Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum / Composição química e efeito inibitório in vitro dos óleos essenciais das cascas dos frutos de três espécies de Citrus e do limoneno sobre o crescimento micelial de Sclerotinia sclerotiorum

Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.(AU)

Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.(AU)

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Chemical composition and in vitro inhibitory effects of essential oils from fruit peel of three Citrus species and limonene on mycelial growth of Sclerotinia sclerotiorum

Abstract Essential oils (EO) from aromatic and medicinal plants generally perform a diverse range of biological activities because they have several active constituents that work in different mechanisms of action. EO from Citrus peel have an impressive range of food and medicinal uses, besides other applications. EO from Citrus reticulata, C. sinensis and C. deliciosa were extracted from fruit peel and analyzed by GC-MS. The major constituent of EO under evaluation was limonene, whose concentrations were 98.54%, 91.65% and 91.27% for C. sinensis, C. reticulata and C. deliciosa, respectively. The highest potential of inhibition of mycelial growth was observed when the oil dose was 300 L. Citrus oils inhibited fungus growth in 82.91% (C. deliciosa), 65.82% (C. sinensis) and 63.46% (C. reticulata). Anti-Sclerotinia sclerotiorum activity of 90% pure limonene and at different doses (20, 50, 100, 200 and 300 L) was also investigated. This monoterpene showed to be highly active by inhibiting 100% fungus growth even at 200 and 300 L doses. This is the first report of the in vitro inhibitory effect of natural products from these three Citrus species and its results show that there is good prospect of using them experimentally to control S. sclerotiorum, in both greenhouse and field conditions.

Resumo Óleos essenciais de plantas aromáticas e medicinais geralmente exibem uma gama diversificada de atividades biológicas, pois possuem vários constituintes ativos que atuam por meio de vários mecanismos de ação. Os óleos essenciais das cascas de Citrus têm uma variedade impressionante de usos em alimentos, medicamentos entre várias outras aplicações. Os óleos essenciais (OE) de Citrus reticulata, C. sinenses e C. deliciosa foram extraídos das cascas dos frutos e analisados por CG-EM. O limoneno foi o constituinte majoritário encontrado nos óleos essenciais avaliados, nas concentrações de 98,54%, 91,65% e 91,27% para C. sinensis, C. reticulata e C. deliciosa, respectivamente. Os maiores potenciais de inibição do crescimento micelial foi observado na dose de 300 µL dos óleos. Os óleos de Citrus inibiram em 82,91% (C. deliciosa), 65,82% (C. sinensis) e 63,46% (C. reticulata) o crescimento do fungo. A atividade anti-Sclerotinia sclerotiorum do limoneno 90% puro e em diferentes doses (20, 50, 100, 200 e 300 µL) foi também investigada e este monoterpeno demonstrou-se altamente ativo inibindo 100% o crescimento do fungo inclusive nas doses de 200 e 300 µL. Este é o primeiro relato sobre o efeito inibitório in vitro dos óleos essenciais destas três espécies de Citrus e os resultados deste estudo mostram que existe uma boa perspectiva de uso destes produtos naturais experimentalmente para controlar o S. sclerotiorum tanto em condições de estufa como em condições de campo.

Determinação da energia metabolizável do farelo residual do milho com e sem enzima em dietas para frangos de corte / Determination of the metabolizable energy of residual corn bran with and without enzyme in diets for broilers

O objetivo deste trabalho foi avaliar o valor nutricional e determinar a energia metabolizável do farelo residual de milho (FRM) sem e com o uso da enzima alfa- amilase. Foi realizado um experimento de metabolismo com 180 pintos machos Cobb com 14 dias, distribuídos em um delineamento inteiramente ao acaso, com seis tratamentos, cinco repetições e seis aves por parcela. As dietas experimentais foram: T1: ração referência (RR), T2: 60% T1 + 40% de FRM, T3: RR + enzima, T4: 60% T1 + 40% de FRM com adição de enzima, T5: RR com substituição de 100% do milho pelo FRM e T6: RR com substituição de 100% do milho pelo FRM com adição de enzima. A composição química do FRM foi: 88,33% de matéria seca (MS), 10,23% de proteína bruta (PB), 15,44% de extrato etéreo (EE), 4,33% de cinzas (CZ) e 4555kcal/kg de energia bruta (EB). Os valores dos coeficientes de metabolizabilidade aparente para o FRM sem e com adição de enzima foram, respectivamente, de 73,37% e 76,33% para MS (p=0,0136), 70,44% e 70,39% para PB (p=0,9595) e de 74,79% e 76,77% para EB (p=0,0128). Os valores da energia metabolizável aparente (EMA) e da EMA corrigida para retenção de nitrogênio (EMAn) para o FRM (na base natural) foram de 3322±19 e 3241±18kcal/kg e de 3334±16 e 3261±17kcal/kg, respectivamente, sem e com adição de enzima. A adição da enzima não teve efeito estatístico significativo sobre os valores de EMA e EMAn, entretanto houve efeito positivo no coeficiente de metabolização da energia.(AU)

The aim of this study was to evaluate the nutritional value and the metabolizable energy of residual corn bran (FRM) with and without the use of alpha amylase enzyme. One metabolism assay was done using 180 Ross male broilers at 14 days of age distributed in an entirely randomized design with six treatments, five replications and six broilers per plot. The assay treatments established were: T1: reference diet (RR), T2: 60% T1 + 40% of FRM, T3: T1 (RR) + enzyme addition, T4: 60% T1 + 40% of FRM with enzyme addition, T5: T1 (RR) with 100% replacement of corn by FRM and T6: T1 (RR) with 100% replacement of corn by FRM with enzyme addition. FRM chemical composition was: 88.33% of dry matter (MS), 10.23% of crude protein (PB), 15.44% of crude fat (EE), 4.33% of ashes (CZ) and 4555Kcal/kg of Gross Energy (EB). The FRM apparent metabolizability coefficients with and without the enzyme addition were, respectively, 73.37% and 76.33% for MS (p=0.0136), 70.44% and 70.39% for PB (p=0.9595) and 74.79% and 76.77% for EB (p=0.0128). The apparent metabolizable energy (EMA) and EMA corrected for nitrogen retention values (as is basis) were 3322±19 and 3241±18Kcal/kg and 3334±16 and 3261±17Kcal/kg, respectively, for with and without the enzyme addition to FRM. The enzyme addition had no statistical significance on the EMA and EMAn values, however, a significant effect on energy metabolizability coefficient was observed.(AU)

Determinação da energia metabolizável do farelo residual do milho com e sem enzima em dietas para frangos de corte / Determination of the metabolizable energy of residual corn bran with and without enzyme in diets for broilers

O objetivo deste trabalho foi avaliar o valor nutricional e determinar a energia metabolizável do farelo residual de milho (FRM) sem e com o uso da enzima alfa- amilase. Foi realizado um experimento de metabolismo com 180 pintos machos Cobb com 14 dias, distribuídos em um delineamento inteiramente ao acaso, com seis tratamentos, cinco repetições e seis aves por parcela. As dietas experimentais foram: T1: ração referência (RR), T2: 60% T1 + 40% de FRM, T3: RR + enzima, T4: 60% T1 + 40% de FRM com adição de enzima, T5: RR com substituição de 100% do milho pelo FRM e T6: RR com substituição de 100% do milho pelo FRM com adição de enzima. A composição química do FRM foi: 88,33% de matéria seca (MS), 10,23% de proteína bruta (PB), 15,44% de extrato etéreo (EE), 4,33% de cinzas (CZ) e 4555kcal/kg de energia bruta (EB). Os valores dos coeficientes de metabolizabilidade aparente para o FRM sem e com adição de enzima foram, respectivamente, de 73,37% e 76,33% para MS (p=0,0136), 70,44% e 70,39% para PB (p=0,9595) e de 74,79% e 76,77% para EB (p=0,0128). Os valores da energia metabolizável aparente (EMA) e da EMA corrigida para retenção de nitrogênio (EMAn) para o FRM (na base natural) foram de 3322±19 e 3241±18kcal/kg e de 3334±16 e 3261±17kcal/kg, respectivamente, sem e com adição de enzima. A adição da enzima não teve efeito estatístico significativo sobre os valores de EMA e EMAn, entretanto houve efeito positivo no coeficiente de metabolização da energia.(AU)

The aim of this study was to evaluate the nutritional value and the metabolizable energy of residual corn bran (FRM) with and without the use of alpha amylase enzyme. One metabolism assay was done using 180 Ross male broilers at 14 days of age distributed in an entirely randomized design with six treatments, five replications and six broilers per plot. The assay treatments established were: T1: reference diet (RR), T2: 60% T1 + 40% of FRM, T3: T1 (RR) + enzyme addition, T4: 60% T1 + 40% of FRM with enzyme addition, T5: T1 (RR) with 100% replacement of corn by FRM and T6: T1 (RR) with 100% replacement of corn by FRM with enzyme addition. FRM chemical composition was: 88.33% of dry matter (MS), 10.23% of crude protein (PB), 15.44% of crude fat (EE), 4.33% of ashes (CZ) and 4555Kcal/kg of Gross Energy (EB). The FRM apparent metabolizability coefficients with and without the enzyme addition were, respectively, 73.37% and 76.33% for MS (p=0.0136), 70.44% and 70.39% for PB (p=0.9595) and 74.79% and 76.77% for EB (p=0.0128). The apparent metabolizable energy (EMA) and EMA corrected for nitrogen retention values (as is basis) were 3322±19 and 3241±18Kcal/kg and 3334±16 and 3261±17Kcal/kg, respectively, for with and without the enzyme addition to FRM. The enzyme addition had no statistical significance on the EMA and EMAn values, however, a significant effect on energy metabolizability coefficient was observed.(AU)

Composição físico-química e valores energéticos de farinhas de silagem de peixe para frangos de corte / Physicochemical composition and energy values of fish silage meal for broilers

Objetivou-se determinar a composição físico-química, os valores energéticos e os coeficientes de digestibilidade de quatro farinhas de silagem de peixe para frangos de corte. Foram produzidas quatro farinhas de silagem de peixe, utilizando-se o resíduo da filetagem de tilápias ensilado com diferentes fontes de carboidratos fermentáveis. Analisou-se a composição físico-química das silagens, e, em seguida, um ensaio de metabolismo com 180 pintos machos da linhagem Cobb de 14 a 25 dias de idade. Também foram avaliados o tempo de trânsito gastrintestinal das rações e o desempenho das aves nas gaiolas. Os animais foram distribuídos em delineamento inteiramente ao acaso, com cinco tratamentos, seis repetições e seis aves por unidade experimental. Os tratamentos consistiram de uma dieta referência e de quatro dietas teste compostas de 60 porcento da ração referência e 40 porcento do resíduo da filetagem de tilápia ensilado com diferentes fontes de carboidratos, sendo a farinha de silagem de peixe com o farelo de algaroba (SFA), com a farinha de varredura de mandioca (SFVM), com o farelo de milho (SFM) e com a casca da mandioca (SCM). A SFM obteve o maior teor de PB, 22,38 porcento, de EE, 27,35 porcento, e o maior tempo de trânsito, com 195,0min; a SCM apresentou o maior valor de MM, 11,12 porcento. Os valores de EMA e EMAn das farinhas de silagem de peixe não diferiram significativamente entre eles. O maior GP e a melhor CA foram apresentados pelos animais do tratamento SFM, e os piores GP e CA pelos frangos de corte alimentados com dietas contendo a SFVM. Com base na composição obtida, estas silagens de peixe têm potencialidade para serem utilizadas em dietas para frangos de corte.(AU)

The objective of this research was to determine the physical and chemical composition energy value and digestibility of four fish silage meal for broilers. Four flours were produced from fish silage using the residue of tilapia filleting ensiled together with different sources of fermentable carbohydrates. We analyzed the physical and chemical composition of silages and then a metabolism trial with 180 male Cobb chicks 14-25 days old, and also evaluated gastrintestinal transit time of feed and performance of birds in cages. The animals were distributed in a completely randomized design with five treatments and six replicates of six birds each. Treatments consisted of a reference diet and four test diets composed of 60 percent of the reference diet with the inclusion of 40 percent of the residue of tilapia silage with different sources of carbohydrates, and the fish silage meal with bran mesquite (SFA) with the scan cassava flour (SFVM) with corn meal (SFM), and peel cassava (SCM). The SFM had the highest content of CP, 22.38 percent, EE, 27.35 percent, and the largest transit time with 195.0 min, the SCM showed the highest MM, 11.12 percent. The AME and AMEn of flour fish silage did not differ significantly between them. The biggest and best GP CA was presented by animals in the SFM treatment and the worse CA GP was for broilers fed diets containing SFVM. Based on the composition obtained, these fish silage have the potential to be used in diets for broilers.(AU)

Neutralization of pharmacological and toxic activities of Bothrops jararacussu snake venom and isolated myotoxins by Serjania erecta methanolic extract and its fractions

Most of the snakebites recorded in Brazil are caused by the Bothrops genus. Given that the local tissue damage caused by this genus cannot be treated by antivenom therapy, numerous studies are focusing on supplementary alternatives, such as the use of medicinal plants. Serjania erecta has already demonstrated anti-inflammatory, antiseptic and healing properties. In the current study, the aerial parts of S. erecta were extracted with methanol, then submitted to chromatographic fractionation on a Sephadex LH20 column and eluted with methanol, which resulted in four main fractions. The crude extract and fractions neutralized the toxic activities of Bothrops jararacussu snake venom and isolated myotoxins (BthTX-I and II). Results showed that phospholipase A2, fibrinogenolytic, myotoxic and hemorrhagic activities were inhibited by the extract. Moreover, the myotoxic and edematous activities induced by BthTX-I, and phospholipase A2 activity induced by BthTX-II, were inhibited by the extract of S. erecta and its fraction. The clotting time on bovine plasma was significantly prolonged by the inhibitory action of fractions SF3 and SF4. This extract is a promising source of natural inhibitors, such as flavonoids and tannins, which act by forming complexes with metal ions and proteins, inhibiting the action of serineproteases, metalloproteases and phospholipases A2.(AU)

Antiophidian properties of plant extracts against Lachesis muta venom

Snakebites comprise a serious health problem in several countries due to their global incidence, which exceeds 2.5 million per year, and the elevated number of victim fatalities. To counteract envenomations, antivenoms have been used regularly for more than a century. Apart from side effects including anaphylactic reactions, antivenoms are not able to efficiently neutralize local tissue damage, which contributes to increasing the severity and morbidity observed in patients. This fact, in turn, may be responsible for economic hardship, particularly in rural populations of developing countries. In the present work, we evaluated the antiophidian properties of 12 Brazilian plant extracts against the hemolytic, coagulant, hemorrhagic and proteolytic effects of Lachesis muta venom. Taken together, our data revealed that most of these aqueous products were capable of inhibiting those activities at different levels, except for Sapindus saponaria extract. In contrast, Stryphnodendron barbatiman extract completely neutralized all the analyzed biological activities. Thus, we may conclude that Brazilian flora may also be useful against L. muta accidents.(AU)