Development and evaluation of topical formulations that contain hydroethanolic extract from Schinus terebinthifolia against HSV-1 infection

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.

Natural compounds based chemotherapeutic against Chagas disease and leishmaniasis: mitochondrion as a strategic target

Over the past years, natural products have been explored in order to find biological active substances to treat various diseases. Regarding their potential action against parasites such as trypanosomatids, specially Trypanosoma cruzi and Leishmania spp., much advance has been achieved. Extracts and purified molecules of several species from genera Piper, Tanacetum, Porophyllum, and Copaifera have been widely investigated by our research group and exhibited interesting antitrypanosomal and antileishmanial activities. These natural compounds affected different structures in parasites, and we believe that the mitochondrion is a strategic target to induce parasite death. Considering that these trypanosomatids have a unique mitochondrion, this cellular target has been extensively studied aiming to find more selective drugs, since the current treatment of these neglected tropical diseases has some challenges such as high toxicity and prolonged treatment time. Here, we summarise some results obtained with natural products from our research group and we further highlighted some strategies that must be considered to finally develop an effective chemotherapeutic agent against these parasites.

Activity of Piperaceae extracts and fractions in the control of Phytomonas serpens / Atividade de extratos e frações de Piperaceae no controle de Phytomonas serpens

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.

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.

Trypanocidal activity of organic extracts from the Brazilian and Spanish marine sponges

Abstract Chagas' disease is a parasitic infection caused by protozoan Trypanosoma cruzi that affect millions of people worldwide. The available drugs for treatment of this infection cause serious side effects and have variable efficacy, especially in the chronic phase of the disease. In this context, natural compounds have shown great potential for the discovery of new chemotherapies for the treatment of this infection and various other diseases. In present study, we evaluated the in vitro antiprotozoal activity of five species of Brazilian and Spanish marine sponges (Condrosia reniformes, Tethya rubra, Tethya ignis, Mycale angulosa and Dysidea avara) against T. cruzi. By GC–MS data, we observed that in these extracts were present the major classes of the following compounds: hydrocarbons, terpenes, steroids and alcohols. The extracts showed activity against the three forms of this parasite and did not induce toxicity in mammalian cells. Better activities were observed with the extracts of marine sponges, C. reniformes (EC50 = 0.6 μg/ml), D. avara (EC50 = 1.1 μg/ml) and M. angulosa (EC50 = 3.8 μg/ml), against trypomastigote forms. In intracellular amastigote forms, the extract of T. ignis showed IC50 of 7.2 μg/ml and SI of 24.65. On this basis, our results indicate that these extracts can be promising chemotherapeutic agents against T. cruzi.

Performance of methods for identification of yeasts isolated fromblotted water: High prevalence of Candida parapsilosis / Desempenho dos métodos de identificação de leveduras de águaengarrafada: alta prevalência deCandida parapsilosis

Yeasts of the Candida genus can colonize epithelium and mucosa of the vertebrate organisms; howeverthese can cause infection in a broad range of body sites. Candida species also can be found in drinking water and they are considered as a potential indicator of water quality. In this study were evaluated three methods to identify yeasts isolated from blotted water (seminested PCR, culture on CHROMagar Candida medium, and Candifast identification system). For this propose, we used 27 isolates fromblotted water and compared with 22 clinical isolates from vaginal fluid. Seminested PCR has shown specificity and sensitivity for identification of the Candida species. Candida albicans and Candida parapsilosis were the prevalent species from vaginal fluid and blotted water, respectively. Culture onCHROMagar and Candifast system had low agreement with snPCR (40.9% and 45.5%, respectively) in the yeasts identification from vaginal fluid. On the other hand, CHROMagar Candida can be used in the presumptive identification of yeasts isolated from bottled water and it had agreement’s percentage of 81.5% with snPCR method.

Leveduras do gênero Candida podem colonizar epitélio e mucosa dos organismos vertebrados, entretanto, podem causar infecções em vários lugares do corpo. Espécies de Candida, também, podem ser encontradas em água e são consideradas um potencial indicador da qualidade de água. Neste trabalho, foram avaliados três métodos de identificação de leveduras isoladas de água engarrafada (seminestedPCR, cultura no meio CHROMagar Candida e sistema de identificação Candifast). Foram utilizados 27 isolados de água engarrafada e comparados com 22 isolados clínicos de fluido vaginal. Seminested PCR tem mostrado especificidade e sensibilidade para a identificação das espécies de Candida. Candida albicans e Candida parapsilosis foram as espécies prevalentes do fluido vaginal e da água engarrafada,respectivamente. Cultura em CHROMagar e o sistema Candifast tiveram baixa concordância com snPCR(40,9% e 45,5%, respectivamente) na identificação de leveduras de fluido vaginal. Em contrapartida, CHROMagar Candida pode ser usado em identificação presuntiva de leveduras de água engarrafada apresentando concordância de 81,5% com o método snPCR.

Characterization of Candida spp. isolated from vaginal fluid: identification, antifungal susceptibility, and virulence profile / Caracterização de Candida spp. isoladas de fluido vaginal: identificação, susceptibilidade antifúngica e perfil de virulência

A total of 101 (20.0%) yeast samples were isolated from vaginal fluids of 504 non-hospitalized patients in Maringá, Paraná State, Brazil and Candida albicans was more frequent specie (93.1%) identified by seminested PCR method. All the isolates were susceptible to amphotericin B and nystatin, and 93.1% of them were susceptible to fluconazole. The acid proteinase, hemolytic and phospholipase activities were observed in 99.0, 90.0, and 88.0% of Candida spp., respectively. Around 67.0% of the strains had adherence indexes of 0.5 to 1.5 yeasts by Vero cell, and most of them showed a hydrophilic profile. Correlation studies indicated hydrophilic yeasts presented higher adherence index, proteinase, and phospholipase activities; and a positive correlation between all enzymes was also observed. In addition, the isolates with high hemolytic activity were less susceptible to fluconazole and amphotericin B. These results of Candida prevalence and antifungal susceptibility corroborate with literature's datas and correlation between virulence factors and MIC values suggest Candida isolates from vaginal fluid less susceptible to antifungal and with higher extracellular enzymes production can be more virulent to cause tissue damage.

Cento e uma leveduras foram isoladas de fluido vaginal de 504 pacientes não hospitalizados em Maringá, Estado do Paraná, Brasil e Candida albicans foi a espécie mais freqüente (93,1%) identificada pelo método seminested PCR. Todos os isolados foram susceptíveis a anfotericina B e a nistatina, e 93,1 % susceptíveis ao fluconazol. Atividades da proteinase ácida, hemolisina, e fosfolipase foram observadas em 99,0, 90,0 e 88,0% dos isolados de Candida spp., respectivamente. Aproximadamente 67,0% das cepas apresentaram índices de aderência de 0,5 a 1,5 leveduras por célula Vero, sendo que a maioria dos isolados mostrou um perfil hidrofílico. Estudos de correlação indicaram que leveduras hidrofílicas apresentaram maiores índices de aderência e atividade da proteinase e fosfolipase; e uma correlação positiva entre todas as enzimas, também, foi observada. Adicionalmente, os isolados com alta atividade hemolítica foram menos susceptíveis aos fluconazol e a anfotericina B. Esses resultados de prevalência de leveduras do gênero Candida e de susceptibilidade aos antifúngicos corroboram com dados da literatura; e as correlações observadas entre os fatores de virulência e valores de CIM sugerem que Candida spp. isoladas de fluido vaginal menos susceptíveis aos antifúngicos e com alta produção de enzimas extracelulares podem ser mais virulentas para causar dano tecidual.

Antileishmanial activity of diterpene acids in copaiba oil

Leishmaniasis is a neglected tropical disease. According to the World Health Organization, there are approximately 1.5-two million new cases of cutaneous leishmaniasis each year worldwide. Chemotherapy against leishmaniasis is based on pentavalent antimonials, which were developed more than a century ago. The goals of this study were to investigate the antileishmanial activity of diterpene acids in copaiba oil, as well as some possible targets of their action against Leishmania amazonensis. Methyl copalate and agathic, hydroxycopalic, kaurenoic, pinifolic and polyaltic acids isolated from Copaifera officinales oleoresins were utilised. Ultrastructural changes and the specific organelle targets of diterpenes were investigated with electron microscopy and flow cytometry, respectively. All compounds had some level of activity against L. amazonensis. Hydroxycopalic acid and methyl copalate demonstrated the most activity against promastigotes and had 50% inhibitory concentration (IC50) values of 2.5 and 6.0 µg/mL, respectively. However, pinifolic and kaurenoic acid demonstrated the most activity against axenic amastigote and had IC50 values of 3.5 and 4.0 µg/mL, respectively. Agathic, kaurenoic and pinifolic acid caused significant increases in plasma membrane permeability and mitochondrial membrane depolarisation of the protozoan. In conclusion, copaiba oil and its diterpene acids should be explored for the development of new antileishmanial drugs.

Antiviral activity of fractions from leaves of Piper regnelli var. pallescens

This study investigated the antiviral potential of fractions and eupomatenoid-5 obtained from Piper regnelli (Miq.) C. DC., Piperaceae, leaves against bovine herpesvirus-1 (BHV-1) and poliovirus. VERO cell monolayers in 96-well cell culture plates were infected with BHV-1 or poliovirus and incubated in the presence and absence of samples for 48 h. The cells were then fixed and stained with sulforhodamine B, and the virus-induced cytopathic effect was measured in a 96-well plate reader at 530 nm. Cytotoxicity was assessed by incubating the cell monolayers with samples for 72 h. The hexane, chloroform, chloroform/ethyl acetate (19:1), and chloroform/ethyl acetate (9:1) fractions showed activity against BHV-1. The chloroform, chloroform/ethyl acetate (19:1), chloroform/ethyl acetate (9:1), chloroform/ethyl acetate (1:1), and ethyl acetate fractions were active against poliovirus. The chloroform/ethyl acetate (9:1) fraction presented the best selectivity index for both viruses. The present study reports the antiviral activity of the extract and fractions of P. regnelli leaves.

The occurrence of Aeromonas spp. in the bottled mineral water, well water and tap water from the municipal supplies

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 percent from the mineral water, 8.3 percent from the artesian water and 6.5 percent 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 percent 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.

A porcentagem de amostras positivas para Aeromonas foi de 12.7 por cento para água mineral, 8.3 por cento para água de poço artesiano e 6.5 por cento 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 por cento, 14.8 por cento e 9.0 por cento 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.

Antimicrobial activity of Brazilian copaiba oils obtained from different species of the Copaifera genus

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.

Yeasts and filamentous fungi in bottled mineral water and tap water from municipal supplies

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 percent and 11.6 percent of the bottled mineral on water dispensers and tap water samples from municipal system, respectively. Twenty-one (35.0 percent) of bottled mineral water and two (3.3 percent) of tap water samples were positive for filamentous fungi. For bottled mineral water 12 (20.0 percent) of 60 samples were positive for total coliform, compared with 3(5.0 percent)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.

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 por cento e 11,6 por cento das amostras de água mineral de garrafão em dispensadores de água e água de torneira do sistema municipal, respectivamente. Vinte e uma (35,5 por cento) das amostras de água mineral de garrafão e duas (3,3 por cento) das amostras de água de torneira foram positivas para fungos filamentosos. Para água mineral de garrafão, 12 (20.0 por cento) das 60 amostras foram positivas para coliforme total, comparado com 3 (5.0 por cento) 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.

Atividade antileishmania do extrato hidroalcoólico e de frações obtidas de folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck / Antileishmanial activity of hydroalcoholic extract and fractions obtained from leaves of Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.)

A atividade biológica do extrato bruto e as várias frações obtidas de folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck foi avaliada em Leishmania amazonensis. Este estudo incluiu o processo de extração e o fracionamento biomonitorado pelo método de cromatografia de adsorção. Um aumento progressivo no efeito antileishmania foi observado durante o processo de purificação. O extrato hidroalcoólico solúvel em água (EBA) apresentou concentração inibitória 50 por cento (IC50) igual a 167 µg/mL enquanto o extrato hidroalcoólico solúvel em acetato de etila (EBAcOEt) mostrou um IC50 de 30 mg/mL sobre o crescimento de formas promastigotas após 48 h de cultivo. A fração hexano (FHex) apresentou uma atividade antileishmania maior que o EBAcOEt com IC50 de 21,5 µg/mL. Análises de citotoxicidade indicaram que as concentrações tóxicas do EBA, EBAcOEt e das frações foram maiores para macrófagos J774G8 do que para os protozoários.

Biological activity of the crude extract and several fractions obtained from Piper regnellii var. pallescens was assessed on Leishmania amazonensis. This study included the extraction process and bioassay-guided fractionation by the adsorption chromatography method. A progressive increase in the antileishmanial effect was observed in the course of the purification process. The hydroalcoholic extract water soluble (EBA) had a 50 percent inhibitory concentration (IC50) at 167 µg/mL whereas the hydroalcoholic extract acetate soluble (EBAcOEt) showed an IC50 of 30 µg/mL against the growth of promastigote forms after 48 h of culturing. The hexan fraction (FHex) showed an antileishmanial activity greater than EBAcOEt with IC50 at 21.5 µg/mL. Analysis of cytotoxicity indicated that the toxic concentrations of the EBA, EBAcOEt, and fractions were higher for J774G8 macrophages than for the protozoans.

Occurrence of Pseudomonas aeruginosain drinking water - DOI: 10.4025/actascibiolsci.v28i1.1053 / Ocorrência de Pseudomonas aeruginosa em água potável - DOI: 10.4025/actascibiolsci.v28i1.1053

A total of 413 samples of drinking water, taken from principal and secondary water systems in the municipality of Bandeirantes, state of Paraná, southern Brazil, were analyzed through membrane filtration for P. aeruginosa and on Plate Count agar for heterotrophic bacteria. Total and fecal coliforms were determined by multiple dilution tube methods. Turbidity, pH, chlorine, fluoride, alkalinity, color, dissolved oxygen, hardness and organic matter were also measured. Of the 413 samples examined, 10.41% were contaminated by P. aeruginosa, including 23,53% of the water from secondary water system, 8,56% of the water from principal water system. No coliform was found in any of the samples. One sample counted above the maximum heterotrophic bacteria level permitted. P. aeruginosa isolated from the water samples were tested for chlorine sensitivity and survived up to three times the minimum chlorine concentration used. It is desirable that Pseudomonas enumeration be performed periodically, in addition to data routinely collected by most water supply systems

Foram analisadas 413 amostras de água potável do sistema principal e do sistema secundário em Bandeirantes, Estado do Paraná, através de membrana filtrante para P. aeruginosa e bactérias heterotróficas em ágar Plate Count. Coliformes totais e fecais foram determinados pelo método dos tubos múltiplos. A qualidade físico-química da água foi determinada em termos de turbidez, pH, cloro, flúor, alcalinidade, cor, oxigênio dissolvido, dureza e matéria orgânica. Das amostras examinadas, 10,41% estavam contaminadas por P. aeruginosa, incluindo 23,53% das amostras do sistema de água secundário, e 8,56% das amostras do sistema de água principal. Somente uma amostra apresentou contagem de bactérias heterotróficas acima do nível máximo recomendado. P. aeruginosa isoladas foram testadas para sensibilidade ao cloro livre e sobreviveram a uma concentração de cloro três vezes acima da concentração mínima usada. É recomendada a determinação periódica de Pseudomonas em adição aos dados rotineiramente coletados na maioria dos sistemas de abastecimento

Biological effects of extracts obtained from Stryphnodendron adstringens on Herpetomonas samuelpessoai

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.

Effects of medicinal plant extracts on growth of Leishmania (L) amazonensis and Trypanosoma cruzi

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.

Bacterial microflora in the gastrointestinal tract of Nile tilapia, Oreochromis niloticus, cultured in a semi-intensive system- DOI: 10.4025/actascibiolsci.v25i2.2007 / Flora bacteriana de tilápia do Nilo, Oreochromis niloticus, cultivada em sistema semi-intensivo - DOI: 10.4025/actascibiolsci.v25i2.2007

This experiment measured total bacterial numbers in the gastrointestinal regions of semi-intensively cultured tilapia, Oreochromis niloticus L. (Cichlidae). Mean bacterial numbers were higher in both anterior and posterior gut than in stomach. The percentage of isolated species and the percentage of isolates from any particular species varied significantly among gastrointestinal tract regions. Aeromonas hydrophila, Aeromonas veronii, Burkholderia cepacia, Chromobacterium violaceum, Citrobacter freundii, Escherichia coli, Flavimonas oryzihabitans and Plesiomonas shigelloides were the most frequently isolated Gram-negative bacilli. From these samples, only Plesiomonas shigelloides was present in both gastrointestinal regions, with higher number in the posterior gut (76%), when compared to anterior gut (4.8%) and stomach (0.6%). Aeromonas hydrophila (0.6%), Escherichia coli (7.4%), and Flavimonas oryzihabitans were isolated only in the stomach, and Citrobacter freundii and Burkholderia cepacia were found only in the posterior gut. Chromobacterium violaceum was the dominant bacteria isolated from stomach and anterior gut, with 90% and 55%, respectively. Unidentified organisms comprised 0 - 39.3% of the gastrointestinal microflora

A flora bacteriana de diferentes partes do trato gastrintestinal de tilápia Oreochomis niloticus L. (Cichlidae) foi determinada. O número médio de bactérias foi maior no intestino anterior e posterior quando comparado ao estômago. A porcentagem total de espécies bacterianas isoladas e a porcentagem de espécies isoladas em uma espécie particular variaram significativamente entre as regiões do trato gastrintestinal. Aeromonas hydrophila, Aeromonas veronii, Burkholderia cepacia, Chromobacterium violaceum, Citrobacter freundii, Escherichia coli, Flavimonas oryzihabitans e Plesiomonas shigelloides foram os bacilos Gram-negativos encontrados com maior freqüência. Destas espécies, somente Plesiomonas shigelloides esteve presente em cada região do trato gastrintestinal, apresentando maior número no intestino posterior (76%), quando comparado com o intestino anterior (4.8%) e o estômago (0.6%). Aeromonas hydrophila (0.6%), Escherichia coli (7.4%), e Flavimonas oryzihabitans foram isoladas somente do estômago, e Citrobacter freundii e Burkholderia cepacia foram encontradas somente no intestino posterior. Chromobacterium violaceum foi a espécie dominante isolada do estômago e do intestino anterior com 90% e 55%, respectivamente. Organismos não identificados compreendem 0 39.3% da microbiota gastrintestinal

Haematological and biochemical values for Nile tilapia Oreochromis niloticus cultured in semi-intensive system - DOI: 10.4025/actascibiolsci.v25i2.2028 / Valores hematológicos e bioquímicos em tilápia do Nilo Oreochromis niloticus cultivada em sistema semi-intensivo - DOI: 10.4025/actascibiolsci.v25i2.2028

The aim of the present study was to investigate the reference ranges for haematological and biochemical values for Nile tilapia, Oreochromis niloticus (Cichlidae), cultivated in a semi-intensive system. Erythrocytes number was 6.93 ± 8.28 x 106/mm3, hemoglobin 10.52 ± 3.09g/dL of blood and hematocrit 31.85 ± 8.45%, mean corpuscular volume (MVC) 148.80 ±. 153.19µ3, mean corpuscular hemoglobin (MCH) 40.74 ± 34.19 pg and mean corpuscular hemoglobin concentration (MCHC) 35.24 ± 14.92%. Total plasma protein and glucose values obtained were 3.06 ± 0.65g/dl and 60.32 ± 20.22mg/dl, respectively. A correlation matrix was established to compare de degree of association among biometric data and hematological parameters, and among each of them and biochemical values. A positive correlation was observed among weight, length, MCV and MHC, as well as between length and hematocrit. The blood glucose level was positively correlated with weight and length, whereas total protein was positively correlated with hemoglobin. The erythrocytes count was positively correlated with hemoglobin and negatively correlated with MCV and MHC. Microscopy examination of blood smears revealed the presence of erythrocytes, neutrophils, lymphocytes, and monocytes. Under the conditions employed here, no basophils or eosinophils, nor their precursors, could be found in blood smears of Nile tilapia

O objetivo do presente estudo foi investigar os valores de referências hematológicos e bioquímicos para a tilápia do Nilo, Oreochromis niloticus (Cichlidae), cultivada em sistema semi-intensivo. A contagem de eritrócitos foi 6,93 ± 8,28 x 106/mm3, hemoglobina 10,52 ± 3,09g/dL de sangue, hematócrito 31,85 ± 8,45%, volume corpuscular médio (MVC) 148,80 ±.153,19µ3, hemoglobina corpuscular média (MCH) 40,74 ± 34,19pg e concentração de hemoglobina corpuscular média 35,24 ± 14,92%. Os valores de proteína plasmática total e da glicose obtidos foram de 3,06 ± 0,65g/dl e 60,32 ± 20,22mg/dl, respectivamente. Uma matriz de correlação foi estabelecida para comparar o grau de associação entre os dados biométricos e parâmetros hematológicos, e entre estes e os valores bioquímicos. Uma correlação positiva foi observada entre peso, comprimento, MCV e MHC, bem como entre comprimento e hematócrito. O nível de glicose sanguínea foi positivamente correlacionado com o peso e o comprimento, enquanto proteína total foi positivamente correlacionada com a hemoglobina. A contagem de eritrócito foi positivamente correlacionada com a hemoglobina e negativamente correlacionada com MCH e MHC. A observação microscópica do esfregaço sangüíneo revelou a presença de eritrócitos, neutrófilos, linfócitos e monócitos. Sob as condições empregadas no estudo não foram encontrados basófilos, ensinófilos ou seus precur

Antibacterial activity of Ocimum gratissimum L. essential oil

The essential oil (EO) of Ocimum gratissimum inhibited Staphylococcus aureus at a concentration of 0.75 mg/ml. The minimal inhibitory concentrations (MICs) for Shigella flexineri, Salmonella enteritidis, Escherichia coli, Klebsiella sp., and Proteus mirabilis were at concentrations ranging from 3 to 12mg/ml. The endpoint was not reached for Pseudomonas aeruginosa (>=24 mg/ml). The MICs of the reference drugs used in this study were similar to those presented in other reports. The minimum bactericidal concentration of EO was within a twofold dilution of the MIC for this organism. The compound that showed antibacterial activity in the EO of O. gratissimum was identified as eugenol and structural findings were further supported by gas chromatography/mass spectra retention time data. The structure was supported by spectroscopic methods.

Fatty Acid and Sterol Composition of Three Phytomonas Species

Fatty acid and sterol analysis were performed on Phytomonas serpens and Phytomonas sp. grown in chemically defined and complex medium, and P. françai cultivated in complex medium. The three species of the genus Phytomonas had qualitatively identical fatty acid patterns. Oleic, linoleic, and linolenic were the major unsaturated fatty acids. Miristic and stearic were the major saturated fatty acids. Ergosterol was the only sterol isolated from Phytmonas sp. and P. serpens grown in a sterol-free medium, indicating that it was synthesized de novo. When P. françai that does not grow in defined medium was cultivated in a complex medium, cholesterol was the only sterol detected. The fatty acids and sterol isolated from Phytomonas sp. and P. serpens grown in a chemically defined lipid-free medium indicated that they were able to biosynthesize fatty acids and ergosterol from acetate or from acetate precursors such as glucose or threonine