Lab partners: oocytes, embryos and company. A personal view on aspects of oocyte maturation and the development of monozygotic twins

The present review addresses the oocyte and the preimplantation embryo, and is intended to highlight the underlying principle of the "nature versus/and nurture" question. Given the diversity in mammalian oocyte maturation, this review will not be comprehensive but instead will focus on the porcine oocyte. Historically, oogenesis was seen as the development of a passive cell nursed and determined by its somatic compartment. Currently, the advanced analysis of the cross-talk between the maternal environment and the oocyte shows a more balanced relationship: Granulosa cells nurse the oocyte, whereas the latter secretes diffusible factors that regulate proliferation and differentiation of the granulosa cells. Signal molecules of the granulosa cells either prevent the precocious initiation of meiotic maturation or enable oocyte maturation following hormonal stimulation. A similar question emerges in research on monozygotic twins or multiples: In Greek and medieval times, twins were not seen as the result of the common course of nature but were classified as faults. This seems still valid today for the rare and until now mainly unknown genesis of facultative monozygotic twins in mammals. Monozygotic twins are unique subjects for studies of the conceptus-maternal dialogue, the intra-pair similarity and dissimilarity, and the elucidation of the interplay between nature and nurture. In the course of in vivo collections of preimplantation sheep embryos and experiments on embryo splitting and other microsurgical interventions we recorded observations on double blastocysts within a single zona pellucida, double inner cell masses in zona-enclosed blastocysts and double germinal discs in elongating embryos. On the basis of these observations we add some pieces to the puzzle of the post-zygotic genesis of monozygotic twins and on maternal influences on the developing conceptus.(AU)

Occurrence of intersexuality in Mugil curema from an estuary of the North coast of Santa Catarina, Brazil - case report / Ocorrência de intersexualidade em Mugil curema em estuário do litoral norte catarinense, Brasil - relato de caso

This study describes the occurrence of reproductive changes in a mullet (Mugil curema) from the Babibtonga Bay, Santa Catarina. Gross and microscopic findings indicated changes in the reproductive system that were characterized by the presence of both male and female gonads, nearing sexual maturity, which was compatible with the diagnosis of intersex. It was not possible to identify predisposing factors that may have contributed to the development of this condition. However, as this species of fish may have their reproductive development affected by anthropogenic influence, it is important to permanently monitor this kind of environment. Considering the number of fish belonging to this species that was examined (n=433), the estimated prevalence for intersexuality was 0.23% (confidence interval: 0.00 to 0.68%, with confidence level of 95%). The findings in this study support the diagnosis of intersexuality in M. curema from the Babitonga Bay, which is the first documented case of this condition in this species on the coast of Santa Catarina, Brazil.

Este estudo descreve a ocorrência de alteração reprodutiva em peixe da espécie Mugil curema, procedente da Baía da Babitonga, Santa Catarina. Macro e microscopicamente, foi identificada alteração no sistema reprodutivo, caracterizada pela presença de gônadas masculina e feminina, próximas da maturidade sexual, caracterizando um caso de intersexo. Não foi possível determinar os fatores que possam ter contribuído para o desenvolvimento dessa alteração reprodutiva, porém, como a espécie animal pode ter suas características de vida e reprodutiva influenciadas pela ação antrópica, é importante um estudo contínuo desses peixes nesse ambiente. Considerando o número de peixes dessa espécie que foram examinados (n=433), a prevalência estimada de intersexualidade foi de 0,23% (intervalo de confiança: 0,00 a 0,68%, com nível de confiança de 95%). Os achados deste estudo suportam o diagnóstico de intersexualidade em M. curema proveniente da Baía da Babitonga, sendo o primeiro caso documentado dessa condição nessa espécie na costa de Santa Catarina, Brasil.

Analysis of nuclear maturation, DNA damage and repair gene expression of bovine oocyte and cumulus cells submitted to ionizing radiation'

Radiotherapy causes destruction of tumor cells, but also threatens the integrity and survival of surrounding normal cells. Then, woman submitted to irradiation for cancer treatment may present permanent ovary damage, resulting in impaired fertility. The objective of this study was to investigate the effects of therapeutic doses of ionizing radiation (IR), used for ovarian cancer treatment in humans, on bovine cumulus-oocyte complexes (COCs) as experimental model. Bovine ovaries were exposed to 0.9 Gy, 1.8 Gy, 3.6 Gy or 18.6 Gy IR, and then COCs were collected and used to evaluate: (a) oocyte nuclear maturation; (b) presence of phosphorylated H2A.X (γH2AX), as an indicator of DNA double-strand breaks (DSBs); and (c) expression of genes involved in DNA repair (TP53BP1, RAD52, ATM, XRCC6 and XRCC5) and apoptosis (BAX). The radiation doses tested in this study had no detrimental effects on nuclear maturation and did not increase γH2AX in the oocytes. However, IR treatment altered the mRNA abundance of RAD52 (RAD52 homolog, DNA repair protein) and BAX (BCL2-associated X protein). We conclude that although IR doses had no apparent effect on oocyte nuclear maturation and DNA damage, molecular pathways involved in DNA repair and apoptosis were affected by IR exposure in cumulus cells.(AU)

Insulin-like growth factor-1 improves in vitro meiotic resumption of dromedary camel (Camelus Dromedarius) oocytes

Despite relatively high maturation rate of in vitro matured oocytes in the dromedary camel, however, blastocyst production is very low after in vitro fertilization (IVF). Herein, the influences of oocyte collection method (follicular aspiration vs slicing; Experiment I), the addition of Insulin-like growth factor I (IGF-I) to the maturation medium (Experiment II) on in vitro maturation (IVM) of oocyte were investigated. Although the nuclear maturation did not differ regardless of collecting method, follicular aspiration led to lower degeneration rates than those in controls (P < 0.05). The percentages of oocytes at MII were greater in the presence of IGF-1 than in its absence (71.9% vs 48.4%, respectively, P<0.05). Additionally, the percentages of degenerated oocytes were higher in the control group compared to oocytes cultured in the presence of IGF-I (23.6% vs 10.4%, respectively, P<0.05). IGF-I treatment improved the quality of MII matured oocytes as evidenced by the decrease of cathepsin B (CTSB) activity, a marker of poor quality oocytes, when compared to control ones (P < 0.05). In conclusion, follicular aspiration decreased the degeneration rate; however, it had no effect on completion of maturation. IGF-I enhanced the IVM of oocyte and decreased degeneration rate.(AU)

Impacto de diferentes métodos de poda da videira 'Niágara Branca' cultivada na região do Planalto Norte Catarinense / Impact of different pruning methods of the 'Niágara Branca' vine cultivated in the region of the Planalto Norte Catarinense

O objetivo deste trabalho foi avaliar o efeito de dois métodos de poda invernal nos índices produtivos e na qualidade da uva 'Niágara Branca' cultivada na região do Planalto Norte Catarinense. O experimento foi realizado nas safras 2019 e 2020, em vinhedo da variedade Niágara Branca, situado em Canoinhas,SC. Os tratamentos consistiram em dois diferentes sistemas de poda: poda curta, deixando-se esporões com duas gemas, e poda mista, deixando-se varas com quatro gemas e esporões contendo duas gemas para renovação no ano seguinte. Avaliou-se índices vegetativos, produtivos, arquitetura de cachos e maturação tecnológica. O método de poda apresenta influência nas características produtivas e qualitativas da videira 'Niágara Branca', sendo que a adoção da poda mista resulta em maior número de ramos planta-¹, maior número de cachos planta-1, maior massa de cacho-1, maior número de bagas cacho-1, o que consequentemente propicia incremento dos índices produtivos. Plantas submetidas a poda mista, produção média de 5,5 e 11,2 kg planta-1, nas safras 2018 e 2019, respectivamente, enquanto plantas submetidas à poda curta obtiveram produção média de 1,9 e 7,2 kg planta-1. Desta forma a poda mista é recomendada para a videira 'Niágara Branca' cultivada na região do Planalto Norte Catarinense, sem que ocorra comprometimento da qualidade da uva.(AU)

The objective of this work was to evaluate the effect of two winter pruning methods on yield rates and quality of the 'Niágara Branca' grape grown in the Planalto Norte region of Santa Catarina. The experiment was carried out in the 2019 and 2020 harvests, in a vineyard on the Niágara Branca variety, located in Canoinhas. The treatments consisted of two different pruning systems: short pruning, leaving spurs with two buds, and mixed pruning, leaving canes with four buds and spurs containing two buds for renewal in the following year. Vegetative and productive indices, cluster architecture and technological maturation were evaluated. The pruning method has an influence on the productive and qualitative characteristics of the Niagara Branca vine, and the adoption of mixed pruning results in a greater number of plant-1branches, a greater number of plant-1bunches, a greater mass of bunch-1,a greater number of berries cluster-1, and consequently provides an increase in production rates.Plants produced mixed pruning, an average of 5.5 and 11.2 kg plant-1, in the 2018 and 2019 harvests, respectively, while plants supplied to short pruning obtained an average production of 1.9 and 7.2 kg plant-1. which is recommended for the 'Niágara Branca' vine grown in the Planalto Norte region of Santa Catarina, without compromising the quality of the grape.(AU)

Composição de cana-de-açúcar para alimentação animal em função da aplicação de bactérias diazotróficas / Composition of sugarcane for animal feeding with the application of diazotrophic bacteria

A cana-de-açúcar pode ser empregada na alimentação animal devido ao seu alto rendimento de matéria seca. No entanto, há muitas recomendações para evitar o emprego como alimento para animais de bom potencial genético, pois apresenta baixo teor proteico e de minerais, bem como e fibra de baixa qualidade. Desta forma, o objetivo deste trabalho foi avaliar os efeitos da aplicação de uma mistura de cinco bactérias diazotróficas (Gluconacetobacter diazotrophicus; Herbaspirillum seropedicae; Herbaspirillum rubrisubalbicans; Paraburkholderia tropica; Nitrospirillum amazonense) e da adubação nitrogenada sobre a produção, maturação e composição químico-bromatológica de cinco variedades de cana-de-açúcar com potencial para serem empregadas na alimentação animal. Os tratamentos foram formados conforme a combinação dos cinco genótipos de cana-de-açúcar (RB867515, RB966928, RB106814, RB106818 e RB106819) com quatro condições de inoculação e adubação nitrogenada em cobertura. Os teores de sólidos solúveis totais foram determinados dos 180 dias após o transplante até a colheita do experimento, onde foi avaliada a produtividade e a amostragem de material foi coletada para as avaliações químico-bromatológicas: matéria seca, nitrogênio acumulado, proteína bruta, fibra em detergente neutro, fibra em detergente ácido, holocelulose, celulose, hemicelulose, lignina, extrativos totais e cinzas. De acordo com os resultados deste estudo, conclui-se que a aplicação de bactérias diazotróficas de forma isolada ou em conjunto com a adubação nitrogenada, aumenta a produtividade e os sólidos solúveis totais, antecipando a maturação, além de melhorar a composição químico-bromatológica das plantas de cana-de-açúcar.

Sugarcane can be used in animal feed due to its high dry matter yield. However, there are many recommendations to avoid its use as food for animals with good genetic potential, since it has a low contents of protein and minerals, as well as fiber of low quality. Therefore, this work aimed to evaluate the effects of applying a mixture of five diazotrophic bacteria (Gluconacetobacter diazotrophicus; Herbaspirillum seropedicae; Herbaspirillum rubrisubalbicans; Paraburkholderia tropica; Nitrospirillum amazonense) and nitrogen fertilization on the production, maturation, and chemical-bromatological composition of five sugarcane varieties with potential to be used in animal feed. The treatments were formed according to the combination of the five sugarcane genotypes (RB867515, RB966928, RB106814, RB106818, and RB106819) with four conditions of inoculation and nitrogen fertilization in coverage. The levels of total soluble solids were determined from 180 days after transplantation until the harvest of the experiment, where productivity was evaluated and the sample material was collected for chemical-bromatological evaluations: dry matter, accumulated nitrogen, crude protein, neutral detergent fiber, acid detergent fiber, holocellulose, cellulose, hemicellulose, lignin, total extracts, and ashes. According to the results of the present study, it is concluded that the application of diazotrophic bacteria alone or in conjunction with nitrogen fertilization, increases productivity and total soluble solids, anticipating maturation, in addition to improving the chemical-bromatological composition of sugarcane plants.

Migratory fishes from rivers to reservoirs: seasonal and longitudinal perspectives

Migratory fishes have high ecological, social, and commercial value, and are strongly affected by river regulation. This study aimed to understand how migratory fishes use the longitudinal gradient, in an upstream to downstream direction of two free-flowing rivers and two reservoirs in a cascade within the Upper Grande River Basin, Brazil. The numeric abundance, biomass, richness, presence of fingerlings and juveniles, and the macroscopic gonadal maturation stage of migratory fishes were ascertained and evaluated bimonthly for two years. Recruitment in migratory fishes seems to rely completely on the free-flowing rivers upstream and their floodplains. Transition areas do not seem to have a significant role in recruitment. Therefore, we highlight the importance of maintaining the free flow of rivers and the integrity of their floodplains.

Population dynamics, vertical distribution and damage characterization of burrower bug in peanut

Although the burrower bug (Cyrtomenus mirabilis) is considered a vitalsoil pest in peanut crops, Arachis hypogaea L., in South America, there is little information on its occurrence and damage characterization. This study aimed to evaluate the vertical distribution and fluctuation of the burrower bug in the soil and the damage this species causes in peanuts. Two peanut cultivars (IAC OL3 and IAC 503) were evaluated in three locations in the state of São Paulo: Pindorama, Ribeirão Preto and Votuporanga, in the 2017/18 and 2018/19 harvests. Trenches were dug 0.5 m long, 0.3 m wide and 0.3 m deep, along plant lines at four spots on each sampling date, and stratified in layers 10 cm deep. Plant samples (0.5 m) were collected and evaluated for number of pods and percentage of kernels with symptoms of insect damage. More than 85 % of the C. mirabilis population was found in depths of up to 10 cm, especially after plant fructification, and an increase in nymphs from 100 days after sowing (DAS) was seen when an increased number of maturing pods was observed. The occurrence of nymphs and adults of C. mirabilis and their damage to peanut kernels was similar in both cultivars (IAC OL3 and IAC 503), when these were harvested according to their developmental cycles.

Detection of coffee fruits on tree branches using computer vision

Coffee farmers do not have efficient tools to have sufficient and reliable information on the maturation stage of coffee fruits before harvest. In this study, we propose a computer vision system to detect and classify the Coffea arabica (L.) on tree branches in three classes: unripe (green), ripe (cherry), and overripe (dry). Based on deep learning algorithms, the computer vision model YOLO (You Only Look Once), was trained on 387 images taken from coffee branches using a smartphone. The YOLOv3 and YOLOv4, and their smaller versions (tiny), were assessed for fruit detection. The YOLOv4 and YOLOv4-tiny showed better performance when compared to YOLOv3, especially when smaller network sizes are considered. The mean average precision (mAP) for a network size of 800 × 800 pixels was equal to 81 %, 79 %, 78 %, and 77 % for YOLOv4, YOLOv4-tiny, YOLOv3, and YOLOv3-tiny, respectively. Despite the similar performance, the YOLOv4 feature extractor was more robust when images had greater object densities and for the detection of unripe fruits, which are generally more difficult to detect due to the color similarity to leaves in the background, partial occlusion by leaves and fruits, and lighting effects. This study shows the potential of computer vision systems based on deep learning to guide the decision-making of coffee farmers in more objective ways.

The effect of Cissus quadrangularis Salisb. extract on maturation of rat mesenchymal stem cells / O efeito do extrato de Cissus quadrangularis Salisb. na proliferação e diferenciação de células-tronco mesenquimais de rato

Degenerative diseases, such as osteoporosis, could be treated by stem cells. The aim of this study was to identify the gene expression of bone marrow mesenchymal stem cells (BM-MSC) derived from Sprague Dawley rats and to assess the effect of Cissus quadrangularis Salisb. extract on their maturation into bone cells. The BM-MSC were divided into three groups: (a) BM-MSCs + osteoblast cell growth basal medium as the positive control; (b) BM-MSCs + Dulbecco's modified eagle's medium (DMEM) + 0.3 mg/mL methanol extract of C. quadrangularis as methanol group; and (c) BM-MSC + DMEM + 0.3 mg/mL ethyl acetate extract of C. quadrangularis as ethyl acetate group. A relative quantification approach using was used to analyze the expression of the alp (alkaline phosphatase) gene, with the beta-actin gene was used to normalize the expression of the alp gene. The intra-assay variation was calculated to validate the RT-qPCR data. Our study found that the intra-assay variation value was acceptable, with most of the coefficients of variability (CV) value <5. Ethyl acetate solvent outperformed methanol solvent in extracting the active compound C. quadrangularis. In the ethyl acetate extract group, the expression of the alp gene increased three times compared to the positive control. In methanol extract group, the expression of alp gene was lower six times compared to positive control. This study suggests that C. quadrangularis extracts using ethyl acetate could induce the maturation of BM-MSCs. However, further studies are warrant to confirm this effect using different indicators.

Doenças degenerativas, como a osteoporose, podem ser tratadas por células-tronco. O objetivo deste estudo foi identificar a expressão gênica de células-tronco mesenquimais da medula óssea (BM-MSCs) derivadas de ratos Sprague Dawley, bem como sua proliferação e diferenciação em células ósseas afetadas pelo extrato de Cissus quadrangularis Salisb. As BM-MSCs foram divididas em três grupos: (1) BM-MSCs + meio basal de crescimento de osteoblastos como controle positivo; (2) BM-MSCs + meio de Eagle modificado por Dulbecco (DMEM) + extrato de metanol; e (3) BM-MSCs + DMEM + extrato de acetato de etila de C. quadrangularis. Uma abordagem de quantificação relativa foi usada para analisar a expressão do gene alp (fosfatase alcalina), com o gene da beta-actina sendo usado para normalizar a expressão do gene alp em cada tratamento. A variação intraensaio foi calculada para validar os dados de RT-qPCR. Os resultados mostraram que o valor da variação intraensaio é aceitável, com a maioria dos valores dos coeficientes de variabilidade (CV) < 5. Além disso, no tratamento T2, a expressão do gene alp aumentou três vezes em relação ao controle positivo. No tratamento T1, o gene alp apresentou nível de expressão seis vezes menor em comparação com o controle positivo. Além disso, o solvente acetato de etila superou o solvente metanol na extração do composto ativo C. quadrangularis Salisb. Este estudo demonstra que extratos de C. quadrangularis Salisb., particularmente com acetato de etila como solvente, podem induzir a proliferação e diferenciação de BM-MSCs. Com base na expressão do gene alp, C. quadrangularis tem o potencial de corrigir a perda óssea.

Early maturation of corpus luteum in rabbits - effect of sildenafil citrate on luteolytic capacity in the early luteal period

Background: Prostaglandin F2 alpha (PGF2 α) binds to the specific receptor (PTGFR) on the corpus luteum (CL) in mammals, inducing regression of the CL structure (luteolysis) and initiating a new cycle. While PGF2 α is effective only on mature CL, the immature CL structure (early luteal phase) resists PGF2 α. In this study, sildenafil citrate, which is used to increase blood flow in the genital organs for treating specific pregnancy issues in women, was administered during the early luteal phase in a rabbit model to test the hypothesis of enhancing blood flow to the CL, thereby promoting earlier maturation and enabling a response to PGF2 α. Materials, Methods & Results: The study was conducted in 2 sub-studies: clinical and molecular. A large number of rabbits were initially included in the sub-studies to ensure a sufficient number of pseudo-pregnant rabbits. Ovulation in rabbits was induced with buserelin acetate and was considered as day 0 of the study. The sub-studies were continued with rabbits whose pseudo-pregnancies were confirmed according to progesterone (P4 ) results. As a result, the studies were continued with a total of 41 pseudo-pregnant New Zealand female rabbits, 21 of which were included in the clinical sub-study and 20 in the molecular sub-study. In both sub-studies, on day 3 of the luteal period, rabbits in the treatment group received 5 mg/kg sildenafil citrate and all rabbits received a single dose of exogenous PGF2 α on day 4 to induce luteolysis. In the clinical sub-study, echotexture and intraovarian blood flow changes in the ovaries were determined by ultrasonography (USG) examination. In the molecular sub-study, the expression changes of Hypoxia Inducible Factor 1 Alpha (HIF1A) and Vascular Endothelial Growth Factor (VEGF) related to angiogenesis, Steroidogenic Acute Regulatory Protein (StAR) related to P4 metabolism, Prostaglandin-Endoperoxide Synthase 2 (PTGS2) related to prostaglandin (PG) mechanism and 15-Hydroxyprostaglandin Dehydrogenase (HPGD) genes at mRNA level were determined using Real Time Polymerase Chain Reaction (RT-PCR) in CL tissues obtained with ovariohysterectomy (OVH) at 1 and 12 h after PGF2 α injection. In addition, blood samples were collected for determine P4 levels from all rabbits. In the clinical sub-study; there was no difference between the groups in mean gray values (MGV), whereas there was a significant decrease in both pulsatile index (PI) and resistance index (RI) values at 40 min after PGF2 α injection (P < 0.05). In the molecular sub-study, it was determined that sildenafil citrate had no significant effect (P > 0.05) on the expression levels 1 and 12 h after PGF2 α injection. According to the results of the molecular sub-study, no significant effect of sildenafil citrate on the mRNA expression levels in the investigated genes was detected (P > 0.05). However, within each group, differences were found according to OVH time after PGF2 α injection. It was observed that PTGS2 and HPGD mRNA expressions decreased at the 12th h compared to the 1st h, while HIF1A expression increased (P < 0.05). Discussion: According to the results obtained from clinical and molecular sub-studies, it was determined that a single dose of sildenafil citrate (5 mg/kg) applied on the 3rd day of the luteal period did not contribute to the maturation process of the CL, did not increase blood flow, and was insufficient to break the resistance of the CL against PGF2 α applied on the 4th day of the luteal period. However, a significant decrease in the PI value at the 40th min after PGF2 α injection suggests that sildenafil citrate has a supportive effect, and that this decrease is also seen in the RI value, suggesting that its effect is insufficient against the vasoconstrictive effect of PGF2 α.

Characterization of colonial cheese: appreciation a traditional product from southern Brazil / Caracterização do queijo colonial: valorização de um produto tradicional do Sul do Brasil

South Brazil produces colonial cheese based on Italian immigration recipes. This study characterized colonial cheese produced in the state of Rio Grande do Sul. The sampling method was by conglomerate. A total of 293 rural producers were interviewed; they also provided cheeses for physicochemical and microbiologicanalyses. For the characterization of colonial cheese, parameters related to milk, processes, social aspects, physical aspects and ingredients were selected and cluster analysis was performed. Results showed that the colonial cheese is made with whole milk, rennet and salt, with a round shape, weighing an average of 1.22 kg, with an average maturation of 9.4 days, high moisture, fat, using raw or pasteurized milk. The recipe is familiar, passed down through generations and the sale is made directly to the consumer.

O Sul do Brasil produz queijo colonial com base nas receitas da imigração italiana. O objetivo deste estudo foi promover a valorização do queijo colonial produzido no estado do Rio Grande do Sul, por meio de sua caracterização. O método de amostragem foi por conglomerado. Foram entrevistados 293 produtores rurais; também forneceram queijos para análises físico-químicas. Para a caracterização do queijo colonial, foram selecionados parâmetros relacionados ao leite, processos, aspectos sociais, aspectos físicos e ingredientes e realizada análise de agrupamento. Os resultados mostram que o queijo colonial é feito com leite integral, coalho e sal, de formato redondo, pesando em média 1,22 kg, com maturação média de 9,4 dias, de alta umidade, gordo, utilizando leite cru ou pasteurizado. A receita é familiar, passada de geração em geração e a venda é feita diretamente ao consumidor.

Corn productivity and topdressing application cost of nitrogen fertilizers / Produtividade de milho e custo da aplicação em cobertura de fertilizantes nitrogenados

Nitrogen(N) is the most required nutrient in the cultivation of corn (Zea maysL.), also representing a significantshare in the production costs. The objective was to evaluate the efficiency for one experimental year of different sourcesof nitrogen fertilizers applied in topdressing on the corn grain yield and their respective fertilization costs. The experiment was carried out under field conditions and without irrigation under no-tillage system, in a Red Oxissol. For sowing, 280 kg ha-1of NPK fertilizer 12-30-20 was used for all treatments. In addition to the control treatment (NPK), four nitrogen formulations wereapplied in topdressing: Urea, Super N, Nitromag, and Sulfammo, both formulations applied 50% in the V3/V4 and 50% V7/V8 stages, in four replicates.Grain yield was determined at the physiological maturation. The type of nitrogen fertilizer did not influence corn grain yield, an average of 9016 kg ha-1. The largest profits were achieved with the application of Super N, Urea and Nitromag. Nitrogen fertilizers with slow and controlled N release does not promote an increase in corn grain yield in relation to Urea. The use of Super N in topdressing has the highest profitability in corn cultivation.(AU)

O nitrogênio (N) é o nutriente mais requerido no cultivo do milho (Zea maysL.), ainda representa significativa participação no custo de produção. O objetivo foi avaliar a eficiência em um ano experimental de diferentes fontes de fertilizantes nitrogenados aplicados em cobertura na produtividade de grãos demilho e os seus respectivos custos da adubação. O experimento foi realizado em condições de campo e sem irrigação sob sistema plantio direto, em Latossolovermelho. Na semeadura foi utilizado 280 kgha-1de adubo NPK 12-30-20 para todos os tratamentos. Além do tratamento testemunha (NPK), quatro formulações nitrogenadas foram aplicadas em cobertura: Ureia, Super N, Nitromag, e Sulfammo, ambas as formulaçõesaplicadas 50% no estádioV3/V4 e 50% V7/V8,em quatro repetições. Na maturação fisiológica foi determinada a produtividade de grãos.O tipo de fertilizante nitrogenado não influenciou na produtividade de grãos de milho, média de 9016 kg ha-1. Os maiores lucros foram com a aplicação de Super N, Ureia e Nitromag. O uso em cobertura de fertilizantes nitrogenados de lenta e controlada liberação de N não promovem aumento na produtividade de grãos de milho em relação a Ureia. O uso de Super N em cobertura apresenta a maior lucratividadeno cultivo do milho.(AU)

Investigation of Egg Weight, Ovarian Follicles Morphology and Growth Differentiation Factor 9 mRNA Expression in Potchefstroom Koekoek Chicken Breed

The growth differentiation factor 9 (GDF9) gene plays a vital role in the growth and maturation of ovarian follicles in laying hens. However, its messenger ribonucleic acid (mRNA) expression levels in preovulatory ovarian follicles of indigenous chickens remain poorly understood. The study aimed to identify the association between egg weight and egg quality traits, ovarian follicles morphology, and mRNA expression levels of the GDF9 gene in pre-ovulatory ovarian follicles of the South African Potchefstroom Koekoek chicken breed. The correlation results showed that egg weight (EW) had a positively high significant correlation (p<0.01) with egg width (EWD), yolk weight (YW), shell surface area (SSA), albumen weight (AW), albumen ratio (AR) and egg volume (EV), and a positive significant correlation (p<0.05) with egg length (EL). The Student's T-test results revealed that the numbers of large yellow follicles were significantly lower (p<0.05) than those of small yellow follicles. ANOVA findings showed that there was a significant difference (p<0.05) in the average weight of the large yellow follicles. The quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR) findings indicated that there were significant differences (p<0.05) in the mRNA expression levels of the GDF9 gene in preovulatory ovarian follicles of the Potchefstroom Koekoek chicken breed. The mRNA expression was more abundant in F1 and F4 than in other ovarian follicles.(AU)

Molecular characteristics and transcriptional regulatory of spermatogenesis-related gene RFX2 in adult Banna mini-pig inbred line (BMI)

RFX2 plays critical roles in mammalian spermatogenesis and cilium maturation. Here, the testes of 12-month-old adult boars of Banna mini-pig inbred line (BMI) were subjected to whole-transcriptome sequencing. The results indicated that the average expression (raw count) of RFX2 gene in BMI testes was 16138.25, and the average expression value of the corresponding transcript ENSSSCT00000043271.2 was 123.1898. The CDS of RFX2 obtained from BMI testes was 2,817 bp (GenBank accession number: OL362242). Gene structure analysis showed that RFX2 was located on chromosome 2 of the pig genome with 19 exons. Protein structure analysis indicated that RFX2 contains 728 amino acids with two conserved domains. Phylogenetic analysis revealed that RFX2 was highly conserved with evolutionary homologies among mammalian species. Other analyses, including PPI networks, KEGG, and GO, indicated that BMI RFX2 had interactions with 43 proteins involving various functions, such as in cell cycle, spermatid development, spermatid differentiation, cilium assembly, and cilium organization, etc. Correlation analysis between these proteins and the transcriptome data implied that RFX2 was significantly associated with FOXJ1, DNAH9, TMEM138, E2F7, and ATR, and particularly showed the highest correlation with ATR, demonstrating the importance of RFX2 and ART in spermatogenesis. Functional annotation implied that RFX2 was involved in 17 GO terms, including three cellular components (CC), six molecular functions (MF), and eight biological processes (BP). The analysis of miRNA-gene targeting indicated that BMI RFX2 was mainly regulated by two miRNAs, among which four lncRNAs and five lncRNAs competitively bound ssc-miR-365-5p and ssc-miR-744 with RFX2, respectively. Further, the dual-luciferase report assay indicated that the ssc-miR-365-5p and ssc-miR-744 significantly reduced luciferase activity of RFX2 3'UTR in the 293T cells, suggesting that these two miRNAs regulated the expression of RFX2. Our results revealed the important role of RFX2 in BMI spermatogenesis, making it an intriguing candidate for follow-up studies.(AU)

Comparison of efficiency between two artificial insemination methods using fresh semen in domestic cat (Felis Catus) / Comparação da eficiência entre dois métodos de inseminação artificial utilizando sêmen fresco em gata doméstica (Felis Catus)

This study aimed to compare domestic cats' pregnancy rates using fresh semen for the bilateral intrauterine insemination (BIUI) method and the novel uterine body insemination (UBI) method. Queens received a single injection of equine chorionic gonadotropin (eCG) (200 IU; IM) to induce ovarian follicular development and, after 83 h, an injection of human chorionic gonadotropin (hCG) (100 IU; IM) for final oocyte maturation and ovulation induction. Thirty-four hours after hCG administration, 3 × 106fresh spermatozoa were used for insemination by the BIUI (n = 8 queens) or by the UBI (n = 7 queens) techniques,respectively. Pregnancy rates were 75.00% (6/8) by BIUI and 42.85% (3/7) by the UBI method. Themean litter size was 3.0 ± 0.86 for the BIUI, and 2.0 ± 1.0 for the UBI method. Spontaneous abortion occurred on day 35 of pregnancy in one queen following the UBI method. Our findings showed that the BIUI of queens with fresh semen resulted in higher pregnancy rates than the novel UBI method; also, acceptable pregnancy rates were achieved following BIUI with fresh semen in the domestic cat.(AU)

O objetivo deste estudo foi comparar as taxas de prenhez em gatas domésticas usando sêmen fresco para o método de inseminação intrauterina bilateral (BIUI) e o novo método de inseminação do corpo uterino (UBI). As gatas receberam uma única injeção de gonadotrofina coriônica equina (eCG) (200 UI; IM) para induzir o desenvolvimento folicular ovariano e, após 83 h, uma injeção de gonadotrofina coriônica humana (hCG) (100 UI; IM) para maturação final do oócito e ovulação indução. Trinta e quatro horas após a administração de hCG, 3 × 106 espermatozoides frescos foram utilizados para inseminação pelas técnicas de BIUI (n = 8 gatas) ou UBI (n = 7 gatas), respectivamente. As taxas de gravidez foram de 75,00% (6/8) pela BIUI e 42,85% (3/7) pelo método UBI. O tamanho médio da ninhada foi de 3,0 ± 0,86 para o método BIUI e 2,0 ± 1,0 para o método UBI. Aborto espontâneo ocorreu no dia 35 de gestação em uma gata seguindo o método UBI. Nossos achados mostraram que a BIUI de gatas com sêmen fresco resultou em maiores taxas de prenhez do que o novo método UBI; também, taxas de prenhez aceitáveis foram alcançadas após BIUI com sêmen fresco no gato doméstico.(AU)

Maternal contributions to pregnancy success: from gamete quality to uterine environment

The establishment and maintenance of a pregnancy that goes to term is sine qua non for the long-term sustainability of dairy and beef cattle operations. The oocyte plays a critical role in providing the factors necessary for preimplantation embryonic development. Furthermore, the female, or maternal, environment where oocytes and embryos develop is crucial for the establishment and maintenance of a pregnancy to term. During folliculogenesis, the oocyte must sequentially acquire meiotic and developmental competence, which are the results of a series of molecular events preparing the highly specialized gamete to return to totipotency after fertilization. Given that folliculogenesis is a lengthy process in the cow, the occurrence of disease, metabolic imbalances, heat stress, or other adverse events can make it challenging to maintain oocyte quality. Following fertilization, the newly formed embryo must execute a tightly planned program that includes global DNA remodeling, activation of the embryonic genome, and cell fate decisions to form a blastocyst within a few days and cell divisions. The increasing use of assisted reproductive technologies creates an additional layer of complexity to ensure the highest oocyte and embryo quality given that in vitro systems do not faithfully recreate the physiological maternal environment. In this review, we discuss cellular and molecular factors and events known to be crucial for proper oocyte development and maturation, as well as adverse events that may negatively affect the oocyte; and the importance of the uterine environment, including signaling proteins in the maternal-embryonic interactions that ensure proper embryo development. We also discuss the impact of assisted reproductive technologies in oocyte and embryo quality and developmental potential, and considerations when looking into the prospects for developing systems that allow for in vitro gametogenesis as a tool for assisted reproduction in cattle.(AU)

Dr Association of bone morphogenetic protein 15 and growth differentiation factor 9 with litter size in livestock: a review study

Litter size is one of the crucial factors in livestock production and is of high economic value, which is affected by ovulation rate, hormones, and growth factors.Growth factors play a multifaceted role in reproductive physiology. This review aims to investigate the association of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9)with litter size in livestock.The transforming growth factor ß(TGF-ß) superfamily includes more than 34 members; GDF9 and BMP15 are among the most significantfactors for regulating fertility and litter size in most livestock species. Ovarian follicles release BMP15 and GDF9 that are involved in the maturation of primary follicles into the basal form, proliferation of granulosa and theca cells, steroidogenesis, ovulation, and formation of the corpus luteum. Besides, these factors are highly expressed in oocytes and are necessary for female fertility and multiple ovulation in several livestock species. Animals with two inactive copies of these factors are sterile, while those with one inactive copy are fertile. Thus, the present reviewprovides valuable information on the association of BMP15 and GDF9with litter size in livestock that can be used as biological markers of multiple ovulation or for improving fertility in livestock.(AU)

Dynamics of nuclear and cytoplasm maturation of bovine oocytes cultivated in vitro in medium supplemented with fulerol / Dinâmica da maturação nuclear e citoplasmática de oócitos bovinos cultivados in vitro em meio suplementado com fulerol

The aim of this work was to evaluate, in vitro, the dynamics of nuclear and cytoplasmic maturation of bovine oocytes in traditional IVM medium (CT) and supplemented with fullerol (MF50), for 36 hours. The nuclear maturation of CT (n=300) and MF50 (n=270) every 6 hours, stained with Hoechst33342 and cytoplasmic, the mitochondrial distribution of CT (n=197) and MF50 (n=159) at every 12 hours, stained with Mitotracker Orange. At 6 hours, CT oocytes (19%) were in MI (metaphase I), while in MF50 they were in GV (germ vesicle) or GVB (GV breakeage), repeating at 12 hours. At 18 hours, 46.3% were matured in CT, and 20% in MF50. At 24 hours, 43.9% of maturation was observed in the MF50 group, and 63.8% in the CT. At 30 and 36 hours, the maturation pattern was stable, but with the onset of oocyte degeneration. There was a delay in cytoplasmic maturation with 36 hours (P < 0.05) in MF50 (53.9% of mature gametes), compared to CT (69.8%). With immature cytoplasm, they were 10.4% and 31.7% for CT and MF50 (P< 0.05), respectively. It was concluded that fullerol possibly interfered in the expansion of cumulus oophorus cells, as well as delayed the meiotic progression and cytoplasmic maturation.

O objetivo deste estudo foi avaliar, in vitro, a dinâmica da maturação nuclear e citoplasmática de oócitos bovinos em meio MIV tradicional (TC) e suplementado com fulerol (MF50), durante 36 horas. Na maturação nuclear do TC (n=300) e do MF50 (n=270) a cada seis horas, corados com Hoechst 33342, e na citoplasmática, avaliou-se a distribuição mitocondrial do TC (n=197) e do MF50 (n=159) a cada 12 horas, corados com Mitotracker Orange (Life® Technologies). Às seis horas, oócitos do TC (19%) se encontravam em MI (metáfase I), enquanto no MF50 estavam em VG (vesícula germinativa) ou QVG (quebra VG), repetindo com 12 horas. Às 18 horas, 46,3% estavam maturados no TC, e 20% no MF50. Com 24 horas, verificaram-se 43,9% de maturação no grupo MF50, e 63,8% no TC. Às 30 e 36 horas, o padrão de maturação foi estável, mas com início de degeneração oócitária. Houve retardo na maturação citoplasmática com 36 horas (P<0,05) no MF50 (53,9% de gametas maduros), comparado ao TC (69,8%). Com citoplasma imaturo, foram 10,4% e 31,7% para TC e MF50 (P<0,05), respectivamente. Conclui-se que o fulerol possivelmente interferiu na expansão das células do cumulus oophorus, bem como retardou a progressão meiótica e a maturação citoplasmática dos oócitos.

Effect of different gonadotropins on in vitro maturation of sheep oocytes / Efeito de diferentes gonadotrofinas na maturação in vitro de oócitos ovinos

The aim of this study was to examine the effect of replacing the use of follicle-stimulating hormone (FSH) with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) on the in vitro maturation (IVM) of sheep oocytes. After sheep ovaries were collected (n=300), the cumulus-oocyte complexes were aspirated, selected, and divided into four groups according to the IVM medium: CON group, in which the basic IVM medium was used; and eCG, hCG, and FSH groups, in which the oocytes were immersed in basic IVM medium with 10 IU/mL eCG, 10 IU/mL hCG, and 10 µg/mL FSH-p, respectively. In vitro maturation of the oocytes was performed at 38.5 °C, in a humidified atmosphere of 5% CO2 in air, for 24 h. Subsequently, the oocytes were evaluated for the degree of cumulus-cell expansion, chromatin configuration, GSH levels, and active mitochondria. There were no significant differences for the rate of cumulus cell expansion. The percentage of oocytes in MII was higher in the eCG group than in the CON and hCG groups (P<0.05) and similar to that of the FSH group. In conclusion, eCG can be used as a substitute for FSH in IVM of sheep oocytes.

O objetivo deste estudo foi avaliar o efeito da gonadotrofina coriônica equina (eCG) e da gonadotrofina coriônica humana (hCG), em substituição ao uso de hormônio folículo estimulante (FSH) na maturação in vitro (MIV) de oócitos ovinos. Após a coleta de ovários (n=300) ovinos, os complexos cúmulus-oócitos (CCOs) foram aspirados, selecionados e divididos em quatro grupos de acordo com o meio de MIV: grupo CON, em que foi utilizado o meio MIV base; e grupos ECG, HCG e FSH, em que os oócitos foram imersos em meio MIV base adicionado de 10 UI/mL de eCG, 10 UI/mL de hCG e 10 µg/mL de FSH-p, respectivamente. A MIV dos oócitos foi realizada a 38,5°C, em atmosfera umidificada de 5% de CO2 em ar, durante 24 horas. Posteriormente, os oócitos foram avaliados, quanto grau de expansão das células do cumulus, configuração da cromatina, níveis de GSH e mitocôndrias ativas. Não foram observadas diferenças significativas com relação à taxa de expansão de células do cumulus. A percentagem de oócitos em MII foi maior no grupo ECG do que no grupo CON e HCG (P<0,05) e semelhante ao grupo FSH. Em conclusão, a eCG pode ser utilizada em substituição ao FSH na MIV de oócitos ovinos.