The multiple roles of trichomes in two Croton species.

Trichomes are common in plants from dry environments, and despite their recognized role in protection and defense, little is known about their role as absorptive structures and in other aspects of leaf ecophysiology. We combine anatomical and ecophysiological data to evaluate how trichomes affect leaf gas exchange and water balance during drought. We studied two congeneric species with pubescent leaves which co-occur in Brazilian Caatinga: Croton blanchetianus (dense trichomes) and Croton adenocalyx (sparse trichomes). We found a novel foliar water uptake (FWU) pathway in C. blanchetianus composed of stellate trichomes and underlying epidermal cells and sclereids that interconnect the trichomes from both leaf surfaces. The water absorbed by these trichomes is redistributed laterally by pectin protuberances on mesophyll cell walls. This mechanism enables C. blanchetianus leaves to absorb water more efficiently than C. adenocalyx. Consequently, the exposure of C. blanchetianus to dew during drought improved its leaf gas exchange and water status more than C. adenocalyx. C. blanchetianus trichomes also increase their leaf capacity to reflect light and maintain lower temperatures during drought. Our results emphasize the multiple roles that trichomes might have on plant functioning and the importance of FWU for the ecophysiology of Caatinga plants during drought.

Antagonism of volatile organic compounds of the Bacillus sp. against Fusarium kalimantanense.

Fusarium kalimantanense is a genetic lineage of Fusarium oxysporum f. sp. cubense (Foc) and belongs to the Fusarium oxysporum species complex (FOSC). This pathogen is a causative agent of Panama disease, an infection that has caused damage to the banana crop worldwide. Bacillus sp. (LPPC170) showed preliminary antagonist activity against F. kalimantanense (LPPC130) in vitro tests from the cultivation of axenic culture and co-culture with inhibition of mycelial growth of phytopathogen of 41.23%. According to these findings, volatile organic compounds (VOCs) emitted from Bacillus sp. were obtained by solid-phase microextraction and identified by gas chromatography coupled with a mass spectrometer (GC-MS). The multivariate data analysis tool (PLS-DA and Heatmap) identified short-chain organic acids as the main antagonistic VOCs responsible for inhibiting the mycelial growth of LPPC130. Acetic acid, propanoic acid, butanoic acid, valeric acid, and isovaleric acid exhibited a strong inhibitory effect on the mycelial growth of LPPC130, with inhibition of 20.68%, 33.30%, 26.87%, 43.71%, and 53.10%, respectively. Scanning electron microscopy revealed that VOCs caused damage to the vegetative and reproductive structures of the fungus. These results suggest Bacillus LPPC170 as an excellent biocontrol tool against the phytopathogen causative agents of Panama disease.

Cashew apple (Anacardium occidentale L.) extract from a by-product of juice processing: assessment of its toxicity, antiproliferative and antimicrobial activities.

Cashew apple extract (CAE) is a product with intense yellow color obtained from residual fibers of juice processing. Although CAE is known to be rich in carotenoids and anacardic acids, the biological activities of this potential natural food colorant remain unexplored. The present study is the first to investigate the toxicity, antiproliferative and antimicrobial activities of the lyophilized CAE (L-CAE) and its encapsulated products, using maltodextrin (M-CAE) or cashew gum (CG-CAE) as carriers. In addition to their high carotenoid content, the phenolic contents in all materials was determined using UPLC-QTOF-MSE. The acute toxicity was performed using adult zebrafish (Danio rerio); antiproliferative activity was assessed using seven different human tumor cell lines [U-251 (glioblastoma), MCF-7 (breast, adenocarcinoma), NCI-ADR/RES (multidrug-resistant ovarian adenocarcinoma), NCI-H-460 (lung, large cell carcinoma), PC-3 (prostate, adenocarcinoma), OVCAR-3 (ovarian adenocarcinoma), and HT-29 (colon, adenocarcinoma)] and an immortalized human keratinocyte (HaCaT) while the antimicrobioal activity was evaluated on Staphylococcus aureus ATCC 25923, Listeria monocytogenes ATCC 19115, Escherichia coli ATCC 25922 and Salmonella Typhimurium ATCC 51812 microorganisms. Both lyophilized and encapsulated CAE samples did not exert acute toxicity against zebrafish neither antiproliferative effect against human tumor and non-tumor cell lines. Further, L-CAE showed potential antimicrobial activity against Listeria monocytogenes, which was confirmed using electron microscopy. The current findings demonstrated that CAE is a potential source of bioactive compounds to use as an additive in the food industry.

Pentacyclic triterpene-loaded emulsion stabilized by Agaricus blazei Murill polysaccharides: Factorial design and cytoprotection study.

In this study, pentacyclic triterpene-loaded emulsions were stabilized by polysaccharides from Agaricus blazei Murill mushroom (PAb). The drug-excipient compatibility results by Fourier Transform Infrared Spectroscopy (FTIR) and Differential Scanning Calorimetry (DSC) showed the absence of physicochemical incompatibilities. The use of these biopolymers at 0.75 % led to obtaining emulsions with droplets smaller than 300 nm, moderate polydispersity, and ζ-potential >30 mV in modulus. The emulsions presented high encapsulation efficiency, suitable pH for topical application, and absence of macroscopic signs of instability during 45 days. Morphological analysis suggested the deposition of thin layers of PAb around the droplets. The encapsulation of pentacyclic triterpene in emulsions, stabilized by PAb, improved the cytocompatibility of this drug against PC12 and murine astrocyte cells. There was a reduction in cytotoxicity, which resulted in a lower accumulation of intracellular reactive oxygen species and maintenance of the mitochondrial transmembrane potential. Based on these results, it is estimated that PAb are promising biopolymers for the emulsions' stabilization by improving their physicochemical and biological properties.

Trichoderma longibrachiatum as a biostimulant of micropropagated banana seedlings under acclimatization.

The use of growth-promoting microorganisms with biostimulant characteristics is an important biological asset for the acclimatization of micropropagated seedlings. The present study aimed to evaluate the efficacy of the application of Trichoderma spp. on the promotion of the growth of micropropagated banana seedlings during acclimatization. The experiment was performed in an 8 × 6 completely randomized design using the following treatments: water, seedlings fertilized with controlled-release fertilizer, commercial biological inputs (A: T. asperellum, B/C: T. harzianum), and LPPC299 and LPPC300 strains. Plant height, pseudostem diameter, number of leaves, total leaf area, root length, fresh and dry mass of the plant, and accumulation of sodium, macronutrients, and micronutrients were evaluated 60 days after inoculation. Strains LPPC299 and LPPC300 were subjected to molecular identification by DNA sequencing of the ITS/5.8S locus. In vitro detection of growth promotion-related mechanisms and mycelial growth of biostimulants were performed using scanning electron microscopy. LPPC299 and LPPC300 had a greater similarity to T. longibrachiatum. LPPC299 was able to promote greater pseudostem diameter, number of leaves, and total leaf area in banana seedlings. T. asperellum (A) favored seedling performance in terms of fresh and dry mass of the plants. The strains were able to produce siderophores, indoleacetic acid, and catalase in vitro. Seedlings inoculated with the strains accumulated Mn, S (LPPC300), and Mg (LPPC299). LPPC299 from the banana rhizosphere was efficient in promoting performance in banana seedlings, showing its potential as a biostimulant for this crop.

In vitro degradability and bioactivity of oxidized bacterial cellulose-hydroxyapatite composites.

Hydroxyapatite-associated bacterial cellulose (BC/HA) is a promising composite for biomedical applications. However, this hybrid composite has some limitations due to its low in vivo degradability. The objective of this work was to oxidize BC and BC/HA composites for different time periods to produce 2,3 dialdehyde cellulose (DAC). The BC and oxidized BC (OxBC) membranes were mineralized to obtain the hybrid materials (BC/HA and OxBC/HA) and their physico-chemical, degradability, and bioactivity properties were studied. The results showed that OxBC/HA was more bioactive and degradable than BC/HA, which isa function of the degree of BC oxidation. High glucose levels in the BC degradation were observed as a function of oxidation degree, and other products, such as butyric acid and acetic acid resulted from DAC degradation. Therefore, this chemical modification reaction favors BC degradation, making it a good biodegradable and bioactive material with a potential for bone regeneration applications.

Resorbable bacterial cellulose membranes with strontium release for guided bone regeneration.

Hybrid materials, based on bacterial cellulose (BC) and hydroxyapatite (HA), have been investigated for guided bone regeneration (GBR). However, for some GBR, degradability in the physiological environment is an essential requirement. The present study aimed to explore the use of oxidized bacterial cellulose (OxBC) membranes, associated with strontium apatite, for GBR applications. BC membranes were produced by fermentation and purified, before oxidizing and mineralizing by immersing in strontium chloride solution and sodium bibasic phosphate for 5 cycles. The hybrid materials (BC/HA/Sr, BC/SrAp, OxBC/HA/Sr and OxBC/SrAp) were characterized for biodegradability and bioactivity and for their physicochemical and morphological properties. In vitro cytotoxicity and hemolytic properties of the materials were also investigated. In vivo biocompatibility was analyzed by performing histopathological evaluation at 1, 3 and 9 weeks in mices. Results showed that the samples presented different strontium release profiles and that oxidation enhances degradation under physiological conditions. All the hybrid materials were bioactive. Cell viability assay indicated that the materials are non-cytotoxic and in vivo studies showed low inflammatory response and increased connective tissue repair, as well as degradation in most of the materials, especially the oxidized membranes. This study confirms the potential use of bacterial cellulose-derived hybrid membranes for GBR.

Utilização da Microscopia Eletrônica de Varredura na detecção de matérias estranhas em águas envasadas / Scanning Electron Microscopy as a tool to detect and quantify microscopic physical hazards in bottled mineral waters

Este trabalho objetiva averiguar a presença de matérias estranhas presentes em águas minerais envasadas e sugerir a utilização da Microscopia Eletrônica de Varredura (MEV), como técnica adequada para detecção e contagem. Oito amostras de água mineral foram obtidas e subamostras de 100 mL foram filtradas em membranas, que foram secas e recobertas com platina e observadas em MEV. Áreas de 21.025 µm2 foram focalizadas e as partículas contadas. Realizaram-se análise de variância e o teste de Regwq para a comparação das médias das contagens. Diferentes partículas visualizadas foram registradas sob distintas magnificações. A análise de variância revelou alta significância entre a quantidade de matérias estranhas das amostras, indicando que as amostras diferiram bastante quanto a essa característica. As médias obtidas obedeceram a uma estratificação, mas foram estimadas com grande precisão, sugerindo-se que a subdivisão em 10 grupos de contagem e a extrapolação para o volume de 100 mL é uma estimativa confiável para a contagem de partículas filtradas nesse volume de água. Foi mostrado que o MEV se apresentou como instrumento auxiliar adequado para a contagem de matérias estranhas em amostras de águas minerais envasadas, especialmente quando suas dimensões não permitem sua identificação convencionalmente. (AU)

This work aims to investigate the presence of microscopic foreign materials in bottled mineral waters and suggest the use of Scanning Electron Microscopy (SEM) as a suitable technique for counting foreign matter. Eight samples of mineral water were obtained, and 100 mL subsamples were filtered through membranes, which were then dried, coated with platinum, and observed under SEM. Areas of 21,025 µm2 were analysed, and the particles present were counted. Variance analysis was carried out, and the Regwq test was applied to compare the counting averages. Samples of the dirt particles were imaged at different magnifications. The analysis of variance revealed high significance among the samples, indicating substantial differences between them. The averages followed a stratification pattern and were estimated with great precision, suggesting that the subdivision into 10 count groups and the extrapolation to 100 mL volume is a reliable estimate for the count of particles filtered in this volume of water. It was shown that SEM was a suitable instrument for counting and observing foreign matter and dirt in samples of bottled natural mineral waters, especially when their dimensions are too small to be detected by conventional methods. (AU)

Eggs hatching and oncomiracidia lifespan of Dawestrema cycloancistrium, a monogenean parasitic on Arapaima gigas.

Within the production chain of the Arapaima gigas (pirarucu), sanitary issues are still faced at the fingerling phase regarding infestations by the monogenean Dawestrema cycloancistrium. The present study had the objectives of describing the morphology and development phases of this parasite's eggs and oncomiracidia and determining the hatching time and lifespan of the oncomiracidia at different temperatures. D. cycloancistrium eggs were oval and elongated and had a single long appendage at one pole. The egg development was divided into four phases: embryonated egg, developing egg, larva appearance and ecloded egg. They were found in four forms: isolated in gill filaments; grouped in clusters that were anchored in a gill filament; grouped in clusters and entangled in part of a dead adult monogenean; and grouped in clusters in the environment, fixed to a substrate. Eclosion occurred after 72-96h, with faster development at 29°C and 32°C than at 24°C. The morphology of the oncomiracidia were rounded and elongated. They had two pairs of pigmented eye-spots as well as two ciliated areas located on each lateral part of the body and another on the anterior part of the body. The lifespan of the D. cycloancistrium oncomiracidia were 50 and 58h at 24°C and 27°C, respectively. Larvae were found in the mucus, indicating that the oncomiracidia penetrated the host integument. Understanding reproductive aspects of the D. cycloancistrium monogenean is important for developing specific prophylactic management strategies in aquaculture and providing valuable data for further studies.

Carnauba wax p-methoxycinnamic diesters: Characterisation, antioxidant activity and simulated gastrointestinal digestion followed by in vitro bioaccessibility.

The beneficial biological effects of cinnamic acid derivatives and the lack of studies on the antioxidant activity and bioavailability of cinnamic esters from carnauba wax, diesters were extracted from carnauba wax powder. Their structural, physical and morphological characteristics, antioxidant activity and in vitro bioaccessibility were measured. p-Methoxycinnamic diester (PCO-C) was identified, which has a crystalline, apolar structure and exhibited significant antioxidant activity (107.27 ± 3.92 µM Trolox/g of dry weight) before and after simulated in vitro gastrointestinal digestion and 32.46% bioaccessibility. In human cells, PCO-C (250 µg/mL) inhibited the production of intracellular reactive oxygen species, with an effect similar to that of Trolox (80 µM). Thermogravimetric analysis showed that PCO-C had high thermal stability and high UV absorption between 250 and 350 nm. These results indicate that this compound is promising as an antioxidant for pharmaceutical and food industry applications, such as the development of active packaging and functional foods.

Chemical constituents of Calotropis procera latex and ultrastructural effects on Haemonchus contortus / Constituintes químicos do látex de Calotropis procera e efeitos ultraestruturais em Haemonchus contortus

Abstract This study aimed to evaluate the anthelmintic and ultrastructural effects of Calotropis procera latex on Haemonchus contortus. C. procera latex was twice centrifuged at 10,000×g and dialyzed to obtain a fraction rich in proteins, named LP (latex protein), and at 3,000 rpm to obtain a fraction rich in secondary metabolites, named LNP (latex non-protein). Specimens of H. contortus exposed to LNP, LP and PBS in the Adult Worm Motility Test (AWMT) were submitted to scanning (SEM) and transmission (TEM) electron microscopy to verify changes in their ultrastructure. Phytochemical tests in the LNP indicated the presence of phenols, steroids, alkaloids and cardenolides. High-Performance Liquid Chromatography (HPLC) characterized the presence of the compounds gallic acid and quercetin in the LNP. The protein content in the LP was 43.1 ± 1.1 mg/mL and 7.7 ± 0.3 mg/mL in LNP. In AWMT, LNP and LP inhibited the motility of 100% of the nematodes, with LNP being more effective than LP and ivermectin more effective than both (p <0.05). Cuticle changes were observed by SEM and TEM in nematodes treated with LP and LNP. Calotropis procera latex has anthelmintic effects against H. contortus, causing damage to its cuticle and other alterations in its ultrastructure.

Resumo Este estudo objetivou avaliar os efeitos anti-helmínticos e ultraestruturais do látex de Calotropis procera sobre Haemonchus contortus. Látex de C. procera foi centrifugado duas vezes à a 10.000xg e dialisado para obter uma fração rica em proteínas, denominada proteínas do látex (LP). E centrifugado e centrifugado a 3.000 rpm, para obter uma fração rica em metabólitos secundários, denominada LNP (látex não proteico). Espécimes de H. contortus expostos à LNP, LP e PBS no Teste de Motilidade dos Nematoides Adultos (TMNA) foram submetidos a microscopia eletrônica de varredura (MEV) e de transmissão (MET), para verificar alterações em sua ultraestrutura. Testes fitoquímicos em LNP indicaram a presença de fenóis, esteroides, alcaloides e cardenolídeos. A presença dos compostos ácido gálico e quercetina em LNP foi caracterizada por Cromatografia Líquida de Alta Eficiência (CLAE). O conteúdo de proteínas em LP foi de 43,1 ± 1,1 mg/mL e de 7,7 ± 0,3 mg/mL em LNP. No TMNA, LNP e LP inibiram a motilidade de 100% dos nematoides, sendo LNP mais eficaz que LP, e a ivermectina mais eficaz que ambos (p <0,05). Alterações na cutícula de nematoides tratados com LP e LNP foram observadas por MEV e MET. O látex de C. procera apresenta efeito anti-helmíntico sobre H. contortus, causando danos à sua cutícula e outras alterações em sua ultraestrutura.

Adsorção de índigo carmim em biomassas mortas de aspergillus niger / Adsorption of indigo carmine on dead biomass of aspergillus niger

As indústrias têxteis geram efluentes ricos em corantes durante toda a sua cadeia produtiva. Neste estudo, ensaios cinéticos foram realizados utilizando biomassas mortas de Aspergillus niger, diferentemente tratadas, como adsorvente para a remoção de índigo carmim a partir de soluções aquosas. Ensaio cinético com carvão ativado comercial também foi realizado para comparação. Os resultados obtidos indicam que a biomassa fúngica sem tratamento algum foi capaz de remover 95% do índigo carmim da solução aquosa, remoção equivalente àquela obtida pelo carvão ativado comercial. Os experimentos mostraram que os tratamentos ácidos melhoraram consideravelmente a cinética da remoção do corante, não influenciando, contudo, a capacidade adsortiva máxima do material. Os tratamentos salinos e alcalinos diminuíram tanto a capacidade máxima quanto a cinética de adsorção.

Textile industries produce effluents rich in dyes throughout its production chain. In this study, kinetic experiments were carried using the dead and treated biomass of Aspergillus niger for indigo carmine removal from the synthetic aqueous solutions of this dye. A kinetic run with activated carbon was also carried for comparison. The obtained results suggest that non-treated fungal biomass has been capable of remove 95% of carmine indigo from the aqueous solutions. This removal was similar to that obtained with activated carbon. The results also indicate that acid treatments considerably improve the kinetic of the dye removal, but they are not influenced the maximum adsorptive capacity of the adsorbent. Moreover, saline and alkaline treatments decreased both the maximum capacity and the kinetic of adsorption.

Qualidade microbiológica de polpas de frutas produzidas e comercializadas nos estados da Paraíba e Pernambuco / Microbiology quality of frozen fruit pulps produced and commercialized in the States of Paraíba and Pernambuco

No período de maio a setembro de 1996, 71 amostras de polpas congeladas de frutas (acerola, cajá e caju) produzidas e comercializadas nos Estados da Paraíba e Pernambuco, foram analisadas quanto ao aspecto microbiológico. Foi avaliado o NMP de coliformes totais e coliformes fecais, contagem de mofos e leveduras, contagem de microrganismos mesófilos e a presença de Salmonella, utilizando metodologia recomendada pela ICMSF. os resultados indicaram que 83,9 por cento das amostras atenderam ao padräo para bolores e leveduras e, que 2,8 por cento das amostras apresentaram Coliformes fecais. Em relaçäo a contagem de bactérias aeróbias mesófilas, observou-se que as amostras apresentaram resultados dentro dos limites aceitáveis pela legislaçäo (1.000.000 ufc/g). Näo foi detectada a presença de Salmonella em nenhuma das amostras analisadas.

Bebidas fermentadas a partir de frutos tropicais / Tropical fruits fermented beverages

Este estudo teve como objetivo elaborar e caracterizar bebidas fermentadas de ata (Annona squamosa L.), cirigüela (Spondias purpurea L.) e mangaba (Hancornia speciosa Gom.), utilizando leveduras comerciais. A partir das polpas dos frutos foram formulados mostos com teores de sólidos solúveis de 16 Brix. Esses foram inoculados com levedura seca ativa, estirpe S. cerevisae var. bayanus e fermentados entre 18 e 21 §C. Alíquotas dos mostos foram coletadas, diariamente, para monitoramento de pH, acidez titulável, teor de sólidos solúveis totais, açúcares totais e teor alcoólico. As bebidas obtidas foram avaliadas quanto às características químicas, físico-químicas e sensoriais. O mosto de ata apresentou fermentaçäo rápida,atingindo estabilizaçäo dos teores alcoólico, sólidos solúveis e açúcares totais no sexto dia de fermentaçäo, tendo o processo terminado no décimo segundo dia. A fermentaçäo do mosto de mangaba foi relativamente rápida, com início da fase tumultuiosa no terceiro dia e término do processo no décimo oitavo dia. O mosto de cirigüela apresentou fermentaçäo lenta, estabilizou-se a partir do décimo dia com término do processo após o vigésimo dia. As bebidas atingiram teores alcoólicos de 8,4 por cento §GL (ata), 9,8 §GL 9mangaba) e 10,0 §GL (cirigüela). Os resultados da análise sensorial demonstraram melhor desempenho da bebida fermentada de mangaba, tanto para a aceitaçäo global quanto para a intençäo de compra, sendo a mais indicada para a obtençäo da bebida fermentada

Microorganismos patogênicos e indicadores em queijo de coalho produzido no estado do Ceará, Brasil / Pathogenic and indicator microorganisms isolated from Ceará, Brazil

Avaliou-se a qualidade higiênico-sanitária de 43 amostras de queijo de coalho produzidas em diferentes microrregiöes do estado do Ceará. Bolores e leveduras foram detectados em 100 por cento das amostras de queijos, com contagem de 1,7 x 10 elevado a quatro a 1,6 x 10 elevado a nove UFC/g. Todas as amostras apresentaram coliformes totais e fecais, confirmaçäo de Escherichia coli em 93,1 por cento das mesmas. Foi verificada a presença de Estafilococos coagulase positiva em 93,1 por cento das amostras de queijos, com contagens variando de 1,0 x 10 elevado a 1 a 2,0 x 10 elevado a nove UFC/G. Apenas 2,3 por cento das amostras encontravam-se de acordo com os padröes microbiológicos vigentes para essa bactéria. A presença de Salmonella foi constatada em 34,9 por cento das amostras de queijos. Listeria sp. foi detectada em 6,9 por cento das amostras, com confirmaçäo de L. monocytogenes em 2,3 por cento dessas. A elevada populaçäo de bolores e leveduras (observada em 100 por cento das amostras de quijos de coalho) indicou deficiência nos procedimentos de higiene e sanitizaçäo das amostras, caracterizando-se como produto em condiçöes higiênicas insatisfatórias. A alta concentraçäo de estafilococos coagulase positiva e de coliformes fecais (aproximadamente 90 por cento das amostras) caracterizou os queijos como produto em condiçöes higiênico-sanitárias insatisfatórias. Queijos de coalho oriundos das cinco microrregiöes do Ceará envolvidas no estudo näo apresentaram segurança alimentar, visto que a maioria continha estafilococos coagulase positiva, L. monocytogenes e Salmonella. A presença dessas duas últimas bactérias permitiu classificar os queijos como produtos impróprios para consumo humano

Alimentos transgênicos: segurança, riscos alimentares e regulamentaçöes / Transgenic foods: safety, dietary risks and regulations

A presente revisäo de literatura aborda o tema alimentos transgênicos, enfocando aspectos de segurança e riscos potenciais ao consumidor, bem como questöes de rotulagem e regulamentaçöes nacionais e internacionais. Os principais alimentos de origem vegetal e animal transgênicos no mundo foram apresentados, assim como as modificaçöes e vantagens decorrentes de suas tranformaçöes. Säo descritos os mecanismosde surgimento de riscos nesses organismos e os procedimentos de avaliaçäo mundialmente adotados. Os parâmetros de alergenicidade, toxicidade, surgimento de patógenos alimentares resistentes e alteraçäo na qualidade nutricional dos alimentos säo destacados pela sua importância em relaçäo à seggurança desses alimentos. A tecnologia do DNA recombinate, associada a outras técnicas convencionais de melhoramento e manejo, apresenta enorme potencial para aumentar a produtividade agrícola, beneficiar o meio ambiente e melhorar a qualidade dos alimentos. No entanto, a imocuidade sob o ponto de vista toxicológico e nutricional deve ser igualmente almejada durante o desenvolvimento de alimentos geneticamente modificados, levando-se em conta, acima de tudo, a segurança final do consumidor.