miscarriage rate, or ongoing pregnancy rate (≥12 weeks) in the study group compared to the control group when stratified by day of biopsy. The limitations are intrinsic to the retrospective nature of the study, and to the fact that the study was conducted in invasive PGT-A patients and not specifically using niPGT-A cases. This study shows that modifying current IVF laboratory protocols to adopt niPGT-A has no impact on the number of blastocysts available for transfer and overall clinical outcomes of transferred embryos. Whether removal of the invasive biopsy step leads to further improvements in pregnancy rates awaits further studies. This study was funded by Igenomix. C.R., L.N.-S., and D.V. are employees of Igenomix. D.S. was on the Scientific Advisory Board of Igenomix during the study. ClinicalTrials.gov (NCT03520933).

Assessing the developmental competence of oocytes matured following rescue in vitro maturation: a systematic review and meta-analysis.

To assess the developmental competence of oocytes matured following rescue in vitro maturation (IVM). PubMed, EmBASE, and SCOPUS were systematically searched for peer-reviewed original papers using relevant keywords and Medical Subject Heading terms. Study quality was assessed using the Newcastle-Ottawa Scale. Odds ratios with a 95% confidence interval were calculated by applying a random effects model. The primary outcomes were fertilization and blastulation rates. Secondary outcomes included abnormal fertilization, cleavage, euploidy, clinical pregnancy, and live-birth rates. Twenty-four studies were included in the meta-analysis. The oocytes matured following rescue IVM showed significantly reduced fertilization, cleavage, blastulation, and clinical pregnancy rates compared to sibling in vivo-matured oocytes. No significant differences were found for the euploidy and live-birth rates in euploid blastocyst transfer. In poor responders, a reduced fertilization rate was observed using in vitro-matured GV but not with in vitro-matured MI. A reduced cleavage rate in MI matured overnight compared to < 6 incubation hours was found. Our results showed compromised developmental competence in oocytes matured following rescue IVM. However, in poor responders, rescue IVM could maximize the efficiency of the treatment. Notably, our data suggests using in vitro MI matured within 6 incubation hours. CRD42023467232.

ЭФФЕКТИВНОСТЬ ПРИМЕНЕНИЯ НОВОКАИНОВОЙ БЛОКАДЫ ПО И.П. ЛИПОВЦЕВУ ДЛЯ ЛЕЧЕНИЯ КОРОВ ПРИ СУБКЛИНИЧЕСКОМ ЭНДОМЕТРИТЕ

The aim of the study is to investigate the effectiveness of using the middle uterine artery puncture according to I.P. Lipovtsev with the introduction of 0.5 % novocaine solution in combination with the antibio¬tic-containing drug Ceftonit Forte in cows with subclinical endometritis. Objectives: to study the therapeutic effectiveness of the introduction of 0.5 % novocaine solution in combination with the antibiotic-containing drug Ceftonit Forte into the middle uterine artery at different times of the estrous cycle; to establish the fertilization rate of cows after the combined use of novocaine blockade according to I.P. Lipovtsev and the drug Ceftonit Forte at different times of the estrous cycle. The object of the study were cows of the red-motley breed with a subclinical form of endometritis. The diagnosis of subclinical endometritis was made according to the method of N.A. Flegmatov. Laboratory studies have established the presence of subclinical endometritis in 29.24–38.21 % of infertile cows. In 16.98–21.69 % of cases, the cervicovaginal mucus did not have visible threadlike or point inclusions of purulent exudate, but the reaction of such mucus with sperm was positive. Therefore, the absence of purulent exudate inclusions in the cervicovaginal mucus cannot indicate the absence of subclinical endometritis in cows. It has been established that a single use of novocaine blockade according to I.P. Lipovtsev in combination with the antibiotic-containing drug Ceftonit Forte of domestic production, regardless of the timing of administration during the estrous cycle, ensures recovery of 95.24–100 % of cows, and the fertilization rate after artificial insemination of females is 76.19–80.95 %. In addition, there is no need to wait for the onset of estrus in cows with subclinical endometritis to administer drugs into the cervical canal, as is generally accepted in veterinary practice, which allows for more rapid treatment of animals with this uterine pathology and significantly reduces the number of days of infertility.

Chromosomal missegregation and aberrant embryo development in repro57 female mice with Rnf212 homozygous mutation.

Repro57 mice, bearing an Rnf212 gene mutation, exhibit infertility in both homozygous mutant males and females, revealing arrested spermatogenesis in males and investigating unclear mechanisms in females. The study highlights aneuploidy and altered kinetochore patterns in repro57 homozygous mutant oocytes, which impact later stages of embryo development. Repro57 mice, induced with N-ethyl-N-nitrosourea and harboring a mutation in the Rnf212 gene, exhibit infertility in both homozygous mutant males and females. Rnf212 plays a crucial role in recombination and crossover designation. In male repro57 homozygous mutants, spermatocytes often degenerate during late prophase, and mature spermatozoa are absent in the seminiferous epithelium, indicating arrested spermatogenesis as the cause of infertility. Despite reports of infertility in Rnf212-knockout female mice, the specific mechanisms underlying infertility in female repro57 homozygous mutants remain elusive. This study investigates the chromosomal and kinetochore patterns of mature oocytes and their developmental potential following in vitro fertilization in female repro57 homozygous mutant mice. While all wild-type oocytes progress to metaphase II and exhibit euploidy, all repro57 homozygous mutant mouse oocytes display aneuploidy. Additionally, kinetochore distances in repro57 homozygous mutant oocytes exceed those observed in wild-type counterparts. Although no significant differences are noted in fertilization and early embryo development rates between wild-type and repro57 homozygous mutant mice, embryos derived from repro57 homozygous mutants exhibit significantly lower morula and blastocyst rates, accompanied by frequent cytokinesis failure and vacuole formation. These findings suggest that the premature segregation of sister chromatids in repro57 homozygous mutant mice adversely impacts the later stages of embryo development.

Enriching NPK Mineral Fertilizer with Plant-Stimulating Peptides Increases Soilless Tomato Production, Grower Profit, and Environmental Sustainability.

The need to increase agricultural production to feed a steadily growing population may clash with the more environmentally friendly but less efficient production methods required. Therefore, it is important to try to reduce the use of chemical inputs without compromising production. In this scenario, natural biostimulants have become one of the most sought-after and researched technologies. In the present study, the results of a greenhouse experiment on hydroponic tomatoes (Solanum lycopersicum L.) are presented, which involved comparing the use of ordinary NPK fertilizer (Cerbero®) with the use of NPK fertilizers enriched with 0.5% protein hydrolysate of plant origin (Cerbero Green®) at both standard (100%) and reduced (70%) fertilization rates. The results highlight how the use of Cerbero Green® fertilizers improves the production performance of tomatoes. More specifically, they show that the use of Cerbero Green® leads to higher marketable yields, especially under reducing fertilizer use, ensuring a positive net change in profit for the grower. In addition, carbon footprint analysis has revealed that the use of Cerbero Green® reduces the environmental impact of hydroponic tomato growing practices by up to 8%. The observed higher yield of hydroponically grown tomatoes even with reduced fertilization rates underlines once again the key role of natural biostimulants in increasing both the economic and environmental sustainability of horticultural production.

Insecticides Cypermethrin and Chlorpyrifos Exposure: Effects on Sperm Motility and Fertility in Cyprinus carpio (Cypriniformes, Cyprinidae)

The agricultural production of the Argentine Pampa is based on intensive cultures, and the most widely used insecticides correspond to cypermethrin (pyrethroid) and chlorpyrifos (organophosphate), among others. Sperm motility and fertility assessments are essential topics for biology and population dynamics studies on fish species. In this study the alterations of these endpoints after exposure to commercial formulation of both insecticides were used to analyze the effects on common carp sperm. After the in vitro exposure at nominal concentrations of 0.25, 0.5, 1, and 2 μg/L of cypermethrin, a significant reduction in motility in 0.5 μg/L concentrations was observed respect control group. The same nominal concentrations were tested for chlorpyrifos, and the same significative effect was detected at 2.0 μg/L. Fertilization rate was significantly reduced at concentrations from 0.5 μg/L cypermethrin exposure, and from 1.0 μg/L in the case of chlorpyrifos. Eggs LC50 / 24 h (95% confidence limit) exposed to cypermethrin was 0.57 μg/L (0.36 - 0.88), and exposed to chlorpyrifos was 2.45 μg/L (1.13-5.29). However, is still debatable how these insecticides impact on fish reproduction, using this in vitro scenario we found a clear reduction in sperm motility and fertility.

The Improvement Effects of a Nutritional Fortifier on the Reproductive Performance, Sex Steroid Hormone Production, and Health of the Striped Bamboo Shark Chiloscyllium plagiosum.

To explore a method of improving the reproductive performance of the striped bamboo shark, three groups (D0, D1, and D2) of mature individuals were fed squid with (D1 and D2) or without (D0) a nutritional fortifier during the breeding seasons of 2022 and 2023. Compared with the D0 group, the D1 and D2 groups had an increase of 20.90% and 31.34% in total eggs, increases of 32.73% and 41.82% in the proportion of lecithal eggs, and a total 119.07% increase in hatching rate, respectively, in 2022. In 2023, the corresponding increase was 17.12% and 9.91% in total eggs, 19.63% and 12.15% in the proportion of lecithal eggs, 43.37% and 43.94% in fertilization rate, 23.94% and 22.22% in hatchability rate, and 66.70% and 8.70% in the survival rate of fry. Moreover, the levels of serum estradiol, testosterone, progesterone, albumin, and total antioxidant capacity and the levels of ARA, EPA, DHA, n-3 PUFA, and n-6 PUFA in both serum and lecithal eggs significantly increased, while the levels of serum triglyceride and total cholesterol were the opposite (p < 0.05). The results demonstrate that feeding the sharks with a nutritional fortifier can increase spawn production and the quality of eggs, regulate the production of sex steroids, and improve the nutrition of eggs and the health of broodstocks.

In Vivo-Matured Oocyte Resists Post-Ovulatory Aging through the Hub Genes DDX18 and DNAJC7 in Pigs.

Assisted reproduction technology (ART) procedures are often impacted by post-ovulatory aging (POA), which can lead to reduced fertilization rates and impaired embryo development. This study used RNA sequencing analysis and experimental validation to study the similarities and differences between in vivo- and vitro-matured porcine oocytes before and after POA. Differentially expressed genes (DEGs) between fresh in vivo-matured oocyte (F_vivo) and aged in vivo-matured oocyte (A_vivo) and DEGs between fresh in vitro-matured oocyte (F_vitro) and aged in vitro-matured oocyte (A_vitro) were intersected to explore the co-effects of POA. It was found that "organelles", especially "mitochondria", were significantly enriched Gene Ontology (GO) terms. The expression of genes related to the "electron transport chain" and "cell redox homeostasis" pathways related to mitochondrial function significantly showed low expression patterns in both A_vivo and A_vitro groups. Weighted correlation network analysis was carried out to explore gene expression modules specific to A_vivo. Trait-module association analysis showed that the red modules were most associated with in vivo aging. There are 959 genes in the red module, mainly enriched in "RNA binding", "mRNA metabolic process", etc., as well as in GO terms, and "spliceosome" and "nucleotide excision repair" pathways. DNAJC7, IK, and DDX18 were at the hub of the gene regulatory network. Subsequently, the functions of DDX18 and DNAJC7 were verified by knocking down their expression at the germinal vesicle (GV) and Metaphase II (MII) stages, respectively. Knockdown at the GV stage caused cell cycle disorders and increase the rate of abnormal spindle. Knockdown at the MII stage resulted in the inefficiency of the antioxidant melatonin, increasing the level of intracellular oxidative stress, and in mitochondrial dysfunction. In summary, POA affects the organelle function of oocytes. A_vivo oocytes have some unique gene expression patterns. These genes may be potential anti-aging targets. This study provides a better understanding of the detailed mechanism of POA and potential strategies for improving the success rates of assisted reproductive technologies in pigs and other mammalian species.

Effects of water and fertilizer regulation on soil microbial community, fruit nutrients, and saponin content of Panax notoginseng: A three years field experiment

In pursuit of maximum yield and economic benefits of Panax notoginseng, farmers usually apply excessive fertilizer and irrigation. Which easily induce the decline of Panax notoginseng quality, fertilizer and water use efficiencies and soil health. From 2021–2023, a field experiment was conducted in Dali Village, Luxi County, Honghe Prefecture, Yunnan Province, to study the response of soil microbial community, root nutrient content, and ginsenoside content of Panax notoginseng to different water and fertilizer regulation. The experiment including three irrigation levels (F1: 120 mm, F2: 240 mm, and F3: 360 mm) four fertilization rates(F1: 48 kg ha−1, F2: 72 kg ha−1, F2: 96 kg ha−1, and 120 kg ha−1),and a CK treatment (no fertilizer and no irrigation) The results showed that the average N, P, and K contents across all treatments were greater in 2023 than those in 2021 and 2022. W3F4 treatment significantly increased N absorption. W2F4 treatment significantly increased P and K absorption. W2F4 treatment significantly increased the Panax notoginseng yield, and W3F2 treatment significantly increased the saponin content. The Panax notoginseng yield ranged from 1165 to 3006 kg∙ha−1, with a maximum average yield in 2023. The Re content averaged 81.5 % in 2023 over 2021, and the highest Re content in W3F2 treatment in 2023 was 108.54 mg∙g−1. In 2023, W2F3 treatment had the highest R1 content of 15.34 mg∙g−1, W2F3 treatment had the highest content of Rg1 of 44.65 mg∙g−1, and W3F2 treatment had the highest Rb1 content of 52.52 mg∙g−1. From 2021–2023, the total number of unique OTUs in W1F1 was the largest, out followed by W3F2, W3F4, and W3F1. The treatment that significantly increased the bacterial Chao1 index was W3F1 and W2F2 in 2021, W3F3, W2F1, W2F1 and W2F3 in 2022, and W2F3 and W2F4 in 2023. The Shannon nor Simpson indices were not varied significantly from CK. At the phylum level, W1F3 in 2021 and W3F1 in 2022 and 2023 improved the relative abundance of Proteobacteria, Acidobacteriota At the genus level, W1F1 in 2021 and 2023 improved the relative abundance of Vicinamibacteraceae_norank and Vicinamibacterales_norank. LEfSe analysis of soil microbial communities showed the most divergent species in 2021 with W3F2, W1F1 in 2022, and W2F1 in 2023. Different water and fertilizer treatments affected the total OTU number of bacterial communities and changed the bacterial community structure. W3F1 and W2F3 treatments significantly increased soil bacterial diversity, and soil microbial diversity regulated plants N as well as R1 and Rg1 content, and the quality of Panax notoginseng is related to microbial diversity. N, P, K, and R1 had a significant positive correlation with microbial community diversity (P<0.05). The W1F1 treatment communities were relatively abundant, and the community species showed high similarity in W3F1 and W2F3.

A hyaluronic acid-containing reagent compatible with glass-bottom dishes and capable of sustained binding of human spermatozoa.

Commercial products currently available for sperm selection utilizing hyaluronic acid (HA) binding prior to intracytoplasmic sperm injection (ICSI) are widely used but have some disadvantages. To potentially circumvent these limitations, we compared ICSI using a self-made hyaluronic acid (smHA) reagent with ICSI using SpermSlow. The binding of the reagents to spermatozoa on plastic- or glass-bottom dishes was quantitated using spermatozoa that were isolated by density-gradient centrifugation and swim-up procedures (N=10/group). Additionally, we investigated the relationship between the HA reagent used prior to ICSI and clinical outcomes after assisted reproduction with HA-ICSI (N=81). The smHA reagent exhibited extremely stable binding to human spermatozoa. The binding time of spermatozoa was significantly longer in the smHA reagent than in SpermSlow on both plastic and glass dishes (plastic: 60.0±0.0 min vs. 2.7±5.9 min, P<0.001; glass: 60.0±0.0 min vs. 2.5±1.8 min, P<0.001). There were no significant differences in the normal fertilization rate between HA-ICSI with the smHA reagent (128/160, 80.0%) and HA-ICSI with SpermSlow (171/231, 74.0%, P=0.184). The frequency of the blastocyst development from the HA-ICSI-derived zygote was significantly higher with the smHA reagent (74/101, 73.3%) than with SpermSlow (76/131, 58.0%, P=0.019). The rates of biochemical pregnancy, clinical pregnancy, fetal heart movement, live birth, and miscarriage were not significantly different between HA-ICSI with the smHA reagent and HA-ICSI with SpermSlow. The blastulation rate was higher for HA-ICSI with the smHA reagent as compared with SpermSlow. Clinical outcomes, excluding blastulation, after HA-ICSI were the same using smHA reagent and using SpermSlow. Spermatozoa binding to the smHA reagent was not attenuated over a 60-min time course. In conclusion, this reagent may shorten and simplify HA-ICSI procedures because smHA can be used with any dish material, making it easier to observe the spindle or assess intracytoplasmic morphology.

A simple method for repeated in vivo sperm collection from laboratory mice

PurposeMouse spermatozoa for archiving laboratory mice or for in vitro fertilization (IVF) are routinely obtained from the cauda epididymis of adult males sacrificed for this purpose. To avoid the death of the donor, we tested whether a precisely timed interruption of the mating act could be used for repeated sperm collection from laboratory mice.MethodsSperm donors (B6D2F1) were mated with a receptive female, and mating behavior was observed. The stud was separated from the female 1–2 s after the onset of the ejaculatory shudder. The ejected copulatory plug with the yellowish viscous ejaculate was carefully removed from the penile cup.ResultsA total of 80 ejaculates were successfully obtained from 100 ejaculations. The latency to first mount was 1.1 ± 1.1 min (mean ± SD) and to ejaculation 8.1 ± 4.7 min. The average number of mounts to ejaculation was 10.5 ± 5.8, and the mean number of spermatozoa per collected ejaculate was 1.86 ± 1.05 × 106. An average fertilization rate of 76% was observed after IVF.ConclusionsSeparating the stud from the female just before ejaculation is feasible, easy to learn, and requires no special equipment. The sperm count of collected ejaculates is lower than natural ejaculations, but higher than previous in vivo sperm collection methods achieved. We recommend this simple sperm collection method in mice, especially when the donor cannot be sacrificed and/or repeated sperm collection from the same animal is required for experimental purposes.