Seasonal and pluviometric effects on the phenolic compound composition and antioxidant potential of Licania macrophylla Benth (Chrysobalanaceae), a medicinal plant from the Amazon rainforest

Abstract Licania macrophylla is a medicinal plant from the Amazon. It is mainly used in the form of a decoction and has been reported to contain several phenolic compounds. However, the effect of seasonality on the phenolic composition and antioxidant potential of this plant has not been well studied, especially in the Amazon region, an area affected by the rainy and less-rainy seasons. Therefore, we evaluated the seasonality of these aromatic compounds and the antioxidant potential of the extracts from L. macrophylla stem bark. We also determined the correlation between the extraction methods used and precipitation levels during each period for 1 year. The total flavonoid and phenolic content, DPPH-scavenging potential, percentage of phosphomolybdenum complex reduction, and iron-reducing power were quantified. The levels of phenolic compounds were the highest in June, whereas those of flavonoids were the highest in September and October; however, these differences were not significant. The extracts from April, November, and June showed the best results for DPPH scavenging, phosphomolybdenum reduction, and iron reduction power, respectively. Significant differences in the phenolic content and DPPH-scavenging activity were observed between the more- and less-rainy seasons. The total phenolic content was positively correlated with FRAP and DPPH, whereas flavonoid levels were negatively correlated.

Comparative leaf morphological analysis of 20 species of Chrysobalanaceae / Morfologia foliar comparativa de 20 espécies de Chrysobalanaceae

Morphological studies focusing on vegetative traits are useful in identifying species when fertile material is not available. The aim of this study was to assess the application of comparative leaf morphology to identify species of the Chrysobalanaceae family. The morphological observations were made with a stereomicroscope. We used the diaphanization technique for viewing venation details. It is shown the descriptions of the leaf morphology, illustrations and an identification key for 20 species from genera Couepia, Licania and Parinari (Chrysobalanaceae) occurring in the Adolpho Ducke Forest Reserve, Manaus, AM, Brazil. The key was constructed using the DELTA (DEscription Language for TAxonomy) software. Leaf traits such as the presence of intersecondary venation and the type of insertion of secondary veins were recorded for each species. These morphological leaf traits are reliable for identifying species of Chrysobalanaceae.

Estudos morfológicos com foco em caracteres vegetativos são úteis na identificação das espécies quando material fértil não está disponível. O objetivo deste estudo foi avaliar a aplicação da morfologia foliar comparativa na identificação de espécies da família Chrysobalanaceae. A caracterização morfológica foi realizada com observação em estereomicroscópio. Foi utilizada a técnica de diafanização para visualização de detalhes da venação. São apresentadas descrições da morfologia foliar, ilustrações e uma chave de identificação de 20 espécies dos gêneros Couepia, Licania e Parinari (Chrysobalanaceae) que ocorrem na Reserva Florestal Adolpho Ducke, Manaus, AM, Brasil. A chave de identificação foi construída com uso do programa DELTA (DEscription Language for TAxonomy). Caracteres foliares como a presença de nervuras intersecundárias e o tipo de inserção das nervuras secundárias foram registrados para cada espécie. Essas características foliares são confiáveis para a identificação de espécies de Chrysobalanaceae.

Couepia paraensis: estudio fitoquímico, ensayos de citotoxicidad y de inhibición de la glucosa-6-fosfatasa / Couepia paraensis: phytochemical study, cytotoxicity assays and inhibition of glucose-6-phosphatase

From the cholroform extract of the aerial parts of Couepia paraensis the triterpenes beta-sitosterol1, betulinic acid acetate 2, and oleanolic acid acetate 3, were isolated. Six triterpenes from the chloroform-methanol, acids: oleanolic 4, pomolic 5, ursolic 6, betulinic 7, 6-beta-hydroxybetulínic 8. Additionally from the methanolic extract three flavonoids were isolated: mricetin 9, quercetin 10 y rutina 11. The chloroform and chloroform-methanol extracts were not citotoxic at concentration of 2,5 and 3,1 ug/ml respectively after 24 hours of incubation. The methanol extract was found to be harmless to a concentration of 50 ug/ml, both at 24 hours (LD50 = 10.77 ug/ml) and 120 hours (LD50 = 28.86 ug/ml) of incubation. Only the methanol extract showed significant inhibition (41 percent) of the activity of G-6-Pase in intact microsomes without affecting the activity of the enzyme in microsomes broken.

Se aislaron e identificaron tres triterpenos: beta-sitosterol 1, acetato del ácido betulínico, 2 y acetato del ácido oleanólico 3 del extracto clorofórmico. Seis triterpenos del extracto cloroformo: metanol (9:1) que fueron identificados como ácidos: oleanólico 4, pomólico 5, ursólico 6, betulínico 7, 6-beta-hidroxibetulínico 8. Mientras que del extracto metanólico se identificaron 3 flavoniodes: miricetina 9, quercetina 10 y rutina 11. Los extractos de cloroformo y cloroformo /metanol resultaron inocuos hasta las concentraciones de 2,5 y 3,1ug/ml respectivamente, después de 24 horas de incubación. El extracto metanólico es inocuo hasta una concentración de 50 ug/ml, tanto a 24 horas (LD50 = 10,77 ug/ml) como a 120 horas (LD50 = 28,86 ug/ml) de incubación. Solamente el extracto metanólico mostró una inhibición significativa (41 por ciento) de la actividad de la G-6-Pasa de microsomas intactos sin afectar la actividad de la enzima en microsomas rotos.

Antimicrobial, antiplasmodial, haemolytic and antioxidant activities of crude extracts from three selected Togolese medicinal plants

OBJECTIVE@#To investigate the antioxidant, antimicrobial, antiplasmodial, acute toxicity and haemolytic activities of methanolic extracts of three plants. Phytochemical analysis to determine the phenolic contents was also carried out.@*METHODS@#The 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging, NCCLS broth microdilution and Plasmodium Lactate Dehydrogenase (pLDH) assays were used to determine antioxidant, antimicrobial and antiplasmodial activities, respectively. Haemolysis assay was conducted on A(+) human red blood cells and acute toxicity on male Swiss albino mice. Phenolics were quantitatively determined using spectrophotometric methods.@*RESULTS@#The DPPH assay yielded interesting antioxidant activities of methanolic extract of Parinari curatellifolia (P. curatellifolia) and Entada africana (E. africana) (IC(50) were 0.20±0.01 μg/mL and 0.47±0.01 μg/mL, respectively). This activity was highly correlated with phenolic contents of extracts. The antimicrobial tests displayed minimal inhibitory concentrations (MICs) values ranging from 0.90 to 1.80 mg/mL for Serratia marcescens (S. marcescens) the most susceptible bacterial strain. MIC value was 1.20 mg/mL for susceptible fungal strains including Mucor rouxi (M. rouxi), Fusarium oxyporum (F. oxyporum) and Rhizopus nigricans (R. nigricans). pLDH assay showed moderate antiplasmodial activity of Balanites aegyptiaca (B. aegyptiaca) (IC(50) = 24.56±3.45 μg/mL), however this extract was highly haemolytic and toxic in mice (LD(50) = 625±128 mg/kg).@*CONCLUSIONS@#Our results support in part the use of the selected plants in the treatment of microbial infections. In addition the plant showed interesting antioxidant activity that could be useful in the management of oxidative stress.

Pollination and breeding system of Couepia uiti (Mart. and Zucc.) Benth (Chrysobalanaceae) in the Pantanal da Nhecolândia

The pollination biology and breeding system of Couepia uiti was studied. In this species, flowers opened at 06:00 AM anthesis, and nectar production began at around 0800 h, reached a maximum volume from 09:30 AM to 10:30 AM, and decreased thereafter. The nectar sugar concentration increased continuously, but showed an abrupt increase from 10:00 AM to 12:00 AM. Pollen release occurred at about 09:30 AM and was quickly collected. The stigmas became receptive at around 12:00 AM. The pollinators of C. uiti included the bees Apis mellifera, Xylocopa sp. and Bombus sp., and three species of wasps. This conclusion was based on the observation that these hymenopterans had C. uiti pollen on their bodies, visited the receptive flowers, and touched the anthers and stigmas, thereby promoting pollination. Of these floral visitors, A. mellifera was considered to be the most efficient pollinator. However, mixed pollination also occurred. The number of C. uiti flowers visited in the morning (n = 52) was three times smaller than in the afternoon (n = 62), and the species richness of floral visitors was also bigger in the afternoon (eight in the afternoon versus five in the morning). This finding indicated that these floral visitors preferred to exploit nectar rather than pollen. Controlled pollination experiments showed that C. uiti was a self-incompatible species that produced fruits only by cross-pollination. Treatments such as agamospermy and spontaneous and self-pollinations did not produce fruits.

Estudei a biologia da polinização e o sistema reprodutivo de Couepia uiti. Esta espécie apresentou antese matutina, produção de néctar iniciou por volta das 08:00 horas, atingindo volume máximo entre 09:30 e 10:30 horas e diminuindo após este horário. A concentração de açúcares no néctar aumentou continuamente, apresentando aumento abrupto entre 10:00 e 12:00 horas. A liberação de pólen ocorreu às 09:30 horas e este foi rapidamente removido. O estigma estava receptivo às 12:00 horas. Considerei polinizadores de C. uiti as abelhas: Apis mellifera, Xylocopa sp., Bombus sp. e três espécies de vespas, pois apresentaram polén de C. uiti em seus corpos. Visitaram flores quando estavam receptivas, tocando as anteras e os estigmas, promovendo a polinização. Dentre os visitantes florais, A. mellifera foi considerado o polinizador mais eficiente, no entanto, polinização mista parece ocorrer. O número de visitas a flores de C. uiti pela manhã (n = 52) foi três vezes menor que o encontrado no período da tarde (n = 162) e o número de espécies de visitantes também foi maior à tarde, sendo encontradas cinco espécies durante a manhã e oito à tarde. Portanto, é possível concluir que estes visitantes florais preferem explorar o néctar em comparação ao pólen. Os tratamentos de polinizações controladas mostraram que C. uiti é uma espécie auto-incompatível, produzindo frutos somente por polinização cruzada. Tratamentos de agamospermia, polinização espontânea e autopolinização não produziram frutos.

A Chrysobalanus icaco extract alters the plasmid topology and the effects of stannous chloride on the DNA of plasmids

Chrysobalanus icaco (C. icaco) leaves are used in folk medicine (known as Abajeru in Brazil) to control the glycaemia in diabetic patients. Stannous chloride (SnCl2) is a powerful reducing agent used for different purposes and presents cytotoxic and genotoxic effects. The aim of this work was to investigate the effect of an aqueous C. icaco extract on the plasmid DNA topology and on the effects of the stannous chloride on DNA plasmid. Plasmid pBSK was incubated with a C. icaco extract in the presence or absence of SnCl2 (200 mg/mL), after that, the agarose gel electrophoresis procedure was carried out. Plasmid incubated only SnCl2 was used as positive control and, as negative control, plasmid incubated with Tris buffer. The gels were stained with ethidium bromide, DNA bands were semiquantified by densitometry. The data showed that C. icaco extract alters the electrophoretic profile and decreases significantly (p < 0.05) the effect of SnCl2 on plasmid DNA. The results obtained in this work could indicate a dose-dependent protective action and a genotoxic effect of C. icaco extract on plasmid DNA.

Folhas de Chrysobalanus icaco (C. icaco) são usadas na medicina popular (conhecido como Abajeru no Brasil) para controlar a glicemia em pacientes diabéticos. Cloreto estanoso (SnCl2) é um agente redutor potente usado para diferentes propostas e apresenta efeitos citotóxico e genotóxico. O objetivo deste trabalho foi investigar os efeitos de um extrato aquoso de C. icaco na topologia de DNA plasmidial e nos efeitos do cloreto estanoso sobre o DNA plasmidial. Plasmídios pBSK foram incubados com um extrato de C. icaco na presença ou ausência do SnCl2 (200 mg/mL), em seguida, o procedimento de eletroforese em gel de agarose foi realizado. Plasmídios incubados somente com SnCl2 foram usados como controle positivo e, como controle negativo, plasmídios incubados com tampão Tris. Os géis foram corados com brometo de etídio e as bandas de DNA foram semiquantificadas por densitometria. Os dados mostraram que o extrato de C. icaco altera o perfil eletroforético e diminui significativamente (p < 0,05) os efeitos do SnCl2 sobre DNA plasmidial. Os resultados obtidos neste trabalho indicam uma ação protetora dependente da dose e um efeito genotóxico de extrato de C. icaco sobre o DNA plasmidial.

Pathological Changes Induced by Chrysobalanus Icaco Seeds

The study was to determine possible pathological changes induced by the consumption of C. icaco and possible health implication. Animal feed tests were carried out using varying amounts of the seed mixed with usual mice meals. The result revealed weight loss; and increase in total bilirubin; protein; albumin; globin; cholesterol level; alkaline phosphate and mean serum glutamic - oxaloacetate transaminase and serum glutamic - pyruvic transaminase activities. Possible health implications are discussed