Anatomical, Histochemical and Biological Studies of Clusia grandiflora Splitg. (Clusiaceae)

Abstract Clusia grandiflora belongs to an important botanical family which is known for its medicinal value, however there are few reports in literature about the species, highlighting the relevance of this study. Anatomical studies with leaves and stems were performed using traditional techniques. In this investigation it was identified particularities of the species such as the presence of vascular system arranged in an opened arc-shaped with the flexed ends towards the inside of the arch with accessory bundles in the petiole. In histochemical studies, performed with different reagents, alkaloids, phenols substances, carbohydrates and lipids were located. The cytotoxic activity of the extracts was performed by tetrazole salt and showed promising results for ethanolic extracts of stems (IC50 human colon cells of 24.30 μg/ mL) and leaves (IC50 ascites gastric cells of 44.15 μg/ mL), without cell membrane disruption of erythrocytes. The antibacterial activity was evaluated by tryptic soy agar and minimal inhibitory concentration assays and showed positive results for Pseudomonas aeruginosa and Escherichia coli, with better result for adventitious roots (32 μg/ mL and 16 μg/ mL, respectively), stems (64 μg/ mL and 32 μg/ mL, respectively) and leaves (64 μg/ mL and 32 μg/ mL, respectively) ethanolic extracts. Thus, these studies were able to characterize the species and show its potential as promising source of active substances.

Clusia criuva Cambess. (Clusiaceae): anatomical characterization, chemical prospecting and antioxidant activity

ABSTRACT This study aims the anatomical description and chemical characterization of aerial parts of Clusia criuva Cambess., Clusiaceae in addition to the evaluation of the antioxidant activity of crude extracts, correlated to the flavonoid content. The morphological characterization was performed using traditional techniques of plant anatomy. For phytochemical studies, crude extracts were obtained by static maceration and analyzed by thin layer chromatography. The antioxidant activity and the flavonoids content were determined by colorimetric methods involving, respectively, 2,2-diphenyl-1-picrylhydrazyl free radical and aluminum chloride. C. criuva has uniseriate epidermis, paracytic stomata, hypostomatic leaves, cuticular flanges and cordiform vascular cylinder with accessory bundles. Chemical prospecting confirmed the abundant presence of terpenes and phenols in the extracts of leaves and of fruits. The methanolic extract of seeds showed the lowest EC50 value, but the methanolic extract of pericarps exhibited the highest maximum antioxidant activity. The results suggested a high percentage of flavonoids in the hexanic extract of pericarps, however, this could represent, in fact, the presence of benzophenones. Secretory ducts and the shape of the midrib are diagnostic for C. criuva. The antioxidant activity is not directly related to the flavonoids. The results indicate the importance of future studies with C. criuva chemical constituents.

Atividade moduladora de extratos etanólico das folhas de Clusia nemorosa G. Mey. (Clusiaceae) sobre drogas antimicrobianas / Actividad moduladora de extractos de las hojas de Clusia nemorosa G. Mey (clusiaceae) sobre drogas antimicrobianas / Modulatory activity of ethanolic extracts of the leaves of Clusia nemorosa G. Mey (Clusiaceae) on antimicrobial drugs

Introduction: the fast development of drug resistance and the slowdown of the development of new active drugs, drew attention to the treatment with the drug combination. Objective: to analyze the antimicrobial activity of the ethanol extract of Cluisa nemorosa (Clusiaceae) against multiresistant strains and standards as well as the modulating action with aminoglycoside amikacin, gentamicin and neomycin. Methods: the plant material (leaves) was collected in Araripe, crushed and put into ethanol 96 % as solvent. Distillation of the solvent was then performed in a rotary evaporator to produce the gross ethanol extrato. A microdilution assay was conducted to verify antibacterial activity and the possible interactions of aminoglycosides associated with the study samples. A sub-inhibitory concentration of 128 µg/mL (Minimum Inhibitory concentration/8) was used. Results: the activity of the extract against the strains isolated patterns had an Minimum Inhibitory concentration > 1024 mg/mL, the action of antibiotics was modulated synergistically by the extract against multidrug-resistant Gram-positive bacteria Staphylococcus aureus and Gram-negative Escherichia coli and Pseudomonas aeruginosa. Conclusions: the synergism of the ethanol extract was verified by microdilution method. Therefore, it is suggested that the extract of Clusia nemorosa be used as a source of natural products for antimicrobial therapy and to combat bacterial multidrug resistance(AU)

Introducción: el rápido desarrollo de resistencia de las drogas y la relentización en el desarrollo de nuevas drogas activas llaman la atención al tratamiento de combinación de drogas. Objetivos: analizar la actividad antimicrobiana del extracto etanólico de Clusia nemorosa (Clusiaceae) frente a cepas multirresistentes y normas, así como la acción modulante con aminoglucósido amikacina, gentamicina y la neomicina. Métodos: el material vegetal (hojas), colectado en el Araripe, fue triturado y sumergido en etanol 96 percent como solvente. De inmediato, fue sometido a destilación del solvente en un evaporador rotativo para la producción de lo extrato etanólico bruto. Se llevó a cabo un ensayo de microdilución para verificar la actividad antibacteriana y las posibles interacciones de aminoglucósidos asociados con las muestras estudiadas, se utilizó una concentración sub-inhibitoria de 128 µg/mL (Concentración mínima inhibitoria/8). Resultados: la actividad del extracto frente a las cepas aisladas patrones tenía una Concentración mínima inhibitoria> 1024mg/mL; la acción de los antibióticos fue modulada por el extracto de forma sinérgica contra multirresistente Gram-positivo Staphylococcus aureus y bacterias Gram-negativas Escherichia coli y Pseudomonas aeruginosa. El sinergismo del extracto de etanol se verificó mediante el método de microdilución. Conclusiones: el extracto de las hojas de C. nemorosa actuaron como un agente modulador. Se sugiere que el extracto de C. nemorosa puede ser utilizado como una fuente de productos naturales en la terapia antimicrobiana y en el combate de la resistencia a múltiples fármacos bacterianos(AU)

Introdução: o rápido desenvolvimento de resistência às drogas e a desaceleração no desenvolvimento de novas drogas ativas, chamaram a atenção para o tratamento com combinação de drogas. Objetivos: analisar a atividade antimicrobiana do extrato etanólico de Clusia nemorosa (Clusiaceae) frente a cepas padrões e multirresistentes bem como a ação moduladora com aminoglicosídeos amicacina, neomicina e gentamicina. Métodos: o material vegetal (folhas), coletado na Chapada do Araripe, foi triturado e submerso em solvente etanol 96 por cento e submetido à destilação do solvente no aparelho evaporador rotativo para a produção do extrato etanólico bruto. Um ensaio de microdiluição foi realizado para verificar a atividade antibacteriana e as possíveis interações dos aminoglicosídeos associados às amostras estudadas, utilizando uma concentração sub-inibitória de 128 µg/mL (concentração inibitória mínima/8). Resultados: a ação do extrato isolado frente às cepas padrões teve uma concentração inibitória mínima > 1024 µg/mL, a ação dos antibióticos foi modulada sinergicamente pelo extrato contra as bactérias multirresistentes Gram-positiva Staphylococcus aureus e Gram-negativas Escherichia coli e Pseudomonas aeruginosa. Conclusões: o extrato das folhas de C. nemorosa atuou como um agente modulador da atividade antimicrobiana. É sugerido que o extrato de Clusia nemorosa pode ser utilizado como uma fonte de produtos naturais na terapêutica antimicrobiana e no combate a multirresistência bacteriana(AU)

Characterization saprobic fungi on leaf litter of two species of trees in the Atlantic Forest, Brazil

Abstract We investigated the composition and structure of fungal communities associated with leaf litter generated by Clusia nemorosa and Vismia guianensis that belong to phylogenetically-related botanical families and exist together in a remnant of the Atlantic Forest in Bahia, Brazil. Samplings were conducted during wet (June 2011) and dry (January 2013) seasons in Serra da Jibóia. The fungi were isolated using particle filtration and the 1,832 isolates represented 92 taxa. The wet season yielded the largest number of isolates (1,141) and taxa (76) compared with the dry season (641 isolates and 37 taxa). The richness and diversity of fungal species associated with C. nemorosa (64 taxa, Simpson=0.95)were higher compared with those of V.guianensis (59 taxa, Simpson =0.90). Analysis of similarity (ANOSIM) revealed significant variations in the composition and community structure of fungi isolated from the two plants as a function of seasons. In contrast, nonmetric multidimensional scaling (NMDS) analysis show that the seasonality was an important influence on the distribution of fungal species. However, the populations of the saprobic fungal communities were dynamic, and several factors may influence such communities in the Atlantic Forest.

Biocontrol evaluation of extracts and a major component, clusianone, from Clusia fluminensisPlanch. & Triana againstAedes aegypti

Studies evaluated the effects of hexanic extracts from the fruits and flowers ofClusia fluminensis and the main component of the flower extract, a purified benzophenone (clusianone), against Aedes aegypti. The treatment of larvae with the crude fruit or flower extracts from C. fluminensis did not affect the survival ofAe. aegypti (50 mg/L), however, the flower extracts significantly delayed development of Ae. aegypti. In contrast, the clusianone (50 mg/L) isolate from the flower extract, representing 54.85% of this sample composition, showed a highly significant inhibition of survival, killing 93.3% of the larvae and completely blocking development of Ae. aegypti. The results showed, for the first time, high activity of clusianone against Ae. aegypti that both killed and inhibited mosquito development. Therefore, clusianone has potential for development as a biopesticide for controlling insect vectors of tropical diseases. Future work will elucidate the mode of action of clusianone isolated from C. fluminensis.

Leaf extract from Clusia nemorosa induces an antinociceptive effect in mice via a mechanism that is adrenergic systems dependent / 中国天然药物

Previous studies on the genus Clusia have shown anti-inflammatory and antiproliferative effects of the leaf extracts, but its antinociceptive activity has never been characterized. In the present study, the antinociceptive activity of the hexane extract of the leaves of Clusia nemorosa G. Mey, called HECn, was examined. Antinociceptive activity was evaluated using acetic acid-induced writhing, formalin, and hot-plate tests. All experiments were carried out on male Swiss mice. The extract (1-400 mg·kg(-1)), given by intraperitoneal route (i.p.) 1 h prior to testing, produced a dose-dependent inhibition on the number of abdominal writhings, with an ID50 of 62 mg·kg(-1). In addition, HECn was able to prevent the visceral pain induced by acetic acid in mice for at least 2 h. In the formalin test, HECn had no effect in the first phase, but produced an analgesic effect on the second phase with the inhibition of licking time. The HECn did not show a significant analgesic effect in the hot plate test. Pretreatment with yohimbine attenuated the antinociceptive effect induced by HECn in the writhing test. However, naloxone, atropine, or haloperidol did not affect antinociception induced by HECn in the writhing test. Together, these results indicate that the extract from the leaves of Clusia nemorosa produces antinociception in models of chemical pain through mechanisms that suggest participation of the adrenergic systems pathway.

Estudo histológico da cicatrização de feridas cutâneas utilizando a banha de bacuri (Platonia insignis Mart.) / Histologic study of skin of wounds healing using the cream of bacuri (Platonia insignis Mart.)

Introdução: O processo de cicatrização é uma sequência de reações para reconstituir o tecido. O bacurizeiro (Platonia insignis Mart.) é nativo da Amazônia e é usado como cicatrizante e antibiótico. Objetivo: Analisar a cicatrização de feridas cutâneas tratadas com a banha de bacuri. Metodologia: Utilizaram-se 46 ratos Wistar, machos. Confeccionou-se uma ferida de 2 cm de diâmetro no dorso dos animais e em seguida foram distribuídos em dois grupos: controle e tratado com banha de bacuri. Foram avaliados no 4º, 7º e 14º dias do pós-operatório. Resultados: A inflamação aguda reduziu a partir do sétimo dia nos dois grupos, sendo mais evidente no grupo experimental. No quarto dia, ambos os grupos não mostraram reepitelização, já no sétimo, os animais tratados com a banha demonstraram reepitelização considerável. Conclusão: A banha de bacuri foi eficiente no sétimo dia de tratamento. Os ácidos graxos presentes nessa substância favorecem o processo cicatricial.

Introduction: The healing process is a sequence of reactions to restore tissue continuity occurs. The bacuri tree (Platonia insignis Mart.) is native to the Amazon, use like antimicrobial and healing action. Objective: To analyze the healing of open wounds and treated with bacuri cream. Methods: We used 46 male Wistar rats. We made up a two-centimeter-diameter wound on the dorsum of all animals which were divided into two groups: the control group and treated and assessed at 4, 7 and 14 days post-operatively. Results: Acute inflammation reduced from seven days in both groups, although it had been more evident in the group treated with the cream. At fourth days both groups did not show any re-epithelialization, the animals treated with the cream on the seventh day showed considerable re-epithelialization. Conclusion: The bacuri cream was effective on the seventh day. The fatty acids present in this substance favor the healing process.

Dinámica del daño foliar en plántulas de Drimys granadensis (Winteraceae) y Clusia multiflora (Clusiaceae) en el bosque altoandino de la Cordillera Oriental colombiana

Leaf damage dynamics of Drimys granadensis (Winteraceae) and Clusia multiflora (Clusiaceae) seedlings in the High-Andean Forest. Because of their slow growth, shade-tolerant species remain exposed to foliar damage long before reproductive age. Foliar damage can be controlled through synchronous leaf production and specialized phytochemical strategies. in this study, the dynamics of foliar damage in different cohorts of seedlings were evaluated to determine if the High-Andean species, Clusia multiflora (Clusiaceae) and Drimys granadensis (Winteraceae) appeal to the synchronous leaf production for controlling the herbivory and pathogenical damage, and to establish the possible relations between the healthy state, growth and mortality of seedlings. Since a recently-emerged seedling could not supply the physiological costs of specialized strategies, we expected the highest synchronous leaf production in the youngest individuals, and differences between cohorts in foliar damage. Four variables were measured to evaluate the health state along time, in three cohorts of seedlings: Proportion of predated leaves (Pp), proportion of healthy leaves (Ps), proportion of leaves with symptoms of disease or punctual damage (Pe) and growing (% increment of height). Both species showed significant differences between periods in the proportion of healthy leaves, but there was not an effect of time-cohort interaction, therefore synchronic production of leaves was not a strategy more used for any particular cohort. Foliar damage oscillated along time, fact that can be explained by the pulses of leaf production. However, this strategy had little efficiency to control the pathogen attack. in general, the unified behavior of all variables was affected by cohort, time, species an all the different interactions. Association between growing and foliar damage were conditioned by climate. The highest mortality ocurred during the dry season, and one fourth of the D. granadensis deaths were caused by the joint action of herbivory and drought. Results suggest that in spite of the synchronous leaf production in all cohorts, this strategy quickly loses the efficiency for controlling foliar damage, because the means of Ps were below of 0.25 at the end of the monitoring period. The differences between species in the dynamics of health state should explain partially their coexistence in the High-Andean forest. Rev. Biol. Trop. 56 (3): 1087-1100. Epub 2008 September 30.

Además de las estrategias fitoquímicas especializadas, las plantas pueden usar la producción sincrónica de hojas como un método de saciado de patógenos y herbívoros. Con el fin de determinar si las especies del bosque altoandino colombiano Clusia multiflora (Clusiaceae) y Drimys granadensis (Winteraceae) recurren a la producción sincrónica de hojas para controlar los efectos de la herbivoría y el ataque microbiano, y establecer que relación existe entre el estado de salud, el crecimiento y la mortalidad de plántulas, se evaluó la dinámica del daño foliar sobre diferentes cohortes. Dado que una plántula tolerante a la sombra recientemente emergida no puede suplir los costos fisiológicos de una estrategia especializada, se esperaba una alta sincronía en la producción de hojas en las plántulas más jóvenes, y diferencias en el daño foliar entre cohortes. Se midieron cuatro variables que evaluaban el estado de salud a lo largo del tiempo, en tres cohortes de plántulas: Proporción de hojas predadas, proporción de hojas sanas, proporción de hojas enfermas o con daño puntual y crecimiento. Ambas especies mostraron diferencias significativas entre épocas, en la proporción de hojas sanas; pero no hubo un efecto de la interacción tiempo-cohorte, por lo tanto la producción sincrónica de hojas no fue una estrategia más usada por alguna cohorte en particular. El daño foliar osciló a través del tiempo, lo cual puede ser explicado por los pulsos en la producción de hojas. Sin embargo, ésta estrategia tuvo poca eficiencia para controlar el ataque por patógenos. En general, el comportamiento unificado de todas las variables fue afectado por la cohorte, el tiempo, la especie y todas las diferentes interacciones. La relación entre crecimiento y daño foliar fue condicionado por el clima. La mayor mortalidad se dio durante la estación seca, y un cuarto de las muertes en D. granadensis fueron causadas por la acción conjunta de herbivoría y sequía. Los resultados sugieren que a pesar de la producción sincrónica de hojas en todas las cohortes, fue poca la eficiencia para controlar el daño foliar, ya que las medias de Ps estuvieron por debajo de 0.25 al final del seguimiento. Las diferencias en la dinámica del estado de salud entre especies podrían explicar parcialmente la coexistencia en el bosque altoandino.

Flavonoids, benzophenones and a new euphane derivative from Clusia columnaris Engl / Flavonóides, benzofenonas e um novo eufano de Clusia columnaris Engl

The polyisoprenylated benzophenones machuone and clusiachromene A have been isolated from the fruits of Clusia columnaris. The hexane extract of the young branches with leaves afforded a new euphane derivative, whose structure was elucidated by spectroscopic methods. On the contrary, the most polar EtOAc and ButOH extracts were constituted of flavonoid C-glucosides (isovitexin, vitexin and vitexin-2"-xyloside) and seven biflavonoids of the so-called Garcinia group.

Dos frutos de Clusia columnaris foram isoladas as benzofenonas poliisopreniladas machuona e clusiacromeno A. Do extrato em hexano obtido de galhos e folhas novas, um novo triterpeno do tipo eufano foi isolado. Sua estrutura foi elucidada através de métodos espectroscópicos. Por outro lado, dos extratos mais polares - em acetato de etila e em butanol, foram isolados os flavonóides C-glicosilados isovitexina, vitexina e vitexina-2"-xilosídeo, além de sete bisflavonóides conhecidos como bisflavonóides do grupo da Garcinia.

Bees visitors of three species of Clusia (Clusiaceae) flowers in Central Amazonia / Abelhas visitantes de flores de três espécies de Clusia (Clusiacea) na Amazônia Central

Observations on bees visitors to three species of Clusia (Clusiaceae) flowers in the Reserva Adolpho Ducke, Manaus, Amazonas, Brazil were made during three two-week periods. The three species of Clusia, namely C. grandiflora, C. panapanari and C. insignis, presented variations regarding the species of bee visitors. A total of 23 bee species visited the three species of Clusia. The Euglossini and Meliponinae bees were the most frequent visitors of the Clusia flowers. Bee collecting behavior of floral resources is described.

Durante seis semanas foram realizadas observações das abelhas visitantes de flores de três espécies de Clusia (Clusiaceae), na Reserva Florestal Adolpho Ducke, em Manaus. As três espécies de Clusia: C. grandiflora Splitg., C. panapanari (Aubl.) e C. insignis Mart. apresentaram diferenças com relação às visitas de espécies de abelhas, tendo sido visitadas por 23 espécies. Abelhas Euglossini e Meliponinae foram os visitantes mais freqüentes em flores de Clusia. O comportamento de coleta de recursos florais é descrito.