Artisanal beer added of Solanum sessiliflorum pulp: total phenolic compounds and antioxidant activity / Cerveja artesanal adicionada de polpa de Solanum sessiliflorum: compostos fenólicos totais e atividade antioxidante

Brazilian beer market presented remarkable increasement last years. Despite of this, Brazilian Amazon states presented discrete growth, with decreasement in some important states as Amazonas. Cubiu (Solanum sessiliflorum) is a fruiting plant which has been used traditionally by people of the upper Amazon and Orinoco basin as artisanal medicine because of its bioactive molecules' richness. Because of world trend of innovative brewing products as fruit-beers, the aim of this work was to evaluate the composition and antioxidant activity of an artisanal beer added of cubiu. A total of 28 g/L of cubiu's pulp were added to the beer in maturation's phase, incubated at 4 °C for 10 days. Compared to a control assay, the resulting beer presented higher turbidity and soluble protein concentration. Total phenolic compounds concentration (435,93 µgGAE/mL) and antioxidant activity (66,36% of free radicals scavenged) were significantly greater than control assay (p <0,001). The results indicates that a low amount of cubiu's pulp is sufficient to makes the beer functionally antioxidant. The subsequent efforts must to evaluate the acceptance of a beer added of S. sessiliflorum pulp to assessing its market potential.

O mercado brasileiro de cerveja apresentou notável crescimento nos últimos anos. Apesar disso, os estados amazônicos brasileiros apresentaram crescimento discreto, com queda em alguns estados importantes como o Amazonas. Cubiu (Solanum sessiliflorum) é uma planta frutífera que tem sido tradicionalmente utilizada pelos povos do alto Amazonas e da bacia do Orinoco como medicamento artesanal devido à sua riqueza em moléculas bioativas. Devido à tendência mundial de produtos cervejeiros inovadores, como as cervejas de frutas, o objetivo deste trabalho foi avaliar a composição e atividade antioxidante de uma cerveja artesanal adicionada de cubiu. Um total de 28 g/L de polpa de cubiu foram adicionados à cerveja em fase de maturação, incubada a 4 °C por 10 dias. Comparada a um ensaio controle, a cerveja resultante apresentou maior turbidez e concentração de proteína solúvel. A concentração total de compostos fenólicos (435,93 µgGAE/mL) e a atividade antioxidante (66,36% dos radicais livres neutralizados) foram significativamente superiores ao ensaio controle (p <0,001). Os resultados indicam que uma pequena quantidade de polpa de cubiu é suficiente para tornar a cerveja funcionalmente antioxidante. Os esforços subsequentes devem avaliar a aceitação de uma cerveja adicionada de polpa de S. sessiliflorum para avaliar seu potencial de mercado.

Efecto antibacteriano del extracto hidroalcohólico de Solanum sessiliflorum Dunal (cocona) sobre Streptococcus mutans

Introducción: La búsqueda de nuevos extractos de origen vegetal con propiedades antibacterianas para mantener la salud bucal, es fundamental para el óptimo desempeño del personal militar. Objetivos: Evaluar el efecto antibacteriano del extracto hidroalcohólico de Solanum sessiliflorum Dunal (cocona) sobre Streptococcus mutans. Métodos: Estudio experimental in vitro y comparativo. Se realizó un ensayo fitoquímico preliminar del extracto hidroalcohólico de Solanum sessiliflorum Dunal. Se emplearon 48 placas de agar Müller-Hinton (Merck®), distribuidas en 6 grupos (n= 8): grupo I (agua destilada), grupo II (etanol al 70 %), grupo III (clorhexidina al 0,12 %), grupo IV (Solanum sessiliflorum Dunal al 25 %), grupo V (Solanum sessiliflorum Dunal al 50 %) y grupo VI (Solanum sessiliflorum Dunal al 75 %). Se utilizó la técnica de difusión con discos descrita por Bauer y Kirby; la cepa empleada fue Streptococcus mutans ATCC 25175 y las mediciones de los halos de inhibición se realizaron a las 24 horas, para determinar la actividad antibacteriana. Resultados: En el ensayo fitoquímico se detectaron compuestos fenólicos, antocianinas, quinonas y glicósidos cardiotónicos. Se comprobó el efecto antibacteriano del grupo VI (Solanum sessiliflorum Dunal al 75 %) con 19,831 ± 0,0553 mm (99,37 %), comparable con el de clorhexidina al 0,12 % (grupo III) 19,956 ± 0,0431 mm (100 %) sobre Streptococcus mutans ATCC 25175. Conclusiones: El extracto hidroalcohólico de Solanum sessiliflorum Dunal al 75 % presenta efecto antibacteriano in vitro sobre cepas de Streptococcus mutans ATCC 25175 con valores similares a clorhexidina al 0,12 % .

Introduction: The search for new extracts of plant origin with antibacterial properties to maintain oral health is essential for the optimal performance of military personnel. Objectives: To evaluate the antibacterial effect of the hydroalcoholic extract of Solanum sessiliflorum Dunal (cocona) on Streptococcus mutans. Methods: In vitro and comparative experimental study. A preliminary phytochemical assay of the hydroalcoholic extract of Solanum sessiliflorum Dunal was performed. Forty-eight Müller-Hinton agar plates (Merck®) were used, distributed in 6 groups (n= 8). Group I (distilled water), group II (70% ethanol), group III (0.12% chlorhexidine), group IV (25% Solanum sessiliflorum Dunal), group V (50% Solanum sessiliflorum Dunal) and group VI (75% Solanum sessiliflorum Dunal); the disc diffusion technique described by Bauer and Kirby was used; the strain used was Streptococcus mutans ATCC 25175 and the inhibition halos were measured at 24 hours to determine the antibacterial activity. Results: Phenolic compounds, anthocyanins, quinones and cardiotonic glycosides were detected in the phytochemical assay. The antibacterial effect of group VI (Solanum sessiliflorum Dunal 75%) was proven with 19,831 ± 0,0553 mm (99,37%), comparable to that of 0,12% chlorhexidine (group III) 19,956 ± 0,0431 mm (100%) on Streptococcus mutans ATCC 25175. Conclusions: The 75% hydroalcoholic extract of Solanum sessiliflorum Dunal shows in vitro antibacterial effect on Streptococcus mutans ATCC 25175 strains with values similar to 0,12% chlorhexidine.

Comparative evaluation of chemical composition and biological activities of tropical fruits consumed in Manaus, central Amazonia, Brazil.

Fruits are widely recognized as sources of biologically active metabolites, such as antioxidant compounds. In this context, fruits commonly consumed in the central Amazonia, especially in its biggest metropolis (Manaus - AM/Brazil), are attractive as potential sources of antioxidant compounds related to biological activities. Most of such fruits are still poorly studied and/or remain unknown outside the Amazon region. Therefore, this study aims to investigate nine fruits (abiu, cubiu, biribá, breadfruit, genipap, peach palm, murici, soursop, and umari) regarding their chemical composition (fixed and volatile), reducing capacity, antioxidant activity, enzyme inhibition, and cytotoxicity. Determination of small organic acids, hydroxycinnamic acids, flavan-3-ols and flavonoid aglycones was done by HPLC-MS/MS, whereas determination of volatile organic compounds (VOCs) was done by HS-SPME/GC-MS. Reducing capacity was determined by the Folin-Ciocalteu method, and antioxidant activities were evaluated by DPPH, ABTS, and H-ORACFL assays. In vitro activities regarding inhibition of enzymes were tested for α-glucosidase, lipase, and α-amylase, and anti-glycation activities were evaluated for methylglyoxal and fructose. Cytotoxicity of fruit extracts was evaluated by cell viability of human fibroblast cell line (MRC-5). A total of 16 antioxidant compounds and 139 VOCs were determined, whose profiles were unique for each studied fruit. Total phenolic contents as well as antioxidant activities found herein were similar or even higher than those reported for several traditional fruits. Some of fruit extracts were able to inhibit α-glucosidase and glycation in methylglyoxal and fructose models, whereas none of them was active for lipase and α-amylase. All of the fruit extracts showed to be non-cytotoxic to MRC-5 cell line.

Cell wall polysaccharides from pulp and peel of cubiu: A pectin-rich fruit.

To investigate the polysaccharides of cubiu (Solanum sessiliflorum D.) fruits, pulp and peel were subjected to sequential extractions using different solvents, giving rise to pectins heterogeneous with respect to their structural characteristics, xylan, arabinogalactoxyloglucans and glucomannans. Chemical, physicochemical and spectroscopic analyses were used to characterize the polysaccharides obtained. Pectins with uronic acid content of commercial grade were obtained from the peel of cubiu fruits using water (25°C and 100°C) and EDTA and from its pulp using water (100°C). The fraction with the highest yield (9.6%) and uronic acid content (79.0%) extracted from the peel using water at 100°C was chemically and structurally characterized. This fraction consists predominantly of a mixture of a homogalacturonan (degree of methyl-esterification 56.9%) and a small amount of rhamnogalacturonan I branched by type I and type II arabinogalactans. The results suggest that cubiu is a promising source of pectins with high uronic acid content extractable by cheaper and environmentally friendly method.

Efeito hipolipemiante da suplementação dietética com a farinha do cubiu (Solanum sessiliflorum Dunal) em ratos hipercolesterolêmicos / Hypolipidemic effect of diet supplementation with cocona (Solanum sessiliflorum Dunal) flour in hypercholesterolemic rats

Estudos com frutas e vegetais ricos em fibra alimentar tem demonstrado efeito redutor das concentrações sanguíneas de colesterol total, atuando de forma preventiva contra a hipercolesterolemia. O objetivo do presente trabalho foi analisar a composição química da farinha de cubiu e avaliar o efeito de seu consumo sobre a lipemia de ratos hipercolesterolêmicos. Foram utilizados 8 animais para cada tratamento, em 7 grupos sendo: 1 controle; 3 experimentais com 5%, 25% e 50% da recomendação de fibras (AIN-93) provenientes da farinha de cubiu; e 3 grupos per feeding tendo celulose e pectina como fonte de fibra alimentar nas mesmas proporções dos grupos experimentais. Ao final do ensaio foi verificado que não houve variação significativa no consumo de ração pelos animais e no ganho de massa corporal no decorrer do experimento. Os tratamentos com farinha de cubiu reduziram as concentrações de colesterol total (-21,6%), LDL-c (-56,8%), aumentaram a excreção de colesterol fecal (+116%), e diminuíram o colesterol hepático (-32,1%). Nenhum dos tratamentos promoveu alteração significativa nas concentrações de triglicerídeos. Os tratamentos com pectina e celulose demonstraram resultados semelhantes aos do cubiu. Os resultados observados servem como base para futuros estudos visando o aproveitamento biotecnológico deste fruto.

Studies with fruit and vegetables rich in dietary fiber have shown lowering effects on the blood concentrations of total cholesterol (TC), acting preventively against hypercholesterolemia. The aim of this study was to analyze the chemical composition of cocona and evaluate the effect of the consumption of cocona flour on lipemia of Wistar rats with hypercholesterolemia. Eight animals were used for each treatment, in 7 groups: control; 3 experimental groups with 5%, 25% and 50% of the recommended fiber (AIN-93) from cocona flour; 3 groups per feeding with pectin and cellulose as source of dietary fiber, in the same proportions of the experimental groups. At the end of the assay, we verified that there were not significant variations in the consumption of the animals and the body weight during the experiment. Treatments with cocna flour were responsible for reducing the concentrations of total cholesterol (-21.6%), LDL-c (-56.8%), increasing the excretion of fecal cholesterol (+116%) and lowering the liver cholesterol (-32.1%). None of the treatments significantly altered the triglyceride concentrations. Treatments using pectin and cellulose show results similar to cocona. The observed results can be used in further studies aimed at the biotechnological exploitation of this fruit.

Biological activities of Solanum sessiliflorum Dunal / Atividades biológicas de Solanum sessiliflorum dunal

This study evaluated the antioxidant, antimicrobial and wound healing activities from the aqueous extract of cubiu (AEC - Solanum sessiliflorum Dunal). The antioxidant activity was evaluated by the scavenging method with 2.2-diphenyl-1.2-picrylhydrazyl radical (DPPH) and, the antimicrobial activity, by the turbidimetric method. The dermal ulcers induced model in rabbits was used in order to study the wound healing activity. Creams containing 5% and 10% AEC and 5% AEC associated with 1% copaiba oil and 1% rosemary oil were tested. The wounds were macroscopically (ulcer aspect and ulcer area contraction) and microscopically examined. The histological study considered the number of inflammatory cells, fibroblasts, blood vessels, collagen and extracellular matrix areas. Analysis of variance (ANOVA) followed by Tukey's post hoc test was performed. The AEC showed IC50=65.12 µg/mL and no antimicrobial activity against all of the strains tested was verified. The formulation containing 5% AEC associated with 1% copaiba oil exhibited the greatest pharmacological potentiality in dermal wound healing due to the increased number of blood vessels (p <0.001) and the largest extracellular matrix area (p <0.001) presented. The results justify further studies about biological activities of cubiu in the search for bioactive compounds.

O presente estudo avaliou as atividades antioxidante, antimicrobiana e cicatrizante do extrato aquoso do cubiu (EAC - Solanum sessiliflorum Dunal). A atividade antioxidante foi avaliada pelo método sequestrante de radicais 2,2-difenil-1,2-picrilhidrazil (DPPH) e, a atividade antimicrobiana, por método turbidimétrico. Para o estudo da atividade cicatrizante foi utilizado o modelo de indução de úlceras dérmicas em coelhos, sendo testados cremes contendo 5% e 10% de EAC e cremes contendo 5% de EAC associado à 1% de óleo de copaíba e à 1% de óleo de alecrim. As lesões foram analisadas macro (aspecto da lesão e contração da área ferida) e microscopicamente. O estudo histológico considerou o número de células inflamatórias, fibroblastos, vasos sanguíneos, e as áreas de colágeno e de matriz extracelular. Foi realizada análise de variância (ANOVA) seguida do teste post hoc de Tukey. O EAC apresentou IC50=65,12 µg/mL e não apresentou atividade antimicrobiana frente a nenhuma das cepas testadas. A formulação contendo 5% de EAC associado a 1% de óleo de copaíba foi a que exibiu maior potencialidade farmacológica na cicatrização de lesões dérmicas, devido ao maior número de vasos sanguíneos (p<0,001) e a maior área de matriz extracelular (p<0,001) apresentados. Os resultados encontrados justificam novos estudos sobre as atividades biológicas do cubiu na busca de compostos bioativos.

Quantificação de macro e micro nutrientes em algumas etnovariedades de cubiu (Solanum sessiliflorum Dunal) / Macro and micro nutrients quantification of some cubiu ethnovarieties (Solanum sessiliflorum Dunal)

Considerando a ampla variabilidade genética de cubiu (Solanum sessiliflorum Dunal), quantificaram-se os macro e micro-nutrientes, objetivando a ampliação da tabela de composição química de alimentos típicos da região amazônica. Os frutos provenientes da Estação Experimental de Hortaliças Alejo von der Pahlen (EEH) do Instituto Nacional de Pesquisas da Amazônia (INPA), localizados no km 14 da Rodovia AM 010 em Manaus, AM, foram processados no Laboratório de Alimentos e Nutrição do INPA. Avaliaram-se oito etnovariedades de cubiu identificados como: 2 I, 3 I, 6, 7, 12, 14, 17, 29 I e III em estádio de maturação comercial. Os teores de elementos minerais foram quantificados pela técnica de Ativação por Nêutrons Instrumental e a fibra alimentar pelo método enzímico-gravimétrico. Os resultados demonstram ser o cubiu um fruto com baixo conteúdo energético (média de 33 kcal), com conteúdo de fibra alimentar total na ordem de 1,6 por cento. Em relação aos macros elementos minerais, a etnovariedade 6, apresentou a maior concentração em potássio (513,5±3,1mg), cálcio (18,9±0,6mg) e a etnovariedade 2 I em Fe (564,4±58,1µg) e Cr (99,3±8,3µg). A menor concentração foi constatada na etnovariedade 12 para os elementos K (229,0±4,5mg), Na (53,7±5,5µg) e Zn (89,3±4,7µg). Apesar das variações em relação as diferentes etno variedades e conseqüentemente concentrações em elementos minerais, o cubiu, pode estar contribuindo para atingir as recomendações desses nutrientes.

Considering the wide genetic variability of cubiu (Solanum sessiliflorum Dunal), its chemical composition was quantified in order to contribute to the chemical composition table of typical Amazonian foods. The cubiu fruit was collected at Alejo von der Pahlen (EEH) experimental station from National Research Institute of Amazonia (INPA). Eight ethno varieties of cubiu identified as 2 I, 3 I, 6, 7, 12, 14, 17, 29 I and III were analyzed. All samples used were ripe. The mineral content was determined by Neutron Activation Analysis technique and alimentary fiber by the enzymatic-gravimetric method. The results showed that cubiu is a hypo caloric food source (mean of 33 kcal and total fiber of about 1.6 percent). Regarding macronutrients, ethnovariety 6 showed high potassium (513.5 ± 3.1 mg) and calcium (18.8 ± 0.6 mg) concentration. Ethnovariety 21, had high Fe (564 ± 58 µg) and Cr (99 ± 8 µg) concentrations. Low concentrations of K (229.0 ± 4.5 mg), Na (53.7 ± 5.5 µg) and Zn (89.3 ± 4.7 µg) were found for ethnovariety 12. Although the cubiu fruit presents great variation of its mineral contents due to different ethnovarieties, it could still contribute as a diet supplement to reach the minimum nutrient requirements for the Amazonian population.

Evaluación genotóxica del extracto acuoso liofilizado de Solanum sessiliflorum Dunal, cocona en células germinales del ratón / Genotoxic evaluation of the lyophilized aqueous extract of Solanum sessiliflorum Dunal, cocona in mouse germ cells

Se ha realizado la evaluación del potencial genotóxico del extracto acuoso liofilizado de los frutos de Solanum sessiliflorum Dunal, mediante el método de anomalías de la cabeza de espermatozoides del ratón. Se utilizaron un total de 21 ratones albinos machos de la cepa Balb/c-53, distribuidos aleatoriamente en tres grupos experimentales con 7 animales por grupo: Solanum sessiliflorum Dunal a dosis de 2000 mg/Kg p.c, ciclofosfamida a dosis de 50 mg/kg p.c. (control positivo) y sohteión salina al 0.9% (control negativo). Se administró el extracto y la solución salina por vía oral y ciclofosfamida por vía intraperitoneal durante 5 días consecutivos. A los 35 días, después de la primera administración, se sacrificaron los animales, se extrajeron los epidídimos, se preparó una solución con tripsina, se coloreó con eosina y se procedió a la extensión de la muestra en láminas porta objetos para su posterior lectura. Se clasificaron los espermatozoides siguiendo el criterio de Wyrobex y Bruce, basado en cabezas normales, amorfas, bananas y sin gancho. No se encontraron diferencias significativas entre el grupo tratado con Solanum sessiliflorum Dunal y el grupo control negativo, por lo que se puede concluir que el extracto acuoso liofilizado de Solanum sessiliflorum Dunal, no posee acción genotóxica a nivel de células germinales.

ANÁLISE MULTIVARIADA DA DIVERGÊNCIA GENÉTICA EM 29 POPULAÇÕES DE CUBIU (Solanum sessiliflorumDunal) AVALIADA NA ZONA DA MATA DO ESTADO DE PERNAMBUCO

The amplitude of the genetic divergence among 29 populations of cubiu from the germplasm collection maintaned by the Amazonian National Research Institute (INPA) was evaluated. The trial was carried out in the Vitória de Santo Antão Experiment Station of the Pernambuco State Research Enterprise (IPA), Pernambuco, Brazil. The experimental design was a randomized complete block, with four replications. The following characters of agronomic importance were evaluated; fruit width (cm); fruit length (cm); stem diameter at soil level (cm); leaf area (cm2); plant height (cm); total fruit number/plant; average fruit weight/ plant (g); estimated fruit yield (Mt/ha); loculi number; pulp thickness (mm) and total soluble solids (%). Cluster analysis (Tocher Optimization Method, using the Mahalanobis Generalized Distance) grouped the 29 cubiu accessions into 9 groups. The accessions from Umariaçú (AM) and São Paulo de Olivença (AM) were most closely related, and are also geographically close, and those from Borba (AM) and São Gabriel da Cachoeira (AM) were most divergent, both genetically and geographically. The accessions from Atalaia do Norte (AM), Borba (AM), Iquitos (Peru), São Gabriel da Cachoeira (AM) and Belém (PA) presented good agronomic traits and were divergent among themselves. Their use as potential progenitors in the cubiu breeding program may provide both good quality and heterosis.

A amplitude da variabilidade genética em 29 populações de cubiu do Programa de Melhoramento Genético de Hortaliças do Instituto Nacional de Pesquisas da Amazônia-INPA, foi avaliada num experimento conduzido na Estação Experimental da Empresa Pernambucana de Pesquisa Agropecuária-IPA, em Vitória de Santo Antão, Estado de Pernambuco. Adotou-se o delineamento experimental em blocos casualizados com 4 repetições. Coletaram-se dados referentes a: largura do fruto (cm); comprimento do fruto (cm); diâmetro do colo (cm); área da folha (cm2); altura da planta (cm); número total de frutos/planta; peso médio de frutos/planta (g); produção estimada de frutos (ton/ha); número de lóculos; espessura da polpa (mm) e de sólidos solúveis totais (%). A análise de agrupamento, pelo Método de Otimização de Tocher, usando a Distância Generalizada de Mahalanobis, agrupou as 29 populações de cubiu em 9 diferentes grupos. Entre os pares de menor e maior divergências genéticas, foram identificadas as populações procedentes de Umariaçu (AM) e de São Paulo de Olivença (AM) e as de Borba (AM) e de São Gabriel da Cachoeira (AM), respectivamente. As populações originárias de Ataláia do Norte (AM), Borba (AM), Iquitos (Peru), São Gabriel da Cachoeira (AM) e de Belém (PA) apresentaram as maiores distâncias genéticas entre os grupos formados. Portanto, podem ser indicadas como progenitores potenciais em programa de melhoramento genético do cubiu.