Phytochemical profile and biological activities of Bromelia antiacantha extracts / Perfil fitoquímico e atividades biológicas de extratos de Bromelia antiacantha

Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

Phytochemical profile and biological activities of Bromelia antiacantha extracts

Abstract Reports from popular medicine usually act as a basis for the development of new drugs from natural compounds with therapeutic actions for serious diseases and prevalence such as cancer. Bromelia antiacantha Bertol. is a species of the Bromeliaceae family, considered an unconventional food plant, found in the south and midwest regions of Brazil. Despite the high nutritional content and pharmacological potential of its fruits, few scientific studies report its biological actions. Thus, this study evaluates the phytochemical profile of aqueous and ethanol extracts obtained from B. antiacantha fruits, as well as their possible antioxidant, antitumor, and cytotoxic activities. The aqueous extract exhibited phenolic compounds and flavonoids, while ethanol extracts indicated the presence of flavonoids and coumarin in their composition, regardless of the region of collection. The ethanolic extract demonstrated a more promising antioxidant effect than the aqueous extract and also induced a significant inhibition in the viability of human cervical cancer cells of the SiHa strain. In addition, treatment with both extracts did not alter the viability of non-tumor cells of the immortalized human keratinocyte lineage (HaCaT). These results bring new data about extracts obtained from a native plant, edible and traditionally used in popular medicine, opening new perspectives for its possible therapeutic application.

Resumo Relatos da medicina popular costumam atuar como referencial para o desenvolvimento de novos fármacos a partir de moléculas naturais com ações terapêuticas para doenças de alta gravidade e prevalência como o câncer. Bromelia antiacantha Bertol. é uma espécie da família Bromeliaceae, considerada uma planta alimentícia não convencional (PANC), encontrada nas regiões sul e centro-oeste do Brasil. Apesar do alto teor nutritivo e potencial farmacológico de seus frutos, poucos estudos científicos relatam suas ações biológicas. Desta forma, este estudo avalia o perfil fitoquímico de extratos aquoso e etanólico obtidos de frutos de B. antiacantha, bem como a sua possível ação antioxidante, antitumoral e citotóxica. O extrato aquoso apresentou compostos fenólicos e flavonoides, enquanto os extratos etanólicos apontam a presença de flavonóides e cumarina em sua composição, independente da região de coleta. O extrato etanólico demonstrou efeito antioxidante mais promissor do que o extrato aquoso e também induziu uma inibição significativa na viabilidade de células humanas de câncer cervical da linhagem SiHa. Além disso, o tratamento com ambos extratos não alterou a viabilidade de células não tumorais da linhagem de queratinócitos humanos imortalizados (HaCaT). Estes dados trazem novas informações sobre extratos obtidos de uma espécie vegetal nativa, comestível e já utilizada tradicionalmente, mas abrindo novas perspectivas quanto a possíveis aplicações terapêuticas.

Anopheles (Kerteszia) cruzii infected by Plasmodium in the Atlantic Forest indicates that the malaria transmission cycle is maintained even after howler monkeys' population decline.

The Atlantic Forests outside of the Amazon region in Brazil are low-frequency malaria hotspots. The disease behaves as a zoonosis maintained by nonhuman primates (NHPs), especially howler monkeys. Between 2016 and 2018, Brazil witnessed the largest yellow fever outbreak since 1980, resulting in massive declines in these NHP populations. However, reports of malaria cases continued in transmission areas. This scenario motivated this survey to determine the frequency of infection of the anophelines by Plasmodium species. Mosquitoes were captured using Shannon traps and CDC light traps and identified as to species based on morphological characters. The screening for malaria parasites targeted only Anopheles species belonging to the subgenus Kerteszia, the proven primary malaria vector. A TaqMan qPCR assay using ribosomal primers (18S rRNA gene) was performed in a Step One Plus Real-time PCR to detect Plasmodium species. Seven hundred sixty field-caught anophelines divided into 76 pools were examined. Out of 76 tested pools, seven (9.21%) were positive. Three pools were Plasmodium malariae-positive, and four were Plasmodium vivax-positive. The anopheline infection was expressed as the maximum infection rate (MIR), disclosing a value of 0.92%, indicative of a steady state. Such stability after the yellow fever outbreak suggests that other species of NHPs could support transmission.

Characterization of the fruit proteolytic system of Bromelia serra Griseb. (Bromeliaceae) and its application in bioactive peptides release.

A crude extract with proteolytic activity was prepared from edible fruits of Bromelia serra, containing cysteine peptidases with molecular masses between 24.1 and 25.9 kDa. The extract presented an optimal pH range of 6.03-9.05, retained more than 80% of activity after thermal pre-treatments at 23, 37, and 45°C (120 min), but it was rapidly inactivated after 10 min at 75°C. These proteases were employed to hydrolyze soybean proteins, bovine casein and bovine whey, achieving degrees of hydrolysis of 18.3 ± 0.6, 29.1 ± 0.7, and 12.6 ± 0.9% (55°C, 180 min), respectively. The casein 180 min-hydrolysate (55°C) presented the maximum value of antioxidant activity (2.89 ± 0.12 mg/mL Trolox), and the whey protein 180 min-hydrolysate (55°C) showed the highest percentage of angiotensin-converting enzyme inhibition (91.9 ± 1.2%). This low-cost enzymatic preparation would be promising for the food industry because it requires mild working conditions and yields hydrolysates with biological activities useful as ingredients for functional food. PRACTICAL APPLICATION: Proteolytic enzymes are employed in the food industry in a wide variety of processes since they modify the properties of proteins causing beneficial effects such as improvement digestibility, diminution of allergenicity, and release of bioactive peptides. Fruits from Bromelia serra possess cysteine peptidases that could be used in food biotechnology because they are capable to hydrolyze soybean and milk proteins by mild working conditions and to provoke the release of bioactive peptides. These hydrolysates containing antioxidative and ACE-inhibitor activities would be useful as ingredients for functional foods or as nutraceuticals, which are nowadays two products highly required by consumers.

Residual malaria of Atlantic Forest systems and the influence of anopheline fauna.

In Brazil, the Amazon region comprises 99.5% of the reported malaria cases. However, another hotspot of the disease is the Atlantic Forest regions, with the sporadic occurrence of autochthonous human cases. In such context, this study sought to investigate the role of anopheline mosquitoes (Diptera: Culicidae) in the residual malaria transmission in Atlantic Forest areas. Two rural areas in the Espírito Santo state were the surveyed sites. Mosquitoes were captured using Shannon trap and CDC light traps and identified into species based on morphological characters. Ecological indexes (Shannon-Wiener diversity, Simpson's dominance, Pielou equability, and Sorensen similarity) were the tools used in the anopheline fauna characterization and comparison along with the two explored areas. The assessment of the sampling adequacy in the studied areas was possible through the generation of a species accumulation curve. A correlation test verified the influence of climatic variables on the anopheline species abundance. A total of 1471 female anopheline mosquitoes were collected from May 2019 to April 2020, representing 13 species. The species richness was higher in Valsugana Velha (hypo-endemic) than in Alto Caparaó (non-endemic). There was a significant variation in the species abundance between Valsugana Velha (n = 1438) and Alto Caparaó (n = 33). The most abundant species was Anopheles (Kerteszia) cruzii complex Dyar and Knab, 1908 representing 87% of the total anophelines collected. These results suggest that the Plasmodium spp. circulation in Brazilian Atlantic Forest areas occurs mainly due to the high frequency of Anopheles (K.) cruzii complex, considered the principal vector of simian and human malaria in the region.

Ethnopharmacology of Fruit Plants: A Literature Review on the Toxicological, Phytochemical, Cultural Aspects, and a Mechanistic Approach to the Pharmacological Effects of Four Widely Used Species.

Fruit plants have been widely used by the population as a source of food, income and in the treatment of various diseases due to their nutritional and pharmacological properties. The aim of this study was to review information from the most current research about the phytochemical composition, biological and toxicological properties of four fruit species widely used by the world population in order to support the safe medicinal use of these species and encourage further studies on their therapeutic properties. The reviewed species are: Talisia esculenta, Brosimum gaudichaudii, Genipa americana, and Bromelia antiacantha. The review presents the botanical description of these species, their geographical distribution, forms of use in popular medicine, phytochemical studies and molecules isolated from different plant organs. The description of the pharmacological mechanism of action of secondary metabolites isolated from these species was detailed and toxicity studies related to them were reviewed. The present study demonstrates the significant concentration of phenolic compounds in these species and their anti-inflammatory, anti-tumor, photosensitizing properties, among others. Such species provide important molecules with pharmacological activity that serve as raw materials for the development of new drugs, making further studies necessary to elucidate mechanisms of action not yet understood and prove the safety for use in humans.

Development, characterization, and transferability of SSR markers for Vriesea carinata (Bromeliaceae) based on RNA sequencing.

PREMISE OF THE STUDY: Expressed sequence tag-simple sequence repeat (EST-SSR) markers were isolated for Vriesea carinata, an endemic bromeliad from the Brazilian Atlantic Forest. These SSR loci may be used to investigate the genetic diversity and population structure of this species and related bromeliads. METHODS AND RESULTS: Based on the transcriptome data of V. carinata, 30 primer pairs were designed and selected for initial validation. Of these primer pairs, 16 generated suitable SSR loci in 69 individuals. The number of alleles per locus ranged from one to 13; the levels of observed and expected heterozygosity per locus ranged from 0.000 to 1.000 and from 0.000 to 0.935, respectively. All loci produced heterologous amplification. Transferability of the loci was tested in 15 species belonging to three Bromeliaceae subfamilies. CONCLUSIONS: The developed EST-SSR markers revealed polymorphism in the four studied populations and could be useful to investigate the genetic diversity of V. carinata and related species. The markers may also be suitable for novel gene annotation and discovery.

Enzyme activity and partial characterization of proteases obtained from Bromelia karatas fruit and compared with Bromelia pinguin proteases.

The enzymatic activity and partial characterization of proteases from Bromelia karatas fruits were evaluated and compared with Bromelia pinguin proteases. The specific activity increased twofold after partial purification in both proteases. Partially purified proteases from Bromelia karatas showed good specific activity at pH 6.0-8.0 and residual activity of 70-100% for 60 min at 37-60 °C, similar to Bromelia pinguin proteases. The K m value of proteases from Bromelia karatas was higher (253.32 µM) than that of Bromelia pinguin proteases (234.94 µM). The use of specific protease inhibitors indicated the presence of cysteine and serine proteases. Proteases with molecular weight of 66.2-97 and 21-31 kDa were detected. Bromelia karatas proteases registered 73% hydrolysis using a soy protein concentrate, similar to the enzyme activity of Bromelia pinguin proteases and commercial bromelain. These results demonstrate that Bromelia karatas proteases could be a potential alternative protease in the food industry.

Toxicological evaluation and anti-inflammatory potential of an ethanolic extract from Bromelia balansae (Bromeliaceae) fruit.

ETHNOPHARMACOLOGICAL RELEVANCE: Bromelia balansae is a relatively unexplored medicinal species that is used for nutritional purposes and in folk medicine to treat cough or wounds. AIM OF THIS STUDY: This study assessed the anti-inflammatory activity of the ethanolic extract obtained from Bromelia balansae fruit (EEBB) as well as the toxicological potential of this extract after single and repeated exposure. MATERIALS AND METHODS: Male rats (Wistar) were gavaged with 2000 mg/kg of extract from the fruit of B. balansae for the acute toxicity test and with 25, 100, or 400 mg/kg of EEBB for the subacute toxicity test. The anti-inflammatory effect of EEBB was evaluated in vivo (30, 100, or 300 mg/kg) by carrageenan (Cg) induced-oedema and pleurisy in Swiss mice. RESULTS: A single oral dose of EEBB did not result in toxicity, demonstrating that the LD50 of this extract was greater than 2000 mg/kg. In the subacute toxicity test, the tested doses produced no significant changes in the haematological, biochemical or histopathological parameters of treated animals. Similarly, there were no statistically significant differences in the sperm parameters. A dose of 300 mg/kg of EEBB significantly reduced oedema formation, Cg-induced mechanical hypersensitivity and cold sensitivity, as well as leukocyte migration in the pleurisy model. CONCLUSION: These results show that EEBB has an anti-inflammatory potential without causing acute or subacute toxicity. These data may contribute to the advancement of biopharmaceutical applications for this species.

Non-Polar Natural Products from Bromelia laciniosa, Neoglaziovia variegata and Encholirium spectabile (Bromeliaceae).

Extensive regional droughts are already a major problem on all inhabited continents and severe regional droughts are expected to become an increasing and extended problem in the future. Consequently, extended use of available drought resistant food plants should be encouraged. Bromelia laciniosa , Neoglaziovia variegata and Encholirium spectabile are excellent candidates in that respect because they are established drought resistant edible plants from the semi-arid Caatinga region. From a food safety perspective, increased utilization of these plants would necessitate detailed knowledge about their chemical constituents. However, their chemical compositions have previously not been determined. For the first time, the non-polar constituents of B. laciniosa , N. variegata and E. spectabile have been identified. This is the first thorough report on natural products from N. variegata , E. spectabile , and B. laciniosa . Altogether, 20 non-polar natural products were characterized. The identifications were based on hyphenated gas chromatography-high resolution mass spectrometry (GC-HRMS) and supported by 1D and 2D Nuclear Magnetic Resonance (NMR) plant metabolomics.

Partial Characterization of the Proteolytic Properties of an Enzymatic Extract From "Aguama" Bromelia pinguin L. Fruit Grown in Mexico.

Plant proteases are capable of performing several functions in biological systems, and their use is attractive for biotechnological process due to their interesting catalytic properties. Bromelia pinguin (aguama) is a wild abundant natural resource in several regions of Central America and the Caribbean Islands but is underutilized. Their fruits are rich in proteases with properties that are still unknown, but they represent an attractive source of enzymes for biotechnological applications. Thus, the proteolytic activity in enzymatic crude extracts (CEs) from wild B. pinguin fruits was partially characterized. Enzymes in CEs showed high proteolytic activity at acid (pH 2.0-4.0) and neutral alkaline (pH 7.0-9.0) conditions, indicating that different types of active proteases are present. Proteolytic activity inhibition by the use of specific protease inhibitors indicated that aspartic, cysteine, and serine proteases are the main types of proteases present in CEs. Activity at pH 3.0 was stable in a broad range of temperatures (25-50 °C) and retained its activity in the presence of surfactants (SDS, Tween-80), reducing agents (DTT, 2-mercapoethanol), and organic solvents (methanol, ethanol, acetone, 2-propanol), which suggests that B. pinguin proteases are potential candidates for their application in brewing, detergent, and pharmaceutical industries.

Ocorrência de Aedes aegypti e Aedes albopictus em bromélias cultivadas no Jardim Botânico Municipal de Bauru, São Paulo, Brasil / Aedes aegypti and Aedes albopictus in bromeliads grown in the Bauru Municipal Botanical Gardens, São Paulo, Brazil / Ocurrencia de Aedes aegypti y Aedes albopictus en bromelias cultivadas en el Jardín Botánico Municipal de Bauru, São Paulo, Brasil

Resumo: O objetivo deste trabalho foi observar a ocorrência de larvas de mosquitos, em especial do Aedes aegypti e Aedes albopictus, no tanque e axilas de bromélias do Jardim Botânico Municipal de Bauru, São Paulo, Brasil, destacando implicações epidemiológicas do uso dessas plantas. A maioria de larvas encontradas foi de mosquitos do gênero Culex, enquanto as de A. aegypti e A. albopictus foi ocasional. O uso de telas para proteção das plantas, a exposição ao sol e a maior quantidade de água no tanque podem ter influenciado a ocorrência e agrupamento das larvas.

The aim of this study was to observe the occurrence of mosquito larvae, especially Aedes aegypti and Aedes albopictus, in the tanks and axillae of bromeliads at the Bauru Municipal Botanical Gardens, São Paulo, Brazil, highlighting the epidemiological implications for the use of these plants. The majority of the larvae belonged to mosquitos from genus Culex, with only occasional findings of A. aegypti and A. albopictus. The use of screens for protection of the plants, exposure to sunlight, and larger amounts of water in the tanks may have influenced the occurrence and grouping of larvae.

El objetivo de este trabajo fue observar la ocurrencia de larvas de mosquitos, en especial las del Aedes aegypti y Aedes albopictus, en el tanque y axilas de bromelias del Jardín Botánico Municipal de Bauru, Sao Paulo, Brasil, destacando las implicaciones epidemiológicas en el cultivo de esas plantas. La mayoría de larvas encontradas fueron de mosquitos del género Culex, mientras que las de A. aegypti y A. albopictus fueron ocasionales. La utilización de telas para la protección de las plantas, su exposición al sol y la mayor cantidad de agua en el tanque pueden haber influenciado la ocurrencia y agrupamiento de larvas.

Volatile compounds profile of Bromeliaceae flowers / Perfil de compuestos volátiles de las flores en las Bromeliaceae

Abstract:Volatile compounds play a vital role in the life cycle of plants, possessing antimicrobial and anti-herbivore activities, and with a significant importance in the food, cosmetic, chemical, and pharmaceutical industry. This study aimed to identify the volatile compounds emitted by flowers of thirteen species belonging to four genera of Bromeliaceae, using headspace solid-phase micro-extraction and detection by gas chromatography-mass spectrometry. A total of 71 volatile compounds belonging to nine chemical groups were identified. The compounds identified represented more than 97 % of the major components in Aechmea bicolor, Ae. bromeliifolia, Ae. distichantha, Ae. fasciata, and Vriesea friburgensis. In the Ananas varieties, over 99 % of the components were identified, and around 90 % in V. simplex. V. friburgensis presented the largest diversity of volatiles with 31 compounds, while Alcantarea nahoumii presented only 14. All three Ananas varieties presented the same 28 compounds in relatively similar abundance, which has been confirmed by principal component analysis. Current taxonomy and pollination syndrome studies available can adequately explain the variation in volatile compounds among species. Rev. Biol. Trop. 64 (3): 1101-1116. Epub 2016 September 01.

ResumenLos compuestos volátiles tienen un papel vital en el ciclo de vida de las plantas. Poseen actividad antimicrobiana y anti-herbivoría biológica y una gran importancia en la industria de alimentos, cosméticos, perfumes, productos químicos y farmacéuticos. Este estudio tuvo como objetivo identificar los compuestos volátiles de trece flores de especies, pertenecientes a cuatro géneros de Bromeliaceae utilizando microextracción en fase sólida mediante cromatografía de gases hifenada con espacio de cabeza acoplada a espectrometría de masas. Se han identificado setenta y un diferentes compuestos volátiles pertenecientes a nueve grupos. Los compuestos identificados representaron más del 97 % de los componentes principales en Aechmea bicolor, Ae. bromeliifolia, Ae. distichantha, Ae. fasciata, Vriesea friburgensis, 99 % en las variedades de Ananas y 90 % en V. Simplex. V. friburgensis mostró la mayor diversidad de compuestos volátiles con 31, mientras que en Al. nahoumii se han encontrado sólo 14 compuestos. Las tres variedades de Ananas presentan los mismos 28 compuestos en cantidades relativamente similares, lo que se confirmó por el análisis de componentes principales. Estudios taxonómicos y síndromes de polinización disponibles podrían explicar la variación de los compuestos volátiles entre especies.

Induction of in vitro shoots of Billbergia euphemiae E. Morren (Bromeliaceae) from leaf explants / Indução in vitro de brotos de Billbergia euphemiae E. Morren (Bromeliaceae) a partir de explantes foliares

Bromeliads are an important group for the maintenance of the Atlantic Forest, with many threatened species due to exacerbated extraction and destruction of their natural habitats. Considering the need of developing protocols for the conservation of these species, the aim of this work was to evaluate the effect of different growth regulators in the in vitro induction of shoots of Billbergia euphemiae. Leaf explants were excised from seedlings derived from in vitro germination and grown on MS medium supplemented with NAA (0, 1 or 2 µM) and BA (0, 2, 4 or 6 µ M) combinations. The evaluation of the number of shoots per explant, shoot length, number of leaves per shoot and longest leaf length average was carried out after 30 and 60 days of culture. The best in vitro responses were observed in the presence of 1 µM NAA after 60 days of culture, which induced the best production of shoots per explant (16.39), as well as the highest rates of shoot length (1.08 cm), number of leaves per shoot (5.00) and the longest leaf length (0.56 cm). This work determined the best conditions for shoot production from leaf explants of B. euphemiae, being the first report on micropropagation of this species.

As bromélias constituem um importante grupo para a manutenção da Floresta Atlântica, com várias espécies ameaçadas de extinção pelo extrativismo exacerbado e a destruição dos habitats naturais. Considerando a necessidade do desenvolvimento de protocolos para a conservação destas espécies, este trabalho teve como objetivo avaliar o efeito de diferentes reguladores de crescimento na indução de brotos de Billbergia euphemiae. Explantes foliares foram excisados de plântulas derivadas da germinação in vitro e inoculados em meio MS suplementado com combinações de ANA (0, 1 ou 2 µM) e BA (0, 2, 4 ou 6 µM). O número de brotos por explantes, comprimento dos brotos, número médio de folhas por broto e comprimento médio da maior folha foram avaliados após 30 e 60 dias de cultura. As melhores respostas foram observadas na presença de ANA a 1 µM, após 60 dias de cultura, que induziu a maior produção de brotos por explante (16,39), assim como as maiores taxas de comprimento dos brotos (1,08 cm), número médio de folhas por broto (5,00) e comprimento da maior folha (0,56 cm). Este trabalho determinou as melhores condições para produção de brotos de B. euphemiae a partir de explantes foliares, representando o primeiro relato de micropropagação desta espécie.

Flagellate protist abundance in phytotelmata of Aechmea distichantha Lem. (Bromeliaceae) in the upper Paraná river basin / Abundância de protozoários flagelados em fitotelmata de Aechmea distichantha Lem. (Bromeliaceae) na bacia do alto rio Paraná

In some Bromeliaceae species the leaf sheaths form a tank or phytotelma, accumulating water and organic detritus, conditions that allow the establishment of different communities. Thus, this study evaluated the relationship between diameter and volume of water in the tank of the bromeliad Aechmea distichantha Lem. and density of flagellates found in the phytotelmata, as well as the influence of the hydrological periods and proximity of sampled strata on the phytotelma patterns of abundance distribution. Samplings were conducted in two hydrological periods in 2010. In each sampling, four plants were collected from three different strata, classified according to the distance from the river level. A one-way ANOVA identified significant differences in flagellate density between hydrological periods. The influence of the water volume in the phytotelmata on the flagellate density was evidenced by regression analyses, as well as the lack of relationship between the density and perimeter of the tank. Moreover, the distance of the strata from the river level did not influence the pattern of flagellate abundance. Thus, our results suggested that the hydrological period and the volume of water in the tanks of Aechmea distichantha are determinant on the abundance of heterotrophic flagellate in the phytotelmata.

Em algumas Bromeliaceae as bainhas foliares formam uma cisterna ou fitotelma, onde ocorre acúmulo de água e detritos orgânicos, que permitem o estabelecimento de diferentes comunidades. Assim, este estudo avaliou a relação entre o diâmetro e volume de água no tanque de Aechmea distichantha Lem. e a abundância de flagelados encontrados nestes fitotelmata, bem como a influência do período hidrológico e a proximidade dos estratos amostrados sobre os padrões de distribuição desse atributo. As coletas foram realizadas em dois períodos hidrológicos no ano de 2010. Foram coletadas quatro plantas em três estratos diferentes de acordo com a distância em relação ao rio. Foram identificadas diferenças significativas entre a densidade dos flagelados e os períodos hidrológicos. Além disso, verificou-se a influência do volume de água presente nos fitotelmata sobre a densidade dos flagelados, bem como a ausência de relação entre a densidade desses protistas e o perímetro do tanque das bromélias. A distância dos distintos estratos em relação ao rio não influenciou a abundância dos flagelados. Assim, os resultados sugerem que o período hidrológico bem como o volume de água presente nos tanques de Aechmea distichantha atuam como um fator determinante sobre a abundância da comunidade de flagelados heterotróficos nos fitotelmata.

Proteolytic enzymes from Bromelia antiacantha as tools for controlled tissue hydrolysis in entomology.

A crude extract with high proteolytic activity (78.1 EU/mL), prepared from ripe fruit of Bromelia antiacantha was used to hydrolyze and remove soft tissues from the epigyne of Apopyllus iheringi. This enzymatic extract presented four actives isoforms which have a broad substrate specificity action. Enzyme action on samples was optimized after evaluation under different conditions of pH, enzyme-substrate ratio and time (parameters selected based on previous studies) of treatment (pH 4.0, 6.0 and 8.0 at 42°C with different amount of enzyme). Scanning electron microscopy was used to evaluate conditions resulting in complete digestion of epigyne soft tissues. Optimal conditions for soft tissue removal were 15.6 total enzyme units, pH 6.0 for 18 h at 42°C.

Hypoglycemic effect of Bromelia plumieri (E. Morren) L.B. Sm., leaves in STZ-NA-induced diabetic rats.

This study confirms the hypoglycemic effects of two extracts obtained from the Bromelia plumieri (BP) plant in streptozotocin-nicotinamide-induced diabetic rats (STZ-NA). BP has been traditionally used in the municipality of Tlanchinol Hidalgo, Mexico, to treat type 2 diabetes. Two different BP extracts were prepared and tested. The first extract was a water extract (WE), similar to that traditionally used to make tea, and the second extract was an ethanol:water extract (EWE). The extracts (WE at 35 and 350 mg/kg, and EWE at 30 and 300 mg/kg) were tested in STZ-NA-induced diabetic rats to determine whether hypoglycemia occurred after oral administration of the extracts. Phytochemistry: Two different extracts were prepared, n-hexane and butanol, to determine the presence of alkaloids, terpenes and flavonoids. The extracts that were administered to the STZ-NA-induced diabetic rats produced a significant hypoglycemic effect as compared with the control group, similar to that achieved with glibenclamide. We also determined that flavonoids were the main components of BP leaves. The results presented here support the hypothesis that extracts obtained from this plant have hypoglycemic effects, which are in agreement with the traditional uses of this plant.

The choice of bromeliads as a microhabitat by Scinax argyreornatus (Anura, Hylidae) / A escolha de bromélias, como micro-habitat, por Scinax argyreornatus (Anura, Hylidae)

The association of anurans to bromeliads presents different degrees of interaction such as: eventual, obligatory and bromeligen. The frog species Scinax argyreornatus shows a regular association with these plants. The goal of this study is to characterise the degree of association between the frog S. argyreornatus to different species of bromeliads. We identified which species of bromeliad is regularly associated with S. argyreornatus and recognised which factors interfere with this association preference. We analysed the Concentration of Relative Dominance of frogs per bromeliad species. As possible criteria for frog association preference to different bromeliads species we established the analysis of leaves number, length and width, number of leaf axils, stored water in the axils, pH and relative humidity, among other organographic components. Our observations were analyzed by ANOVA followed by the Tukey test. We also evaluated the preference for association by the constancy of Bodenheimer. The correlation matrix indicated that the relative humidity is the factor responsible for the frog-bromeliads association, except for Aechmea sp.. However Aechmea sp. was the species with greater constancy of occupation followed by Quesnelia arvensis and Neoregelia johannis. According to our statistical results, Aechmea sp. and Q. arvensis are not different regarding organographic parameters, but differ from N. johannis. Our observations suggest that the bromeliads organographic structure and the relative humidity are key conditions which influence the preferences of S. argyreornatus to bromeliad species, while the other features showed no correlation.

A associação dos anuros às bromeliáceas pode apresentar diferentes graus de interação, como: bromelículas eventuais, bromelículas obrigatórios e bromelígenas. Scinax argyreornatus apresenta uma associação frequente a essas plantas. Propõe-se, neste estudo, qualificar esse grau de associação, buscando identificar qual espécie de bromélia é mais ocupada, reconhecendo os fatores que interferem nessa escolha de ocupação. Analisou-se a Concentração de Dominância Relativa dos anuros por espécie de bromélia, sendo considerados como prováveis critérios de escolha: número, largura e comprimento das folhas, número de axilas foliares, volume de água acumulado, pH e umidade relativa, entre outros componentes organográficos, que foram submetidos à ANOVA e, sempre que observadas diferenças significativas, ao teste de Tukey. Avaliou-se também a escolha de ocupação pela constância de Bodenheimer. A matriz de correlação apontou a UR% como o fator responsável pela ocorrência do animal nas bromélias, exceto para Aechmea sp.; entretanto, esta foi a planta com maior Constância de ocupação, seguida por Quesnelia arvensis e Neoregelia johannis. Segundo a ANOVA, Aechmea sp. e Q. arvensis não apresentam diferenças entre si do ponto de vista organográfico, mas diferem de N. johannis. Foi possível inferir que a estrutura organográfica da planta e a UR% são as condições que interferem na escolha de ocupação das bromélias por S. argyreornatus, enquanto as outras características não demonstraram nenhuma correlação.

Avaliação das atividades antimicrobiana, citotóxica, moluscicida e antioxidante de Bromelia antiacantha Bertol: (Bromeliaceae) / Evaluation of antimicrobial, cytotoxic, molluscicidal and antioxidant activities of Bromelia antiacantha Bertol: (Bromeliaceae)

A Bromelia antiacantha Bertol. (Bromeliaceae), conhecida como gravatá ou caraguatá, apresenta características medicinais, alimentícias e ornamentais. Neste trabalho foram avaliadas as atividades antifúngica, antibacteriana, citotóxica, moluscicida e antioxidante de extratos alcoólicos das folhas e frutos desta espécie. Na avaliação da atividade antimicrobiana pelo método de microdiluição os extratos alcoólicos não foram eficientes frente à cepa referência do fungo Candida albicans (ATCC 90028), cepas clínicas de C. albicans e C. glabrata, e frente às bactérias S. aureus (ATCC 6538), P. aeruginosa (ATCC 9027) e E. coli (ATCC 8739). A atividade citotóxica, avaliada pelo bioensaio de toxicidade sobre náuplios de Artemia salina Leach (TAS), foi positiva para ambos extratos, com valores de DL50 variando em torno de 620 µg mL-1 e 360 µg mL-1, para os extratos alcoólicos dos frutos e das folhas, respectivamente. A atividade moluscicida, avaliada frente a caramujos da espécie Biomphalaria glabrata (Molusca, Gastropoda), não mostrou toxicidade aguda para soluções de 400 µg mL-1. As propriedades antioxidantes dos extratos investigadas in vitro mediante o método de DPPH• foram consideradas modestas (~ 35 por cento de inibição na concentração de 1000 µg mL-1). Dentre as atividades biológicas que foram testadas a atividade citotóxica foi a mais pronunciada.

Bromelia antiacantha Bertol. (Bromeliaceae), popularly known as "gravatá" or "caraguatá", shows medicinal, nutritional and ornamental properties. In the present study, the alcoholic extracts of leaves and fruits of this species were evaluated for antifungal, antibacterial, cytotoxic, molluscicidal and antioxidant activities. The alcoholic extracts evaluated by microdilution method were not efficient against the standard strain of Candida albicans (ATCC 90028), clinical strains of C. albicans and C. glabrata, and S. aureus (ATCC 6538), P. aeruginosa (ATCC 9027) and E. coli (ATCC 8739). The cytotoxic activity evaluated by toxicity bioassay on nauplii of Artemia salina Leach (TAS) was positive for both extracts, with LD50 values ranging around 620 µg mL-1 and 360 µg mL-1 for alcoholic extracts from leaves and fruits, respectively. The molluscicidal activity evaluated against Biomphalaria glabrata (Mollusca, Gastropoda) did not present acute toxicity for 400 µg mL-1 solutions. The antioxidant properties of the extracts investigated in vitro using the DPPH• method were considered modest (~ 35 percent of inhibition at 1000 µg mL-1). Among the evaluated biological activities, cytotoxic activity was the most marked.

Características germinativas de sementes de Ananas ananassoides (Baker) L. B. Sm. (Bromeliaceae) / Germination characteristics of seeds of Ananas ananassoides (Baker) L. B. Sm. (Bromeliaceae)

Ananas ananassoides é uma planta perene com hábito de crescimento herbáceo e que tem enorme potencial paisagístico porque apresenta flores e brácteas com coloração exuberante e folhas com margens espinescentes que estão distribuídas em roseta. O objetivo deste experimento foi o de avaliar as características germinativas e o efeito de tratamentos pré-germinativos nas sementes da espécie. Primeiro, em um delineamento inteiramentecasualizado, foram investigados seis tratamentos em quatro repetições de 25 sementes. Os tratamentos constaram da imersão em água destilada à temperatura ambiente (25 ± 1,5°C) durante 24, 48 e 72h, imersão em água quente a 90°C durante 2 min. e imersão em ácido sulfúrico p.a. durante 2 min. e com posterior lavagem em água corrente por 24h que foram comparados com um controle. Depois de quatro meses de armazenamento, sementes intactas (controle) foram comparadas com sementes imersas em água a 90°C por 2 min. Assim, a maior porcentagem de germinação (92%) foi obtida com sementes recém-colhidas e imersas em água a 90oC durante 2 min., mas o armazenamento por quatro meses melhorou a porcentagem de germinação (G ≥ 96,0%). Os tempos médios de germinação não diferiram entre as sementes recém-colhidas (de 21,0 a 26,8 dias) e as armazenadas (entre 18,4 e 19,5 dias). A germinação das sementes recém-colhidas e armazenadas foi lenta e distribuída ao longo do tempo experimental.

Ananas ananassoides is a perennial plant with ornamental potential because of flowers and bracts with exuberant colors and herbaceous development where the leaves have spiny margins that are distributed in rosettes. The objective of this experiment with six completely randomized treatments and four replicates of 25 seeds was to evaluate the effects of seed soaking into distilled water at environmental temperature (25 ± 1.5°C) for 24, 48, and 72h; seed soaking into water at 90°C for 2 minutes; seed soaking into sulfuric acid p.a. for 2 minutes followed by tap water washing for 24h, and the control. Thereafter, 4-month-old seeds were immersed into water at 90°C for 2 minutes and compared to the intact seeds (control). Initially, the highest percentage of germination (G=92%) was evaluated in newly collected seeds that were immersed in water at 90°C for 2 minutes. No increases in the percentage of seed germination were detected by seed soakinginto water or sulfuric acid in comparison to the seeds that were immersed in water at 90°C for 2 minutes. However, the percentage of germination was higher than 96% when the seeds were stored for four months. No difference in the mean time to germination was detected between newly collected (between 21.0 and 26.8 days) and stored seeds (between 18.4 and 19.5 days). The germination of newly collected and stored seeds was slow and distributed throughout the experimental time