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Somatic cell biobanking is a promising strategy for developing reproductive techniques. Although cryopreservation, a technique used for creating biobanks, has been performed on Galea spixii, structural and physiological damage to its cells highlight the need to optimize the cryoprotective solution being used. Therefore, the osmoprotective activity of 5 mM L-proline was evaluated as an alternative cryoprotectant for G. spixii fibroblast conservation. The concentration was defined based on previous studies conducted on mammalian cells. Cells derived from the skin of six individuals were cultured until the fifth passage were cryopreserved under the following treatments: (i) control (non-cryopreserved); (ii) a solution with 10% dimethyl sulfoxide (Me2SO), 10% fetal bovine serum (FBS), and 0.2 M sucrose; (iii) a solution with 10% Me2SO, 10% FBS, and 5 mM L-proline; and (iv) a solution with 10% Me2SO, 10% FBS, 0.2 M sucrose, and 5 mM L-proline. Tests were conducted to analyze cell morphology, viability, metabolism, proliferation, and apoptosis; reactive oxygen species (ROS) levels; and mitochondrial membrane activity (ΔΨm). A reduction in the number of viable cells (72.3% ± 1.2%) was observed in the sucrose-containing group compared to the control (86.7% ± 2.0%) and L-proline (88.4% ± 1.8% and 87.8% ± 2.1%) groups. After apoptotic analysis, a reduction in the number of viable cells was observed in the group with sucrose alone (74.6% ± 4.1%) compared to the control group (88.2% ± 1.1%). The ROS levels (1.03 ± 0.5 and 1.07 ± 0.5, respectively) and ΔΨm values (0.99 ± 0.42 and 1.22 ± 0.73, respectively) observed in the groups with L-proline were similar to that observed in the control group (1.00 ± 0.5 and 1.00 ± 0.4, respectively). Moreover, no difference was observed between groups for cell morphology, metabolism, or proliferation. Thus, L-proline is a cryoprotectant agent that can be used during G. spixii fibroblast cryopreservation, alone or with sucrose. In addition, we developed an adequate biobank for G. spixii, whereby stored cells could be used for reproductive techniques.

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The biobanks from dermal biopsies represent an interesting strategy for biodiversity conservation. Nevertheless, the morphological and cellular patterns of the dermis can be influenced by the age and sex of the individual. Therefore, evaluating these factors is interesting for forming biobanks of Antillean manatees. These animals, representatives of marine fauna, have had their population reduced, and biobanks are essential for their conservation. Then, we evaluated the effects of age (3.5 years vs. 3.6-16 years vs. 23.6 years) and sex (males vs. females) on morphological and cellular parameters using histological and in vitro culture techniques. Regardless of age, no differences were observed for dermal thickness, collagen fibres, tissue proliferative activity and viable cell recovery. Nonetheless, fibroblast reduction was observed in groups aged 23.6 years compared to other animals (p < 0.05). Additionally, cells from animals aged 3.6-16 years showed more significant mitochondrial damage than the other groups (p < 0.05). Regardless of sex, no differences were observed for dermal thickness, collagen fibres, tissue proliferative activity and viable cell recovery; however, females had fewer fibroblasts than males (p < 0.05). Cells from females showed lower mitochondrial damage when compared to cells from males. In summary, although age and sex do not influence dermal thickness and cell recovery, variations in the number of fibroblasts and mitochondrial characteristics were observed among the groups. These differences may be significant for understanding the dermis aspects to be correlated to biobank systems.

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Although oocyte in vitro maturation (IVM) is routinely used for in vitro embryo production in mice and rats, its use in wild rodents remains unexplored. Evidence suggests that hormone and growth factor supplementation influence oocyte meiotic resumption. This study evaluated the synergistic effects of follicle-stimulating hormone (FSH) and epidermal growth factor (EGF) on the IVM and parthenogenetic development of red-rumped agouti oocytes. Initially, we evaluated the IVM rates, mature oocyte quality, oocyte morphometry, and early embryonic development during IVM in the presence of 10, 50, and 75 mIU/mL FSH. No differences among the FSH concentrations were observed for IVM rates, oocyte morphometry, cumulus cell expansion, and viability. Although oocytes matured with 50 mIU/mL FSH showed a higher rate of cumulus expansion index (CEI), only oocytes matured with 10 mIU/mL FSH resulted in morulae after chemical activation (7.9% ± 4.2%). Thus, 10 mIU/mL FSH was used for further experiments. We subsequently evaluated the synergistic effects of 10, 50, and 100 ng/mL EGF and 10 mIU/mL FSH on the same parameters. No differences among the groups were observed in IVM rates, oocyte morphometry, and cumulus viability. Nevertheless, FSH with 10 ng/mL EGF showed a CEI superior to that of the other groups. Furthermore, oocytes matured with FSH alone or with both FSH and 10 or 50 ng/mL EGF developed morulae after activation (5.8%-8.3%). In conclusion, oocytes matured with 10 mIU/mL FSH and 10 ng/mL EGF are recommended for use in red-rumped agouti oocyte IVM, as they positively influence embryonic development.

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Methods for seminal plasma (SP) removal and the selection of collared peccary sperm for fertilization were compared. The experiments evaluated the following: the (I) impact of centrifugation for SP removal before swim-up for sperm selection and (II) a comparison of different Percoll® gradient densities (PG 45-90% and PG 35-70%). Non-selected sperm served as the control. Sperm quality was assessed based on motility patterns, morphology, membrane functional integrity, viability, reactive oxygen species (ROS), glutathione (GSH), and DNA integrity. Subsequently, the most successful group in the previous experiment and washing by centrifugation (WC) were compared for motility patterns and fertilization using pig oocytes. Swim-up decreased motility and enhanced ROS compared to the control. Centrifugation before swim-up harmed integrity and viability compared to the control. PG 45-90% (96.8 vs. 69.7 vs. 40.7 µm/s) allowed for a better velocity average pathway (VAP), a better velocity straight line, and better linearity (LIN) than those of the control and PG 35-70% (88.4 vs. 56.0 vs. 27.3 µm/s). Thus, PG 45-90% was used for fertilization. PG 45-90% obtained a higher VAP, a higher amplitude of the lateral head, straightness, and higher LIN than those of the control and WC. Cleavage (25.2-26.3%) and morula (8.1-10.5%) rates did not differ between the groups. Therefore, PG 45-90% and WC were efficient in isolating collared peccary sperm capable of fertilizing pig oocytes.

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Cell lines are valuable tools to safeguard genetic material from species threatened with extinction that is mainly due to human action. In this scenario, the puma constitutes a species whose population is being rapidly reduced in the ecosystems it inhabits. For the first time, we characterized puma skin-derived cell lines and assessed these cells after extended culture (experiment 1) and cryopreservation (experiment 2). Initially, we identified and characterized four dermal fibroblast lines using morphology, ultrastructure, and immunofluorescence assays. Moreover, we evaluated the effects of culture time (1st, 3rd, and 10th passages) and cryopreservation on their morphology, ultrastructure, viability, metabolism, proliferative activity, reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔΨm), and apoptosis. The cells showed a typical spindle-shaped morphology with centrally located oval nuclei. The cells were identified as fibroblasts by staining for vimentin. In vitro culture after the 1st, 3rd, and 10th passages did not alter most of the evaluated parameters. Cells in the 3rd and 10th passages showed a reduction in ROS levels (p < 0.05). The ultrastructure revealed morphological damage in the prolongments, and nuclei of cells derived from the 3rd and 10th passages. Moreover, cryopreservation resulted in a reduction in ΔΨm compared with that of noncryopreserved cells, suggesting that the optimization of cryopreservation methods for puma fibroblasts is essential. In conclusion, we found that viable fibroblasts could be obtained from puma skin, with slight changes after the 10th passage in in vitro culture and cryopreservation. This is the first report on the development of cell lines derived from pumas.

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Somatic resource banks play a crucial role in the conservation of genetic diversity, allowing for the preservation of biological samples from different populations. Puma somatic cells can be recovered from these banks and used in assisted techniques toward enhancing their multiplication and conservation. In response to the population reduction of this ecologically importance species, we aimed to evaluate the capacity of cryopreservation of somatic tissues on the maintenance of the integrity and quality of the cells recovered after culture, with the aim of establishing a somatic tissue bank that will allow for the safeguarding of a wide genetic sampling of pumas. Cryopreservation increased the thickness of the corneum layer in the tissues, and the number of perinuclear halos and empty gaps. Nevertheless, cryopreservation was able to maintain normal fibroblast patterns, even showing an increase in the percentage of collagen fibers. Cryopreservation maintained the proliferative potential of the tissues and the parameters evaluated during in vitro culture, mainly regarding the viability, proliferative activity, and apoptosis levels. Nevertheless, cells from cryopreserved tissues showed decreased metabolism and mitochondrial membrane potential when compared to cells from non-cryopreserved tissues. In summary, we demonstrated for the first time that puma somatic tissues subjected to cryopreservation are viable and maintain tissue integrity, featuring minimal changes after warming. Although viable somatic cells are obtained from these tissues, they undergo alterations in their metabolism and mitochondrial membrane potential. Improvements in the conservation conditions of somatic samples are needed to increase the quality of somatic tissue banks in this species.

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Biological resource banks represent valuable tools for the conservation of species vulnerable to extinction, such as the jaguar. Cryobanks of skins have the potential to safeguard rare genotypes, allowing the potential exploitation of biological samples in animal multiplication technologies and the study of genetic variability. Determination of the most suitable skin regions for tissue conservation can help increase the efficiency of cryobanks and the storage of biological samples. To this end, we evaluated the effects of vitrification of skin tissues from the ear, caudal, and femoral regions of a post-mortem jaguar belonging to a zoo in Brazil. Non-vitrified and vitrified samples were evaluated and compared using quantitative methods, focusing on skin thickness, cell quantification, number of perinuclear halos, collagen and elastic density, and proliferative activity. No differences were observed in skin thickness, number of perinuclear halos, elastic density, and proliferative activity between non-vitrified and vitrified tissues in skin from any region. However, vitrified tissues derived from femoral skin showed a reduction in the number of fibroblasts, epidermal cells and collagen density compared to non-vitrified tissues. In summary, the ear and caudal regions provided the best conservation of somatic tissues derived from jaguars, and skin samples from these regions are therefore the most suitable for the formation of cryobanks.

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Distintas estratégias de conservação têm sido adotadas visando a manutenção e recuperação da biodiversidade, especialmente de espécies que se encontram em diferentes níveis de ameaça à extinção. Neste grupo de espécies, encontram-se os grandes felídeos, os quais em virtude das ações antrópicas, como a destruição e a fragmentação de habitat, necessitam de esforços voltados para a conservação de seu material genético. Uma estratégia empregada para essa finalidade consiste nos bancos de recursos somáticos, os quais são definidos como a criopreservação de tecidos e células somáticas oriundas de diferentes populações. Essas amostras biológicas, quando adequadamente conservadas, são elementoschave para a multiplicação das espécies por meio de seu emprego na clonagem por transferência nuclear e indução de células à pluripotência. Assim, o objetivo desta revisão é abordar os aspectos relevantes envolvidos na formação de bancos de recursos somáticos em grandes felídeos, destacando os estudos desenvolvidos pelo Laboratório de Biotecnologia Animal (LBA), da Universidade Federal Rural do Semi-Árido (UFERSA) em onças-pintadas e onças-pardas.

Different conservation strategies have been adopted to maintain and recover biodiversity, especially for species that are at different levels of threat to extinction. In this group of species are the large felids, which, due to anthropic actions, such as habitat destruction and fragmentation, require efforts aimed at the conservation of their genetic material. A strategy employed for this purpose consists of somatic resource banks, which are defined as the cryopreservation of tissues and somatic cells from different populations. These biological samples, when properly preserved, are key elements for the multiplication of species through their use in cloning by nuclear transfer and induction of cells to pluripotency. Thus, the aim of this review is to address the relevant aspects involved in the formation of somatic resource banks in large felids, highlighting the studies carried out by the Laboratory of Animal Biotechnology (LBA) of the Federal Rural University of the Semi-Arid (UFERSA) in jaguars and pumas.

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Skin and cartilage have been the main source for the recovery of somatic cells to be used in conservation strategies in wild mammals. In this sense, an important step for the cryopreservation of these samples is to recognize the properties of the skin and cartilage. Thus, knowing that the skin may differ among species and aiming to contribute to the establishment of cryobanks, the study examined the differences in the ear skin and cartilage of wild rodents from South America, agouti (Dasyprocta leporina) and spix's yellow-toothed cavy (Galea spixii). Ultrastructural and quantitative methods were used to measure skin and cartilage thickness, density of collagen and elastic fibers, cell type number and distribution, and proliferative activity. Although ultrastructural analysis revealed a similar pattern between species, morphometric analysis of the skin and cartilage showed differences between agoutis and cavies regarding thickness of epidermis layers (corneum: 5.3±2.5μm vs. 3.9±0.6μm; intermediate: 16.4±6.2μm vs. 23.4±8.1μm; basal: 9.9±2.1μm vs. 4.8±0.5μm), dermis (183.1±44.0μm vs. 258.2±22.9μm), total skin (211.8±46.0μm vs. 290.3±23.7μm) and perichondrium (27.6±6.1μm vs. 10.5±1.8μm). A greater number of epidermal cells (61.7±15.2 vs. 24.8±7.6) and chondrocytes (32.7±9.0 vs. 27.5±4.7) were observed in agouti, while the cavy presented a greater number of melanocytes (12.6±4.7 vs. 29.9±6.2), keratinocytes (14.7±4.2 vs. 29.8±7.6), and fibroblasts (103.6±24.7 vs. 112.2±11.3). Moreover, a higher percentage of collagen fibers and proliferative activity was observed in the skin of cavies, when compared to the skin of agoutis. Therefore, there are differences between agouti and cavy for ear skin and cartilage, requiring the establishment of species-specific cryopreservation protocols.(AU)

A pele e cartilagem têm sido uma importante fonte de recuperação de células somáticas a serem utilizadas em estratégias de conservação em mamíferos silvestres. Nesse contexto, uma importante etapa para criopreservação é conhecer, inicialmente, as propriedades que compõem a pele e cartilagem. Sabendo, então, que a pele pode diferir-se entre espécies e com o objetivo de contribuir para o estabelecimento de criobancos, o estudo evidenciou as diferenças da pele e da cartilagem do pavilhão auricular apical de cutias (Dasyprocta leporina) e preás (Galea spixii) que são roedores silvestres presentes na América do Sul. Para tanto, métodos ultraestruturais e quantitativos foram utilizados para mensurar a espessura da pele e da cartilagem, densidade de fibras colágenas e elásticas, número e distribuição dos tipos celulares e atividade proliferativa. Embora as propriedades ultraestruturais em cutias e preás tenham se mostrado semelhantes, avaliações acerca da morfometria da pele e da cartilagem demonstrou diferenças, especialmente nas camadas epidérmicas (córnea: 5,3±2,5μm vs. 3,9±0,6μm; espinhosa: 16,4±6,2μm vs. 23,4±8,1μm; basal: 9,9±2,1μm vs. 4,8±0,5μm), derme (183,1±44,0μm vs. 258,2±22,9μm), pele total (211,8±46,0μm vs. 290,3±23,7μm) e pericôndrio (27,6±6,1μm vs. 10,5±1,8μm). Além disso, um número maior de células epidérmicas (61,7±15,2 vs. 24,8±7,6) e condrócitos (32,7±9,0 vs. 27,5±4,7) foram observados em cutias, enquanto em preás um maior número de melanócitos (12,6±4,7 vs. 29,9±6,2), queratinócitos (14,7±4,2 vs. 29,8±7,6) e fibroblastos (103,6±24,7 vs. 112,2±11,3) foram evidenciados. Ainda, em preás, uma maior porcentagem de fibras colágenas e da atividade proliferativa foram observadas quando comparadas a pele de cutias. Portanto, existem diferenças entre cutias e preás para pele e cartilagem do pavilhão auricular, exigindo desta forma um estabelecimento de protocolos de criopreservação específica para cada uma destas espécies.(AU)

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Skin and cartilage have been the main source for the recovery of somatic cells to be used in conservation strategies in wild mammals. In this sense, an important step for the cryopreservation of these samples is to recognize the properties of the skin and cartilage. Thus, knowing that the skin may differ among species and aiming to contribute to the establishment of cryobanks, the study examined the differences in the ear skin and cartilage of wild rodents from South America, agouti (Dasyprocta leporina) and spix's yellow-toothed cavy (Galea spixii). Ultrastructural and quantitative methods were used to measure skin and cartilage thickness, density of collagen and elastic fibers, cell type number and distribution, and proliferative activity. Although ultrastructural analysis revealed a similar pattern between species, morphometric analysis of the skin and cartilage showed differences between agoutis and cavies regarding thickness of epidermis layers (corneum: 5.3±2.5μm vs. 3.9±0.6μm; intermediate: 16.4±6.2μm vs. 23.4±8.1μm; basal: 9.9±2.1μm vs. 4.8±0.5μm), dermis (183.1±44.0μm vs. 258.2±22.9μm), total skin (211.8±46.0μm vs. 290.3±23.7μm) and perichondrium (27.6±6.1μm vs. 10.5±1.8μm). A greater number of epidermal cells (61.7±15.2 vs. 24.8±7.6) and chondrocytes (32.7±9.0 vs. 27.5±4.7) were observed in agouti, while the cavy presented a greater number of melanocytes (12.6±4.7 vs. 29.9±6.2), keratinocytes (14.7±4.2 vs. 29.8±7.6), and fibroblasts (103.6±24.7 vs. 112.2±11.3). Moreover, a higher percentage of collagen fibers and proliferative activity was observed in the skin of cavies, when compared to the skin of agoutis. Therefore, there are differences between agouti and cavy for ear skin and cartilage, requiring the establishment of species-specific cryopreservation protocols.(AU)

A pele e cartilagem têm sido uma importante fonte de recuperação de células somáticas a serem utilizadas em estratégias de conservação em mamíferos silvestres. Nesse contexto, uma importante etapa para criopreservação é conhecer, inicialmente, as propriedades que compõem a pele e cartilagem. Sabendo, então, que a pele pode diferir-se entre espécies e com o objetivo de contribuir para o estabelecimento de criobancos, o estudo evidenciou as diferenças da pele e da cartilagem do pavilhão auricular apical de cutias (Dasyprocta leporina) e preás (Galea spixii) que são roedores silvestres presentes na América do Sul. Para tanto, métodos ultraestruturais e quantitativos foram utilizados para mensurar a espessura da pele e da cartilagem, densidade de fibras colágenas e elásticas, número e distribuição dos tipos celulares e atividade proliferativa. Embora as propriedades ultraestruturais em cutias e preás tenham se mostrado semelhantes, avaliações acerca da morfometria da pele e da cartilagem demonstrou diferenças, especialmente nas camadas epidérmicas (córnea: 5,3±2,5μm vs. 3,9±0,6μm; espinhosa: 16,4±6,2μm vs. 23,4±8,1μm; basal: 9,9±2,1μm vs. 4,8±0,5μm), derme (183,1±44,0μm vs. 258,2±22,9μm), pele total (211,8±46,0μm vs. 290,3±23,7μm) e pericôndrio (27,6±6,1μm vs. 10,5±1,8μm). Além disso, um número maior de células epidérmicas (61,7±15,2 vs. 24,8±7,6) e condrócitos (32,7±9,0 vs. 27,5±4,7) foram observados em cutias, enquanto em preás um maior número de melanócitos (12,6±4,7 vs. 29,9±6,2), queratinócitos (14,7±4,2 vs. 29,8±7,6) e fibroblastos (103,6±24,7 vs. 112,2±11,3) foram evidenciados. Ainda, em preás, uma maior porcentagem de fibras colágenas e da atividade proliferativa foram observadas quando comparadas a pele de cutias. Portanto, existem diferenças entre cutias e preás para pele e cartilagem do pavilhão auricular, exigindo desta forma um estabelecimento de protocolos de criopreservação específica para cada uma destas espécies.(AU)

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Growth measurements such as leaf area (LA) and dry matter (DM) are important in experiments about plants population, fertilization, irrigation and others parameters of cultivation, in garlic crop. The LA and DM are commonly defined as destructive, lengthy and cause loss of plants in the experimental units. The objective of this study was to fit mathematical models using linear models that estimate the leaf area and dry matter of garlic plants - variety Ito. For that, garlic plants were collected at 30, 45, 60, 75, 90, 115 and 120 days after planting. The measurements of width (W), length (L) of leaves, LA, DM, pseudostem diameter (PD), number of leaves per plant (NL) and height (H) were determined in each time. The models were fitted to estimate the LA or DM as function of the variables W, L, L*W, PD and LA. The statistical analysis of the linear regression, coefficient of determination of the linear regression (R2), root mean square error (RMSE), modified concordance index (d1) and the BIAS index were verified to determine the most representative models. It`s possible to estimate the LA and the leaf DM of garlic plants using the variables: length, width, pseudostem diameter and height of plants.

Medidas de crescimento como área foliar (AF) e matéria seca (MS) são importantes em experimentos com população de plantas, adubação, irrigação e outros parâmetros de cultivo, na cultura do alho. Muitas vezes a AF e MS são definidas por avaliações destrutivas, demoradas e com perdas de plantas nas unidades experimentais. Objetivou-se, com este trabalho, definir modelos matemáticos através de medidas lineares, que estimem a área foliar e a matéria seca das folhas de plantas de alho da variedade Ito. Para isso, quinze plantas foram coletadas aos 30, 45, 60, 75, 90, 115 e 120 dias após o plantio (DAP). As medidas de largura (L), comprimento (C) das folhas, AF, MS, diâmetro do pseudocaule (DP), número de folhas por planta (NF) e altura da planta (AP) foram determinadas em cada época. Ajustaram-se modelos para estimar a AF ou MS em função das variáveis L, C, L*C, DP e AF. Para determinar os modelos mais representativos foram verificados a análise estatística da regressão linear, o coeficiente de determinação da regressão linear (R2), raiz do quadrado médio do erro (RQME), índice de concordância modificado (d1) e o índice BIAS. É possível estimar a AF e MS foliar de plantas de alho através das variáveis comprimento, largura, diâmetro do pseudocaule e altura de plantas.

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The objective of this study was to evaluate the effect of nitrogen sources and rates on the physicochemical characteristics and yield of tomato plants. Forty hybrids were cultivated at 100 and 400 kg ha-1of N, combined with four sources (urea, ammonium sulfate, ammonium nitrate and calcium), plus a treatment without N application in a randomized complete block design four replicates. Size, stem diameter, number of leaves, SPAD (Soil Plant Analysis Development), leaf nitrogen, number of fruits / plants, fruit firmness, bark thickness and average fruit size, ° BRIX, pH, calcium, potassium and sodium in fruits. There was an increase in SPAD index, Brix and longitudinal diameter of fruits as a function of the N dose. The use of ammonium nitrate and calcium provided stronger fruits. Urea and ammonium nitrate provided the highest pH value in tomato fruits. The application of the 100 kg ha-1 dose of N resulted in the highest potassium content in fruits. The highest productivity was obtained with the application of sources containing ammonium and the lowest in the control treatment. Sources and doses of nitrogen fertilizers influenced growth, productivity and parameters related to tomato quality.

Objetivou-se avaliar o efeito das fontes e doses de nitrogênio nas características físico-químicas e produtividade do tomateiro. Foi cultivado o híbrido Forty com doses de 100 e 400 kg ha-1 de N, combinadas com quatro fontes (ureia, sulfato de amônio, nitrato de amônio e de cálcio), mais um tratamento sem aplicação de N em delineamento em blocos casualizados com quatro repetições. Avaliou-se altura, diâmetro do caule, número de folhas, SPAD (Soil Plant Analysis Development), nitrogênio na folha, número de frutos/plantas, firmeza do fruto, espessura da casca e tamanho médio dos frutos, °BRIX, pH, cálcio, potássio e sódio nos frutos. Houve incremento do índice SPAD, ºBrix e diâmetro longitudinal dos frutos em função da dose de N. A utilização do nitrato de amônio e de cálcio proporcionou frutos mais firmes. A ureia e o nitrato de amônio propiciaram o maior valor de pH em frutos de tomate. A aplicação da dose de 100 kg ha-1 de N acarretou o maior teor de potássio nos frutos. A maior produtividade foi obtida com a aplicação de fontes que continham amônio e a menor no tratamento controle. As fontes e as doses de fertilizantes nitrogenados influenciaram o crescimento, produtividade e os parâmetros relacionados à qualidade do tomate.

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The cryobanks of agouti somatic tissues represent a promising tool for the conservation of this species and of those that are phylogenetically related and endangered. For these purposes, one strategy to guarantee the quality of samples after warming would be to choose the appropriate tissue vitrification technique. Therefore, we evaluated the effects of two different techniques, direct vitrification in cryovials (DVC) and solid-surface vitrification (SSV), on the preservation of ear somatic tissues derived from agoutis kept in a scientific center of creation. Noncryopreserved somatic tissues were used as controls. Although SSV reduced the thickness of the dermis and cartilage (p < 0.05), the epidermal thickness of these samples was observed to be similar to controls (p > 0.05). Notably, the number of fibroblasts was not altered with either technique. However, both vitrification methods led to an increase in the number of perinuclear halos, with a particularly strong increase observed in DVC-derived fragments (p < 0.05). Compared with the DVC group, SSV showed a larger number of normal chondrocytes and smaller number of degenerate chondrocytes. Furthermore, the number of empty lacunae in SSV-derived fragments remained similar to controls (p > 0.05). In summary, SSV was found to be a more efficient method for vitrifying agouti somatic tissues compared with DVC. These results are important for the proper formation of agouti somatic banks, an essential step in the study of biological resources in this species.

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Objetivo: investigar a associação entre o polimorfismo TP53 Arg72Pro em pacientes diagnosticados com acidente vascular encefálico hemorrágico (AVEH) ou aneurisma intracerebral em uma amostra do Distrito Federal. Método: Tratou-se de um estudo caso-controle, com 162 indivíduos equitativamente divididos nos grupos, com anotações das características clínicas do prontuário e análise da genotipagem por meio da estratégia de PCR. As frequências genotípicas foram estimadas por contagem direta. O nível de significância adotado foi de 5%. Resultados: Foi verificado que a presença do alelo Arg do polimorfismo do TP53 Arg72Pro atuou como fator do risco para a ocorrência do AVEH/aneurisma intracerebral. A presença do genótipo Arg/Arg aumentou o risco para o prognóstico ruim (ERM >3) em pacientes portadores do AVEH/aneurisma (P<0,01; OR= 6,07). Não houve associação estatística entre HAS, diabetes, tabagismo e etilismo e a presença do polimorfismo TP53 Arg72Pro no grupo estudado. Conclusão: Concluiu-se que a presença do alelo Arg do polimorfismo do TP53 Arg72Pro está associada ao aumento do risco de ocorrência do AVEH/aneurisma intracerebral.

Objective: to investigate the association between the TP53 Arg72Pro polymorphism in patients diagnosed with hemorrhagic stroke (HS) or intracerebral aneurysm in a sample from Distrito Federal. Method: This was a case-control study, with 162 individuals equally divided into groups, with notes on the clinical characteristics of the medical record and analysis of genotyping using the PCR strategy. Genotype frequencies were estimated by direct counting. The level of significance adopted was 5%.Results: we found that the presence of the Arg allele of the TP53 Arg72Pro polymorphism acted as a risk factor for the occurrence of HS/intracerebral aneurysm. The presence of the Arg/Arg genotype increased the risk for poor prognosis (ERM> 3) in patients with HS/aneurysm (P <0.01; OR = 6.07). There was no statistical association between SAH, diabetes, smoking and alcohol consumption and the presence of the TP53 Arg72Pro polymorphism in the studied group. Conclusion: It was concluded that the presence of the Arg allele of the TP53 Arg72Pro polymorphism is associated with an increased risk of occurring HS/intracerebral aneurysm.

Objetivo: investigar la asociación entre el polimorfismo TP53 Arg72Pro en pacientes diagnosticados de accidente cerebrovascular hemorrágico (AVEH) o aneurisma intracerebral en una muestra del Distrito Federal. Método: Este fue un estudio de casos y controles, con 162 individuos igualmente divididos en grupos, con notas sobre las características clínicas de la historia clínica y el análisis de genotipado utilizando la estrategia de PCR. Las frecuencias de genotipo se estimaron por conteo directo. El nivel de significación adoptado fue del 5%. Resultados: Se encontró que la presencia del alelo Arg del polimorfismo TP53 Arg72Pro actuó como un factor de riesgo para la aparición de AVEH/aneurisma intracerebral. La presencia del genotipo Arg / Arg aumentó el riesgo de mal pronóstico (ERM> 3) en pacientes con AVEH/aneurisma (P <0.01; OR = 6.07). No hubo asociación estadística entre SAH, diabetes, tabaquismo y consumo de alcohol y la presencia del polimorfismo TP53 Arg72Pro en el grupo estudiado. Conclusión: se concluyó que la presencia del alelo Arg del polimorfismo TP53 Arg72Pro se asocia con un mayor riesgo de aparición de AVEH/aneurisma intracerebral.

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RESUMO

The study aimed to evaluate the effect of strontium chloride (SrCl2) with cytochalasin B (CB) on the activation of agouti oocytes matured in vitro for embryo production. Thus, ovaries were used for oocyte recovery by slicing. Subsequently, viable oocytes were destined for in vitro maturation (IVM) and after 24 h evaluated for the expansion and viability of the cumulus cells and presence of the first polar body (1PB). After IVM, oocytes were activated with a combination of 10 mM SrCl2 and 5 μg/mL CB for 6 h and evaluated for embryo development kinetics. Hence, 93.3% of cumulus cell expansion was observed, with 91.2% viability and 37.3% of oocytes with the presence of 1PB. Regarding embryonic development, 43.2% (19/44) of cleaved structures and 6.8% (3/44) of morulae were observed in relation to the number of oocytes and 18.8% (3/16) morulae in relation to the number of cleaved structures. Thus, the combination of SrCl2 with CB promoted the activation of oocytes matured in vitro from agouti resulting in morulae. Finally, with this study, fundamental steps for in vitro conservation through reproductive biotechniques were developed in agoutis.

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RESUMO

Although widely used as reproductive biotechnology in cattle, in vitro embryo production (IVEP) has variable efficiency. During in vitro maturation (IVM), supplementing the medium with antioxidant potential could be an affordable alternative to increase the efficiency of IVEP, requiring the evaluation of new components, such as eugenol, β-caryophyllene, and acetyl eugenol. Thus, bovine oocytes were matured according to different antioxidants. Metaphase II oocytes were identified by Hoechst staining. The oxidative status was measured by evaluation of reactive oxygen species (ROS), and glutathione (GSH). After eight repetitions, no difference was observed among groups containing antioxidants, being all these groups superior to negative control (p<0.05). The ROS levels decreased and GSH increased in oocytes matured with EUG (p<0.05). These results indicate apositive effect of eugenol on the oxidative status of oocytes matured with this compound, showing that eugenol can be an efficient supplement in the IVM of bovine oocytes.

Assuntos

RESUMO

Although widely used as reproductive biotechnology in cattle, in vitro embryo production (IVEP) has variable efficiency. During in vitro maturation (IVM), supplementing the medium with antioxidant potential could be an affordable alternative to increase the efficiency of IVEP, requiring the evaluation of new components, such as eugenol, β-caryophyllene, and acetyl eugenol. Thus, bovine oocytes were matured according to different antioxidants. Metaphase II oocytes were identified by Hoechst staining. The oxidative status was measured by evaluation of reactive oxygen species (ROS), and glutathione (GSH). After eight repetitions, no difference was observed among groups containing antioxidants, being all these groups superior to negative control (p<0.05). The ROS levels decreased and GSH increased in oocytes matured with EUG (p<0.05). These results indicate apositive effect of eugenol on the oxidative status of oocytes matured with this compound, showing that eugenol can be an efficient supplement in the IVM of bovine oocytes.(AU)

Assuntos

RESUMO

The study aimed to evaluate the effect of strontium chloride (SrCl2) with cytochalasin B (CB) on the activation of agouti oocytes matured in vitro for embryo production. Thus, ovaries were used for oocyte recovery by slicing. Subsequently, viable oocytes were destined for in vitro maturation (IVM) and after 24 h evaluated for the expansion and viability of the cumulus cells and presence of the first polar body (1PB). After IVM, oocytes were activated with a combination of 10 mM SrCl2 and 5 μg/mL CB for 6 h and evaluated for embryo development kinetics. Hence, 93.3% of cumulus cell expansion was observed, with 91.2% viability and 37.3% of oocytes with the presence of 1PB. Regarding embryonic development, 43.2% (19/44) of cleaved structures and 6.8% (3/44) of morulae were observed in relation to the number of oocytes and 18.8% (3/16) morulae in relation to the number of cleaved structures. Thus, the combination of SrCl2 with CB promoted the activation of oocytes matured in vitro from agouti resulting in morulae. Finally, with this study, fundamental steps for in vitro conservation through reproductive biotechniques were developed in agoutis.(AU)

Assuntos

RESUMO

This study aimed to determine the extraction curves and accumulation order of macronutrients of two onion cultivars. In commercial fields in the region of Alto Paranaíba, MG, Brazil, onion plants of the cultivars Optima and Sirius were collected during the cycle in 2014 and 2015 to determine dry matter and macronutrient accumulation. The yield was determined at harvest time. Three phases of dry matter and nutrient accumulations were observed. The first phase had a slow accumulation and was followed by a fast increment and subsequent stabilization in phases two and three, respectively, for most of the nutrients in both cultivars and years of cultivation. Duration and accumulated at each phase were different between cultivars and years of cultivation. The total and commercial bulb yield did not vary between cultivars but only between years of cultivation, with average values of 117 and 60 Mg ha-1 in 2014 and 2015, respectively. The partitioning of dry matter and macronutrients followed the same model for both cultivars and years of cultivation. Macronutrients with the largest partitioning for bulbs are P and S. The decreasing extraction order for both cultivars was K > Ca > N > P > Mg > S in 2014 and K > N > Ca > P > S > Mg in 2015.

Objetivou-se determinar as curvas de extração e a ordem de acúmulo de macronutrientes de dois cultivares de cebola. Em cultivos comerciais na região do Alto Paranaíba - MG, plantas de cebola das cultivares Optima e Sirius foram coletadas durante o ciclo, nos anos de 2014 e 2015, para determinação do acúmulo de matéria seca e macronutrientes. A produtividade foi determinada na colheita. Observaram-se três fases nos acúmulos de matéria seca e nutrientes: uma primeira, de acúmulo lento, foi seguida por incremento rápido e posterior estabilização nas fases dois e três, respectivamente, para maioria dos nutrientes em ambas cultivares e anos de cultivo. A duração das fases e o acumulado em cada uma foram distintos entre as cultivares e anos de cultivo. A produtividade total e comercial de bulbos não variou entre as cultivares, apenas entre os anos de cultivo, obtendo-se média de 117 e 60 Mg ha-1, nos anos de 2014 e 2015, respectivamente. A partição de matéria seca e macronutrientes pela cebola seguiu o mesmo modelo para ambas cultivares e anos de cultivo. Os macronutrientes com maior partição para os bulbos são o P e o S. A ordem decrescente de extração, para ambas cultivares, foi K > Ca > N > P > Mg > S em 2014, e K > N > Ca > P > S > Mg em 2015.

Assuntos

RESUMO

This study aimed to determine the extraction curves and accumulation order of macronutrients of two onion cultivars. In commercial fields in the region of Alto Paranaíba, MG, Brazil, onion plants of the cultivars Optima and Sirius were collected during the cycle in 2014 and 2015 to determine dry matter and macronutrient accumulation. The yield was determined at harvest time. Three phases of dry matter and nutrient accumulations were observed. The first phase had a slow accumulation and was followed by a fast increment and subsequent stabilization in phases two and three, respectively, for most of the nutrients in both cultivars and years of cultivation. Duration and accumulated at each phase were different between cultivars and years of cultivation. The total and commercial bulb yield did not vary between cultivars but only between years of cultivation, with average values of 117 and 60 Mg ha-1 in 2014 and 2015, respectively. The partitioning of dry matter and macronutrients followed the same model for both cultivars and years of cultivation. Macronutrients with the largest partitioning for bulbs are P and S. The decreasing extraction order for both cultivars was K > Ca > N > P > Mg > S in 2014 and K > N > Ca > P > S > Mg in 2015.(AU)

Objetivou-se determinar as curvas de extração e a ordem de acúmulo de macronutrientes de dois cultivares de cebola. Em cultivos comerciais na região do Alto Paranaíba - MG, plantas de cebola das cultivares Optima e Sirius foram coletadas durante o ciclo, nos anos de 2014 e 2015, para determinação do acúmulo de matéria seca e macronutrientes. A produtividade foi determinada na colheita. Observaram-se três fases nos acúmulos de matéria seca e nutrientes: uma primeira, de acúmulo lento, foi seguida por incremento rápido e posterior estabilização nas fases dois e três, respectivamente, para maioria dos nutrientes em ambas cultivares e anos de cultivo. A duração das fases e o acumulado em cada uma foram distintos entre as cultivares e anos de cultivo. A produtividade total e comercial de bulbos não variou entre as cultivares, apenas entre os anos de cultivo, obtendo-se média de 117 e 60 Mg ha-1, nos anos de 2014 e 2015, respectivamente. A partição de matéria seca e macronutrientes pela cebola seguiu o mesmo modelo para ambas cultivares e anos de cultivo. Os macronutrientes com maior partição para os bulbos são o P e o S. A ordem decrescente de extração, para ambas cultivares, foi K > Ca > N > P > Mg > S em 2014, e K > N > Ca > P > S > Mg em 2015.(AU)

Assuntos