Maturation Medium Research Articles

Investigation of the effect of cow estrous serum when added to the maturation medium of oocyte-cumulus complexes <i>IN VITRO</i>

Saving and multiplying the existing livestock population is an important strategic objective that ensures the country's food security and levelling the dependence on imports of genetically valuable highly productive animals. Obtaining farm animal embryos in vitro is one of the methods of assisted reproductive technologies (ART), which can be used both lifetime and post-mortem. This method allows to optimise breeding work with livestock in a short period of time due to the large number of offspring obtained from highly productive individuals. Aim and objectives: to investigate the effect of bovine estrous serum on meiotic oocyte maturation and early embryogenesis prior to blastocyst development, when added to oocyte-cumulus complexes (OCC) maturation medium. A blind randomised controlled trial was conducted on the effect of bovine estral serum on blastocyst development when added to OCC maturation medium. Estral serum was obtained from the blood of clinically healthy cows with pronounced signs of puberty. The study was carried out in conditions of industrial livestock breeding. The material for the study were oocyte-cumulus complexes obtained by the method of lifetime aspiration from 89 Holstein cows with milk productivity of more than 10 thousand kg per lactation. Oocyte-cumulus complexes were divided into 2 groups: Group 1 (control) - oocyte maturation was carried out in commercial IVM medium without addition of estrous serum, Group 2 (experiment) - oocyte maturation was carried out in commercial IVM medium with addition of es trous serum in concentration from 5 to 15%. The hormones, growth factors and biologically active substances contained in estrous serum have no effect on the achievement of OCC at the stage of metaphase II of meiosis, as well as at the stage of fractions; moreover, a negative effect at these stages of development can be traced. However, substances contained in estrous serum are able to saturate and accumulate in OCC at the stage of maturation to metaphase II of meiosis, thus enhancing the potential of oocytes to embryonic development and further positively affect their achievement of the blastocyst stage. The difference in the formation of blastocysts from crushed embryos reaches more than 25%, and the difference in the formation of blastocysts from viable JCCs matured in media without serum and modified with estrous serum is 37.5% in favour of the latter.

In vitro maturation of mouse oocytes in dose-dependent manner of platelet concentrate-derived lysates

Platelet concentrates (PCs) including platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) are extensively used as autologous products in regenerative medicine. In this study, the effects of different concentrations of platelet concentrate-derived lysates (PC-Ls) including PRP-derived lysate (PRP-L) and PRF-derived lysate (PRF-L) were demonstrated separately on the oocyte maturation. Since then, the effectiveness and appropriate concentration of PC-Ls will be confirmed which will be the premise for studies on improving the composition of oocyte maturation medium utilizing active components of autologous origin. The complexes of oocyte-cumulus-granulosa (COCGs) were collected from mature female mice (6-8 weeks old, Mus musculus var. Albino). The maturation medium (M-199 medium) for the COCGs was supplemented with 5%, 10%, 15% PRP-L and 5%, 10% and 15% PRF-L. Oocytes were assessed for maturity after 24 hours and 48 hours of culture through morphological (polar body appearance and developmental stage of oocyte, follicular cell morphology) and functional (in vitro fertility) evaluation. First, all groups showed the same oocyte morphology which included the first polar body. However, the development of follicular cells only happened in the groups treated with 10%, 15% PRP-L and 10%, 15% PRF-L during the culture phase. The 15% PRF-L group had the most significant proportion of mature oocytes after 24 hours of culture which was 22.88%. After 48 hours of culture, this number rose to 50.67% in the 10% PRF-L treatment group (P-value 0.05). The percentage of developed oocytes from the 10% PRF-L treatment that could be fertilized was 41.67%. The platelet concentrates-derived lysates promoted oocyte maturation independently and PRF-L supported oocyte development better than PRP-L. This study suggested a novel role for second-generation platelet concentrates (PRF) in in vitro oocyte maturation. The culture time should not be extended to 48 hours.

Impact of media supplements FGF2, LIF and IGF1 on the genome activity of porcine embryos produced in vitro

In this article, we focused on the impact of precisely chemically modified FLI maturation medium enriched with fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), insulin-like growth factor 1 (IGF1), and polyvinyl alcohol (PVA) and its potential to improve the efficiency of in vitro production of porcine embryos. We hypothesized that enhancing the composition of the maturation medium could result in an elevated production of embryos in vitro and can affect EGA. FLI medium resulted in a significantly higher rate of oocyte blastocyst maturation and formation compared to the control DMEM medium. In addition, immunocytochemical labelling confirmed the detection of UBF in 4-cell FLI parthenogenic embryos, suggesting similarities with natural embryo development. Through RNAseq analysis, upregulated genes present in 4-cell FLI embryos were found to play key roles in important biological processes such as cell proliferation, cell differentiation, and transcriptional regulation. Based on our findings, we demonstrated the positive influence of FLI medium in the evaluation of in vitro embryo production, EGA detection, transcriptomic and proteomic profile, which was confirmed by the positive activation of the embryonal genome in the 4-cell stage of parthenogenetically activated embryos.

Effect of different concentrations of resveratrol on nuclear maturation and in-vitro development competence of oocytes of Nili Ravi buffalo.

Buffaloes are considered animals of the future with the ability to survive under unfavorable conditions. However, the lack of access to superior germplasm poses a significant challenge to increasing buffalo production. Resveratrol has been shown to improve oocyte quality and developmental competence in various animals during in vitro embryo development. However, limited information is available on the use of resveratrol to improve the in vitro maturation and development competence of Nili Ravi buffalo oocytes. Therefore, the current study aimed to investigate the influence of different concentrations of resveratrol on the maturation, fertilization, and development of buffalo oocytes under in vitro conditions. Oocytes were collected from ovaries and subjected to in vitro maturation (IVM) using varying concentrations of resveratrol (0 µM, 0.5 µM, 1 µM, 1.5 µM, and 2 µM), and the maturation process was assessed using a fluorescent staining technique. Results indicated no significant differences in oocyte maturation, morula rate, and blastocyst rate among the various resveratrol concentrations. However, the cleavage rate notably increased with 1 µM and 1.5 µM concentrations of resveratrol (p < 0.05). In conclusion, the study suggests that adding 1 µM of resveratrol into the maturation media may enhance the cleavage and blastocyst hatching of oocytes of Nili Ravi buffaloes. These findings hold promise for advancing buffalo genetics, reproductive performance, and overall productivity, offering potential benefits to the dairy industry, especially in Asian countries.

Evaluation of seed yield, essential oil compositions, and fatty acid profiles in advanced fennel (Foeniculum vulgare Mill) breeding populations

Metabolites derived from bitter fennel serve as flavoring and healing agents in the pharmaceutical, food, and cosmetic sectors. Previously, we developed three fennel breeding populations (Early, Medium, and Late Maturity Synthetic Cultivars) with the potential to produce higher seed yield. This investigation aimed to assess the quality and quantity of the important metabolites in these three cultivars alongside their elite parental populations. Our results showed that the highest amounts of essential oil yield and seed yield were produced by, two breeding populations, Medium and Late Maturity Synthetic Cultivars. These cultivars showed positive seed yield heterosis rates of 28% and 20%, respectively, above the averages of their parental populations. GC-MS analysis of the essential oil samples identified 24 components in total. The major compounds in all genotypes were trans-anethole (34.09–77.84%), limonene (4.99–12.66%), methyl chavicol (1.35–38.69%), and fenchone (6–13.07%). The highest amount of trans-anethole belonged to Fasa and the Medium Maturity Synthetic with 77.84 % and 73.10%, respectively. The analysis of fatty acid compositions in the genotypes was conducted using GC-FID, which revealed the presence of 19 unique fatty acids, of which ten were saturated, and nine were unsaturated fatty acids. Petroselinic acid (67.11–78.44%), and linoleic acid (9.14–11.60%) were the major unsaturated fatty acids in fennel seeds. The Early Maturity Synthetic exhibited the least saturated fatty acid content (7.24%), while the highest levels of unsaturated fatty acids were found in both the Early Maturity Synthetic (89%) and Hashtgerd (88.55%). The genotypes were categorized into four groups using a heat map analysis with hierarchical clustering. Overall, the results showed that the three synthetic cultivars of fennel have outperformed the elite parental populations for seed and essential oil yields, concentration of the major industrial secondary metabolites such as trans-anethole, and higher proportions of unsaturated fatty acids. These cultivars show potential for usage in the pharmaceutical and healthcare industries.

Bio-efficacy of certain new insecticides against pod borer complex in pigeonpea (Cajanus cajan L.)

A study on evaluation of certain new insecticides against pod borer complex in pigeonpea was conducted at Regional Agricultural Research Station, Warangal for three consecutive years from Kharif 2019-20 to Kharif 2021 22 in medium maturity pigeonpea cultivar ‘Rudreswara’ (WRG-65). Seven treatments including untreated control were evaluated for their relative efficacy towards suppression of pod borer larval population and reduction of percent pod damage. The insecticide treatments along with untreated control were applied two times during the crop season viz., 50% flowering and pod formation stage. The results revealed that chlorantraniliprole 18.5 SC @ 0.3 ml/L followed by emamectin benzoate 5 SG proved significantly superior against gram pod borer (Helicoverpa armigera) and spotted pod borer (Maruca vitrata) with highest mean percent larval reduction and cumulative least percent pod damage (10.8 and 14.7%, respectively) over control. Further, chlorantraniliprole 18.5 SC followed by emamectin benzoate 5 SG @ 0.4 g/l recorded higher grain yields and best ICBR ratio (1:4.6; 1:4.5). Thus, the effective chemicals observed in the study could be used as basis for selection of label recommended insecticides for successful management of pod borers in pigeonpea.

Meiotic and developmental competence of growing pig oocytes derived from small antral follicles is enhanced in culture medium containing FGF2, LIF, and IGF1 (FLI medium)

BackgroundOocytes of large animal species isolated from small ovarian follicles (< 2 mm) are less competent to support early embryonic development after in vitro maturation and fertilization than their counterparts isolated from medium-sized and preovulatory follicles. This study aimed to assess the effect of a new maturation medium containing FGF2, LIF, and IGF1 (FLI medium) on the meiotic and developmental competence of pig cumulus-oocytes complexes (COCs) derived from the small and medium-sized follicles.MethodsThe growing oocytes were isolated from 1 to 2 (small follicle; SF) and the fully-grown ones from 3 to 6 (large follicle; LF) mm follicles and matured in a control M199 medium with gonadotropins and EGF and the FLI medium enriched by the triplet of growth factors. The matured oocytes were parthenogenetically activated and cultured to the blastocyst stage. Chromatin configuration before and during the culture and MAP kinase activity were assessed in the oocytes. Finally, the expression of cumulus cell genes previously identified as markers of oocyte quality was assessed.ResultsThe maturation and blastocyst rates of oocytes gained from LF were significantly higher than that from SF in the control medium. In contrast, similar proportions of oocytes from LF and SF completed meiosis and developed to blastocysts when cultured in FLI. Most of the oocytes freshly isolated from SF possessed germinal vesicles with fine filaments of chromatin (GV0) or chromatin surrounding the nucleolus (GVI; 30%); the oocytes from LF were mainly in GVI (or GVII) exhibiting a few small lumps of chromatin beneath the nuclear membrane. When cultured in the FLI medium for 16 h, an acceleration of the course of maturation in oocytes both from SF and LF compared to the control medium was observed and a remarkable synchrony in the course of chromatin remodeling was noticed in oocytes from SF and LF.ConclusionsThis work demonstrates that the enrichment of culture medium by FGF2, LIF, and IGF1 can enhance the meiotic and developmental competence of not only fully-grown, but also growing pig oocytes and significantly thus expanding the number of oocytes available for various assisted reproductive technology applications.

The Impact of Follicular Fluid on the in vitro Maturation of Camel Oocytes

Background: The objective of this research is to find out how much of an impact adding camel follicular fluid (CFF) to camel oocytes in vitro maturation (IVM) medium has on getting meiosis back on track. Methods: Ovaries from camels were obtained at the central slaughterhouse in Riyadh, Kingdom of Saudi Arabia (KSA). Oocytes were extracted from follicles ranging in size from 2 to 8 mm. Camel oocytes were cultivated in a maturation medium containing varying amounts of camel follicular fluid: 0% (control), 2.5% and 5% for a duration of 32-36 hours. Result: According to the result, the maturation rate of oocytes was considerably (p.05) lower in the 2.5% CFF and 5% CFF groups compared to the control group. It is advised that greater CFF concentrations of camel oocytes be added to the IVM culture in order to improve nuclear and cytoplasmic maturation and boost oocyte competency.

Generation of Functional and Mature Sympathetic Neurons from Human Pluripotent Stem Cells via a Neuroepithelial Route.

The sympathetic nervous system (SNS) is a crucial branch of the autonomic nervous system (ANS) that is responsible for regulating visceral function and various physiological processes. Dysfunction of the SNS can lead to various diseases, such as hypertension and metabolic disorders. However, obtaining sympathetic neurons from human tissues for research is challenging. The current research aimed at recapitulating the process of human sympathetic neuron development and achieved the successful establishment of a stepwise, highly efficient in vitro differentiation protocol. This protocol facilitated the generation of functional and mature sympathetic neurons from human pluripotent stem cells (hPSCs) using a chemical-defined induction medium. Initially, each differentiation stage was refined to derive sympathoadrenal progenitors (SAPs) from hPSCs through neural epithelial cells (NECs) and trunk neural crest stem cells (NCSCs). hPSC-derived SAPs could be expanded in vitro for at least 12 passages while maintaining the expression of SAP-specific transcription factors and neuronal differentiation potency. SAPs readily generated functional sympathetic neurons (SymNs) when cultured in the neuronal maturation medium for 3-4 weeks. These SymNs expressed sympathetic markers, exhibited electrophysiological properties, and secreted sympathetic neurotransmitters. More importantly, we further demonstrated that hPSC-derived SymNs can efficiently regulate the adipogenesis of human adipose-derived stem cells (ADSCs) and lipid metabolism in vitro. In conclusion, our study provided a simple and robust protocol for generating functional sympathetic neurons from hPSCs, which may be an invaluable tool in unraveling the mechanisms of SNS-related diseases.

Accelerated startup of full-scale drinking water biofilters using mature media: Insights into microbial communities and manganese oxide coating

This study addressed the critical challenge of expediting biofilter commencement in groundwater treatment. By incorporating 8 cm-thick mature media, the start-up period was reduced to approximately 7 days. Significantly, NH4+-N and Mn2+ removal efficiencies reached 96.5% and 95.7%, with effluent concentrations at 0.07 mg/L and 0.04 mg/L, respectively. The loose and porous manganese oxide coating (MnOx) emerged as a pivotal factor in facilitating rapid biofilter maturation, providing attachment points for microorganisms. Analysis revealed variations in microbial communities associated with Fe2+ and Mn2+ removal between primary and secondary filters. The convergence of microbial communities in the mature and original medium of the secondary filter highlights the critical role of MnOx. Bacteria's essential contribution during the startup phase, driving MnOx formation, proves crucial for contaminant removal in the stable phase. This research provides concise insights into improving groundwater treatment system efficiency.

A multicriteria approach for assessing the maturity of supply chains regarding the implementation of circular economy practices in Brazil

ABSTRACT This study investigates the extent to which circular economy (CE) practices are adopted within supply chains in the Brazilian context. An expert survey was conducted to assess eight critical factors for the integration of CE practices in supply chains. The collected data was analyzed using a modified version of the Technique for Order Preference by Similarity to Ideal Solution (TOPSIS), specifically the Fuzzy TOPSIS class. Sensitivity analyses were performed to validate the robustness of the results. The findings indicated that practices related to economic and risk management, cleaner production, top management support, and knowledge management were categorized as having medium maturity. Conversely, practices associated with governance, product development, infrastructure, and social and cultural changes are still in an early stage. This research provides insights for organizations by identifying areas of strength and areas requiring attention in the adoption of CE practices within Brazilian supply chains. Policymakers can use the findings to design targeted policies and incentives that encourage and facilitate the development of circularity in supply chains, thus promoting CE principles. This research highlights the importance of addressing specific factors related to CE practices within supply chains, providing a framework for further investigations and strategies to bridge the gap between CE and supply chain management. The findings suggest that there are social and cultural challenges associated with the adoption of CE practices, implying a need for awareness campaigns, education, and community involvement to drive positive social change and adoption of CE principles.

Monitoring swine virus transmission in embryos derived from commercial abattoir oocytes.

Pigs are pivotal in agriculture and biomedical research and hold promise for xenotransplantation. Specific-pathogen-free (SPF) herds are essential for commercial swine production and xenotransplantation research facilities. Commercial herds aim to safeguard animal health, welfare, and productivity, and research facilities require SPF status to protect immunocompromised patients. Somatic cell nuclear transfer (SCNT) embryos are the norm for producing cloned and genetically edited animals. Oocytes for embryo reconstruction are most conveniently sourced from commercial abattoirs with unclear disease statuses. However, research on viral clearance from donor oocytes during embryo reconstruction remains limited. SCNT has previously been shown to reduce the transmission of Porcine reproductive and respiratory syndrome virus, Bovine viral diarrhea virus, Porcine Circovirus type 2, and Porcine parvovirus. Still, it is lacking for other pathogens, including endogenous viruses. This project contains two preliminary studies investigating the polymerase chain reaction (PCR) assay detection of common swine viruses through the phases of producing parthenogenic and SCNT embryos. Exogenous pathogens detected in oocyte donor tissue or the oocyte maturation media were not detected in the produced embryos. Porcine endogenous retrovirus type C (PERVC) was not removed by parthenogenic embryo activation and was detected in 1 of the 2 tested SCNT embryos reconstructed using a PERVC-negative cell line. SCNT and parthenogenic embryo construction similarly reduced exogenous virus detection. SCNT embryo construction helped reduce endogenous virus detection. This project demonstrates the importance of screening embryos for endogenous viruses and shows the usefulness of parthenogenic embryos in future exogenous virus clearance studies.

FGF2, LIF, and IGF-1 supplementation improves mouse oocyte in vitro maturation via increased glucose metabolism†.

Chemically defined oocyte maturation media supplemented with FGF2, LIF, and IGF-1 (FLI medium) enabled significantly improved oocyte quality in multiple farm animals, yet the molecular mechanisms behind such benefits were poorly defined. Here, we first demonstrated that FLI medium enhanced mouse oocyte quality assessed by blastocyst formation after in vitro fertilization and implantation and fetal development after embryo transfer. We then analyzed the glucose concentrations in the spent media; reactive oxygen species concentrations; mitochondrial membrane potential; spindle morphology in oocytes; and the abundance of transcripts of endothelial growth factor-like factors, cumulus expansion factors, and glucose metabolism-related genes in cumulus cells. We found that FLI medium enabled increased glucose metabolism through glycolysis, pentose phosphate pathway, and hexosamine biosynthetic pathway, as well as more active endothelial growth factor-like factor expressions in cumulus cells, resulting in improved cumulus cell expansion, decreased spindle abnormality, and overall improvement in oocyte quality. In addition, the activities of MAPK1/3, PI3K/AKT, JAK/STAT3, and mTOR signaling pathways in cumulus cells were assessed by the phosphorylation of MAPK1/3, AKT, STAT3, and mTOR downstream target RPS6KB1. We demonstrated that FLI medium promoted activations of all these signaling pathways at multiple different time points during in vitro maturation.

Dual isolation of primary neurons and oligodendrocytes from guinea pig frontal cortex.

Primary cell culture is a technique that is widely used in neuroscience research to investigate mechanisms that underlie pathologies at a cellular level. Typically, mouse or rat tissue is used for this process; however, altricial rodent species have markedly different neurodevelopmental trajectories comparatively to humans. The use of guinea pig brain tissue presents a novel aspect to this routinely used cell culture method whilst also allowing for dual isolation of two major cell types from a physiologically relevant animal model for studying perinatal neurodevelopment. Primary neuronal and oligodendrocyte cell cultures were derived from fetal guinea pig's frontal cortex brain tissue collected at a gestational age of 62 days (GA62), which is a key time in the neuronal and oligodendrocyte development. The major advantage of this protocol is the ability to acquire both neuronal and oligodendrocyte cellular cultures from the frontal cortex of one fetal brain. Briefly, neuronal cells were grown in 12-well plates initially in a 24-h serum-rich medium to enhance neuronal survival before switching to a serum-free media formulation. Oligodendrocytes were first grown in cell culture flasks using a serum-rich medium that enabled the growth of oligodendrocyte progenitor cells (OPCs) on an astrocyte bed. Following confluency, the shake method of differential adhesion and separation was utilized via horizontally shaking the OPCs off the astrocyte bed overnight. Therefore, OPCs were plated in 12-well plates and were initially expanded in media supplemented with growth hormones, before switching to maturation media to progress the lineage to a mature phenotype. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on both cell culture types to analyze key population markers, and the results were further validated using immunocytochemistry. Primary neurons displayed the mRNA expression of multiple neuronal markers, including those specific to GABAergic populations. These cells also positively stained for microtubule-associated protein 2 (MAP2; a dendritic marker specific to neurons) and NeuN (a marker of neuronal cell bodies). Primary oligodendrocytes expressed all investigated markers of the oligodendrocyte lineage, with a majority of the cells displaying an immature oligodendrocyte phenotype. This finding was further confirmed with positive oligodendrocyte transcription factor (OLIG2) staining, which serves as a marker for the overall oligodendrocyte population. This study demonstrates a novel method for isolating both neurons and oligodendrocytes from the guinea pig brain tissue. These isolated cells display key markers and gene expression that will allow for functional experiments to occur and may be particularly useful in studying neurodevelopmental conditions with perinatal origins.

ANIMAL COMPONENT-FREE WORKFLOWS TO DERIVE TENOCYTE-LIKE CELLS FROM HUMAN MESENCHYMAL STROMAL CELLS AND PLURIPOTENT STEM CELLS

Cell-based therapies offer a promising strategy to treat tendon injuries and diseases. Both mesenchymal stromal cells (MSCs) and pluripotent stem cells (PSCs) are good candidates for such applications due to their self-renewing and differentiation capacity. However, the translation of cell-based therapies from bench to bedside can be hindered by the use of animal-derived components in ancillary materials and by the lack of standardised media and protocols for in vitro tenogenic differentiation. To address this, we have optimized animal component-free (ACF) workflows for differentiating human MSCs and PSCs to tenocyte-like cells (TLCs) respectively. MSCs isolated from bone marrow (n = 3) or adipose tissue (n = 3) were expanded using MesenCult™-ACF Plus Culture Kit for at least 2 passages, and differentiated to TLCs in 21 days using a step-wise approach. Briefly, confluent cultures were treated with an ACF tenogenic induction medium for 3 days, followed by treatment with an ACF maturation medium for 18 days. Monolayer cultures were maintained at high density without passaging for the entire duration of the protocol, and the medium was changed every 2 – 3 days. In a similar fashion, embryonic (n = 3) or induced PSCs (n = 3) were first differentiated to acquire a mesenchymal progenitor cell (MPC) phenotype in 21 days using STEMdiff™ Mesenchymal Progenitor Kit, followed by the aforementioned tenogenic protocol for an additional 21 days. In all cases, the optimized workflows using ACF formulations consistently activated a tenogenic transcriptional program, leading to the generation of elongated, spindle-shaped tenomodulin-positive (TNMD+) cells and deposition of an extracellular matrix predominantly composed of collagen type I. In summary, here we describe novel workflows that can robustly generate TLCs from MSCs and hPSC-derived MPCs for potential translational applications.

Synergistic impact of α-linolenic acid and α-tocopherol on in vitro maturation and culture of buffalo oocytes.

The objective of the current study was to investigate the synergistic impact of α-Tocopherol and α-Linolenic acid (100 µM) on IVM and IVC of Nili Ravi buffalo oocytes. Oocytes were obtained from the ovaries of slaughtered buffaloes within two hours after slaughter and brought to laboratory. Buffalo cumulus oocyte complexes were placed randomly in the five experimental groups included; GROUP 1: Maturation media (MM) + 100 µM ALA (control), GROUP 2: MM + 100 µM ALA + 50μM α-Tocopherol, GROUP 3: MM + 100 µM ALA + 100μM α-Tocopherol, GROUP 4: MM + 100 µM ALA + 200 μM α-Tocopherol and GROUP 5: MM + 100 µM ALA + 300 μM α-Tocopherol under an atmosphere of 5% CO2 in air at 38.5 °C for 22-24 h. Cumulus expansion and nuclear maturation status was determined (Experiment 1). In experiment 2, oocytes were matured as in experiment 1. The matured oocytes were then fertilized in Tyrode's Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of α-Linolenic acid (100 µM) and α-Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of α-Linolenic acid (100 µM) in IVM media and increasing concentration of α-tocopherol in the culture media on early embryo development (Experiment 3), the presumptive zygotes were randomly distributed into the five experimental groups with increasing concentration of α-tocopherol in culture media. Higher percentage of MII stage oocytes in experiment 1(65.2±2.0), embryos at morula stage in experiment 2 (30.4±1.5) and experiment 3 (22.2±2.0) were obtained. However, overall results for cumulus cell expansion, maturation of oocyte to MII stage and subsequent embryo development among treatments remain statistically similar (P > 0.05). Supplementation of α-tocopherol in maturation media having α-Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.

Irrigation Response, Water Use, and Lint Yield of Upland Cotton Cultivars

Highlights In-season crop water use and canopy temperatures after first square did not differ among cultivars. Lint yield was significantly affected by cultivar in all study years. Cultivar effects on lint yield were significant only at higher irrigation levels. Crop water productivity increased with increasing irrigation in two of the three study years. Abstract. Upland cotton (Gossypium hirsutum L.) production requires less irrigation compared with other crops and thus provides an opportunity to reduce risk and maintain profitability in areas where water is limited. Water use, canopy temperature, lint yield, and crop water productivity were evaluated for four early to medium maturity upland cotton cultivars under three levels (100%, 66%, and 33%) of alternate furrow subsurface drip irrigation (SDI) in a thermally limited environment. Crop evapotranspiration (ET) across cultivars and years averaged 627, 547, and 467 mm for the 100%, 66%, and 33% irrigation levels, respectively, and did not differ among cultivars (P &amp;gt; 0.05). Changes in stored soil water within each irrigation level were similar among cultivars, with significant differences occurring infrequently. Measured canopy temperatures from first white flower to two weeks after cutout did not significantly differ among cultivars (P &amp;gt; 0.10) within each irrigation level. Crop water use during boll maturation, as inferred from the developed crop coefficient curve, was considerably less than reported by other studies, signifying that irrigation could be terminated earlier without reducing lint yield. The cultivar effect on lint yield was significant in all study years (P &amp;lt; 0.001), but only at the 66% and 100% irrigation levels, with one cultivar exceeding the average yields of all evaluated cultivars by 13% across the three study years. Medium maturity cultivars usually yielded less than early maturity cultivars, especially for a year with less accumulation of thermal energy. Crop selection and late season irrigation water management were both key to improving cotton water productivity. Keywords: Upland cotton, Subsurface drip irrigation, Water productivity, Root water uptake, Canopy temperature.

MicroRNAs associated with the quality of follicular fluids affect oocyte and early embryonic development.

Oocyte and embryo quality differs significantly among individuals. Follicular fluid (FF) is a solo environment of oocyte maturation and may flux into the oviduct. Supplementation of invitro maturation (IVM) and culture (IVC) medium with extracellular vesicles of FFs supports oocyte maturation and embryonic development. We addressed a hypothesis that miRNA profiles in FFs are crucial background of oocyte maturation and embryonic development. FFs were collected from the ovaries of individual cows, and the FFs were classified into Good or Poor FF based on the developmental rate to the blastocyst stage of enclosed oocytes. miRNAs associated with the Good FFs were explored using small RNA sequencing. In addition, FFs were classified using the concentration of Good-FF-associated miRNAs. These classified FFs or miRNA were added to the IVM or IVC mediums. Supplementation of IVM and IVC medium with Good FF improved embryonic development. Good FFs contained miR-151-3p and miR-425-5p at a high concentration compared with those in Poor FFs. FFs selected by the concentration of miR-151-3p and miR-425-5p improved oocyte maturation and embryonic development. Supplementation of IVM or IVC medium with either miR-151-3p or miR-425-5p improved embryonic development to the blastocyst stage. miRNAs were associated with the Good FFs determined oocyte maturation and embryonic development.

Adaptability of sunflower hybrids of the medium maturity group to abiotic factors in the conditions of Southern Ukraine

Adaptability of sunflower hybrids of the medium maturity group to abiotic factors in the conditions of Southern Ukraine

Registration of “Maya” Variety of Soybean [Glycine max (L.) Merrill)

Maya (PI-567190) soybean was developed by the Fedis Agricultural Research Center and was released in 2023. It is a high yielding medium maturity group variety adapted to the eastern Ethiopia particularly east Oromia. Then the trial was developed to evaluate and release stable high yielding and disease resistant/tolerant soybean variety for the east Oromia and other similar agro-ecologies. “Maya” soybean variety with pedigree of PI-567190 was developed through selection. It was evaluated at three locations; Fedis, Mechara and Babile for two years (2021 and 2022). Similarly, during verification trial “Maya” was evaluated with two standard checks (Awasa 04 and SC-status) and one candidate variety (PI-230970). “Maya” recorded 26.22 % of oil content while recently released variety “SC-STATUS” recorded 25.23. It has high oil contents that commercial variety Awasa-04 and recently released variety SC- STATUS. “Maya” soybean variety had higher seed yield and better stability performances with comparable oil content. It is resistant to common soybean diseases viz., bacterial blight, rust and bacterial pustule than commercial variety Awasa-04.