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.

Bromélias (Bromeliaceae) como criadouros de mosquitos (Diptera: Culicidae) em condomínios residenciais no município de Paulínia/SP, Brasil / Bromeliads (Bromeliaceae) as breeding sites for mosquitoes (Diptera: Culicidae) in gated community in the municipality of Paulínia/SP, Brazil

Embora as bromélias sejam identificadas como os principais criadouros de algumas espécies de culicídeos dos gêneros Anopheles, Culex, Wyomyia e Runchomyia em áreas preservadas ou com pouca interferência humana, ainda não há consenso sobre sua relevância como habitat para mosquitos das espécies Aedes aegypti e Aedes albopictus em áreas urbanas. Isso representa um desafio para a vigilância entomológica e para o sucesso das ações que visam orientar a população sobre as medidas de prevenção e controle vetorial nessas plantas. Foi realizada a caracterização a fauna de culicídeos presentes nesses reservatórios e a caracterização de aspectos morfológicos das bromélias domesticadas presentes em condomínios residenciais. A pesquisa ocorreu entre os meses de outubro de 2021 e julho de 2022, foram inspecionadas 660 bromélias domesticadas presentes em condomínios residenciais situados em área urbana do município de Paulínia/SP. Os imaturos de culicídeos foram coletados e identificados através de chaves dicotômicas. Os caracteres morfológicos altura, diâmetro, líquido coletado e capacidade de armazenar líquido das bromélias foram coletados e analisados. O teste de Mann-Whitney e o Coeficiente de Spearman resultaram em p<0,001 para todas as variáveis, quando avaliada a presença ou ausência de imaturos. Este último apresentou correlação positiva moderada com a quantidade de imaturos coletados. A frequência de bromélias positivas para imaturos de culicídeos foi de 47,12% entre as plantas que continham líquido armazenado. Foram coletados 6.721 imaturos, destes, 81,80% pertencem à espécie Aedes aegypti e 5,19% à espécie Aedes albopictus. Em áreas urbanizadas, onde há uma concentração maior de bromélias em condomínios residenciais, a proliferação desses vetores pode ser ainda mais preocupante devido à frequência de regas e à redução dos predadores e competidores da espécie. As bromélias do gênero Alcantarea apresentaram maior frequência quanto à presença de culicídeos (77,4% de positividade). Portanto, é fundamental o monitoramento desses criadouros nesse cenário. Essa análise pode fornecer informações valiosas sobre a extensão do problema e auxiliar na implementação de estratégias de controle mais eficazes. A compreensão da distribuição e da densidade das bromélias em diferentes regiões urbanizadas é essencial para direcionar ações de prevenção e controle de doenças transmitidas por culicídeos, tais como a dengue, zika e chikungunya.

Although bromeliads are identified as the main breeding sites for some species of culicids in the genera Anopheles, Culex, Wyomyia, and Runchomyia in preserved, there is still no consensus on their relevance as habitats for mosquitoes of the species Aedes aegypti and Aedes albopictus in urban areas. This represents a challenge for entomological surveillance and the success of actions aimed at guiding the population on prevention and vector control measures in these plants. The fauna of culicids present in these reservoirs and the morphological aspects of domesticated bromeliads in gated community were characterized. Between October 2021 and July 2022, 660 domesticated bromeliads present in gated community located in the urban area of Paulínia, São Paulo, were inspected. Culicid immatures were collected and identified using dichotomous keys. The morphological characteristics of height, diameter, collected liquid, and liquid storage capacity of the bromeliads were collected and analyzed. The Mann-Whitney test and Spearman's coefficient resulted in p<0.001 for all variables when evaluating the presence or absence of immatures. The latter showed a moderate positive correlation with the quantity of immatures collected. The frequency of bromeliads positive for culicid immatures was 47.12% among the plants that contained stored liquid. A total of 6,721 immatures were collected, of which 81.80% belonged to the species Aedes aegypti and 5.19% to the species Aedes albopictus. In urbanized areas, where there is a higher concentration of bromeliads in gated community, the proliferation of these vectors can be even more concerning due to the frequency of watering and the reduction of predators and competitors of the species. Bromeliads of the genus Alcantarea showed a higher frequency of culicid presence (77.4%). Therefore, monitoring these breeding sites in this scenario is fundamental. This analysis can provide valuable information about the extent of the problem and assist in the implementation of more effective control strategies. Understanding the distribution and density of bromeliads in different urbanized regions is essential to guide actions for the prevention and control of diseases transmitted by culicids, such as dengue, Zika, and chikungunya.

Proteolytic characterization of a novel enzymatic extract from Bromelia serra leaves.

Bromelia serra leaves collected from Corrientes, Argentina, were assessed to analyze and characterize the proteolytic system and to evaluate its potential use as an industrial catalyst. The specific activity of the enzymatic extract (EE), which was prepared using acetone as a precipitating agent of the crude extract (CE), increased 2-3 folds with different substrates (hemoglobin, azocasein and casein). The proteins present in the EE have isoelectric points between 4.55-8.15 and they were significant inhibited by pepstatin A (50%) and E-64 (15%). Proteolytic activity in EE presented high activity in acidic pH (2.7-4), and low activity in neutral alkaline pH (6-11.75). The EE optimum activity was reached at 60ºC, and referring to the thermal stability, it retained over 97% of the proteolytic activity after incubation at a temperature range of 37‒60 ºC for 60 min. The effect of reducing agents and ionic strength were also measured, and it showed that the EE had its maximum activity with 5mM of cysteine, and it was inactivated with 2.5 M of NaCl. The chromatography procedures presented two purified enzymes of 21 and 54 KDa with proteolytic activity. The characteristics of the EE suggest that it is a potential candidate as an industrial catalyst.

Phytochemical profile and biological activities of Bromelia antiacantha extracts.

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.

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.

Protein extract of Bromelia karatas L. rich in cysteine proteases (ananain- and bromelain-like) has antibacterial activity against foodborne pathogens Listeria monocytogenes and Salmonella Typhimurium.

Bromelia karatas L. is a plant species from the Americas. The presence of proteases in fruits of B. karatas has been reported but scarcely studied in detail. Proteolytic enzymes from Ananas comosus have displayed antifungal and antibacterial activity. Thus, novel proteases present in B. karatas may be useful as a source of compounds against microorganisms in medicine and food production. In this work, the protein extract from the fruits of B. karatas was characterized and its antibacterial activity against Salmonella Typhimurium and Listeria monocytogenes was determined for the first time. Proteins highly similar to ananain and the fruit bromelain from A. comosus were identified as the main proteases in B. karatas fruits using liquid chromatography with tandem mass spectrometry (LC-MS/MS). The soluble protein extract (SPE) at a concentration of 2.0 mg/mL displayed up to 80% of antibacterial activity against S. Typhimurium. Complete inhibition of L. monocytogenes was reached with up to 1.65 mg/mL of SPE. Plant protease extract containing ananain-like enzyme inhibited up to 90% against S. Typhimurium and up to 85% against L. monocytogenes using only 10 µg/mL of the partial-purified enzyme.

Seed mass maturity in the terrestrial bromeliad Hechtia perotensis (Bromeliaceae), endemic to Mexico / Madurez de las semillas en la bromelia terrestre Hechtia perotensis (Bromeliaceae), endémica de México

Abstract Introduction: H. perotensis is a plant with a high potential for ecological restoration because it yields thousands of seeds and grows under low levels of rain, poor soils and contrasting temperatures. However, little is known of the seed mass maturity (high seed germination, low seed fresh weight and low seed moisture content) in this species. Objective: Assess seed germination in the laboratory of H. perotensis during seed development and along the floral stalk (infructescence) in two sites one in rocky location and another near a lake. The hypothesis was that there is a time after flowering in which seeds have highest germination and fresh weight and that the apical, centre and base of the infructescence are different in seed germination and fresh weight in both sites. Methods: Capsules were collected in two sites one in rocky land (Frijol Colorado, Perote, Veracruz) and another near one lake (Alchichica, Puebla), in the months of August, September and November 2016 and January 2017. A repeated measure design (RMD) was used to analyze the effects of infructescence section on seed weight, moisture content and seed germination (41, 87, 152 and 215 days after flowering). Each evaluation time comprised five replicates, each one with 15 seeds. Results: Difference in seed germination, seed weight and moisture content between sections of the infructescence was not significant. However, significant differences were found not only between first and last sample dates, but mainly between first and second dates. Eighty-seven days after flower pollination seed moisture content was lower than 20 % and up to 80 % of seed germinated in both sites of sampling. Conclusions: In this study it was found that the moisture content of H. perotensis seed can be used as an indicator of the physiological maturity of the seed and it is also related to germination of the seed.

Resumen Introducción: H. perotensis es una planta con un alto potencial para la restauración ecológica debido a que produce miles de semillas y crece con niveles bajos de lluvia, suelos pobres y temperaturas contrastantes. Sin embargo, el desarrollo de la madurez de las semillas (germinación alta, baja humedad y bajo peso fresco de semilla) en esta especie es muy poco conocido. Objetivo: Evaluar la germinación en el laboratorio de las semillas de H. perotensis, durante el desarrollo de la semilla y a lo largo del tallo floral (infrutescencia) en dos sitios; uno en un área rocosa y otro cerca de un lago. La hipótesis fue que hay un tiempo después de la floración, en el que las semillas presentan la mayor germinación y menor peso fresco, y que las secciones apical, central y base de la infrutescencia, la germinación y el peso fresco de la semilla son diferentes en los dos sitios. Métodos: se recolectaron las cápsulas en dos sitios uno en un área rocosa (Frijol Colorado, Perote, Veracruz) y otro cerca de un lago (Alchichica, Puebla), en los meses de Agosto, Septiembre y Noviembre de 2016 y Enero de 2017. Se usó un diseño de medidas repetidas (DMR) para analizar los efectos de la sección de la infrutescencia sobre el peso fresco de la semilla, el contenido de humedad y la germinación de la semilla (41, 87, 152 y 215 días, después de la floración). Cada tiempo de evaluación comprendió cinco réplicas y 15 semillas. Resultados: La diferencia en la germinación de la semilla, el peso de la semilla y el contenido de humedad entre las secciones de la infrutescencia no fue significativa. Sin embargo, se encontraron diferencias significativas entre las fechas de muestreo inicial y final, pero sobre todo entre la primera y la segunda fecha. Asimismo, ochenta y siete días después de la polinización de las flores, el contenido de humedad de las semillas fue inferior al 20 % y superior al 80 % en la germinación de las semillas en ambos sitios de muestreo. Conclusiones: en este estudio se encontró que el contenido de humedad de la semilla de H. perotensis, puede usarse como un indicador de la madurez fisiológica de la semilla y también está relacionado con la germinación de la semilla.

Go with the flow: The extent of drag reduction as epiphytic bromeliads reorient in wind.

Vascular epiphytes represent almost 10% of all terrestrial plant diversity. Being structurally dependent on trees, epiphytes live at the interface of vegetation and atmosphere, making them susceptible to atmospheric changes. Despite the extensive research on vascular epiphytes, little is known about wind disturbance on these plants. Therefore, this study investigated the wind-epiphyte mechanical interactions by quantifying the drag forces on epiphytic bromeliads when subjected to increasing wind speeds (5-22 m s-1) in a wind tunnel. Drag coefficients (Cd) and Vogel exponents (B) were calculated to quantify the streamlining ability of different bromeliad species. Bromeliads' reconfiguration occurred first via bending and aligning leaves in the flow direction. Then leaves clustered and reduced the overall plant frontal area. This reconfiguration caused drag forces to increase at a slower rate as wind velocity increased. In the extreme case, drag force was reduced by 50% in a large Guzmania monostachia individual at a wind velocity of 22 m s-1, compared to a stiff model. This species had one of the smallest Cd (0.58) at the highest wind velocity, and the largest negative mean B (-0.98), representing the largest reconfiguration capacity amongst the tested bromeliads. The streamlining ability of bromeliads was mainly restricted by the rigidity of the lower part of the plant where the leaves are already densely clustered. Wind speeds used in this study were generally low as compared to storm force winds. At these low wind speeds, reconfiguration was an effective mechanism for drag reduction in bromeliads. This mechanism is likely to lose its effectiveness at higher wind speeds when continuous vigorous fluttering results in leaf damage and aspects such as root-attachment strength and substrate stability become more relevant. This study is a first step towards an understanding of the mechanical bottleneck in the epiphyte-tree-system under wind stress.

Diet of a bromeligenous species of treefrog in the Brazilian semiarid / Dieta de uma espécie bromelígena de anuro do semiárido brasileiro

Phyllodytes tuberculosus is a bromeligenous anuran species whose geographic distribution is restricted to areas of the Caatinga and to transition zones between Caatinga and Atlantic Forest in the central-south of the state of Bahia. Its existence in close relationship with bromeliads and its occurrence in dry and highly seasonal environments drove us to the supposition that species is opportunistic/generalist regard to diet. We collected the items present in the stomach of 43 P. tuberculosus individuals with stomach-flushing method. Of these, 18 stomach contents were obtained during the dry period (April­October), and 25 during the rainy period (November­March). We recorded nine classes of prey, finding the greatest relative importance for the categories Formicidae and Coleoptera in dry season and Blattodea (Termites), Formicidae and Coleoptera in the rainy season. Phyllodytes tuberculosus presented a varied, possibly opportunistic diet in semiarid environments. Additionally, we report an apparent temporal differentiation related to the typical Caatinga seasonality. This study adds to the data on the natural history of an anuran species adapted to a semiarid environment with a narrow geographic distribution.

Phyllodytes tuberculosus é uma espécie de anuro bromelígena cuja distribuição geográfica está restrita a áreas de Caatinga e zonas de transição entre Caatinga e Mata Atlântica no centro-sul do estado da Bahia. Sua relação próxima com bromélias e sua ocorrência em ambientes secos e fortemente sazonais nos levou a supor que a espécie apresenta uma dieta generalista/oportunista. Por meio de lavagem estomacal, coletamos os itens alimentares presentes no estômago de 43 indivíduos de P. tuberculosus. Destes, 18 estômagos foram obtidos durante a estação seca (Abril-Outubro) e 25 durante a estação chuvosa (Novembro­Março). Nós registramos nove classes de presas, sendo que as maiores importâncias relativas foram observadas para os itens Formicidae e Coleoptera na estação seca e Blattodea (Termites), Formicidae e Coleoptera no conteúdo recuperado na estação chuvosa. Phyllodytes tuberculosus apresentou uma dieta variada, possivelmente oportunista, condizente com a vida em ambientes semiáridos. Ademais, também observamos uma aparente diferenciação temporal relacionada a sazonalidade típica da Caatinga. Este estudo adiciona dados a respeito da história natural de uma espécie de anuro adaptada a ambientes semiáridos e com distribuição geográfica restrita.

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.

Extreme rainfall events alter the trophic structure in bromeliad tanks across the Neotropics.

Changes in global and regional precipitation regimes are among the most pervasive components of climate change. Intensification of rainfall cycles, ranging from frequent downpours to severe droughts, could cause widespread, but largely unknown, alterations to trophic structure and ecosystem function. We conducted multi-site coordinated experiments to show how variation in the quantity and evenness of rainfall modulates trophic structure in 210 natural freshwater microcosms (tank bromeliads) across Central and South America (18°N to 29°S). The biomass of smaller organisms (detritivores) was higher under more stable hydrological conditions. Conversely, the biomass of predators was highest when rainfall was uneven, resulting in top-heavy biomass pyramids. These results illustrate how extremes of precipitation, resulting in localized droughts or flooding, can erode the base of freshwater food webs, with negative implications for the stability of trophic dynamics.

Transcriptional foliar profile of the C3-CAM bromeliad Guzmania monostachia.

Guzmania monostachia is an epiphytic tank bromeliad that displays the inducible CAM photosynthesis under stressful conditions and had the highest stomata density in the leaf apex, while the base portion has the highest density of trichomes, which are specialized structures used to acquire water and nutrients from the tank solution. In order to correlate the genetic factors behind these morpho-physiological characteristics along the leaf blade of G. monostachia, a comparative transcriptome analysis was performed to identify the functional enriched pathways and unigenes that could play a role in the apical, middle and basal leaf portions. A total of 653 million reads were used for de novo transcriptome assembly, resulting in 48,051 annotated unigenes. Analysis of differentially expressed genes (DEGs) among distinct leaf regions revealed that 806 DEGs were upregulated in the apex compared to the middle portion, while 9685 DEGs were upregulated in the apex and 9784 DEGs were upregulated in the middle portions compared to the base. Our outcomes correlated some DEGs and identified unigenes with their physiological functions, mainly suggesting that the leaf apex was related to the regulation of stomatal movement, production of chlorophyll, cellular response to stress, and H2O2 catabolic process. In contrast, the middle portion showed DEGs associated with the transport of amino acids. Furthermore, DEGs from the leaf base were mainly correlated with responses to nutrients and nitrogen compounds, regulation of potassium ion import, response to water deprivation, and trichome branching, indicating that, at least in part, this leaf portion can replace some of the root functions of terrestrial plants. Therefore, possibly candidate unigenes and enriched pathways presented here could be prospected in future experimental work, opening new possibilities to bioengineer non-inducible CAM plants and/or improve the fertilization use efficiency by increasing leaf nutrient acquisition of crop plants.

Reproductive site selection in a bromeliad breeding treefrog suggests complex evolutionary trade-offs.

Reproductive site selection is a key determinant of fitness in many taxa. However, if the site characteristics that enhance offspring survival are detrimental to the parent's survival or mating success, then complex evolutionary trade-offs occur. In the Brazilian Atlantic Forest, males of the treefrog species Aparasphenodon arapapa use the temporary water bodies in forest-floor bromeliads to court and mate. Males fit tightly into the plant with the head blocking the access and after mating, stay in the bromeliad with the offspring. Since evaporation of the temporary water body inside the bromeliad results in reproductive failure, we expected that males would simply choose the largest bromeliad tanks with the most water. We found that although this was generally true, males seemed to avoid both very large bromeliads and very high water volumes. Field observations suggested a trade-off mechanism for this pattern, whereby very large and water-filled tanks would reduce the male's ability to effectively seal the tank entrance, avoid predation, or call to mating females. Males also avoided bromeliads with leaf litter and preferred slightly inclined plants. Our results indicate that during reproductive site selection, this bromeliad-breeder needs to engage in complex trade-offs between selection pressures, balancing water requirements against the need for defense and potentially, the ability to attract a mate.

Bromeliad targeted foliar treatments with Aqualuer® 20-20 for hot spot mitigation using three hand sprayers against Aedes albopictus (Diptera: Culicidae).

Bromeliads serve as a key environmental resource for Aedes albopictus (Skuse) in some peridomestic landscapes and are not addressed through conventional source reduction. As an alternative, three handheld sprayers were evaluated in point-source applications of permethrin to bromeliads in Saint Augustine, FL. Each machine was screened for its utility in point-source harborage treatments, while we simultaneously evaluated bromeliad-focused treatments for reducing Ae. albopictus egg abundance in bromeliad plants. Semi-field tests against caged Ae. albopictus produced 100% mortality at 24 h with each sprayer. Three field sites were assigned to each machine and a control, and bromeliad residual applications of Aqualuer 20-20® were assessed through egg surveillance using ovitraps both within and beyond 30 m of the treated bromeliad harborage. The treatment decreased Ae. albopictus egg abundance across the field sites for three weeks post-treatment with no significant differences among the equipment. Functional differences among machines, such as automated pressure control or leak guards on the nozzle, depend on the accessories that offer the greatest logistical benefit to the user. The field results suggest bromeliads are a viable target for harborage treatments. Selective treatments to bromeliads could offer a possible solution to containers that homeowners refuse to eliminate.

Insights into the evolutionary dynamics of Neotropical biomes from the phylogeography and paleodistribution modeling of Bromelia balansae.

PREMISE OF THE STUDY: Historical abiotic and biotic factors have strongly affected species diversification and speciation. Although pre-Pleistocene events have been linked to the divergence of several Neotropical organisms, studies have highlighted a more prominent role of Pleistocene climatic oscillations in shaping current patterns of genetic variation of plants. METHODS: We performed phylogeographic analyses based on plastidial markers and modeled the current distribution and paleodistribution of Bromelia balansae (Bromeliaceae), an herbaceous species with a wide geographical distribution in South America, to infer the processes underlying its evolutionary history. KEY RESULTS: Combined molecular and paleodistributional modeling analyses indicated retraction during the Last Glacial Maximum followed by interglacial expansion. Populations occurring in the semideciduous Atlantic Forest and the Cerrado formed two distinct genetic clusters, which have been historically or ecologically isolated since late Pliocene to early Pleistocene. Populations located in the transition zone had higher levels of genetic diversity, as expected by the long-term climatic stability in the region detected in our ecological niche models. CONCLUSIONS: Our study adds important information on how herbaceous species have been affected by past climate in Central and Southeast Brazil, helping to disentangle the complex processes that have triggered the evolution of Neotropical biota.

Ciliate species from tank-less bromeliads in a dry tropical forest and their geographical distribution in the Neotropics.

The study of ciliate diversity in tropical environments remains scarce. In Neotropical forests, bromeliads are a common component of the vascular flora; bromeliads with tank morphology intercept rain water and detritus, resulting in the formation of a phytotelm, where heterotrophic protist communities like ciliates can establish. However, it is not known if ciliates inhabit tank-less bromeliads. For this reason, the goal of the present study was to investigate if ciliates can establish between the leaf axils of five terrestrial and epiphytic tank-less bromeliad species in a dry tropical forest in west Mexico. We collected samples of rain water and detritus from the leaf axils during the humid season of years 2015 and 2016. For ciliate taxonomical identification, we used optical microscopy, in vivo observation, and silver impregnation techniques. To summarize information about geographical distribution of ciliates identified at species level in the Neotropics, we provided their records from previous works, at country level with locality and georeferenciation. We recorded 27 taxa of ciliates, where the class Oligohymenophorea contained the largest richness of taxa. Drepanomonas revoluta, Leptopharynx bromeliophilus, and Tetrahymena sp. were recorded from all the species of bromeliads. Bromelia karatas was the species that hosted the largest number of ciliate taxa (22). Our results indicated that Glaucomides bromelicola, Gonostomum bromelicola, Leptopharynx bromelicola and L. bromeliophilus, species which are considered endemic to tank bromeliads, can also inhabit tank-less bromeliads. We provided previous records of 19 ciliate species from eleven countries within the Neotropical region, and Bromeliothrix metopoides was the species most frequently recorded in Neotropical countries (9). Therefore, tank-less bromeliads can constitute a temporal habitat for ciliates, and function as cysts reservoirs in environments with a pronounced seasonality like dry tropical forests.

Characterization of nitrogen deposition in a megalopolis by means of atmospheric biomonitors.

An increase of nitrogen deposition resulting from human activities is not only a major threat for global biodiversity, but also for human health, especially in highly populated regions. It is thus important and in some instances legally mandated to monitor reactive nitrogen species in the atmosphere. The utilization of widely distributed biological species suitable for biomonitoring may be a good alternative. We assessed the suitability of an ensemble of atmospheric biomonitors of nitrogen deposition by means of an extensive sampling of a lichen, two mosses, and a bromeliad throughout the Valley of Mexico, whose population reaches 30 million, and subsequent measurements of nitrogen metabolism parameters. In all cases we found significant responses of nitrogen content, C:N ratio and the δ15N to season and site. In turn, the δ15N for the mosses responded linearly to the wet deposition. Also, the nitrogen content (R2 = 0.7), the C:N ratio (R2 = 0.6), and δ15N (R2 = 0.5) for the bromeliad had a linear response to NOx. However, the bromeliad was not found in sites with NOx concentrations exceeding 80 ppb, apparently of as a consequence of excess nitrogen. These biomonitors can be utilized in tandem to determine the status of atmospheric nitrogenous pollution in regions without monitoring networks for avoiding health problems for ecosystems and humans.

Ornamental bromeliads of Miami-Dade County, Florida are important breeding sites for Aedes aegypti (Diptera: Culicidae).

BACKGROUND: A major public health concern is the emergence and geographical spread of vector-borne diseases such as Zika and yellow fever. Ornamental bromeliads retaining water in their leaf axils represent potential breeding sites for mosquitoes. As the role of ornamental bromeliads in breeding Aedes aegypti in Miami-Dade County, Florida is unknown, we hypothesize that ornamental bromeliads are important breeding sites for Ae. aegypti. Our objective was to survey bromeliads in areas with high densities of adult Ae. aegypti, including those with 2016 local transmission of Zika virus. METHODS: Ornamental bromeliads were surveyed for the presence of immature mosquitoes at 51 locations of Miami-Dade County, Florida. Bromeliads were sampled for the presence of immature stages of mosquitoes, their reservoirs were drained and screened for the presence of immature mosquitoes. Immature mosquitoes were stored in plastic containers and preserved in 70% ethanol until morphological identification. Biodiversity of species assemblages was assessed by Shannon's and Simpson's indices, and individual rarefaction curves and plots of cumulative abundance, Shannon's index and evenness profiles. RESULTS: Ornamental bromeliads were present in all surveyed areas, yielding a total of 765 immature mosquitoes, comprising five taxonomic units: Ae. aegypti, Wyeomyia mitchellii, Wyeomyia vanduzeei, Culex quinquefasciatus and Culex biscaynensis. The biodiversity indices point to a low diversity scenario with a highly dominant species, Ae. aegypti. DISCUSSION: Our findings suggest that ornamental bromeliads are contributing for the proliferation of Ae. aegypti in the County of Miami-Dade, which may indicate a shift in the paradigm of usage of bromeliads as breeding sites, highlighting that ornamental phytotelmata bromeliads are to be considered in future vector-control strategies to control Zika and other arboviruses.

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.

Influence of N-P-K fertilization at the acclimatization stage on micropropagated seedlings of Tillandsia bulbosa Hook / Influência da adubação com N-P-K na fase de aclimatização em mudas micropropagadas de Tillandsia bulbosa Hook

Bromeliads are known worldwide for their ornamental potential. In Brazil, species of the genus Tillandsia occur in the Atlantic rainforest, Amazon rainforest, and rocky fields. This work aimed to evaluate the influence of nitrogen, phosphorus, and potassium fertilization on micropropagated seedlings of Tillandsia bulbosa, at the acclimatization stage, and their leaf anatomy. The experiment was carried out in a completely randomized design, in a 4x4+1 factorial scheme, using coconut coir: earthworm humus: sand mixture as substrate (2:1:1). Urea, single superphosphate, and potassium chloride were used as nitrogen, phosphorus and potassium sources, respectively, at proportions of 50, 100, 200, and 400% of the dose recommended. The doses were distributed in four applications, testing total application at planting (1); ½ application at planting and ½ at 80 DAP (2); application at planting, at 50 DAP, and at 100 DAP (3); and » application at planting, » at 30 DAP, » at 60 DAP, and » at 120 DAP (4); and a control (without fertilization). Leaves anatomy was analyzed at 180 days after planting. Fertilization did not significantly influence the development of seedlings during acclimatization. The doses of 50, 100, and 200% provided thicker parenchyma of chlorophyll and aquifer and leaf blade. Tillandsia bulbosa can be acclimatized without fertilizer application.

As bromélias são conhecidas mundialmente por seu potencial ornamental. No Brasil, as espécies de Tillandsia podem ser encontradas na Mata Atlântica, Floresta Amazônica e campos rupestres. O objetivo deste trabalho foi verificar a influência de adubação com nitrogênio, fósforo e potássio em plântulas micropropagadas de Tillandsia bulbosa, na fase de aclimatização e na sua anatomia foliar. O experimento foi implantado em delineamento inteiramente casualizado, em esquema fatorial 4x4+1, utilizando como substrato a mistura de pó de coco: húmus de minhoca: areia (2:1:1). Ureia, super fosfato simples e cloreto de potássio foram utilizados como fontes de nitrogênio, fósforo e potássio, nas proporções de 50, 100, 200 e 400% da dose recomendada. As doses foram distribuídas em quatro aplicações, testando aplicação total no plantio (1); ½ no plantio e ½ aos 80 DAP (2); no plantio, aos 50 DAP e aos 100 DAP (3) e » no plantio, » aos 30 DAP, » aos 60 DAP e » aos 120 DAP (4), e uma testemunha (sem adubação). Foi realizado o estudo anatômico das folhas aos 180 dias. A adubação não influenciou significativamente o desenvolvimento das plântulas durante a aclimatização. As doses de 50, 100 e 200% proporcionaram maior espessura de parênquimas clorofiliano e aquífero e do limbo foliar. A aclimatização de Tillandsia bulbosa pode ser realizada sem a necessidade de adubação.