Fertilization Program Research Articles

The Consequence of Short Insemination Strategy on Sperm Biological Characteristics, Embryo Morphokinetics, and Clinical Outcomes in the IVF Program

Overnight coincubation of the cumulus–oocyte complexes (COCs) with spermatozoa may be accompanied by overproduction of reactive oxygen species (ROS) and possible determinant effects on sperm cellular status in the in vitro fertilization (IVF) program. The COCs from 100 cases were divided into two groups of short (2 hr) and long (18 hr) inseminations. The malondialdehyde (MDA) level as by-product of ROS was assessed in the insemination medium. The sperm DNA integrity, the mitochondrial membrane potential (MMP), and acrosome reaction (AR) were evaluated using sperm chromatin dispersion (SCD), fluorescence stain of JC-1, and double staining, respectively. Normally, fertilized oocytes (n = 525) were assessed via time-lapse monitoring (TLM) for assessment of the time of fading (tPNF), 2 until 8 timing (t2-t8), s1 (t2-tPNF), s2 (t4-t3), s3 (t8-t5), and duration of cell cycles. Finally, the best embryos were transferred, and clinical outcomes were assessed. Higher rates of MDA concentration, DNA fragmentation, and AR and low rate of MMP were noticed in long compared to short insemination groups ( p ≤ 0.0001 ). The morphokinetic parameters showed five-cell stage (t5) and cell cycle 3 (cc3) that were significantly different between the groups ( p = 0.04 and p = 0.03 , respectively). A high level of ROS observed in the long insemination group might have a detrimental effect on sperm status. Although, similar embryo quality and clinical outcomes were noticed in two insemination groups, but the trend was toward the short insemination.

Micronutrient Fe and Mn uptake from peat by marigold and petunia

Peat substrates have the potential to function as micronutrient sources such as iron (Fe) and manganese (Mn). Typically, when planning fertilization program for crops, the components of peat substrates are not taken into account. The objective was to quantify potentially plant-available micronutrients of Fe and Mn from peat substrates by floriculture species when no additional Fe and/or Mn were applied in the fertilizer solution. Greenhouse experiments were conducted using Tagetes erecta ‘Atlantis Primrose’ marigold (iron-efficient) and Petunia x hybrida ‘Celebrity Pink Morn’ petunia (iron-inefficient) to compare extractable Fe and Mn from two peat sources with actual plant uptake of these nutrients by the plants. In Experiment 1, marigold was grown hydroponically in plastic tubs using four different nutrient solutions: complete, without Fe, without Mn, and without both Fe and Mn. Within a period of less than two weeks after transplant, the absence of Fe and Mn severely limited marigold growth. The hydroponic study highlighted the importance of providing both Fe and Mn nutrients immediately after sowing to support normal marigold growth. In Experiment 2, both marigold and petunia were grown from seed in two peat substrates (Blackmore seedling peat “BM peat” and Sunshine Professional Black Bale peat “SG peat”) and an acid-washed sand substrate. Each peat was limed at 3 g.L−1 of reagent grade CaCO3 resulting in an initial substrate-pH of 5.0 for SG peat and 5.5 for BM peat. Four nutrient solutions [(a) complete, or complete with (b) no Fe, (c) no Mn, and (d) no Fe or Mn] were applied with each irrigation without leaching. Initial substrate nutrient levels were extracted using the SME (saturated medium extract) method using either water (SMEW) or 0.005 M DTPA (diethylenetriamine-pentaacetic acid, SMED) as extractants. Over a 5-week period (during the vegetative, non-blooming stage), peat substrates provided sufficient Fe and Mn for marigold, but iron deficiency symptoms were observed in marigolds grown in sand without Fe fertilizer. Petunia plants developed iron deficiency symptoms in all substrates without Fe fertilizer. The absence of Mn fertilizer, however, did not have a significant impact on petunia growth and health, indicating that peat substrates likely provided adequate Mn for non-symptomatic petunia plant growth. The micronutrient Fe and Mn budget analysis suggested that the SMEW method underestimated the availability of Fe and Mn to the plants, whereas the SMED method may have overestimated the extractable Fe and Mn levels in relation to the actual tissue uptake. Overall, the peat substrates showed a high potential for providing Fe and Mn in quantities sufficient for the initial growth of iron-efficient marigold, but was inadequate for iron-inefficient petunia.

Influence of Soil and Nutrient Management Practices on Crop Productivity and Quality in High Tunnel Organic Leafy Green Production

Despite the growing interest in high tunnel organic vegetable production, limited information is available regarding optimizing nutrient management for organic leafy greens. This 3-year study examined the impacts of cowpea cover crop as well as different organic fertilizers and composts on yield, leaf mineral nutrient content, and phytochemical properties of organic leafy greens produced in high tunnels under Florida sandy soil conditions. The experiment was arranged in a split-split-plot design with three replications. The whole plots consisted of a cowpea (Vigna unguiculata ‘Iron & Clay’) cover crop and a weedy fallow control, with fertilization treatments in the subplots, including preplant application of granular fertilizer vs. weekly injection of liquid fish fertilizer at the same seasonal rates of nitrogen (112 kg/ha), phosphorus (9.8 kg/ha), and potassium (74.4 kg/ha). The sub-subplots included yard waste-based compost (22.4 Mt/ha), cow manure-based compost (22.4 Mt/ha), vermicompost (5.6 Mt/ha), and no compost control. Cowpea was broadcasted (112 kg/ha) in early July or mid-August and terminated 51 to 53 days after seeding. Pac choi (Brassica rapa var. chinensis ‘Mei Qing Choi’) was transplanted in mid-Sep. or mid-Oct. and harvested after 33 to 36 days. Baby spinach (Spinacia oleracea ‘Corvair’) or baby leaf lettuce (Lactuca sativa ‘Outredgeous’) was direct seeded subsequently as a catch crop. Each experimental unit remained in the same location across the 3 years of the study. Cover cropping had little influence on yields, leaf mineral nutrients, ascorbic acid content, total phenolics, and total antioxidant capacity of pac choi and baby spinach/lettuce. Compared with preplant application of the granular organic fertilizer, weekly liquid organic fertigation improved pac choi marketable yield and dry weight by 16.8% and 5.4% on average, respectively, and enhanced leaf nitrogen and phosphorus contents on a dry weight basis. Relative to the no compost control, yard waste compost consistently improved marketable yields of pac choi by 11.6% on average and led to higher yields of the baby spinach/lettuce catch crop in years 1 and 3, suggesting that compost applications may enhance seasonal nutrient availability to better meet crop demand. However, compost application exhibited inconsistent effects on crop mineral nutrient and phytochemical contents across the years, which could be attributed to the different nutrient compositions of the composts applied in each season, as well as the legacy effects from the previous season. Furthermore, the compost benefits may be influenced by the fertilization program as indicated by their interaction effects observed in this study.

#278 : Relationship Between Level Expression of miRNA-191 and miRNA-548c-3p in Embryo Culture Medium and Embryo Chromosomal Status

Background: microRNA (miRNA) is expressed by the human embryos and shows the difference between euploid and aneuploid embryos. miRNA which is secreted in culture media is expected to be a biomarker for embryo chromosomal status testing. This research aims to analyse the relationship between expression level of miRNA-191 and miRNA-548c-3p in embryo culture media and embryo chromosomal status. Methods: This cross-sectional research was done on 30 embryos from 12 patients aged 28-40 years old who underwent an in vitro fertilization (IVF) program in three clinics. Embryos were cultured until blastocyst stage. PGT-A was done after blastocyst biopsy on day 5 or 6, and chromosomal analysis was done using the NGS method. The expression levels of miRNA-191 and miRNA-548c-3p were analysed from culture media sample of the embryo using qPCR method. Then the relationship between miRNA-191 and miRNA-548c-3p expression level with embryo chromosomal status was analysed. Results: There is no significant difference in subjects’ characteristics between the euploid and aneuploid embryo groups (p>0.05) [Table 1]. There is no significant difference in the expression level of miRNA-191 of the euploid and aneuploid embryo groups. (Median quantification 8.69 vs 1.064 with p=0.351) [Table 2]. There is no significant difference in the expression level of miRNA-548c-3p of the euploid and aneuploid embryo groups. (Median quantification 9.06 vs 6.635 with p=0.709) [Table 3]. Conclusions: miRNA-191 expression level and miRNA-548c-3p expression level has no relationship with embryo chromosomal status. The level expression of miRNA-191 and miRNA-548c-3p in embryo culture has not been a good biomarker candidate for embryo chromosomal status.

#166 : Investigating the Effect of Parental Age on Intracytoplasmic Sperm Injection Reproductive Outcomes in Indonesia: Prognostic Tools and Prediction Approaches to Maximize Human Embryo Implantation

Background and Aims: Reproductive issues impact one in eight Indonesian married couples. Intra Cytoplasmic Sperm Injection (ICSI) is still expensive; hence many couples don’t get evaluated or enroll fertility programs. We decided to study the correlation between age, hormonal status, and ICSI embryo quality because Indonesia’s infertility rate is rising. This is the first investigation into the factors behind ICSI outcomes in Indonesia. Method: In 2022, two fertility centers in Surabaya and Tangerang, Indonesia, performed ICSI on couples. 150 couples receiving ICSI at two research sites comprise the sample. Surgically retrieved or frozen sperm cycles and incomplete medical records were excluded. The Spearman test was used to investigate the correlation between parental age, hormonal status, and ICSI embryo quality on day 1 and day 3. (D1 and D3). Results: 150 ICSI cycles were studied. The hormonal baseline varied from normal to subnormal, while paternal and maternal age ± SDs were 37.03 ± 6.08 and 34.28 ± 5.11. The Spearman test demonstrated a significant negative connection between paternal age and embryo quality D1 (p<0.005, r=-0.363) and D3 (good category: p<0.005, r=-0.172; intermediate category: p<0.005, r=-0.250; poor category: p<0.005, r=-0.221). The Spearman test showed a significant negative connection between maternal age and embryo quality D1 (p<0.005, r=-0.413) and D3 (good category: p<0.005, r=-0.231; intermediate category: p<0.005, r=-0.294; poor category: p<0.005, r=-0.291). Conclusion: This study reveals that married couples have a substantial impact on their hormonal condition, the rate of fertilization, the quality of D1 and D3 embryos, and the number of embryos generated following ICSI. To increase the success rate of ICSI procedures, it is necessary to maximize fertility at a younger age.

#242 : Correlation of Sperm DNA Fragmentation with Laboratory Performance in Good and Poor Prognosis Patient Groups at Morula IVF Jakarta

BACKGROUND AND AIMS: Infertility in males contributes to about 40% of cases of infertility in couples, either as the main factor or along with female factors. Fragmentation of sperm DNA is known as a cause of male infertility, which impacts the failure of couples to undergo assisted reproductive programs, such as embryo development and recurrent miscarriage. However, some literature studies are still debating this. This research aims to determine the correlation between sperm DNA fragmentation and IVF laboratory performance. METHODS: Infertile couples undergoing an in vitro fertilization program at Morula IVF Jakarta in 2022 are the samples for this study. We divided them into two groups: good prognosis (maternal age [Formula: see text]38 years, AFC[Formula: see text]7; AMH[Formula: see text]1.1) and poor prognosis (maternal age > 38 years, AFC <7, AMH<1.1). Sperm DNA fragmentation was examined using the sperm chromatin dispersion (SCD) assay. Correlation statistical analysis using the Spearman test (p-value <0.05). RESULTS: Based on receiver operating characteristic (ROC) curve analysis, it was found that the sperm DNA fragmentation rate >14.9% had a significant negative correlation with IVF laboratory performance, particularly in the fifth-day embryo quality and blastocyst rate in the good prognosis group. Whereas in the group with poor prognosis, all laboratory performance parameters were not affected by high sperm DNA fragmentation values. CONCLUSIONS: Sperm DNA fragmentation rate > 14.9% correlated with fifth-day embryo quality and a lower blastocyst rate in the good prognosis group.

Favorable Fertility Outcome in a Large Cohort of Sickle Cell Disease Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation

Introduction: Fertility preservation is an important consideration for individuals with sickle cell disease (SCD) who may face reproductive challenges. While SCD itself does not directly affect fertility, certain complications and its treatment such as hematopoietic cell transplantation (HCT) can impact the reproductive health in SCD patients. HCT is a potential curative therapy for SCD, however, one of the negative outcomes is the premature gonadal failure and infertility. Both male and female patients have different risk factors that may impact the fertility post-transplant. Therefore, these patients should undergo fertility preservation counselling pre transplant. In this study, we report the fertility outcome of large transplant cohort undergoing Non-myeloablative (NMA) conditioning regimen. Methods: We retrospectively collected the data for all SCD patients who were ≥ 14 years old and who underwent allogeneic HCT in our center from 2015 until 2022 with a minimum of 1 year follow up post-transplant. All patients received hydroxyurea for 2 to 3 months pre- transplant at maximum tolerated dose. The conditioning regimen was alemtuzumab 1 mg /kg divided over 5 days from day - 7 to day -3 and Total Body Irradation 200 cGy on day -2 (with testicular shielding for male patients). Graft versus host disease prophylaxis with sirolimus started from day -1 and continued for at least 1 year post-transplant. We started the fertility program in October 2019 where all patients get counselled for fertility preservation methods. Male patients undergo sperm cryopreservation after holding hydroxyurea for 3 months and female patients were referred to a specialized oncology fertility clinic for either oocyte cryopreservation or treatment with Gonadotropin-releasing hormone (GnRH) agonists pre transplant. Primary objective of the study was to assess post-transplant fertility (defined by either ability to conceive for married patients, or development of azoospermia for male patient or secondary amenorrhea for female patients). Secondary objectives were to assess pre transplant fertility, assess the success of different fertility preservations methods and finally to assess for risk factors (pre- transplant and transplant) that affect the fertility potential. Results: We identified a total of 200 SCD patients who underwent HCT. Out of these 200 patients, 107 (54%) were included in our fertility program. The median post-transplant follow up was 3.5 years (1 - 7.5). One hundred and twelve patients (56%) were male, and eighty eight (44%) were female patients. All patients were on hydroxyurea pre transplant with average dose 1000 mg and a duration of less than 2 years in 47 (23.5%) patients, 2 to 5 years in 94 (47%) patients and more than 5 years in 59 (39.5%) patients. Other baseline characteristics are shown in table 1. In the pre-transplant fertility assessment; 58 patients were married and 48 (82%) of them had children pre transplant. 36 male patients attempted sperm cryopreservation and were successful in 33 (91%). 39 female patients received Leuprolide (GNRH agonists) pre-transplant with no complications and 6 female patients attempted oocyte cryopreservation and all were successful. One patient developed cerebral venous thrombosis during the oocyte cryopreservation course. For the post-transplant fertility assessment; out of 57 married patients 26 (45%) were able to conceive. From these 26 pregnancies; 20 ended with successful live-births, 1 ended with abortion and 5 are still pregnant. 6 patients were able to conceive twice and one patient had one twin babies. For male patients 74 did semen analysis at minimum 1 year post-transplant; 35 (47%) had azoospermia and 39 (53%) had viable sperms. Majority recovered spermatogenesis after 1 year (24 patients), at 2 years (9 patients) and after 3 years (3 patients). Three male patients had Azoospermia at one year but was able to conceive successfully without repeating the test. For female patients 71 (80.7%) were able to regain menses and 17 (19.3%) developed secondary amenorrhea. Table 2 shows the pre and post-transplant fertility measures outcome. In conclusion In this study, we have shown the fertility outcomes of male and female patients with SCD undergoing NMA conditioning HCT can be preserved in a significant proportion of patients. All adult patients should be counselled for fertility preservation methods before undergoing curative allogeneic HCT.

Enhancement of preimplantation mouse embryo development with optimized in vitro culture dish via stabilization of medium osmolarity.

We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.

Studies on Potassium Fractions in Central and Southern Dry Zones of Coconut Growing Areas of Hassan District of Karnataka, India

The experiment was conducted in an order to get an insight about the potassium (K) fractions in Central and Southern dry zone of coconut growing areas of Hassan district, Karnataka during 2022-23. The soil samples from surface (0-30 cm) and sub-surface (30-60 cm) were collected from the soils of selected coconut gardens of Central and Southern dry zones of Hassan district for the study. Results revealed that the average water soluble K, hot water soluble K, exchangeable K, non-exchangeable K, lattice K and total K in surface soils of coconut gardens of Central dry zone of Hassan district registered was 2.76, 9.25, 54.65, 555.35, 5896.00 and 6509.00 mg kg-1, respectively. Whereas the above potassium fractions in sub-surface soil recorded were 2.67, 7.70, 47.36, 699.66, 7282.00 and 8032.00 mg kg-1, respectively. The various forms of potassium in surface soils of Southern dry zone revealed that water soluble K, hot water-soluble K, exchangeable K, non-exchangeable K, lattice K and total K was 2.87, 8.85, 53.51, 546.61, 5603.00 and 6206.00 mg kg-1, respectively. Whereas the above potassium fractions in sub-surface soil recorded were 2.21, 7.20, 45.21, 709.20, 7431.00 and 8188.00 mg kg-1, respectively. The trend of decrease in water soluble K and exchangeable K from surface to sub-surface layers was noticed in soils of Central and Southern dry zone whereas non-exchangeable K, lattice K and total K exhibited an increasing trend with depth. The contribution of each form of potassium to total potassium was in the order of water soluble < hot water-soluble K< exchangeable < non-exchangeable < lattice K. Knowledge of different forms of potassium in soil together with their distribution has greater relevance in assessing the long-term K supplying power of soil to coconut palm and is important in formulating a sound fertilizer program for a given set of soil.

Agronomic biofortification with magnesium nanofertilizer and its effect on the nutritional quality of beans

Crop quality has been compromised due to nutrient deficiencies. The macronutrient magnesium (Mg) is essential; however, it has not been considered in agronomic fertilization programs, affecting human health. The objective of the study was to increase the Mg content in the fruits of green beans cv. ‘Strike’ by applying Mg-nanofertilizer, as well as evaluating its effect on growth, performance and nutritional quality, versus magnesium sulfate (MgSO4). The experiment was carried out under shade mesh conditions in Delicias, Chihuahua, Mexico during the period August-October 2022. A completely randomized experimental design was used, with two Mg sources: Mg nanofertilizer (NanoMg) and MgSO4 at doses of 50, 100 and 200 ppm and a control without application, forming seven treatments with six repetitions each. The results indicate that the maximum yield was provided by NanoMg and MgSO4 at 200 ppm, with values greater than 300%. The Mg distribution pattern for the NanoMg treatments presented the following concentration order: root>leaf>stem>fruit; while, for the MgSO4 treatments it was: leaf>root>stem>fruit. The most efficient treatment in increasing the Mg content in the fruit was NanoMg at 200 ppm, which achieved a biofortification of more than 120% with respect to the control. Therefore, when consuming 100 g of green beans cv. ‘Strike’ biofortified by NanoMg, the recommended daily needs of the human being could be satisfied. Finally, it is concluded that nanofertilizers are the best option for a biofortification program since they offer a sustainable alternative by increasing productivity and quality in green bean fruits.

Intra-uterine programming of future fertility.

The developmental origins of health and disease (DOHaD) shows that a relationship exists between parental environment at large, foeto-placental development and the risk for the offspring to develop non-transmittable disease(s) in adulthood. This concept has been validated in both humans and livestock. In mammals, after fertilization and time spent free in the maternal reproductive tract, the embryo develops a placenta that, in close relationship with maternal endometrium, is the organ responsible for exchanges between dam and foetus. Any modification of the maternal environment can lead to adaptive mechanisms affecting placental morphology, blood flow, foetal-maternal exchanges (transporters) and/or endocrine function, ultimately modifying placental efficiency. Among deleterious environments, undernutrition, protein restriction, overnutrition, micronutrient deficiencies and food contaminants can be outlined. When placental adaptive capacities become insufficient, foetal growth and organ formation is no longer optimal, including foetal gonadal formation and maturation, which can affect subsequent offspring fertility. Since epigenetic mechanisms have been shown to be key to foetal programming, epigenetic modifications of the gametes may also occur, leading to inter-generational effects. After briefly describing normal gonadal development in domestic species and inter-species differences, this review highlights the current knowledge on intra-uterine programming of offspring fertility with a focus on domestic animals and underlines the importance to assess transgenerational effects on offspring fertility at a time when new breeding systems are developed to face the current climate changes.

Minimizing Carbon Dioxide Emissions with Clinoptilolite Zeolite in Moris Pineapple Cultivation on Drained Sapric Soils

Drained tropical peat soils for agriculture emit more carbon dioxide (CO2) into the atmosphere from their stored carbon compared with their pristine state. Field and laboratory experiments were conducted to assess whether the natural zeolite of the clinoptilolite (ZeoC) species could be included in the pineapple fertilization program to decrease the CO2 emissions from tropical sapric soils. The static closed-chamber and laboratory incubation methods were used to determine the effects of ZeoC on the CO2 emitted from a drained sapric soil planted with Moris pineapple. The treatments assessed were as follows: (a) suggested ratio of ZeoC (5 g, 10 g, 14 g, and 20 g of ZeoC) and 20 g compound NPK 30:1:32 fertilizer, enumerated based on the pineapple plant requirement; (b) 20 g of compound NPK 30:1:32 fertilizer only; and (c) unfertilized sapric soils. The drained sapric soils amended with ZeoC (rate of 5 g to 20 g/plant) minimized the CO2 emissions compared with those without the ZeoC, because of the physical and chemical sorption of organic compounds and polar CO2 onto the lattices of ZeoC, which inhibited organic matter decomposition. ZeoC fertilization reduces sapric soil acidity and improves Moris pineapple fruit quality attributes and yield. Monthly pineapple fertilization with ZeoC at the vegetative and flowering phases is an alternative agronomic strategy to reduce CO2 emissions. This approach does not reduce pineapple yield on drained tropical sapric soils.

Actors and Actor Relations in Governance of Subsidized Fertilizer Policy in Indonesia

The study investigates actor relationships in subsidized fertilizer programs within Indonesian rice production hubs. It aims to comprehend roles and interactions among actors in budget allocation, fertilizer distribution, policy oversight, and farmer involvement. The research employs a descriptive qualitative approach, analyzing how institutional practices and hierarchies impact the subsidized fertilizer market. The study focuses on Indramayu and Karawang Regencies in West Java and Sidenreng Rappang (Sidrap) and Wajo Regencies in South Sulawesi, all significant rice production centers. Data was gathered through in-depth interviews involving central, regional, distributor, store actors, and farmers. Results reveal collaboration and competition as primary relationship modes in the subsidized fertilizer program. Cooperation exists among institutional players at the central level during budget deliberations, while relationships emerge when actors unite to advocate for budget adjustments or specific fertilizer types. Factors like discourse, institutional practices, and capitalist interests shape these inter- actions. Discourse might favor chemical or organic fertilizers. Farmers occupy a relatively marginalized position in actor conflicts, warranting increased policymaker attention. Although the government seeks improvements, farmers still face challenges such as inadequate doses, delays in distribution, and financial constraints

Deep machine learning applied to support clinical decision-making in the treatment of infertility using assisted reproductive technologies

Introduction. Machine learning (ML) applied to data analysis allows to more accurately and targetedly determine the most significant correctable and non-correctable predictors of onset of pregnancy in assisted reproductive technology (ART) programs in patients of different age groups. Analysis of data using various techniques and comparison of results obtained via two models will determine the most significant factors for onset of pregnancy in the ART program.Aim. To determine the most significant clinical and embryological predictors of onset of pregnancy using standard regression analysis and a decision tree algorithm to predict pregnancy in the ART program.Materials and methods. A total of 1,021 married couples were included in the retrospective study. The study analysed clinical and laboratory test findings and stimulated cycle parameters depending on the effectiveness of the ART program. A regression analysis was carried out and a decision tree algorithm was built using the Gini criterion to determine the most significant factors.Results. We identified “general” signs that require further validation on other models, including ML: the presence/absence of a history of pregnancies, stimulated cycle parameters (oocyte cumulus complex, number of metaphase II (MII) oocytes, number of zygotes), spermogram indicators on the day of puncture, number of high and good quality embryos, as well as the embryo grading.Conclusion. rFSH (follitropin-alpha, Gonal-f) gives a significant result in two of the five available age groups, follitropin-beta, corifollitropin alfa – in one of the five groups only. Building a model that includes not only the couple’s medical history data, but also molecular markers using machine learning methods will not only allow us to most accurately determine the most promising groups of patients for in vitro fertilization (IVF) programs, but also increase the efficiency of ART programs by selecting the highest quality embryo to be transferred.

In vitro fertilization program in white rhinoceros.

To save endangered rhinoceros species, assisted reproductive technologies are warranted. We here report in vitro blastocyst generation of the Near-Threatened Southern white rhinoceros and, for the first time, also of the technically Extinct Northern white rhinoceros. The Anthropocene is marked by a dramatic biodiversity decline, particularly affecting the family Rhinocerotidae. Three of five extant species are listed as Critically Endangered (Sumatran, Javan, black rhinoceros), one as Vulnerable (Indian rhinoceros), and only one white rhino (WR) subspecies, the Southern white rhinoceros (SWR), after more than a century of successful protection is currently classified as Near Threatened by the IUCN, while numbers again are declining. Conversely, in 2008, the SWR's northern counterpart and second WR subspecies, the Northern white rhinoceros (NWR), was declared extinct in the wild. Safeguarding these vanishing keystone species urgently requires new reproductive strategies. We here assess one such strategy, the novel in vitro fertilization program in SWR and - for the first-time NWR - regarding health effects, donor-related, and procedural factors. Over the past 8 years, we performed 65 procedures in 22 white rhinoceros females (20 SWR and 2 NWR) comprising hormonal ovarian stimulation, ovum pick-up (OPU), in vitro oocyte maturation, fertilization, embryo culture, and blastocyst cryopreservation, at an efficiency of 1.0 ± 1.3 blastocysts per OPU, generating 22 NWR, 19 SWR and 10 SWR/NWR hybrid blastocysts for the future generation of live offspring.

First report of frequencies of Y chromosome microdeletions at a reproductive medicine center in Peru

ObjectiveY chromosome Microdeletions are the second genetic cause of infertility in men. Despite its importance for infertility treatment, there is no previous research in Peru. The aim of this study was to determine the frequencies and characteristics of Y chromosome microdeletions in a group of men who sought infertility consultation at a specialized reproductive medicine center in Peru. MethodsIn this study, 201 semen samples were analyzed. The samples were obtained from Niu Vida's fertility program. Each seminal sample was analyzed according to the recommendations of the Laboratory Manual of the World Health Organization (WHO) 2010. A buccal swab and a 500 μL aliquot of seminal sample were used for the molecular study of Y chromosome microdeletions in each patient. The frequencies and the type of Y chromosome microdeletion in the AZFa, AZFb and AZFc regions were evaluated. ResultsThe prevalence of Y chromosome microdeletions in the AZF region was 6.45% in oligozoospermic and azoospermic patients, and a prevalence of 20% was observed specifically in azoospermic patients. No microdeletions of AZFb type were detected. A partial region microdeletion of AZFa was detected in a teratozoospermic patient with a normal sperm count. ConclusionsThe study represents the first report on the incidence of Y chromosome microdeletions in Peru. Our results indicate a high prevalence of microdeletions in azoospermic patients compared to similar studies. It is suggested to assess the presence of AZFa microdeletions and to evaluate additional genetic markers in this region to identify specific mutations that may cause impaired sperm production and male infertility in the Peruvian male population.

Development of banana (Musa AAA) under different doses of biochar and chemical fertilizers

The main objective of the research was to evaluate the application of different doses of biochar as organic fertilizer to a banana (Musa AAA) crop. The biochar was derived from cocoa pod husks (CPH-B) and banana stalk (BC) and applied to banana plants between 130 and 140 cm tall. The treatments included doses of 32 kg ha-1 (T1) and 48 kg ha-1 (T2) of CPH-B, 32 kg ha-1 (T3) and 48 kg ha-1 (T4) of BC. In all experiments, 100 kg ha-1 potassium nitrate and 100 kg ha-1 phosphorus (P2O5) were applied. Significant differences (p≤0.05) were found in root weight, height and pseudostem circumference at T4. Bunch weights ranged from 16.44 kg (T2) to 21.10 kg (T4). The number of clusters per bunch increased from 6.6 (T1) to 8.5 (T2), with significant differences. Treatments T2 and T4 positively affected plant growth compared to the control plot, suggesting they could be used as an alternative in fertilization programs. Keywords: banana stalk; cocoa pod husk; nutrients

Remoción de macronutrientes de la fruta de aguacate cv. Hass en Antioquia y Caldas, Colombia

In recent years, Colombia have become globally as one the major producer of Hass avocado. Contributions of all nutrients are made in the fertilization plans, but it is not known if the additions made have been adequate and opportune. Thus, the objective of this study was to determine the effects of site and year of productions on the removal of nutrients during the harvest stage of different tissues of the Avocado fruit cv. Hass in two farms (El-Sinaí and La-Escondida) located in the high-Andean tropical zone of Colombia in three years of production (2018-2020). For this purpose, fruit samples were taken at the harvest stage of highly productive trees from two sites in Colombia (i) El-Sinaí (Anserma, Caldas; altitude 2,000 m, average annual temperature and precipitation 18.65°C and 1,837 mm) and (ii) La-Escondida (Rionegro, Antioquia; altitude 2,200 m, average annual temperature and precipitation 16.84°C and 2,091 mm). The fruit tissues (peel, pulp, seed-coat and seed) were subjected to elemental composition analysis. Significant (P<0.05) differences were detected for each element between tissues, years, sites and their interactions. The total removal range for macronutrients was: N 1.53-2.57, P 0.21-0.38, K 2.53-3.51, Ca 0.20-0.56, Mg 0.15-0.24 and S 0.13-0.18 (kg t-1). These results indicate that the order of nutrient accumulation in the fruit was K > N > Ca > P >Mg > S and they may be used as a reference for elaboration of fertilization programs in the regions studied, particularly in terms of the doses and ratios of nutrients. Also, the nutrient concentration in the different fruit tissues may be used as a reference of fruit nutritional status of productive trees for farmers interested in monitoring and improving nutritional of low-yield trees.

Evaluation of the Effect of Biostimulation on the Yielding of Golden Delicious Apple Trees

The aim of the study was to assess the impact of N Pro and Seactiv Complex technologies based on sea algae extracts on the yield and quality of Golden Delicious apple trees compared to the standard fertilization program. Research conducted in the years 2008–2017 showed that the tree fertilization technology and the year of research had a significant impact on the quality and yield parameters. The abundance of seaweed extracts had a positive effect on the greater number of fruits per tree (pcs.), their weight (t × ha−1; kg tree−1), the percentage of fruits exceeding 7.5 cm in size (%) and the average weight of fruits (g) compared to the control combination. In most of the analyzed study years, total yield exceeded the 90 t × threshold regardless of the fertilization program. The use of technology based on sea algae extracts during 10 years of research had a positive effect on the quality and weight compared to the standard fertilization technology commonly used in cultivation.

Studies on the hormone and cytokine producing function of human cumulus cells and its interrelation with fertility in polycystic ovarian syndrome

Cumulus cells (CC) are the specialized layer of follicular cells that are in close contact with the oocyte. They are considered as indirect markers of the oocyte quality. Changes of these cells suggest a damage of the ovary. Determination of cytokines in cumulus cell culture may predict the chance for the conception and development of pregnancy. The objective of the present study was to obtain a primary culture of CC from healthy donors and patients with polycystic ovarian syndrome (PCOS), to identify the most significant differences in production of key cytokines in the CC monocultures of patients and healthy individuals in order to predict the results of in vitro fertilization. Materials and methods: the cell culture technique was used, i.e., cumulus cells of healthy donors and patients with polycystic ovary syndrome were obtained by transvaginal puncture of follicles in the in vitro fertilization (IVF) program. This procedure does not affect the rights of embryo, since the CC are not used at the stage of IVF procedure and intracytoplasmic sperm injection (Ethical Committee Protocol No. 9 of May 16, 2022). IL-6, IL-10, IFN and progesteroneparameters were tested in adhesive cultures of CC by ELISA technique on days 1, 3, 7 of in vitro experiments. Results: We revealed a continuous secretion of progesterone, IL-6, IL-10, IFN in adhesive monocultures of CC. In the patients with PCOS, we have found a sharp increase of progesterone level in cultural media (p 0.01) on the 7th day, By contrast, the initially increased progesterone levels proved to be significantly decreased in donors on the 7th day of culture. Moreover, in the culture of CC from patients with PCOS (7th day of the experiment), the values of IL-6, IL-10 increased only two-fold compared with 30-fold increase of these cytokines in healthy donors (p 0.01). At the same term, we have observed a threefold decrease in IFN in the CC cultures of PCOS patients (p1-7 0.01; p 0.05) compared with cultured controls, which showed a 20-fold increase (p1-7 0.01), thus determining differences in total cytokine balance and, probably, influencing the pregnancy prognosis. Conclusion: Significant multidirectional changes of cytokine levels in the culture of cumulus cells of the patients with PCOS and in healthy individuals may be regarded as determining factors in formation of blastocyst and preservation of the embryo. A further in vitro research on the production of cytokines and sex steroids by CC is especially important on day 5 to 7, when the oocytes are selected for entry into the in vitro fertilization cycle. The study of morpho-functional properties of little-studied cumulus cells using the cell culture technique will enable us for a deeper study on the mechanisms of disturbed folliculogenesis in PCOS, and, thereby, improve the reproductive prognosis in this disorder.