Abstract The therapeutic drugs to treat Herpes simplex virus (HSV) infections have toxic side effects and there has been an emergence of drug-resistant strains. Therefore, the search for new treatments for HSV infections is mounting. In the present study, semi-solid formulations containing a crude hydroethanolic extract (CHE) from Schinus terebinthifolia were developed. Skin irritation, cutaneous permeation, and in vivo therapeutic efficacy of the formulations were investigated. Treatment with the ointment formulations did not result in any signs of skin irritation while the emulsions increased the thickness of the epidermis in Swiss mice. The cutaneous permeation test indicated that the CHE incorporated in the formulations permeated through the skin layers and was present in the epidermis and dermis even 3 h after topical application. In vivo antiviral activity in BALB/c mice treated with the CHE ointments was better than those treated with the CHE emulsions and did not significantly differ from an acyclovir-treated group. Taken together, this suggests that the incorporation of CHE in the ointment may be a potential candidate for the alternative topical treatment of herpetic lesions.

RESUMO: Protozoários do gênero Phytomonas são parasitas prejudiciais a várias culturas agrícolas de importância econômica. Devido a sua atividade biológica reconhecida, extratos brutos de folhas de Piper aduncum, P. crassinervium, P. hispidum e P. amalago, foram testadas pela técnica de microdiluição em placa para avaliar o seu potencial antiparasitário contra Phytomonas serpens. Os resultados mostraram que o extrato bruto etanólico de P. crassinervium e P. amalago apresentaram as melhores concentrações inibitórias para 50% das células (IC50), 16,5 µg mL-1 na fase clorofórmio e 18 µg mL-1 na fase aquosa, respectivamente, após 48 h de tratamento. Análises de citotoxicidade foram realizadas através do método colorimétrico da sulforodamina-B, em células de mamíferos LLCMK2. A fase clorofórmio de P. crassinervium foi submetida ao processo de fracionamento, no qual as frações acetato de etila e diclorometano obtiveram melhores valores de IC50. Imagens de microscopia eletrônica de varredura (MEV) mostraram alterações na membrana celular dos parasitas tratados com fase aquosa de P. amalago. Os dados obtidos indicam potencial efeito antiparasitário das espécies de Piper analisadas contra P. serpens, sendo consideradas candidatas promissoras para formulações de bioprodutos para controle do parasito.

ABSTRACT: Protozoa of the genus Phytomonas are harmful parasites to several agricultural crops of economic importance. Due to their recognized biological activity, crude extracts of Piper aduncum, P. crassinervium, P. hispidum, and P. amalago leaves, were tested using the microdilution plate technique to assess the antiparasitic potential against Phytomonas serpens. Results showed that the ethanolic crude extract of P. crassinervium and P. amalago presented the best inhibitory concentration for 50% of the cells (IC50), 16.5 µg mL-1 in chloroform phase, and 18 µg mL-1 in aqueous phase, respectively, after 48 h treatment. Cytotoxicity analyses were performed using the colorimetric method of sulforhodamine-B in LLCMK2 mammalian cells. The chloroform phase of P. crassinervium was subjected to the fractionation process, in which the ethyl acetate and dichloromethane fractions obtained better IC50 values. Scanning electron microscopy (SEM) images showed alterations in the cell membrane of the treated parasites. The data obtained indicate a potential antiparasitic effect of the Piper species analyzed against P. serpens, being considered promising candidates for formulations of bioproducts to control the parasite.

In vitro activity of the essential oil from Piper diospyrifolium leaves was tested using disk diffusion techniques. The antifungal assay showed significant potencial antifungal activity: the oil was effective against several clinical fungal strains. The majority compounds in the essential oil were identified as sesquiterpenoids by GC-MS and GC-FID techniques.

A porcentagem de amostras positivas para Aeromonas foi de 12.7% para água mineral, 8.3% para água de poço artesiano e 6.5% para água do sistema público de abastecimento. O isolamento de Aeromonas spp. foi significativamente maior em água mineral e água de poço artesiano do que em água do sistema público. A ocorrência de Aeromonas spp. não teve correlação significativa com os indicadores de contaminação tradicionalmente utilizados (coliformes totais) em amostras de água de poço artesiano. No entanto, esta correlação foi positiva e significativa em água mineral e água do sistema público. A presença ou ausência de correlação entre bactérias indicadoras e a presença de Aeromonas pode refletir o aparecimento ocasional de patógenos em águas para consumo humano e as diferentes taxas de sobrevivência e isolamento destes agentes comparados com os indicadores fecais de contaminação. A constatação de que 41.6%, 14.8% e 9.0% respectivamente amostras de água de poço, água mineral e água do abastecimento público utilizadas neste estudo apresentaram índices de coliformes maiores do que os aceitáveis pela legislação brasileira é um fato preocupante. Estes números mostram a necessidade de melhoria nos sistemas de monitoramento para a indústria de águas minerais e o sistema público de abastecimento. As cepas isoladas pertencentes ao gênero Aeromonas foram identificadas ao nível de espécie como A. hydrophila e A. jandaei. A significância do grande número de isolamentos de espécies de Aeromonas em saúde pública ainda não está clara. É necessário o estudo dos efeitos de cepas específicas utilizando modelos animais de infecção. Estes resultados podem contribuir para a reavaliação dos critérios empregados para a análise da qualidade microbiológica da água e a definição de limites de densidades para o gênero Aeromonas em águas destinadas ao consumo humano.

The aim of this work was to study the occurrence of Aeromonas sp in the bottled mineral water, well water and tap water from the municipal supplies. Positive samples were found for Aeromonas spp. 12.7% from the mineral water, 8.3% from the artesian water and 6.5% from the tap water. The recovery of Aeromonas spp. was significantly higher in the bottled mineral and artesian water than in the tap water from municipal supplies. The occurrence of the Aeromonas spp. did not correlate significantly with the contamination indicator bacteria (i.e. total coliforms) in the artesian water samples. However, a significant correlation was found between Aeromonas spp. and total coliforms in the both mineral water and tap water samples. The presence or absence of a correlation between the indicator bacteria and Aeromonas could reflect the occasional appearance of the pathogen in the drinking water and the different rates of survival and recovery of these agents compared with those fecal indicators. The finding that 41.6, 14.8 and 9.0 % of the artesian water, bottled mineral water and tap water, respectively, sampled in the current study failed to meet the Brazilian standard for total coliforms in the drinking water should therefore be of concern.

Terpenos e aromas voláteis das flores de Calendula officinalis cultivados no sudoeste do Brasil foram isolados por arraste a vapor (SD), dedo frio (HS-CF) e micro extração em fase sólida (HS-SPME) acoplada à espectrometria de massas. As flores secas da C. officinalis contêm 0,1% de óleo essencial e foram identificadas 27 substâncias químicas através do cálculo do índice de Kováts e interpretação dos espectros de massas. As substâncias majoritárias presentes no óleo essencial das flores de C. officinalis, obtido por SD, HS-SPME e HS-CF foram δ-cadinene (22,5; 22,1 e 18,4 %) γ-cadinene (8,9, 25,4 e 24,9 %) e 20.4 % de α-cadinol foi observado apenas na extração por arraste a vapor.

Terpenes and aroma volatiles from flowers of Calendula officinalis cultivated in southeastern Brazil were obtained by steam distillation (SD), headspace-cold finger (HS-CF) extraction and headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography and mass spectrometric analysis. The dried flowers contained 0.1% oil. Kovats indices and mass spectra were used to identify 27 individual components in the various volatile fractions. The main components present in the volatile fractions of the C. officinalis flowers, obtained by SD, HS-SPME, and HS-CF, were δ-cadinene (22.5, 22.1, and 18.4 %) and γ-cadinene (8.9, 25.4, and 24.9 %) while 20.4 % of α-cadinol was seen only after SD extraction.

The antimicrobial activity of copaiba oils was tested against Gram-positive and Gram-negative bacteria, yeast, and dermatophytes. Oils obtained from Copaifera martii, Copaifera officinalis, and Copaifera reticulata (collected in the state of Acre) were active against Gram-positive species (Staphylococcus aureus, methicillin-resistant S. aureus, Staphylococcus epidermidis, Bacillus subtilis, and Enterococcus faecalis) with minimum inhibitory concentrations ranging from 31.3-62.5 µg/ml. The oils showed bactericidal activity, decreasing the viability of these Gram-positive bacteria within 3 h. Moderate activity was observed against dermatophyte fungi (Trichophyton rubrum and Microsporum canis). The oils showed no activity against Gram-negative bacteria and yeast. Scannning electron microscopy of S. aureus treated with resin oil from C. martii revealed lysis of the bacteria, causing cellular agglomerates. Transmission electron microscopy revealed disruption and damage to the cell wall, resulting in the release of cytoplasmic compounds, alterations in morphology, and a decrease in cell volume, indicating that copaiba oil may affect the cell wall.

A Calendula officinalis L., pertence à família Asteraceae, é originária da Europa e está aclimatada na região Sul do Brasil, onde vem crescendo o seu cultivo comercial para fins farmacêuticos e cosméticos. Suas inúmeras propriedades terapêuticas entre as quais antiinflamatória, cicatrizante e anti-séptica, justificam a grande procura desta planta pela população. O presente trabalho compara a composição química da fração volátil da cultura de calêndula aclimatada na região Noroeste do Paraná, com os dados da literatura da cultura na França. As substancias químicas majoritários identificadas por cromatografia gasosa acoplada à espectrometria de massas (CG/MS), nas duas culturas, foram sesquiterpenos hidrocarbonetos e álcoois; d-cadineno (22,53%) e a-cadinol (20,40%) para a cultura de calêndula produzida na região Noroeste do Paraná, Brasil, e d-cadineno (12,10%) e a-cadinol (25,50%) distinto do registrado para a cultura de calêndula produzida na região de Massif Central, França. Adicionalmente, para garantir a qualidade da planta na comercialização, foram realizadas análises físico-químicas (perda por secagem = 85,0% e perda por dessecação = 11,6%).

Calendula officinalis L., belongs to the Asteraceae family from Europe and is acclimatized in the south region of Brazil where its commercial cultivation has been increasing for pharmaceutical and cosmetic purposes. Its several therapeutic properties (anti-inflammatory, healing, antiseptic) justify the great demand for this plant by the population. The present study compares the chemical composition of the volatile fraction of two cultivations of calendula, one from Massif Central, France, and the other acclimatized in the Northeast region of Paraná State, Brazil. Most compounds identified by Gas Chromatography-mass spectrometry (GC-MS) in both cultivations included the sesquiterpenes hydrocarbons and alcohols; d-cadinene (22.53%) and á-cadinol (20.40%) for the cultivation of the calendula produced in the Northeast region of Paraná State, and d-cadinene (12.10%) and a-cadinol(25.50%) for the cultivation of the calendula produced in Massif Central, France. In addition, physical-chemical analyses (determination of water at room temperature = 85.0% and loss on drying = 11.6%), were carried out in order to assure the quality of the plant for commercialization.

O principal objetivo do presente estudo foi avaliar a prevalência de leveduras e fungos filamentosos em água potável, bem como investigar suas correlações com bactérias indicadoras de contaminação fecal. Leveduras foram detectadas em 36,6% e 11,6% das amostras de água mineral de garrafão em dispensadores de água e água de torneira do sistema municipal, respectivamente. Vinte e uma (35,5%) das amostras de água mineral de garrafão e duas (3,3%) das amostras de água de torneira foram positivas para fungos filamentosos. Para água mineral de garrafão, 12 (20.0%) das 60 amostras foram positivas para coliforme total, comparado com 3 (5.0%) das 60 amostras de água de torneira. A água coletada de garrafões de água mineral dos dispensadores foi marcadamente mais contaminada que as amostras de água de torneira. Candida spp identificadas ao nível de espécie foram C. parapsilosis, C. glabrata e C. albicans. Como está sendo reportado, água mineral de garrafão em dispensador e água de torneira pode ser considerada como possíveis vias de transmissão de fungos filamentosos e leveduras, e podem constituir um potencial risco para a saúde, principalmente de pessoas imunocomprometidas.

The main objective of this study was to analyse the occurrence of yeasts and filamentous fungi in drinking water as well as to investigate their correlation with the indicator bacteria of faecal pollution. Yeasts were detected in 36.6% and 11.6% of the bottled mineral on water dispensers and tap water samples from municipal system, respectively. Twenty-one (35.0%) of bottled mineral water and two (3.3%) of tap water samples were positive for filamentous fungi. For bottled mineral water 12 (20.0%) of 60 samples were positive for total coliform, compared with 3(5.0%)out of 60 samples from tap water. The mineral water from dispensers was more contaminated than tap water. Strains belonging to the genera Candida identified to the species level were C. parapsilosis, C. glabrata and C. albicans. Thus, bottled mineral water from water dispensers and tap water could be considered a possible transmission route for filamentous fungi and yeasts, and could constitute a potential health hazard, mainly to immunocompromised indivuals.

Extratos de folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck (Piperaceae), uma planta medicinal utilizada no Brasil para tratar doenças infecciosas, foram testados para a atividade antifúngica sobre as leveduras Candida albicans, C. krusei, C. parapsilosis e C. tropicalis. O extrato em acetato de etila apresentou boa atividade contra C. albicans com uma CIM de 125 mg µL-1, moderada atividade contra C. krusei e C. parapsilosis (CIM de 500 mg µL-1) e foi inativo contra C. tropicalis (CMI> 1000 mg µL-1). Com base nestes resultados, o extrato em acetato de etila foi fracionado em nove frações em cromatografia de coluna de sílica gel. As frações hexano e clorofórmio mostraram variados níveis de atividade antifúngica contra as leveduras testadas. Posterior separação da fração hexânica em cromatografia de coluna resultou nas substâncias puras eupomatenóide-6, eupomatenóide-5, eupomatenóide-3 e conocarpano. A elucidação estrutural das substâncias foi baseada em dados espectrais (RMN de ¹H e de 13C, HSQC, HMBC, gNOE, IV e EM). O conocarpano foi a única substância isolada, com atividade contra as leveduras. As propriedades antifúngicas do extrato de P. regnellii demonstraram preliminar validação científica do uso da planta na medicina popular.

Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck (Piperaceae) is a medicinal plant traditionally used in Brazil to treat infectious diseases. The extracts obtained from the leaves of P. regnellii were investigated for their antifungal activities against the yeasts Candida albicans, C. krusei, C. parapsilosis, and C. tropicalis. The EtOAc extract presented a significant activity against Candida albicans with MIC at 125 µg mL-1, and a moderate activity against both C. krusei and C. parapsilosis with MIC at 500 µg mL-1. Candida tropicalis was not inhibited by this extract at concentrations as high as 1000 µg mL-1. Based on these findings, the EtOAc extract was fractionated by silica gel column chromatography into nine fractions. The hexane and CHCl3 fractions showed varied levels of antifungal activity against all test yeasts. Further column chromatography separation of the hexane fraction afforded the pure compounds eupomatenoid-6, eupomatenoid-5, eupomatenoid-3 and conocarpan. The structure of the compounds was based on spectral data (¹H and 13C NMR, HSQC, HMBC, gNOE, IR and MS). Conocarpan was the only active compound on the yeasts. The antifungal property of P. regnellii extract provides preliminary scientific validation for the traditional medicinal use of this plant.

Chemical studies of the leaves of L. divaricata afforded 3beta-p-hydroxybenzoyl-tormentic acid, a triterpene with an ursene-type skeleton, a mixture whose main compound was an oleanene derivative, the maslinic acid, a C-glycoside flavone, vitexin and glucopyranosylsitosterol. A flavonoid, characterized as (-)-epicatechin, which belongs to the flavan-3-ol class, was isolated from the stem's bark. The structures of the compounds were elucidated by spectroscopic analysis. The antibacterial, antifungal and antiproliferative activities of the crude methanolic extracts of leaves and bark were evaluated and the antibacterial properties of the fractions of the barks were also investigated.

The bioassay-guided fractionation against Biomphalaria glabrata of hydroalcoholic extracts of Calophyllum brasiliense aerial parts led to the isolation of the coumarin, named (-) mammea A/BB. The compound had its structure determined by both spectroscopic techniques (NMR ¹H, NMR 13C, gHSQC, gHMBC and MS) and some literature comparison data. The probit analysis of (-) mammea A/BB showed LD50 = 0.67 ppm and LD90 = 1.47 ppm. In addition, the dichloromethane extract obtained from C. brasiliense leaves with significant molluscicidal activity against Biomphalaria glabrata was analyzed by HPLC-UV.

We report the effect of Stryphnodendron adstringens on the trypanosomatid Herpetomonas samuelpessoai. The parasites were grown at 28ºC in a chemically defined medium containing crude extract and fractions at concentrations from 100 to 5000 µg/ml obtained from S. adstringens. Concentrations of 500, 1000, 2500, and 5000 µg/ml both crude extract and semi-purified fraction progressively inhibited the protozoans' growth. At a concentration of 100 µg/ml, crude extract or a semi-purified (F3) fraction did not affect the growth of the protozoans. The F3-9 - F3-12 sub-fractions, at a concentration of 1000 µg/ml, also showed increased inhibitory activity on H. samuelpessoai. The IC50 of the crude extract and the F3 fraction were 538 and 634 µg/ml, respectively. Ultrastructural and enzymatic alterations in the trypanosomatids were also evaluated. H. samuelpessoai cultivated in the presence of IC50 crude extract showed considerable ultrastructural alterations, such as marked mitochondrial swelling with a large number of cristae and evident Golgi complex vesiculation, as observed by transmission electron microscopy. Cells exposed to 538 µg/ml of crude extract at 28ºC for 72 h, showed decreased activity of the enzyme succinate cytochrome c reductase, a typical mitochondrion marker, as compared to untreated cells

O presente estudo foi conduzido para avaliar a atividade antibacteriana dos extratos etanol:água e aquoso das folhas, raízes e casca do caule de Psidium guajava L. As atividades antibacterianas dos extratos contra as bactérias foram testados usando o ensaio de microdiluição em caldo. O extrato aquoso das folhas, raízes e casca do caule de P. guajava foram ativos contra as bactérias Gram-positivas Staphylococcus aureus (CIMs=500, 125 e 250 µg/ml, respectivamente) e Bacillus subtilis (CIMs=500 µg/ml), e foram inativas contras as bactérias Gram-negativas Escherichia coli e Pseudomonas aeruginosa (CIMs >1000 µg/ml). Os extratos etanol:água apresentaram maior atividade quando comparados com os extratos aquosos. Com base nestes resultados, o extrato de folhas de P. guajava foi fracionado em cromatografia em coluna de sílica gel em um bioensaio de fracionamento direcionado, produzindo uma mistura de flavonoides, uma mistura de alfa e beta-amirina e beta-sitosterol. A mistura de flavonoides mostrou boa atividade sobre S. aureus com CIM de 25 µg/ml. O beta-sitosterol foi inativo para todas as bactérias testadas.

The present study was designated to evaluate the antibacterial activities of aqueous and ethanol:water extracts from leaves, roots and stem bark of Psidium guajava L. The antibacterial activities of the extracts against bacteria were tested by using both microdilution assay. The aqueous extracts of P. guajava leaves, roots and stem bark were active against the gram-positive bacteria Staphylococcus aureus (MICs=500, 125 and 250 µg/ml, respectively) and Bacillus subtilis (MICs=500 µg/ml), and virtually inactive against the gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa (MICs >1000 µg/ml). The ethanol:water extracts showed higher antimicrobial activity as compared to aqueous extracts. Based on this finding, the ethanol:water extract of P. guajava leaves was fractionated on silica gel column chromatography in a bioassay-guided fractionation affording flavonoid mixture, triterpenes (alpha- and beta-amyrin) and sterol (beta-sitosterol). Flavonoid mixture showed good activity on S. aureus with MIC of 25 µg/ml. beta-sitosterol was inactive for all the bacteria tested.

Este estudo descreve a triagem de extratos obtidos de 19 espécies de plantas usadas na medicina tradicional brasileira para o tratamento de várias doenças. Os extratos foram testados contra formas amastigota axênica e promastigota de Leishmania (L.) amazonensis, e formas epimastigota de Trypanosoma cruzi in vitro na concentração de 100 mg/ml. Baccharis trimera, Cymbopogon citratus, Matricaria chamomilla, Mikania glomerata, Ocimum gratissimum, Piper regnellii, Prunus domestica, Psidium guajava, Sambucus canadensis, Stryphnodendron adstringens, Tanacetum parthenium, e Tanacetum vulgare apresentaram efeito significante contra um ou ambos parasitas, com a porcentagem de inibição de crescimento entre 49,5 e 99%. Os extratos não mostraram efeito citotóxico em hemácias de carneiro. Essas plantas medicinais podem ser fontes alternativas de novos compostos clinicamente ativos contra L. amazonensis e T. cruzi.

This study describes the screening of extracts obtained from 19 species of plants used in Brazilian traditional medicine for treatment of a variety of diseases. The extracts were tested against axenic amastigote and promastigote forms of Leishmania (L.) amazonensis, and epimastigote forms of Trypanosoma cruzi in vitro at a concentration of 100 mg/ml. Baccharis trimera, Cymbopogon citratus, Matricaria chamomilla, Mikania glomerata, Ocimum gratissimum, Piper regnellii, Prunus domestica, Psidium guajava, Sambucus canadensis, Stryphnodendron adstringens, Tanacetum parthenium, and Tanacetum vulgare showed significant effects against one or both parasites, with a percentage of growth inhibition between 49.5 and 99%. The extracts showed no cytotoxic effect on sheep erythrocytes. These medicinal plants may be sources of new compounds that are clinically active against L. amazonensis and T. cruzi.

Atividade antioxidante de extrato, frações, subfrações e substâncias isoladas das cascas de Stryphnodendron obovatum Benth. foi avaliada através da redução do radical 1,1-difenil-2-picrilhidrazila (método DDPH·) em cromatografia em camada delgada. O extrato bruto (CE, acetona:água), as frações acetato de etila (EtOAc) e aquosa (FW), as subfrações (F1-F12) e as substâncias isoladas I, II e III apresentaram a capacidade de reduzir o radical DDPH·. A atividade antifúngica foi determinada pela concentração inibitória mínima (CIM) e concentração fungicida mínima (CFM) frente às amostras de leveduras Candida albicans, Candida parapsilosis, Candida krusei e Candida tropicalis. O extrato bruto (CE), as frações (EtOAC e FW), e os compostos isolados I, II e III, como também as subfrações cromatográficas (F1-F12) foram inativos frente a todas as leveduras testadas. Por outro lado, as subfrações cromatográficas F13-15 e F20 apresentaram atividade antifúngica moderada. Já as subfrações F16-19 e F21-22 mostraram boa atividade antifúngica frente às cepas de C. albicans e C. parapsilosis. As substâncias I, II, e III, isoladas da fração EtOAc por cromatografia e recromatografia em coluna de Sephadex® LH-20, foram identificadas como sendo os monômeros de flavan-3-ol, epigalocatequina e galocatequina, e um dímero, epigalocatequina-(4<FONT FACE=Symbol>b®</FONT>8)-galocatequina, respectivamente.

The antioxidant activity of stem-bark extracts from Stryphnodendron obovatum Benth., including fractions and isolated compounds, was evaluated by DPPH in thin-layer chromatography. All the fractions and isolated compounds showed antioxidant activity. Antifungal activity was determined by the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) against the yeasts Candida albicans, Candida parapsilosis, Candida krusei and Candida tropicalis. All extracts (CE, EtOAc and FW), subfractions (F1-F12) and the compounds I, II and III were inactive against the yeasts. Against C. parapsilosis and C. albicans, fractions F13-15 and F20 showed moderate antifungal activity, and fractions F16-19 and F21-22 showed good activity. Chemical isolation of the ethyl-acetate fraction resulted in the identification of three compounds: epigallocatechin, gallocatechin and epigallocatechin-(4<FONT FACE=Symbol>b®</FONT>8)-gallocatechin.