Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit

Abstract Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Resumo As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit / Compostos fenólicos e atividade antifúngica dos extratos acetato de etila e metanólico dos frutos maduros de Capsicum chinense Jacq

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

As perdas de alimentos por contaminação causada por fungos são de grande impacto negativo para a agricultura, gerando altos prejuízos econômicos. Para evitar as perdas de diversos produtos alimentícios pela contaminação fúngica são utilizados fungicidas sintéticos e naturais. As espécies do gênero Capsicum são usadas há muitos anos para auxiliar na conservação de alimentos por possuírem substâncias químicas com ação antifúngica entre outras. Neste contexto, o objetivo deste estudo foi identificar alguns compostos fenólicos por cromatografia líquida de alta eficiência acoplada à espectrometria de massas sequencial (LC-ESI-MS/MS) presentes nos extratos acetato de etila (EAE) e metanólico (ME) dos frutos maduros da pimenta biquinho (C. chinense) e avaliar atividade antifúngica de EAE e ME contra os fungos Sclerotinia sclerotiorum, Rhizopus stolonifer e Colletotrichum gloeosporioides. Os extratos foram obtidos de forma sequencial, utilizando o procedimento de maceração. A atividade antifúngica foi avaliada seguindo a metodologia de difusão em disco, nas doses de 25 µL, 50 µL, 100 µL e 200 µL de cada extrato diluído. A análise química evidenciou a presença de ácido protocatequico, ácido gentisico, ácido vanílico, kaempferol-3-O-robinobiosídeo e naringenina em ambos os extratos. EAE revelou maior poder de inibição do crescimento micelial nas doses de 100µL e 200µL contra os três fungos testados, enquanto ME exibiu fraca atividade inclusive na maior dose investigada. Entretanto, estudos mais aprofundados ainda são necessários para consolidar seu uso e aplicação prática na área agronômica.

Phenolic compounds and antifungal activity of ethyl acetate extract and methanolic extract from Capsicum chinense Jacq. ripe fruit.

Food loss due to contamination caused by fungi has much impact on agriculture and leads to significant economic losses. Synthetic and natural fungicides have been used for avoiding losses of several food products due to fungal contamination. As a result, species of the genus Capsicum have been used for preserving food because of their chemical compounds with antifungal activity. Therefore, this study aimed at identifying some phenolic compounds found in both ethyl acetate extract (EAE) and methanolic extract (ME) from habanero pepper (C. chinense) ripe fruit by liquid chromatography tandem mass spectrometry with electrospray ionization (LC-ESI-MS/MS) and at evaluating their antifungal activities against fungi Sclerotinia sclerotiorum, Rhizopus stolonifer and Colletotrichum gloeosporioides. Extracts resulted from a sequential process of maceration. Antifungal activity was evaluated by the disk diffusion method (DDM) at the following doses of both diluted extracts: 25 µL, 50 µL, 100 µL and 200 µL. The chemical analysis showed that there were protocatechuic acid, gentisic acid, vanillic acid, kaempferol-3-O-robinobiosideo and naringenin in both extracts. EAE showed high inhibition of mycelial growth at both doses 100µL and 200µL against the three fungi while methanolic exhibited weak activity even at the highest dose under investigation. However, further in-depth studies are needed to reinforce their uses and practical applications to the agricultural field.

Chemical composition and biological activities of essential oil from flowers of Psidium guajava (Myrtaceae) / Composição química e atividades biológicas do óleo essencial das flores de Psidium guajava (Myrtaceae)

Abstract Xylella fastidiosa is a plant-pathogenic bacterium that lives inside host xylem vessels, where it forms biofilm which is believed to be responsible for disrupting the passage of water and nutrients. Pectobacterium carotovorum is a Gram-negative plant-specific bacterium that causes not only soft rot in various plant hosts, but also blackleg in potato by plant cell wall degradation. Chagas disease, which is caused by Trypanosoma cruzi, has been commonly treated with nifurtimox and benzonidazole, two drugs that cause several side effects. As a result, the use of natural products for treating bacterial and neglected diseases has increased in recent years and plants have become a promising alternative to developing new medicines. Therefore, this study aimed to determine, for the first time, the chemical composition of essential oil from Psidium guajava flowers (PG-EO) and to evaluate its in vitro anti-Xylella fastidiosa, anti-Pectobacterium carotovorum, anti-Trypanosoma cruzi and cytotoxic activities. PG-EO was obtained by hydrodistillation in a Clevenger apparatus while its chemical composition was determined by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Major compounds identified in PG-EO were α-cadinol (37.8%), β-caryophyllene (12.2%), nerolidol (9.1%), α-selinene (8.8%), β-selinene (7.4%) and caryophyllene oxide (7.2%). Results showed that the PG-EO had strong trypanocidal activity against the trypomastigote forms of Trypanosoma cruzi (IC50 = 14.6 μg/mL), promising antibacterial activity against X. fastidiosa (MIC = 12.5 μg/mL) and P. carotovorum (MIC = 62.5 μg/mL), and moderate cytotoxicity against LLCMK2 adherent epithelial cells in the concentration range (CC50 = 250.5 μg/mL). In short, the PG-EO can be considered a new source of bioactive compounds for the development of pesticides and trypanocide drugs.

Resumo Xylella fastidiosa é uma bactéria patogênica que vive dentro dos vasos do xilema hospedeiro, onde forma um biofilme responsável por interromper a passagem de água e nutrientes. Pectobacterium carotovorum é uma bactéria Gram-negativa que causa não só podridão macia em várias plantas hospedeiras, mas também canela-preta na batata por degradação da parede celular da planta. A doença de Chagas, causada pelo Trypanosoma cruzi, é comumente tratada com nifurtimox e benzonidazol, duas drogas que causam vários efeitos colaterais. Como resultado, o uso de produtos naturais para o tratamento de doenças bacterianas e negligenciadas aumentou nos últimos anos e as plantas continuam sendo uma alternativa promissora para o desenvolvimento de novos medicamentos. Portanto, este estudo teve como objetivo determinar, pela primeira vez, a composição química do óleo essencial de flores de Psidium guajava (PG-EO) e avaliar suas propriedades anti-Xylella fastidiosa, anti-Pectobacterium carotovorum, anti-Trypanosoma cruzi e citotóxica in vitro. PG-EO foi obtido por hidrodestilação em um aparelho Clevenger, enquanto sua composição química foi determinada por cromatografia em fase gasosa com detecção por ionização por chama (CG-DIC) e por cromatografia em fase gasosa acoplada à espectrometria de massa (CG-EM). Os principais compostos identificados no PG-EO foram α-cadinol (37,8%), β-cariofileno (12,2%), nerolidol (9,1%), α-selineno (8,8%), β-selineno (7,4%) e óxido de cariofileno (7,2%). Os resultados mostraram que o PG-EO apresentou forte atividade tripanocida contra as formas tripomastigotas de T. cruzi (CI50 = 14,6 μg/mL), promissora atividade antibacteriana contra X. fastidiosa (MIC = 12,5 μg/mL) e P. carotovorum (MIC = 62,5 μg/mL) e citotoxicidade moderada contra células epiteliais aderentes (LLCMK2) na faixa de concentração (CC50 = 250,5 μg/mL). Em suma, o PG-EO pode ser considerado uma nova fonte de compostos bioativos para o desenvolvimento de pesticidas e drogas tripanocidas.

Chemical composition and biological activities of essential oil from flowers of Psidium guajava (Myrtaceae).

Xylella fastidiosa is a plant-pathogenic bacterium that lives inside host xylem vessels, where it forms biofilm which is believed to be responsible for disrupting the passage of water and nutrients. Pectobacterium carotovorum is a Gram-negative plant-specific bacterium that causes not only soft rot in various plant hosts, but also blackleg in potato by plant cell wall degradation. Chagas disease, which is caused by Trypanosoma cruzi, has been commonly treated with nifurtimox and benzonidazole, two drugs that cause several side effects. As a result, the use of natural products for treating bacterial and neglected diseases has increased in recent years and plants have become a promising alternative to developing new medicines. Therefore, this study aimed to determine, for the first time, the chemical composition of essential oil from Psidium guajava flowers (PG-EO) and to evaluate its in vitro anti-Xylella fastidiosa, anti-Pectobacterium carotovorum, anti-Trypanosoma cruzi and cytotoxic activities. PG-EO was obtained by hydrodistillation in a Clevenger apparatus while its chemical composition was determined by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Major compounds identified in PG-EO were α-cadinol (37.8%), ß-caryophyllene (12.2%), nerolidol (9.1%), α-selinene (8.8%), ß-selinene (7.4%) and caryophyllene oxide (7.2%). Results showed that the PG-EO had strong trypanocidal activity against the trypomastigote forms of Trypanosoma cruzi (IC50 = 14.6 µg/mL), promising antibacterial activity against X. fastidiosa (MIC = 12.5 µg/mL) and P. carotovorum (MIC = 62.5 µg/mL), and moderate cytotoxicity against LLCMK2 adherent epithelial cells in the concentration range (CC50 = 250.5 µg/mL). In short, the PG-EO can be considered a new source of bioactive compounds for the development of pesticides and trypanocide drugs.

Metabolic stress and reproductive features in post-partum goats supplemented for a long period with detoxified castor meal as the source of dietary nitrogen / Estresse metabólico e características reprodutivas em cabras pós-parto suplementadas por longo período com mamona desintoxicada como fonte de nitrogênio na dieta

This study evaluated the effect of detoxified castor meal on the reproductive performance, metabolic stress, milk production, and kid development in peripartum goats. The diet of the animals were with (DCM, n= 20) or without (WDCM, n= 21) detoxified castor meal during the entire gestation and until weaning, 60 days post-birth. No differences were observed in the gestation period, litter size, rate of multiple births, and mortality between the two groups. The postpartum plasma concentrations of progesterone remained below 1ng/mL in all animals, thus, confirming the absence of active corpora lutea. The thickness of sternum adipose tissue and loin area, levels of urea and cholesterol, milk production, and daily weight gain in the kids were low in the DCM group when compared to those in the WDCM group (P< 0.05). To conclude, the use of detoxified castor meal in peripartum goats resulted in lower level of performance in the kids because of reductions in the amount of milk received from their mothers during lactation. In addition, the diet containing detoxified castor meals was not efficient in recovering from the loss of stored body reserves able to initiate the recovery of the cyclic activity of the goats.(AU)

Este estudo avaliou o efeito da torta de mamona desintoxicada na reprodução, no estresse metabólico, na produção de leite e no desenvolvimento de cabritos no periparto de cabras. Um grupo foi alimentado com torta de mamona (DCM, n=20), e o outro (WDCM, n=21) não recebeu tal suplemento , durante a gestação até o desmame, 60 dias pós-parto. Não foram observadas diferenças significativas no período de gestação, no número de cabritos, na taxa de partos múltiplos e na mortalidade entre os dois grupos. Em todos os animais, a concentração plasmática de progesterona ficou abaixo de 1ng/mL, confirmando a ausência de atividade lútea. A espessura da gordura subcutânea do esterno e da área de olho-de-lombo, a concentração de ureia e colesterol, a produção de leite e o ganho de peso dos cabritos foram menores no grupo DCM (P<0,05). Conclui-se que o uso de torta de mamona desintoxicada no periparto de cabra resultou em cabritos mais leves devido à redução na produção de leite das matrizes e as cabras não retornaram ao cio, pois não recuperaram a massa corporal.(AU)

Chemical composition and biological activities of essential oil from flowers of Psidium guajava (Myrtaceae)

Abstract Xylella fastidiosa is a plant-pathogenic bacterium that lives inside host xylem vessels, where it forms biofilm which is believed to be responsible for disrupting the passage of water and nutrients. Pectobacterium carotovorum is a Gram-negative plant-specific bacterium that causes not only soft rot in various plant hosts, but also blackleg in potato by plant cell wall degradation. Chagas disease, which is caused by Trypanosoma cruzi, has been commonly treated with nifurtimox and benzonidazole, two drugs that cause several side effects. As a result, the use of natural products for treating bacterial and neglected diseases has increased in recent years and plants have become a promising alternative to developing new medicines. Therefore, this study aimed to determine, for the first time, the chemical composition of essential oil from Psidium guajava flowers (PG-EO) and to evaluate its in vitro anti-Xylella fastidiosa, anti-Pectobacterium carotovorum, anti-Trypanosoma cruzi and cytotoxic activities. PG-EO was obtained by hydrodistillation in a Clevenger apparatus while its chemical composition was determined by gas chromatography-flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Major compounds identified in PG-EO were -cadinol (37.8%), -caryophyllene (12.2%), nerolidol (9.1%), -selinene (8.8%), -selinene (7.4%) and caryophyllene oxide (7.2%). Results showed that the PG-EO had strong trypanocidal activity against the trypomastigote forms of Trypanosoma cruzi (IC50 = 14.6 g/mL), promising antibacterial activity against X. fastidiosa (MIC = 12.5 g/mL) and P. carotovorum (MIC = 62.5 g/mL), and moderate cytotoxicity against LLCMK2 adherent epithelial cells in the concentration range (CC50 = 250.5 g/mL). In short, the PG-EO can be considered a new source of bioactive compounds for the development of pesticides and trypanocide drugs.

Resumo Xylella fastidiosa é uma bactéria patogênica que vive dentro dos vasos do xilema hospedeiro, onde forma um biofilme responsável por interromper a passagem de água e nutrientes. Pectobacterium carotovorum é uma bactéria Gram-negativa que causa não só podridão macia em várias plantas hospedeiras, mas também canela-preta na batata por degradação da parede celular da planta. A doença de Chagas, causada pelo Trypanosoma cruzi, é comumente tratada com nifurtimox e benzonidazol, duas drogas que causam vários efeitos colaterais. Como resultado, o uso de produtos naturais para o tratamento de doenças bacterianas e negligenciadas aumentou nos últimos anos e as plantas continuam sendo uma alternativa promissora para o desenvolvimento de novos medicamentos. Portanto, este estudo teve como objetivo determinar, pela primeira vez, a composição química do óleo essencial de flores de Psidium guajava (PG-EO) e avaliar suas propriedades anti-Xylella fastidiosa, anti-Pectobacterium carotovorum, anti-Trypanosoma cruzi e citotóxica in vitro. PG-EO foi obtido por hidrodestilação em um aparelho Clevenger, enquanto sua composição química foi determinada por cromatografia em fase gasosa com detecção por ionização por chama (CG-DIC) e por cromatografia em fase gasosa acoplada à espectrometria de massa (CG-EM). Os principais compostos identificados no PG-EO foram -cadinol (37,8%), -cariofileno (12,2%), nerolidol (9,1%), -selineno (8,8%), -selineno (7,4%) e óxido de cariofileno (7,2%). Os resultados mostraram que o PG-EO apresentou forte atividade tripanocida contra as formas tripomastigotas de T. cruzi (CI50 = 14,6 g/mL), promissora atividade antibacteriana contra X. fastidiosa (MIC = 12,5 g/mL) e P. carotovorum (MIC = 62,5 g/mL) e citotoxicidade moderada contra células epiteliais aderentes (LLCMK2) na faixa de concentração (CC50 = 250,5 g/mL). Em suma, o PG-EO pode ser considerado uma nova fonte de compostos bioativos para o desenvolvimento de pesticidas e drogas tripanocidas.