Exogenous application of salicylic acid on the mitigation of salt stress in Capsicum annuum L / Aplicação exógena de ácido salicílico na mitigação do estresse salino em Capsicum annuum L

ABSTRACT: Salinity has limited conventional vegetable cultivation, especially in semi-arid regions. In this regard, the use of elicitors that act to induce tolerance to salt stress, such as salicylic acid, has emerged as a promising alternative. This study evaluated the effects of foliar spraying with salicylic acid on the mitigation of salt stress on the morphophysiology and production of bell pepper cv. All Big. The study was conducted in a greenhouse in Campina Grande - PB, Brazil, adopting a completely randomized design, in a 4 x 4 factorial arrangement, corresponding to four levels of electrical conductivity of irrigation water (0.8; 1.6; 2.4; and 3.2 dS m-1) and four concentrations of salicylic acid (0; 1.2; 2.4 and 3.6 mM), with three replicates. The foliar application of salicylic acid at a concentration of 1.6 mM attenuated the effects of salt stress in gas exchange, growth, mean fruit weight, and total production per plant, and decreased the percentage of intercellular electrolyte leakage of sweet pepper cv. All Big plants, at 80 days after sowing.

RESUMO: A salinidade tem afetado negativamente o cultivo de hortaliças, sobretudo em regiões semiáridas. Neste sentido, o uso de elicitores que atuem na indução de tolerância ao estresse salino, como ácido salicílico, tem se destacado como uma alternativa promissora. Objetivou-se com esse estudo, avaliar os efeitos da pulverização foliar do ácido salicílico na mitigação do estresse salino na morfofisiologia e produção do pimentão cv. All Big. O estudo foi conduzido em casa de vegetação em Campina Grande - PB, utilizando-se o delineamento inteiramente casualizado, em arranjo fatorial 4 x 4, sendo quatro níveis de condutividade elétrica da água de irrigação (0,8; 1,6; 2,4; e 3,2 dS m-1) e quatro concentrações de ácido salicílico (0; 1,2; 2,4 e 3,6 mM), com três repetições. A aplicação foliar de ácido salicílico na concentração de 1,6 mM atenuou os efeitos do estresse salino nas trocas gasosas, no crescimento, no peso médio de fruto e na produção total por planta, e diminuiu a porcentagem de extravasamento de eletrólitos intercelulares das plantas de pimentão cv. All Big aos 80 dias após o semeio.

Potassium fertilization as salt stress attenuator in sour passion fruit / Adubação potássica como atenuante do estresse salino em maracujazeiro-azedo

Water scarcity associated with irregular rainfall in the semi-arid region of Northeastern Brazil stands out as a limiting factor for agricultural production. Thus, the use of waters with high concentration of salts is an alternative to expand irrigated agriculture in this region. In this context, this study evaluated the water status, intercellular electrolyte leakage, photosynthetic pigments, and gas exchange of 'BRS SC1' sour passion fruit as a function of irrigation with water of different levels of salinity and potassium doses. The experiment was carried out in pots adapted as drainage lysimeters under field conditions at the Experimental Farm of the Federal University of Campina Grande in São Domingos - PB, Brazil. The experimental design was randomized blocks, in a 5 × 4 factorial scheme, whose treatments were obtained by combining two factors: five levels of electrical conductivity of irrigation water - ECw (0.3, 1.1, 1.9, 2.7, and 3.5 dS m-1), associated with four potassium doses (60, 80, 100, and 120% of the recommendation), with three replicates. Water with electrical conductivity greater than 0.3 dS m-1 reduced the relative water content, chlorophyll a and chlorophyll b contents, CO2 assimilation rate, and instantaneous water use efficiency of 'BRS SC1' sour passion fruit plants. The estimated potassium dose of 85% of the recommendation (equivalent to 293 g per plant per year) mitigated the deleterious effects of salt stress on stomatal conductance, transpiration, internal CO2 concentration, and instantaneous carboxylation efficiency of passion fruit 'BRS SC1'.

Water scarcity associated with irregular rainfall in the semi-arid region of Northeastern Brazil stands out as a limiting factor for agricultural production. Thus, the use of waters with high concentration of salts is an alternative to expand irrigated agriculture in this region. In this context, this study evaluated the water status, intercellular electrolyte leakage, photosynthetic pigments, and gas exchange of 'BRS SC1' sour passion fruit as a function of irrigation with water of different levels of salinity and potassium doses. The experiment was carried out in pots adapted as drainage lysimeters under field conditions at the Experimental Farm of the Federal University of Campina Grande in São Domingos - PB, Brazil. The experimental design was randomized blocks, in a 5 × 4 factorial scheme, whose treatments were obtained by combining two factors: five levels of electrical conductivity of irrigation water - ECw (0.3, 1.1, 1.9, 2.7, and 3.5 dS m-1), associated with four potassium doses (60, 80, 100, and 120% of the recommendation), with three replicates. Water with electrical conductivity greater than 0.3 dS m-1 reduced the relative water content, chlorophyll a and chlorophyll b contents, CO2 assimilation rate, and instantaneous water use efficiency of 'BRS SC1' sour passion fruit plants. The estimated potassium dose of 85% of the recommendation (equivalent to 293 g per plant per year) mitigated the deleterious effects of salt stress on stomatal conductance, transpiration, internal CO2 concentration, and instantaneous carboxylation efficiency of passion fruit 'BRS SC1'.

Genistein and Daidzein effects on the physiological indices of Soybean Cyst Nematodes

ABSTRACT: Soybean cyst nematode (SCN, Heterodera glycines) is a chief plantparasitic nematode of soybean. Application of synthetic chemical nematicides poses negative side effects to human health and the environment. Therefore, the search for a safe and effective approach is more relevant. This study evaluated the effects of Genistein and Daidzein on the physiological index of soybean cyst nematodes, individual morphology, reversal frequency, respiration, and body fluid leaking of secondstage juveniles (J2s) of soybean cyst nematode. The results showed that body length of J2s decreased, while stylet and tail transparent area elongated. Additionally, after the treatment, the body became hollow and shrunken, the J2s stiffened, whereas the reversal frequency decreased dramatically after 24 h of treatment. Moreover, the body fluid leakage was intensified and respiration was inhibited. Oxygen consumption decreased by 86.7 % and 70.1 %, while, in contrast, electrical conductivity increased by 40.1 % and 36 % at 100 g mL1 of Genistein and Daidzein, respectively, after 48 h of exposure. The smaller number of J2s in soybean roots, the slower development rate, and the abnormal sexual differentiation were found in greenhouse assay. Thus, Genistein and Daidzein, especially Genistein, have strong effects on the physiological index of soybean cyst nematodes. Moreover, the effects were time and dosagedependent.

Antiadhesion and antibiofilm potential of Fagonia indica from Cholistan desert against clinical multidrug resistant bacteria

Abstract High resistance to antimicrobials is associated with biofilm formation responsible for infectious microbes to withstand severe conditions. Therefore, new alternatives are necessary as biofilm inhibitors to control infections. In this study, the antimicrobial and antibiofilm activities of Fagonia indica extracts were evaluated against MDR clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica has antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against multidrug resistant (MDR) clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica had antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against MDR isolates. The maximum inhibitory effects of Fagonia indica chloroform extract on biofilm formation was observed on Staphylococcus aureus (71.84%) followed by Klebsiella pneumoniae (70.83%) after 48 hrs showing that inhibition is also time dependent. Our results about bacterial cell protein leakage indicated that MDR isolates treated with chloroform extract of Fagonia indica showed maximum protein leakage of K. pneumoniae (59.14 µg mL-1) followed by S. aureus (56.7 µg mL-1). Cell attachment assays indicated that chloroform extract resulted in a 43.5-53.5% inhibition of cell adherence to a polystyrene surface. Our results revealed that extracts of Fagonia indica significantly inhibited biofilm formation among MDR clinical isolates, therefore, could be applied as antimicrobial agents and cost effective biofilm inhibitor against these MDR isolates.

Resumo A alta resistência aos antimicrobianos está associada à formação de biofilme responsável por micróbios infecciosos para suportar condições severas. Portanto, novas alternativas são necessárias como inibidores de biofilme para controlar infecções. Neste estudo, as atividades antimicrobiana e antibiofilme dos extratos de Fagonia indica foram avaliadas contra isolados clínicos MDR. O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica tem efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-1, e valor de concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados clínicos multirresistentes (MDR). O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica teve efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-1, e concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados MDR. Os efeitos inibitórios máximos do extrato de clorofórmio Fagonia indica na formação de biofilme foi observada em Staphylococcus aureus (71,84%), seguido por Klebsiella pneumoniae (70,83%) após 48 horas, mostrando que a inibição também é dependente do tempo. Nossos resultados sobre extravasamento de proteínas de células bacterianas indicaram que isolados MDR tratados com extrato clorofórmico de Fagonia indica apresentaram vazamento máximo de proteínas de K. pneumoniae (59,14 µg mL-1), seguido por S. aureus (56,7 µg mL-1). Ensaios de fixação de células indicaram que o extrato de clorofórmio resultou em uma inibição de 43,5-53,5% da aderência das células a uma superfície de poliestireno. Nossos resultados revelaram que extratos de Fagonia indica inibiram significativamente a formação de biofilme entre isolados clínicos MDR, portanto, poderiam ser aplicados como agentes antimicrobianos e inibidores de biofilme de baixo custo contra esses isolados MDR.

Ionic homeostasis, biochemical components and yield of Italian zucchini under nitrogen forms and salt stress

Abstract This research was carried out aiming at evaluating the effects of nitrate and ammonium ions on nutrient accumulation, biochemical components and yield of Italian zucchini (cv. Caserta) grown in a hydroponic system under salt stress conditions. The experiment was carried out in a greenhouse utilizing an experimental design in randomized blocks, arranged in a 2 x 5 factorial scheme, with 4 replications. The treatments consisted of two forms of nitrogen (nitrate - NO3- and ammonium - NH4+) and 5 electrical conductivity levels of irrigation water (ECw) (0.5, 2.0, 3.5, 5.0 and 6.5 dS m-1). The analysis of the results indicated that supply of N exclusively in NH4+ form promotes greater damage to the leaf membrane and reduction in accumulation of macronutrients and higher Na+/K+, Na+/Ca++ and Na+/Mg++ ratios in the shoots of zucchini plants. Electrical conductivity of irrigation water above 2.0 dS m-1 reduces the accumulation of nutrients in shoot and yield of Italian zucchini plant. The toxicity of NH4+ under Italian zucchini plants overlap the toxicity of the salinity, since its fertilization exclusively with this form of nitrogen inhibits its production, being the NO3- form the most suitable for the cultivation of the species.

Resumo Este trabalho foi desenvolvido com o objetivo de avaliar os efeitos dos íons nitrato e amônio sobre o acúmulo de nutrientes e produção da abobrinha italiana (cv. Caserta) cultivada em sistema hidropônico sob estrese salino. O experimento foi conduzido em casa de vegetação utilizando o delineamento experimental em blocos casualizados, arranjados em esquema fatorial 2 x 5, com 4 repetições. Os tratamentos foram constituídos de duas formas de nitrogênio (nitrato - NO3- e amônio - NH4+) e cinco níveis de condutividade elétrica da água de irrigação (CEa) (0,5; 2,0; 3,5; 5,0 e 6,5 dS m-1). As análises dos resultados indicaram que suprimento de N exclusivamente em forma de NH4+ promove maiores danos na membrana foliar e redução no acúmulo de macronutrientes e maiores relações Na+/K+, Na+/Ca++ e Na+/Mg++ na parte aérea das plantas de abobrinha. A irrigação com água a cima de 2,0 dS m-1 reduz o acúmulo de nutrientes na parte aérea das plantas e a produção de abobrinha. A toxicidade do NH4+ sob abobrinha italiana sobrepõe-se à toxicidade da salinidade, pois a fertilização exclusiva com esta forma de nitrogênio inibe sua produção, sendo a forma NO3- a mais adequada para o cultivo da espécie.

Antiadhesion and antibiofilm potential of Fagonia indica from Cholistan desert against clinical multidrug resistant bacteria / Potencial de antiadesão e antibiofilme de Fagonia indica do deserto do Cholistão contra bactérias multirresistentes clínicas

High resistance to antimicrobials is associated with biofilm formation responsible for infectious microbes to withstand severe conditions. Therefore, new alternatives are necessary as biofilm inhibitors to control infections. In this study, the antimicrobial and antibiofilm activities of Fagonia indica extracts were evaluated against MDR clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica has antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-¹ against multidrug resistant (MDR) clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica had antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-¹ and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-¹ against MDR isolates. The maximum inhibitory effects of Fagonia indica chloroform extract on biofilm formation was observed on Staphylococcus aureus (71.84%) followed by Klebsiella pneumoniae (70.83%) after 48 hrs showing that inhibition is also time dependent. Our results about bacterial cell protein leakage indicated that MDR isolates treated with chloroform extract of Fagonia indica showed maximum protein leakage of K. pneumoniae (59.14 µg mL-¹) followed by S. aureus (56.7 µg mL-1). Cell attachment assays indicated that chloroform extract resulted in a 43.5-53.5% inhibition of cell adherence to a polystyrene surface. Our results revealed that extracts of Fagonia indica significantly inhibited biofilm formation among MDR clinical isolates, therefore, could be applied as antimicrobial agents and cost effective biofilm inhibitor against these MDR isolates.

A alta resistência aos antimicrobianos está associada à formação de biofilme responsável por micróbios infecciosos para suportar condições severas. Portanto, novas alternativas são necessárias como inibidores de biofilme para controlar infecções. Neste estudo, as atividades antimicrobiana e antibiofilme dos extratos de Fagonia indica foram avaliadas contra isolados clínicos MDR. O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica tem efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-¹, e valor de concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados clínicos multirresistentes (MDR). O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica teve efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-¹, e concentração bactericida mínima (MBC) de 500-3000 µg mL-¹ contra isolados MDR. Os efeitos inibitórios máximos do extrato de clorofórmio Fagonia indica na formação de biofilme foi observada em Staphylococcus aureus (71,84%), seguido por Klebsiella pneumoniae (70,83%) após 48 horas, mostrando que a inibição também é dependente do tempo. Nossos resultados sobre extravasamento de proteínas de células bacterianas indicaram que isolados MDR tratados com extrato clorofórmico de Fagonia indica apresentaram vazamento máximo de proteínas de K. pneumoniae (59,14 µg mL-¹), seguido por S. aureus(56,7 µg mL-¹). Ensaios de fixação de células indicaram que o extrato de clorofórmio resultou em uma inibição de 43,5-53,5% da aderência das células a uma superfície de poliestireno. Nossos resultados revelaram que extratos de Fagonia indica inibiram [...].

Antiadhesion and antibiofilm potential of Fagonia indica from Cholistan desert against clinical multidrug resistant bacteria / Potencial de antiadesão e antibiofilme de Fagonia indica do deserto do Cholistão contra bactérias multirresistentes clínicas

High resistance to antimicrobials is associated with biofilm formation responsible for infectious microbes to withstand severe conditions. Therefore, new alternatives are necessary as biofilm inhibitors to control infections. In this study, the antimicrobial and antibiofilm activities of Fagonia indica extracts were evaluated against MDR clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica has antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-¹ against multidrug resistant (MDR) clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica had antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-¹ and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-¹ against MDR isolates. The maximum inhibitory effects of Fagonia indica chloroform extract on biofilm formation was observed on Staphylococcus aureus (71.84%) followed by Klebsiella pneumoniae (70.83%) after 48 hrs showing that inhibition is also time dependent. Our results about bacterial cell protein leakage indicated that MDR isolates treated with chloroform extract of Fagonia indica showed maximum protein leakage of K. pneumoniae (59.14 µg mL-¹) followed by S. aureus (56.7 µg mL-1). Cell attachment assays indicated that chloroform extract resulted in a 43.5-53.5% inhibition of cell adherence to a polystyrene surface. Our results revealed that extracts of Fagonia indica significantly inhibited biofilm formation among MDR clinical isolates, therefore, could be applied as antimicrobial agents and cost effective biofilm inhibitor against these MDR isolates.(AU)

A alta resistência aos antimicrobianos está associada à formação de biofilme responsável por micróbios infecciosos para suportar condições severas. Portanto, novas alternativas são necessárias como inibidores de biofilme para controlar infecções. Neste estudo, as atividades antimicrobiana e antibiofilme dos extratos de Fagonia indica foram avaliadas contra isolados clínicos MDR. O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica tem efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-¹, e valor de concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados clínicos multirresistentes (MDR). O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica teve efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-¹, e concentração bactericida mínima (MBC) de 500-3000 µg mL-¹ contra isolados MDR. Os efeitos inibitórios máximos do extrato de clorofórmio Fagonia indica na formação de biofilme foi observada em Staphylococcus aureus (71,84%), seguido por Klebsiella pneumoniae (70,83%) após 48 horas, mostrando que a inibição também é dependente do tempo. Nossos resultados sobre extravasamento de proteínas de células bacterianas indicaram que isolados MDR tratados com extrato clorofórmico de Fagonia indica apresentaram vazamento máximo de proteínas de K. pneumoniae (59,14 µg mL-¹), seguido por S. aureus(56,7 µg mL-¹). Ensaios de fixação de células indicaram que o extrato de clorofórmio resultou em uma inibição de 43,5-53,5% da aderência das células a uma superfície de poliestireno. Nossos resultados revelaram que extratos de Fagonia indica inibiram [...].(AU)

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos

Abstract This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Resumo Este estudo investigou o uso da melatonina para conter os efeitos da apoptose em embriões vitrificados de zebrafish (D. rerio). Embriões descorionados no estágio de 22-24 somitos foram divididos (n = 60 / tratamento) em tratamento não vitrificado (Grupo Controle, melatonina 0 M) e tratamentos vitrificados com 0 M (T1), 1 µM (T2) e 1 mM de melatonina (T3). Para os tratamentos vitrificados, utilizou-se uma solução à base de metanol/propilenoglicol e os embriões foram armazenados em -196 °C por uma semana. Após o descongelamento, foram realizadas análises de taxa de sobrevivência, microscopia eletrônica de varredura, expressão dos genes anti (bcl-2) e pró-apoptóticos (bax/caspase-3), formação de espécies reativas de oxigênio (EROS) e análises de fragmentação de DNA. Não foram obtidos embriões vivos a partir dos tratamentos vitrificados, observando uma rápida degeneração imediatamente após o descongelamento, com ruptura da camada vitelina e vazamento de seu conteúdo, seguida de quebra das células epiteliais e melanização do tecido. Em relação ao processo apoptótico. T3 apresentou expressão gênica relativa alta para os três genes (P 0,05), além disso, T2 apresentou expressão semelhante as dos genes pró-apoptóticos de GC (P 0,05). A formação de EROS revelou que GC apresentou menor percentual de área de superfície embrionária afetada (3,80 ± 0,40%) (P 0,05), ao contrário, não foram encontradas diferenças entre os outros grupos. T1 foi mais significativamente (P 0,05) danificado pela fragmentação do DNA. Os grupos vitrificados com melatonina apresentaram níveis de dano semelhantes ao do GC (P> 0,05). A inclusão de 1 µM de melatonina na solução de vitrificação, contrariou os efeitos do processo apoptótico em embriões pós-descongelamento, sugerindo sua utilidade na criopreservação de embriões de peixes.

Genistein and Daidzein effects on the physiological indices of Soybean Cyst Nematodes

Soybean cyst nematode (SCN, Heterodera glycines) is a chief plant-parasitic nematode of soybean. Application of synthetic chemical nematicides poses negative side effects to human health and the environment. Therefore, the search for a safe and effective approach is more relevant. This study evaluated the effects of Genistein and Daidzein on the physiological index of soybean cyst nematodes, individual morphology, reversal frequency, respiration, and body fluid leaking of second-stage juveniles (J2s) of soybean cyst nematode. The results showed that body length of J2s decreased, while stylet and tail transparent area elongated. Additionally, after the treatment, the body became hollow and shrunken, the J2s stiffened, whereas the reversal frequency decreased dramatically after 24 h of treatment. Moreover, the body fluid leakage was intensified and respiration was inhibited. Oxygen consumption decreased by 86.7 % and 70.1 %, while, in contrast, electrical conductivity increased by 40.1 % and 36 % at 100 μg mL−1 of Genistein and Daidzein, respectively, after 48 h of exposure. The smaller number of J2s in soybean roots, the slower development rate, and the abnormal sexual differentiation were found in greenhouse assay. Thus, Genistein and Daidzein, especially Genistein, have strong effects on the physiological index of soybean cyst nematodes. Moreover, the effects were time and dosage-dependent.

Antiadhesion and antibiofilm potential of Fagonia indica from Cholistan desert against clinical multidrug resistant bacteria / Potencial de antiadesão e antibiofilme de Fagonia indica do deserto do Cholistão contra bactérias multirresistentes clínicas

High resistance to antimicrobials is associated with biofilm formation responsible for infectious microbes to withstand severe conditions. Therefore, new alternatives are necessary as biofilm inhibitors to control infections. In this study, the antimicrobial and antibiofilm activities of Fagonia indica extracts were evaluated against MDR clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica has antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against multidrug resistant (MDR) clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica had antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against MDR isolates. The maximum inhibitory effects of Fagonia indica chloroform extract on biofilm formation was observed on Staphylococcus aureus (71.84%) followed by Klebsiella pneumoniae (70.83%) after 48 hrs showing that inhibition is also time dependent. Our results about bacterial cell protein leakage indicated that MDR isolates treated with chloroform extract of Fagonia indica showed maximum protein leakage of K. pneumoniae (59.14 µg mL-1) followed by S. aureus (56.7 µg mL-1). Cell attachment assays indicated that chloroform extract resulted in a 43.5-53.5% inhibition of cell adherence to a polystyrene surface. Our results revealed that extracts of Fagonia indica significantly inhibited biofilm formation among MDR clinical isolates, therefore, could be applied as antimicrobial agents and cost effective biofilm inhibitor against these MDR isolates.

A alta resistência aos antimicrobianos está associada à formação de biofilme responsável por micróbios infecciosos para suportar condições severas. Portanto, novas alternativas são necessárias como inibidores de biofilme para controlar infecções. Neste estudo, as atividades antimicrobiana e antibiofilme dos extratos de Fagonia indica foram avaliadas contra isolados clínicos MDR. O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica tem efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-1, e valor de concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados clínicos multirresistentes (MDR). O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica teve efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-1, e concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados MDR. Os efeitos inibitórios máximos do extrato de clorofórmio Fagonia indica na formação de biofilme foi observada em Staphylococcus aureus (71,84%), seguido por Klebsiella pneumoniae (70,83%) após 48 horas, mostrando que a inibição também é dependente do tempo. Nossos resultados sobre extravasamento de proteínas de células bacterianas indicaram que isolados MDR tratados com extrato clorofórmico de Fagonia indica apresentaram vazamento máximo de proteínas de K. pneumoniae (59,14 µg mL-1), seguido por S. aureus (56,7 µg mL-1). Ensaios de fixação de células indicaram que o extrato de clorofórmio resultou em uma inibição de 43,5-53,5% da aderência das células a uma superfície de poliestireno. Nossos resultados revelaram que extratos de Fagonia indica inibiram significativamente a formação de biofilme entre isolados clínicos MDR, portanto, poderiam ser aplicados como agentes antimicrobianos e inibidores de biofilme de baixo custo contra esses isolados MDR.

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos / Uso da melatonina como inibidor do processo apoptotico para a criopreservação de embriões de zebrafish (Danio rerio)

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P < 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P < 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P < 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P < 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Este estudo investigou o uso da melatonina para conter os efeitos da apoptose em embriões vitrificados de zebrafish (D. rerio). Embriões descorionados no estágio de 22-24 somitos foram divididos (n = 60 / tratamento) em tratamento não vitrificado (Grupo Controle, melatonina 0 M) e tratamentos vitrificados com 0 M (T1), 1 µM (T2) e 1 mM de melatonina (T3). Para os tratamentos vitrificados, utilizou-se uma solução à base de metanol/propilenoglicol e os embriões foram armazenados em -196 °C por uma semana. Após o descongelamento, foram realizadas análises de taxa de sobrevivência, microscopia eletrônica de varredura, expressão dos genes anti (bcl-2) e pró-apoptóticos (bax/caspase-3), formação de espécies reativas de oxigênio (EROS) e análises de fragmentação de DNA. Não foram obtidos embriões vivos a partir dos tratamentos vitrificados, observando uma rápida degeneração imediatamente após o descongelamento, com ruptura da camada vitelina e vazamento de seu conteúdo, seguida de quebra das células epiteliais e melanização do tecido. Em relação ao processo apoptótico. T3 apresentou expressão gênica relativa alta para os três genes (P <0,05), além disso, T2 apresentou expressão semelhante as dos genes pró-apoptóticos de GC (P <0,05). A formação de EROS revelou que GC apresentou menor percentual de área de superfície embrionária afetada (3,80 ± 0,40%) (P <0,05), ao contrário, não foram encontradas diferenças entre os outros grupos. T1 foi mais significativamente (P <0,05) danificado pela fragmentação do DNA. Os grupos vitrificados com melatonina apresentaram níveis de dano semelhantes ao do GC (P> 0,05). A inclusão de 1 µM de melatonina na solução de vitrificação, contrariou os efeitos do processo apoptótico em embriões pós-descongelamento, sugerindo sua utilidade na criopreservação de embriões de peixes.

Quantum yield, chlorophyll, and cell damage in yellow passion fruit under irrigation strategies with brackish water and potassium / Rendimento quântico, clorofila e dano celular em maracujazeiro-amarelo sob estratégias de irrigação com água salobra e potássio

The occurrence of water sources with high concentrations of salts in the semiarid region of Northeast Brazil stands out as a limiting factor in the cultivation of irrigated yellow passion fruit. Thus the search for irrigation strategies with brackish water is fundamental for the sustainability of irrigated crops. The objective of the present study was to determine the quantum yield, chlorophyll levels, and cell damage in yellow passion fruit cultivated under different irrigation strategies with brackish water and potassium fertilization. The experiment was conducted under field conditions in São Domingos - PB, Brazil, adopting a randomized block design in a 6×2 factorial scheme, consisting of six strategies of use of brackish water applied in the different phenological stages of crop and two doses of potassium (60 and 100% of the recommendation), with four replicates. Two levels of irrigation water salinity were used, with low (1.3 dS m-1) and high electrical conductivity (4.0 dS m-1). The potassium dose of 100% recommendation corresponds to the application of 345 g of K2O per plant per year. The use of brackish water (4.0 dS m-1) increased chlorophyll fluorescence and negatively affected the photosynthetic pigments of yellow passion fruit, regardless of the phenological stage. Salt stress intensified intercellular electrolyte leakage in passion fruit plants under higher dose of potassium. Our results show that fertilization equivalent to 100% of the potassium recommendation is excessive for yellow passion fruit 'BRS GA1' in the tropical semiarid region under salt stress conditions.

A ocorrência de mananciais com altas concentrações de sais no semiárido nordestino destaca-se como fator limitante no cultivo do maracujazeiro-amarelo irrigado. Assim, a busca por estratégias de irrigação com água salobra é fundamental para a sustentabilidade da agricultura irrigada. O objetivo deste trabalho foi determinar o rendimento quântico, os teores de clorofila e dano celular em maracujazeiro-amarelo cultivado sob diferentes estratégias de irrigação com água salobra e adubação potássica. O experimento foi conduzido em condições de campo em São Domingos - PB, Brasil, adotando-se o delineamento em blocos casualizados em esquema fatorial 6×2, correspondendo a seis estratégias de uso de água salobra aplicadas nos diferentes estádios fenológicos da cultura e duas doses de potássio (60 e 100% da recomendação), com quatro repetições. Foram utilizados dois níveis de salinidade da água de irrigação, com baixa (1,3 dS m-1) e alta condutividade elétrica (4,0 dS m-1). A dose de potássio de 100% corresponde à aplicação de 345 g de K2O por planta por ano. O uso de água salobra (4,0 dS m-1) aumentou a fluorescência da clorofila e afetou negativamente os pigmentos fotossintéticos do maracujá-amarelo, independente do estádio fenológico. O estresse salino intensificou o extravasamento de eletrólitos em plantas de maracujazeiro adubadas sob alta dose de potássio. Os resultados mostram que a adubação equivalente a 100% da recomendação de potássio é excessiva para o cultivo do maracujazeiro-amarelo 'BRS GA1' no semiárido tropical, sob condições de estresse salino.

Ionic homeostasis, biochemical components and yield of Italian zucchini under nitrogen forms and salt stress / Homeostase iônica, componentes bioquímicos e produção da abobrinha italiana sob formas de nitrogênio e estresse salino

This research was carried out aiming at evaluating the effects of nitrate and ammonium ions on nutrient accumulation, biochemical components and yield of Italian zucchini (cv. Caserta) grown in a hydroponic system under salt stress conditions. The experiment was carried out in a greenhouse utilizing an experimental design in randomized blocks, arranged in a 2 x 5 factorial scheme, with 4 replications. The treatments consisted of two forms of nitrogen (nitrate - NO3 - and ammonium - NH4 + ) and 5 electrical conductivity levels of irrigation water (ECw) (0.5, 2.0, 3.5, 5.0 and 6.5 dS m-1). The analysis of the results indicated that supply of N exclusively in NH4 + form promotes greater damage to the leaf membrane and reduction in accumulation of macronutrients and higher Na+ /K+ , Na+ /Ca++ and Na+ /Mg++ ratios in the shoots of zucchini plants. Electrical conductivity of irrigation water above 2.0 dS m-1 reduces the accumulation of nutrients in shoot and yield of Italian zucchini plant. The toxicity of NH4 + under Italian zucchini plants overlap the toxicity of the salinity, since its fertilization exclusively with this form of nitrogen inhibits its production, being the NO3 - form the most suitable for the cultivation of the species.

Este trabalho foi desenvolvido com o objetivo de avaliar os efeitos dos íons nitrato e amônio sobre o acúmulo de nutrientes e produção da abobrinha italiana (cv. Caserta) cultivada em sistema hidropônico sob estrese salino. O experimento foi conduzido em casa de vegetação utilizando o delineamento experimental em blocos casualizados, arranjados em esquema fatorial 2 x 5, com 4 repetições. Os tratamentos foram constituídos de duas formas de nitrogênio (nitrato - NO3 - e amônio - NH4 + ) e cinco níveis de condutividade elétrica da água de irrigação (CEa ) (0,5; 2,0; 3,5; 5,0 e 6,5 dS m-1). As análises dos resultados indicaram que suprimento de N exclusivamente em forma de NH4 + promove maiores danos na membrana foliar e redução no acúmulo de macronutrientes e maiores relações Na+ /K+ , Na+ /Ca++ e Na+ /Mg++ na parte aérea das plantas de abobrinha. A irrigação com água a cima de 2,0 dS m-1 reduz o acúmulo de nutrientes na parte aérea das plantas e a produção de abobrinha. A toxicidade do NH4 + sob abobrinha italiana sobrepõe-se à toxicidade da salinidade, pois a fertilização exclusiva com esta forma de nitrogênio inibe sua produção, sendo a forma NO3 - a mais adequada para o cultivo da espécie.

Ionic homeostasis, biochemical components and yield of Italian zucchini under nitrogen forms and salt stress / Homeostase iônica, componentes bioquímicos e produção da abobrinha italiana sob formas de nitrogênio e estresse salino

This research was carried out aiming at evaluating the effects of nitrate and ammonium ions on nutrient accumulation, biochemical components and yield of Italian zucchini (cv. Caserta) grown in a hydroponic system under salt stress conditions. The experiment was carried out in a greenhouse utilizing an experimental design in randomized blocks, arranged in a 2 x 5 factorial scheme, with 4 replications. The treatments consisted of two forms of nitrogen (nitrate - NO3- and ammonium - NH4+) and 5 electrical conductivity levels of irrigation water (ECw) (0.5, 2.0, 3.5, 5.0 and 6.5 dS m-1). The analysis of the results indicated that supply of N exclusively in NH4+ form promotes greater damage to the leaf membrane and reduction in accumulation of macronutrients and higher Na+/K+, Na+/Ca++ and Na+/Mg++ ratios in the shoots of zucchini plants. Electrical conductivity of irrigation water above 2.0 dS m-¹ reduces the accumulation of nutrients in shoot and yield of Italian zucchini plant. The toxicity of NH4+ under Italian zucchini plants overlap the toxicity of the salinity, since its fertilization exclusively with this form of nitrogen inhibits its production, being the NO3- form the most suitable for the cultivation of the species.

Este trabalho foi desenvolvido com o objetivo de avaliar os efeitos dos íons nitrato e amônio sobre o acúmulo de nutrientes e produção da abobrinha italiana (cv. Caserta) cultivada em sistema hidropônico sob estrese salino. O experimento foi conduzido em casa de vegetação utilizando o delineamento experimental em blocos casualizados, arranjados em esquema fatorial 2 x 5, com 4 repetições. Os tratamentos foram constituídos de duas formas de nitrogênio (nitrato - NO3- e amônio - NH4+) e cinco níveis de condutividade elétrica da água de irrigação (CEa) (0,5; 2,0; 3,5; 5,0 e 6,5 dS m-¹). As análises dos resultados indicaram que suprimento de N exclusivamente em forma de NH4+ promove maiores danos na membrana foliar e redução no acúmulo de macronutrientes e maiores relações Na+/K+, Na+/Ca++ e Na+/Mg++ na parte aérea das plantas de abobrinha. A irrigação com água a cima de 2,0 dS m-¹ reduz o acúmulo de nutrientes na parte aérea das plantas e a produção de abobrinha. A toxicidade do NH4+ sob abobrinha italiana sobrepõe-se à toxicidade da salinidade, pois a fertilização exclusiva com esta forma de nitrogênio inibe sua produção, sendo a forma NO3- a mais adequada para o cultivo da espécie.

Urethral reconstruction combined with modified urethrostomy in a cat after prepubic urethrostomy

Background: In cats, urethral obstruction is generally caused by various reasons, including feline idiopathic cystitis, urethral crystals, urethral trauma, mucous plug, congenital or acquired anatomical deformity, and urolithiasis, especially in male cats. Depending on the severity and duration of clinical signs, immediate management including conservative or surgical therapy for restoration of urethral patency is required. Repetitive urethral obstruction due to intraluminal plugs, obstruction that cannot be resolved by medical management, and urethral strictures, trauma, or neoplasia should be managed by surgery. When the penile or pelvic urethra is ruptured or not long enough to mobilize the pelvic urethra to the perineal skin by repetitive perineal urethrostomy, prepubic urethrostomy is indicated. Potential complications of prepubic urethrostomy include urinary incontinence, peristomal dermatitis. In such cases, management of the peristomal site or placement of an artificial urethral sphincter have been reported previously. However, to date, urethral reconstruction using pre-existing penis has not been reported after prepubic urethral stricture following prepubic urethrostomy. The objective of this report is to describe surgical procedure of the urethral reconstruction combined with modified perineal urethrostomy in a cat with prepubic urethral stricture after prepubic urethrosotmy. Case: A 3-year-old castrated Scottish straight cat presented with dysuria after prepubic urethrostomy. The owner reported that the patient was diagnosed with urethral rupture, had undergone prepubic urethrostomy 2 months prior to presentation, and had persistent dysuria despite repetitive surgical revision of the prepubic urethral stoma at a local hospital. On physical examination, the preputial orifice and the penis appeared grossly normal. On ultrasonography, pericystic and periurethral fat had an edematous, striated appearance with alternating hyperechoic and anechoic regions. A small amount of free fluid was visible around the urethra. Abdominal radiography revealed a mildly distended bladder and loss of serosal detail around the area of the urinary bladder neck, consistent with inflammation and free fluid observed on ultrasonography. The retrograde urethrogram showed no leakage in either sites of the prepubic urethral orifice or the penis. Hence, complete urethral reconstruction with modified perineal urethrostomy was performed. The patient had normal urination at the 15-month follow-up. Discussion: Prepubic urethrostomy is beneficial for the patients whose penile or pelvic urethra is ruptured or not long enough to mobilize the pelvic urethra to the perineum. Complications of prepubic urethrostomy include skin necrosis around the stoma; urinary incontinence; stricture of the urethral stoma due to several reasons such as surgical-site irritation; poor mucosa-skin apposition; failure to provide tension-free stoma; and failure to expose wider pelvic urethra. In these cases, surgical management of the level of prepubic urethral stoma has been recommended by previous studies. However, no studies have been reported urethral reconstruction using pre-existing penile urethra in a patient with prepubic urethral stricture so far. To the author's knowledge, this case is the first report describing urethral reconstruction in a cat with urethral stricture after prepubic urethrostomy.

Primary perianal malignant melanoma in a dog - combination therapy

Background: Melanocytic neoplasm can arise from melanocytes in any location of the body. Malignant melanoma (MM)has a poor prognosis in dogs and presence of lymphvascular invasion, distant metastasis, or mitotic activity present prognostic value. Primary melanoma affecting the gastrointestinal tract has been rarely reported in veterinary literature, thusthe prognosis affecting gastrointestinal tract is unknown. Electrochemotherapy (ECT) is an effective local treatment whichcombines chemotherapeutic drugs mainly bleomycin or cisplatin followed by the delivery of permeabilizing electricalpulses However, other hydrophilic drugs seem to present an increase cytotoxic effect such as carboplatin.Case: A 9-year-old mixed-breed neutered dog was referred to a private clinic with a mass in the perianal region diagnosedas perianal melanoma. No metastasis was observed on abdominal ultrasound nor chest x-ray (3 views). Clinical signs notedwere tenesmus, hemorrhagic discharge, weight loss and hyporexia. Considering the tumor volume (16.0 x 10.0 cm), a neoadjuvant ECT session was proposed. The authors opted for carboplatin (300 mg/m2, intravenously), administered over 20min and cisplatin intratumorally (1 mg/cm3, equivalent to 1 mL/1cm3 total volume 20 mL) administered in the upper partsof the mass that could be reached while avoiding drug leakage. After administration, sequences of eight biphasic pulses,(100 microseconds), with a voltage ranging from 650-1,000V/cm (pulse generator Onkodisruptor®) using a hexagonal/single pair and plate electrode were delivered. At day 30th, a partial response was observed accordingly to RECIST system,with tumor size of 5.0 x 5.0 cm (65.4 cm3). A second ECT session was performed with the same previous protocol, butwith a decreased dosage of carboplatin (240 mg/m2 consistent with 20% reduction) due to adverse effects in the first session, resulting in stable disease at day 60th (30 days after second ECT). Then...

Primary perianal malignant melanoma in a dog - combination therapy

Background: Melanocytic neoplasm can arise from melanocytes in any location of the body. Malignant melanoma (MM)has a poor prognosis in dogs and presence of lymphvascular invasion, distant metastasis, or mitotic activity present prognostic value. Primary melanoma affecting the gastrointestinal tract has been rarely reported in veterinary literature, thusthe prognosis affecting gastrointestinal tract is unknown. Electrochemotherapy (ECT) is an effective local treatment whichcombines chemotherapeutic drugs mainly bleomycin or cisplatin followed by the delivery of permeabilizing electricalpulses However, other hydrophilic drugs seem to present an increase cytotoxic effect such as carboplatin.Case: A 9-year-old mixed-breed neutered dog was referred to a private clinic with a mass in the perianal region diagnosedas perianal melanoma. No metastasis was observed on abdominal ultrasound nor chest x-ray (3 views). Clinical signs notedwere tenesmus, hemorrhagic discharge, weight loss and hyporexia. Considering the tumor volume (16.0 x 10.0 cm), a neoadjuvant ECT session was proposed. The authors opted for carboplatin (300 mg/m2, intravenously), administered over 20min and cisplatin intratumorally (1 mg/cm3, equivalent to 1 mL/1cm3 total volume 20 mL) administered in the upper partsof the mass that could be reached while avoiding drug leakage. After administration, sequences of eight biphasic pulses,(100 microseconds), with a voltage ranging from 650-1,000V/cm (pulse generator Onkodisruptor®) using a hexagonal/single pair and plate electrode were delivered. At day 30th, a partial response was observed accordingly to RECIST system,with tumor size of 5.0 x 5.0 cm (65.4 cm3). A second ECT session was performed with the same previous protocol, butwith a decreased dosage of carboplatin (240 mg/m2 consistent with 20% reduction) due to adverse effects in the first session, resulting in stable disease at day 60th (30 days after second ECT). Then...(AU)

Ionic homeostasis, biochemical components and yield of Italian zucchini under nitrogen forms and salt stress / Homeostase iônica, componentes bioquímicos e produção da abobrinha italiana sob formas de nitrogênio e estresse salino

This research was carried out aiming at evaluating the effects of nitrate and ammonium ions on nutrient accumulation, biochemical components and yield of Italian zucchini (cv. Caserta) grown in a hydroponic system under salt stress conditions. The experiment was carried out in a greenhouse utilizing an experimental design in randomized blocks, arranged in a 2 x 5 factorial scheme, with 4 replications. The treatments consisted of two forms of nitrogen (nitrate - NO3- and ammonium - NH4+) and 5 electrical conductivity levels of irrigation water (ECw) (0.5, 2.0, 3.5, 5.0 and 6.5 dS m-1). The analysis of the results indicated that supply of N exclusively in NH4+ form promotes greater damage to the leaf membrane and reduction in accumulation of macronutrients and higher Na+/K+, Na+/Ca++ and Na+/Mg++ ratios in the shoots of zucchini plants. Electrical conductivity of irrigation water above 2.0 dS m-¹ reduces the accumulation of nutrients in shoot and yield of Italian zucchini plant. The toxicity of NH4+ under Italian zucchini plants overlap the toxicity of the salinity, since its fertilization exclusively with this form of nitrogen inhibits its production, being the NO3- form the most suitable for the cultivation of the species.(AU)

Este trabalho foi desenvolvido com o objetivo de avaliar os efeitos dos íons nitrato e amônio sobre o acúmulo de nutrientes e produção da abobrinha italiana (cv. Caserta) cultivada em sistema hidropônico sob estrese salino. O experimento foi conduzido em casa de vegetação utilizando o delineamento experimental em blocos casualizados, arranjados em esquema fatorial 2 x 5, com 4 repetições. Os tratamentos foram constituídos de duas formas de nitrogênio (nitrato - NO3- e amônio - NH4+) e cinco níveis de condutividade elétrica da água de irrigação (CEa) (0,5; 2,0; 3,5; 5,0 e 6,5 dS m-¹). As análises dos resultados indicaram que suprimento de N exclusivamente em forma de NH4+ promove maiores danos na membrana foliar e redução no acúmulo de macronutrientes e maiores relações Na+/K+, Na+/Ca++ e Na+/Mg++ na parte aérea das plantas de abobrinha. A irrigação com água a cima de 2,0 dS m-¹ reduz o acúmulo de nutrientes na parte aérea das plantas e a produção de abobrinha. A toxicidade do NH4+ sob abobrinha italiana sobrepõe-se à toxicidade da salinidade, pois a fertilização exclusiva com esta forma de nitrogênio inibe sua produção, sendo a forma NO3- a mais adequada para o cultivo da espécie.(AU)

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos / Uso da melatonina como inibidor do processo apoptotico para a criopreservação de embriões de zebrafish (Danio rerio)

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Este estudo investigou o uso da melatonina para conter os efeitos da apoptose em embriões vitrificados de zebrafish (D. rerio). Embriões descorionados no estágio de 22-24 somitos foram divididos (n = 60 / tratamento) em tratamento não vitrificado (Grupo Controle, melatonina 0 M) e tratamentos vitrificados com 0 M (T1), 1 µM (T2) e 1 mM de melatonina (T3). Para os tratamentos vitrificados, utilizou-se uma solução à base de metanol/propilenoglicol e os embriões foram armazenados em -196 °C por uma semana. Após o descongelamento, foram realizadas análises de taxa de sobrevivência, microscopia eletrônica de varredura, expressão dos genes anti (bcl-2) e pró-apoptóticos (bax/caspase-3), formação de espécies reativas de oxigênio (EROS) e análises de fragmentação de DNA. Não foram obtidos embriões vivos a partir dos tratamentos vitrificados, observando uma rápida degeneração imediatamente após o descongelamento, com ruptura da camada vitelina e vazamento de seu conteúdo, seguida de quebra das células epiteliais e melanização do tecido. Em relação ao processo apoptótico. T3 apresentou expressão gênica relativa alta para os três genes (P 0,05). A inclusão de 1 µM de melatonina na solução de vitrificação, contrariou os efeitos do processo apoptótico em embriões pós-descongelamento, sugerindo sua utilidade na criopreservação de embriões de peixes.

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos / Uso da melatonina como inibidor do processo apoptotico para a criopreservação de embriões de zebrafish (Danio rerio)

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P < 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P < 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P < 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P < 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.(AU)

Este estudo investigou o uso da melatonina para conter os efeitos da apoptose em embriões vitrificados de zebrafish (D. rerio). Embriões descorionados no estágio de 22-24 somitos foram divididos (n = 60 / tratamento) em tratamento não vitrificado (Grupo Controle, melatonina 0 M) e tratamentos vitrificados com 0 M (T1), 1 µM (T2) e 1 mM de melatonina (T3). Para os tratamentos vitrificados, utilizou-se uma solução à base de metanol/propilenoglicol e os embriões foram armazenados em -196 °C por uma semana. Após o descongelamento, foram realizadas análises de taxa de sobrevivência, microscopia eletrônica de varredura, expressão dos genes anti (bcl-2) e pró-apoptóticos (bax/caspase-3), formação de espécies reativas de oxigênio (EROS) e análises de fragmentação de DNA. Não foram obtidos embriões vivos a partir dos tratamentos vitrificados, observando uma rápida degeneração imediatamente após o descongelamento, com ruptura da camada vitelina e vazamento de seu conteúdo, seguida de quebra das células epiteliais e melanização do tecido. Em relação ao processo apoptótico. T3 apresentou expressão gênica relativa alta para os três genes (P <0,05), além disso, T2 apresentou expressão semelhante as dos genes pró-apoptóticos de GC (P <0,05). A formação de EROS revelou que GC apresentou menor percentual de área de superfície embrionária afetada (3,80 ± 0,40%) (P <0,05), ao contrário, não foram encontradas diferenças entre os outros grupos. T1 foi mais significativamente (P <0,05) danificado pela fragmentação do DNA. Os grupos vitrificados com melatonina apresentaram níveis de dano semelhantes ao do GC (P> 0,05). A inclusão de 1 µM de melatonina na solução de vitrificação, contrariou os efeitos do processo apoptótico em embriões pós-descongelamento, sugerindo sua utilidade na criopreservação de embriões de peixes.(AU)