Parkia from Cerrado: phytochemical bioprospection, toxicity and in vitro bioactivities of bark and flower extracts.

Parkia platycephala is the only species of the genus Parkia that is endemic to the brazilian Cerrado and the tree symbol of the state of Tocantins, but there are still few studies regarding its bioprospecting. In this study, we aimed to investigate the phytochemical composition, toxicity and bioactivities of the bark and flower of Parkia platycephala. Hot sequential extractions (Soxhlet) were performed using methanol and hydroethanolic solution (70%), after degreasing the sample (hexane). The presence of flavonoids, tannins, steroids and alkaloids was detected in the preliminary screening. Trilinolein, (Z)-9-octadecenamide, 3-O-methyl-d-glucose were detected by Gas Chromatography coupled to Mass Spectrometry (GC-MS). In the Liquid Chromatography with Diode Array Detector (LC-PDA) analysis, it was detected exclusively ferulic acid (bark) and ellagic acid (flower). The ethanolic extract of the bark (IC50=10.69 ± 0.35 µgmL-1) has an antioxidant potential (DPPH• radical) higher than that of the rutin standard (IC50=15.85 ± 0.08 µgmL-1). All extracts showed excellent anticholinesterase potential (Ellman), with emphasis on the ethanol extract of the flower (IC50 =5.34 ± 0.12 µgmL-1). Regarding toxicity (Artemia salina), the methanolic extract of the bark and the ethanolic extract of the flower presented high and moderate levels, respectively. Such results limit the concentrations of biological activities in this study, however, the antioxidant and anticholinesterase indices fall short of toxicity. The results demonstrated promising antioxidant and anticholinesterase activities of both the bark and the flower of Parkia platycephala.

Oocyte in vitro maturation with eugenol improves the medium antioxidant capacity and total cell number per blastocyst.

This study investigates the impact of eugenol (EU) supplementation on bovine oocyte in vitro maturation (IVM) and antioxidant capacity, as well as in vitro embryo production and quality after conventional in vitro fertilization (IVF). A total of 1077 cumulus oocyte complexes were cultured in TCM-199+ without EU supplementation (control treatment) or supplemented with EU at the concentrations of 10 µM (EU-10), 20 µM (EU-20), or 40 µM (EU-40). After IVM, the oocytes were subjected to IVF and embryo culture. The addition of EU at 40 µM to the IVM medium improved (P < 0.05) the antioxidant capacity and cleavage rate when compared to the control treatment. Moreover, a positive correlation (r = 0.61, P < 0.03) was observed between cleavage rate and EU concentration. The addition of EU at concentrations of 10 and 20 µM decreased (P < 0.05) the calreticulin (CALR) levels in expanded blastocysts when compared to the control treatment and EU-40 treatment. However, the EU-10 and EU-20 treatments had a greater (P < 0.05) mean total cell number (TCN) per expanded blastocyst when compared to the control treatment and EU-40 treatment. In conclusion, the addition of EU to the enriched culture medium during IVM of bovine oocytes improved the antioxidant capacity of the spent medium, as well as the cleavage rate and embryonic quality (i.e., TCN/expanded blastocyst), and reduced the endoplasmic reticulum stress (i.e., CALR levels) in the embryos. Thus, we recommend enriching the IVM medium with 10 µM EU for in vitro bovine embryo production.

Efeito do extrato etanólico bruto e das frações da Hyptidendron canum (Pohl ex Benth. ) Harley em brânquias de Oreochromis niloticus L / Effect of the crude ethanol extract and fractions of Hyptidendron canum (Pohl ex Benth. ) Harley on the gills of Oreochromis niloticus L

Alguns problemas relacionados à criação intensiva de peixes estão mais evidentes devido ao aumento da aquicultura em todo o mundo, destacando-se os distúrbios nutricionais e o aumento de doenças nos sistemas de produção. Com isso as pesquisas envolvendo produtos derivados de plantas vem ganhando espaço nessa área. A Hyptidendron canum é uma planta utilizada na medicina popular como antimalárica, anti-inflamatória, antiulcerativa e anti-hepatotóxica. Objetivou-se, neste trabalho, avaliar os efeitos celulares e teciduais do extrato etanólico bruto e das frações hexano, clorofórmio e acetato de etila da H. canum nas brânquias de Oreochromis niloticus L. Para isso, o extrato etanólico e as frações foram administrados no peixe através da ração. Após 24 horas da ingestão da ração, os peixes foram sacrificados e o segundo arco branquial de cada um foi processado histologicamente e corado com tricômico de Masson e Hematoxilina e Eosina (HE). Pela análise qualitativa das brânquias ao microscópio de luz, observou-se que o extrato bruto e as três frações promoveram, em diferentes intensidades nas lamelas, descamação e destacamento do epitélio respiratório, alteração da curvatura, desorganização do eixo, hiperplasia celular do tecido epitelial interlamelar e vasodilatação nas lamelas e no vaso central do filamento. Os resultados deste experimento mostraram que o extrato etanólico bruto e as três frações de H. canum promoveram processos inflamatórios e/ou lesões sistêmicas, dose dependente para O. niloticus.

Some problems related to intensive fish farming are more evident because of the increased aquaculture worldwide, and we highlight the nutritional disorders and the increase of diseases in the production systems. Thus, research involving plant-derived products has been increasing in this area. The Hyptidendron canum is a plant used in folk medicine as an antimalarial, anti-inflammatory, anti-ulcer and antihepatotoxic agent. This study proposes to evaluate the cellular and tissue effects of the crude ethanol extract and the hexane, chloroform and ethyl acetate fractions of H. canum on the gills of Oreochromis niloticus L. The ethanol extract and fractions were administered through the feed of the fish. Twenty-four hours later, the fish were sacrificed and their second gill arch dissected. Histological analyses were performed using Masson's trichrome and Haematoxylin and Eosin (H&E). The qualitative analysis of the material showed that the crude extract and the three fractions caused varying degrees of peeling and detachment of the lamellar respiratory epithelium, alterations in the curvature and axis of the gill, cell hyperplasia in the interlamellar epithelium and lamellar vasodilation, as well as vasodilation in the central vessel of the filament. The results of this experiment showed that the crude ethanol extract and the three fractions of H. canum promoted inflammation and/or systemic lesions, dose dependent, for O. niloticus.

Activity of cycloartane-type triterpenes and sterols isolated from Musa paradisiaca fruit peel against Leishmania infantum chagasi.

UNLABELLED: The aim of the study was to evaluate in vitro the antileishmanial activity of triterpenes and sterols isolated from Musa paradisiaca (banana) fruit peel used traditionally to treat leishmaniasis. The compounds were isolated from the ethanolic extract of the peel of the banana fruit by column chromatography. The chemical structure of compounds was determined by (1)H and (13)C - nuclear magnetic resonance spectroscopy. The cytotoxicity was measured in RAW 264.7 cells and LLC-MK2. Leishmanicidal activity against L. infantum chagasi promastigotes was performed by the MTT colorimetric method and activity against amastigotes was assayed in mammalian cells using in situ ELISA method. Five compounds were identified, consisting of three triterpenes: cycloeucalenone, 31-norcyclolaudenone and 24-methylene-cicloartanol and a mixture of two sterols: beta-sitosterol and stigmasterol. With the exception of cycloeucalenone, all compounds showed statistically similar activity against promastigote to pentamidine. While, acting against amastigotes, excluding 31-norcyclolaudenone, other compounds showed activity similar to amphotericin B. All compounds showed low cytotoxicity in mammalian cells. CONCLUSION: This study partially confirms the use of Musa paradisiaca in folk medicine against leishmaniasis. Further in vivo studies are necessary to evaluate the efficacy.

Correlação entre as atividades antiradical, antiacetilcolinesterase e teor de fenóis totais de extratos de plantas medicinais de farmácias vivas / Correlation between the anti-radical, anti-acetylcholinesterase activities and phenolic content of some extracts of live herbal pharmacies in the Northeast of Brazil

O objetivo deste trabalho foi avaliar a correlação entre o conteúdo de fenóis de extratos de 18 plantas medicinais comuns nas Farmácias Vivas com a atividade sequestradora do radical livre DPPH e com a inibição da enzima acetilcolinesterase. Maiores conteúdos de fenóis totais foram encontrados nas plantas: Eugenia uniflora, Lippia alba, Lippia microphylla, Mentha x villosa, Ocimum gratissimum e Ocimum selloi. As espécies que apresentaram maior atividade antioxidante, semelhante a quercetina, foram: Spondias mombim, Malphigia glabra, L. alba e Croton zenhtneri,. As plantas Cecropia pachystachia, L. alba, L. microphylla, M. glabra, O. gratissimum e Plectranthus ornatus mostraram maiores halos de inibição da enzima acetilcolinesterase. Combinando as duas ações, as plantas mais ativas foram a M. glabra e a L. alba e somente a segunda correlaciona-se diretamente com o maior teor de fenóis totais. Através deste estudo, M. glabra e L. alba foram consideradas as plantas mais promissoras para estudos subsequentes a fim de encontrar novos compostos com ação potencial contra a doença de Alzheimer.

The purpose of this study was to evaluate the phenolic content of 18 medicinal plant extracts of "Live Pharmacies" and correlate them with their free radical scavenging activity and inhibition of the acetylcholinesterase enzyme. Higher amounts of phenolic compounds were found in the following plants: Eugenia uniflora, Lippia alba, Lippia microphylla, Mentha x villosa, Ocimum gatissimum, and Ocimum selloi. The species that showed higher anti-radical activity were Croton zenhtneri, Lippia alba, Malphigia glabra, and Spondias mombim, similar to quercetin. The plants Cecropia pachystachia, L. alba, L. microplylla, M. glabra, O. gratissimum, and Plectranthus ornatus showed larger inhibition zones of the acetylcholinesterase enzyme. By correlating the two actions L. alba and M. glabra are common as more active to both tests and only L. alba correlates directly with the phenolic content. Through this study, L. alba and M. glabra are considered the most promising plants for further studies to find new compounds with potential action against Alzheimer's disease.

Toxicity of Brazilian plant seed extracts to two strains of Aedes aegypti (Diptera: Culicidae) and nontarget animals.

Seed ethanolic extracts of 21 Brazilian plants were evaluated for ovicidal, larvicidal, and pupicidal activities against insecticide-susceptible (SS) and field-collected (FC) strains of Aedes aegypti (L.) (Diptera: Culicidae), as well as for their effects on nontarget organisms. Myracrodruon urundeuva Fr. Allemao extract was highly toxic to both mosquito strains. Schinopsis brasiliensis Engler extract showed low toxicity and was 38-68 times less toxic to Ae. aegypti larvae than was M. urundeuva extract. The pupicidal activity (LC50) of 14 plant seed extracts ranged between 9 and 433/g/ml, and toxicities were comparable to both mosquito strains. Piptadenia moniliformis Benth. and Luetzelburgia auriculata (Allemao) Ducke extracts showed the highest activities against pupae of FC and SS strains. None of the extracts showed 100% ovicidal activity. In addition, the active extracts did not show high acute toxicity to mice (LD50 > 1.5 g/kg), except that of Enterolobium contortisiliquum (Vell.) Morong. Most of the active extracts exhibited low toxicity against brine shrimp (Artemia sp.) nauplii. The extracts of M. urundeuva, P. moniliformis, and L. auriculata are promising sources of recognized classes of insecticidal compounds with good selectivity against immature stages of Ae. aegypti.

Análise tecidual e celular das brânquias de Oreochromis niloticus L. tratadas com extrato etanólico bruto e frações das folhas da pitanga (Eugenia uniflora L. ) - Myrtaceae / Tissue and cell analysis of Oreochromis niloticus L. gill treated with crude ethanol extract and fractions from pitanga (Eugenia uniflora L. ) leaves Myrtaceae

Eugenia uniflora L. (Myrtaceae) é uma planta que ocorre no bioma Cerrado e é utilizada popularmente no tratamento de diarréias, inflamações, hiperglicemia e hipertensão. Estudos prévios revelaram atividade antimicrobiana da E. uniflora in vitro. Tendo em vista o uso popular, este trabalho objetivou avaliar as possíveis atividades celulares e teciduais sistêmicas do extrato bruto e das frações das folhas dessa planta em brânquias de Oreochromis niloticus L. (tilápia nilótica). Para isso, o extrato etanólico e as frações das folhas dessa planta foram administrados no peixe, por via oral, adicionadas à ração. Após um período de 24 horas, os peixes foram sacrificados e o segundo arco branquial de cada peixe foi dissecado, fixado em formalina neutra, desidratado, incluído em parafina e cortado. Nas análises histológicas, utilizaram-se tricômico de Masson e hematoxilina e eosina (HE). Pelas análises qualitativas na microscopia de luz, concluiu-se que o extrato etanólico bruto e as frações das folhas da E. uniflora apresentaram efeito sistêmico nas tilápias nilóticas atingindo as brânquias. As ações tóxicas como destacamento e descamação do epitélio respiratório e hiperplasia das células do epitélio interlamelar, foram mais pronunciadas nas tilápias que ingeriram maiores concentrações. Este trabalho colaborou para identificar o efeito vasodilatador dessa planta, e contribuiu para estabelecer a tilápia nilótica como sistema-modelo para testes com princípios ativos de plantas. Espera-se, com esses testes, viabilizar o uso de plantas como medicamentos para tratamentos de peixes, a manutenção da saúde de animais em cultivo intensivo e extensivo, a partir do qual se possibilite emprego alternativo aos medicamentos sintéticos.

Eugenia uniflora L. (Myrtaceae) is a plant found in the Cerrado biome and traditionally used in the treatment of diarrheas, inflammations, hyperglycemia and hypertension. Previous studies have revealed in vitro antimicrobial activity of E. uniflora. Considering its popular use, this study aimed to assess possible systemic cellular and tissue activities of the crude extract and the fractions from the leaves of this plant on Oreochromis niloticus L. (Nile tilapia) gill. Thus, ethanol extract and fractions from the leaves of this plant were orally administered to the fish in their rations. After 24 hours, the fish were sacrificed and the second gill arch of each fish was dissected, fixed in neutral formalin, dehydrated, embedded in paraffin and sectioned. Masson's trichome and hematoxylin and eosin (HE) were used in the histological analyses. Qualitative analyses using a light microscope led to the conclusion that the crude ethanol extract and the fractions from E. uniflora leaves presented systemic effect on Nile tilapias, affecting the gills. Toxic actions such as respiratory epithelium detachment and lifting, and hyperplasia of interlamellar epithelial cells were more pronounced in the tilapias that ingested higher concentrations. This study helped to identify the vasodilator effect of this plant and contributed to the definition of the Nile tilapia as a model system for testing plant active principles. These tests are expected to make feasible not only the use of plants as fish medication but also the maintenance of the health of animals in intensive and extensive cultures through the possible use of alternatives to synthetic medication.

Antiviral activities of extracts and phenolic components of two Spondias species against dengue virus

In recent years, the search for natural plant products to fight viral diseases has been increasing. In this work, two Spondias species, namely S. mombin and S. tuberosa, found in Ceará state (Brazil), and their main phenolic components were evaluated against dengue virus. In vitro antiviral tests were performed against type-2 dengue virus by the MTT method and standard cytopathic effect reduction assay in C6/36 cells. Cytotoxicity was also evaluated by MTT. The presence of phenolic compounds quercetin, rutin, and ellagic acid in plant extracts was characterized by HPLC analysis. Both Spondias species extracts and components were nontoxic to the cells whereas rutin and quercetin displayed relevant antiviral activity with IC50 of 362.68 µg/mL and 500 µg/mL, respectively.

Extratos vegetais usados no controle de dípteros vetores de zoonoses / Plant extracts used in the control of dipteran vectors of zoonosis

Os insetos são importantes transmissores de doenças que cada vez mais ocorrem em áreas periurbanas ou urbanas, devido à introdução ou reaparecimento de seus respectivos vetores. As estratégias de controle das doenças transmissíveis por vetores biológicos são de difícil execução, principalmente quando associadas à existência de reservatórios domésticos e silvestres e aos aspectos ambientais. O uso de inseticidas direcionado ao combate de formas adultas de dípteros vetores tem sido freqüente. Entretanto, o amplo uso dos inseticidas sintéticos desde a descoberta do DDT para o controle de pragas domésticas e da agricultura, como também de vetores que transmitem doenças ao homem, levou a uma maior preocupação em relação à toxicidade e impacto ambiental destes agentes. Além disso, a resistência a inseticidas tornou-se uma preocupação crescente na agricultura, economia e na saúde pública. Um grande número de diferentes espécies de plantas representando diferentes áreas geográficas ao redor do mundo tem se mostrado capaz de causar efeitos letais e subletais sobre insetos. Esta revisão aborda a utilização de extratos de plantas com potencial no controle de dípteros vetores de zoonoses.

Insects are important transmitters of diseases increasingly occurring in peri-urban or urban areas due to the introduction or the reemergence of their respective vectors. Strategies to control diseases transmitted by biological vectors are difficult to implement, especially when associated with the existence of domestic and wild reservoirs and environmental aspects. The use of insecticides against adult forms of dipteran vectors has been frequent. However, the widespread use of synthetic insecticides since the discovery of DDT to control domestic and agricultural pests, as well as vectors that transmit diseases to humans, has led to greater concern for the toxicity and environmental impact of these agents. Moreover, resistance to insecticides has become an increasing concern in agriculture, economy and public health. A large number of different plant species representing different geographical areas around the world have been capable of causing lethal and sublethal effects on insects. This review addresses the use of potential plant extracts to control dipteran vectors of zoonosis.

Croton sonderianus essential oil samples distinctly affect rat airway smooth muscle.

Plants of the genus Croton have been used extensively in the northeast of Brazil for treating various clinical conditions. Previous studies have demonstrated that the essential oil of some specimens of Croton sp. have a relaxing effect on tracheal smooth muscle. Our study aimed to characterize the effects of Croton sonderianus essential oil samples, collected at 1:00 pm (EO-13) and 9:00 pm (EO-21), on rat tracheal smooth muscle. The two samples were submitted to gas chromatography (GC) and mass spectrometry (MS) analysis to identify their components. Rat tracheal smooth muscle strips were used to assess the biological activity. The major constituents of EO-21 were: spathulenol (18.32%), beta-caryophyllene (14.58%) and caryophyllene oxide (8.54%) and the major constituents of EO-13 were bicyclogermacrene (16.29%), beta-phellandrene (15.42%) and beta-caryophyllene (13.82%). These samples showed an antispasmodic effect on tracheal smooth muscle strips pre-contracted with high K+ concentration (80 mM) or with acetylcholine. EO-21 increased baseline tonus while EO-13 provoked a decrease. These results demonstrated that EO-13 and EO-21 have different chemical composition and showed myorelaxant activity. In conclusion, EO-13 and EO-21 may have potential therapeutic use in the treatment of bronchospasm.

Chemical composition of Eucalyptus spp. essential oils and their insecticidal effects on Lutzomyia longipalpis.

The chemical composition of essential oils from three species of plants belonging to the Eucalyptus genus was determined and, their insecticidal effects on egg, larva and adult phases of Lutzomyia longipalpis were assessed. The insects were collected in the municipality of Sobral in the State of Ceará, Brazil. Five treatments with different concentrations were performed along with two negative controls, distilled water and Tween 80 (3%), and a positive control, cypermethrin (0.196mg/ml). The tests were carried out in plastic pots internally coated with sterile plaster and filled with a substrate made of rabbit feces and crushed cassava leaves. The eggs, larvae and adults were sprayed with the oils. The hatched larvae were counted for 10 consecutive days and observed until pupation. Insect mortality was observed after 24, 48 and 72h. E. staigeriana oil was the most effective on all three phases of the insect, followed by E. citriodora and E. globulus oils, respectively. The major constituents of the oils were Z-citral and alpha-citral (E. staigeriana), citronellal (E. citriodora) and 1,8-cineole (E. globulus). The Eucalyptus essential oils constitute alternative natural products for the control of L. longipalpis since the median effective concentration (EC(50)) values revealed relevant action as compared with other natural products, some of their chemical constituents are already known for their insecticidal activity and these oils are produced in commercial scale in Brazil.

Anthelmintic activity of Cocos nucifera L. on intestinal nematodes of mice.

In this study, we evaluated the anthelmintic activity of the liquid extracted from the bark of the green coconut (LBGC), as well as butanol extract obtained from LBGC, on mouse intestinal nematodes. Thirty-six naturally infected mice were distributed into six groups receiving the following treatments: Group I: 1000 mg/kg of LBGC; Group II: 2000 mg/kg of LBGC; Group III: 500 mg/kg of butanol extract; Group IV: 1000 mg/kg of butanol extract; Group V: 0.56 mg/kg febendazole; and Group VI: 3% dimethylsulfoxide. The chemical composition of the LBGC and its butanol extract was determined by phytochemical tests. A dose of 1000 mg/kg of butanol extract had 90.70% efficacy in reducing the mouse worm burden (p<0.05). Phytochemical tests revealed the presence of triterpens, saponnins and condensed tannins in the LBGC and butanol extracts. These results suggest that Cocos nucifera extracts may be useful in the control of intestinal nematodes.

Anthelmintic activity of Cocos nucifera L. against sheep gastrointestinal nematodes.

The development of anthelmintic resistance has made the search for alternatives to control gastrointestinal nematodes of small ruminants imperative. Among these alternatives are several medicinal plants traditionally used as anthelmintics. This work evaluated the efficacy of Cocos nucifera fruit on sheep gastrointestinal parasites. The ethyl acetate extract obtained from the liquid of green coconut husk fiber (LGCHF) was submitted to in vitro and in vivo tests. The in vitro assay was based on egg hatching (EHT) and larval development tests (LDT) with Haemonchus contortus. The concentrations tested in the EHT were 0.31, 0.62, 1.25, 2.5 and 5 mg ml(-1), while in the LDT they were 5, 10, 20, 40 and 80 mg ml(-1). The in vivo assay was a controlled test. In this experiment, 18 sheep infected with gastrointestinal nematodes were divided into three groups (n=6), with the following doses administered: G1-400 mg kg(-1) LGCHF ethyl acetate extract, G2-0.2 mg kg(-1) moxidectin (Cydectin) and G3-3% DMSO. The worm burden was analyzed. The results of the in vitro and in vivo tests were submitted to ANOVA and analyzed by the Tukey and Kruskal-Wallis tests, respectively. The extract efficacy in the EHT and LDT, at the highest concentrations tested, was 100% on egg hatching and 99.77% on larval development. The parameters evaluated in the controlled test were not statistically different, showing that despite the significant results of the in vitro tests, the LGCHF ethyl acetate extract showed no activity against sheep gastrointestinal nematodes.

Anthelmintic activity of Lippia sidoides essential oil on sheep gastrointestinal nematodes.

Medicinal plants have been investigated for their anthelmintic properties and shown to be effective against eggs and larvae of gastrointestinal nematodes. The aim of this study was to evaluate the efficacy of the Lippia sidoides essential oil (LsEO) on sheep gastrointestinal nematodes. Initially, 44 naturally infected sheep were divided and treated with 200 microg kg(-1) ivermectin and 230 and 283 mg kg(-1) LsEO, respectively, plus the control. Fecal samples were collected from each animal to determine epg at 7, 14 and 21 days after treatment. In another test, 21 sheep were distributed and treated with 200 microg kg(-1) ivermectin, 283 mg kg(-1) LsEO and the control, respectively. Seven days after treatment, they were euthanized and necropsied to count and identify the nematodes from the abomasum, small and large intestines. In the first test, the efficacy of 230 and 283 mg kg(-1) LsEO and ivermectin was 38%, 45.9% and 40.2%, respectively, 7 days after treatment, and 30%, 54% and 39.6%, respectively, 14 days after treatment. In the second experiment, the respective efficacy of 283 mg kg(-1) LsEO and ivermectin was 56.9% and 34.4% against Haemonchus spp., and 39.3% and 63.6% against Trichostrongylus spp.

Antifungal activity of essential oils of Croton species from the Brazilian Caatinga biome.

AIMS: To find new antifungal agents among essential oils from Brazilian Croton species. METHODS AND RESULTS: Plant leaves were steam distilled and the obtained essential oils were analyzed by gas chromatography/mass spectroscopy. The main constituents were estragole and anethole for Croton zehntneri, methyl-eugenol and bicyclogermacrene for Croton nepetaefolius and spathulenol and bicyclogermacrene for Croton argyrophylloides. The antifungal activity of essential oils was evaluated against Candida albicans, Candida tropicalis and Microsporum canis by the agar-well diffusion method and the minimum inhibitory concentration (MIC) by the broth microdilution method. Essential oils of Croton species demonstrated better activity against M. canis. Among the three plants C. argyrophylloides showed the best results, with MIC ranging from 9 to 19 microg ml(-1). The acute administration of the essential oil up to 3 g kg(-1) by the oral route to mice was devoid of overt toxicity. CONCLUSIONS: The studied essential oils are active in vitro against the dermatophyte M. canis and present relative lack of acute toxicity in vivo. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of its antifungal activity and low toxicity, the essential oils of studied Croton species are promising sources for new phytotherapeutic agents to treat dermatophytosis.

Evaluation of antimicrobial activity of the crude ethanol extract of Eugenia uniflora L. leaves

Eugenia unifl ora L. is a tree whose leaves are used in popular medicine as an antihypertensive, antimicrobial drug, in the treatment of bronchitis, influenza and as an antipyretic. This paper reports the antimicrobial activity of a crude ethanol extract of E. unifl ora L. leaves. The crude extract was prepared from material collected in Goiânia, Goiás (Brazil), dried, pulverized and subjected to phytochemical screening. The antimicrobial activity was tested against spore-forming and non-sporing Gram-positive and Gram-negative bacteria as well as Candida albicans, using the well-diffusion test and the agar dilution method to determine the minimum inhibitory concentration (MIC). The phytochemical screening showed the presence of tannins, steroids, triterpenes, heterosides, anthraquinones, flavonoids and saponins. Antimicrobial activity testing showed that the crude E.uniflora L. leaf extract inhibited all the Gram-positive bacteria (MICs from 0.273 to 8.75 mg/mL), among which the spore-formers exhibited MICs from 1.094 to 2.187 mg/mL. The MIC for most Gram-negative bacteria varied from 4.375 to 17.5 mg/mL. C. albicans NTC 2010 (MIC of 0.547 mg/mL) inhibition was also noted. The antimicrobial activity found in this in vitro study of E. uniflora L. may justify its popular use as a medicine.

Anthelmintic activity of Croton zehntneri and Lippia sidoides essential oils.

Because of the development of anthelmintic resistant populations, the search for new drugs is essential to maintain the productivity of small ruminants. The aim of this study was to evaluate the anthelmintic activity of Croton zehntneri and Lippia sidoides essential oils and their major constituents, anethole and thymol. The effects of these oils and their constituents were determined by in vitro assays with the eggs and larvae of the sheep gastrointestinal nematode Haemonchus contortus. The two essential oils were evaluated on intestinal nematodes of mice at 800 mg kg(-1) dose. In the last experiment, the mice were treated with larger doses of L. sidoides, 1200 and 1600 mg kg(-1). The essential oils and their constituents prevented more than 98% of the H. contortus eggs from hatching at a concentration of 1.25 mg ml(-1) and inhibited more than 90% of H. contortus larval development at a concentration of 10 mg ml(-1). At a concentration of 800 mg kg(-1), the two essential oils were 46.3% and 11.64% effective against Syphacia obvelata and Aspiculuris tetraptera. At 1200 and 1600 mg kg(-1), L. sidoides essential oil's efficacy on the mouse worm burden was 57.6% and 68.9%, respectively. The fact that L. sidoides essential oil was almost 70% effective against mouse intestinal nematodes indicates it should be evaluated against gastrointestinal nematodes of sheep and goats.

Essential oil of croton nepetaefolius and its main constituent, 1,8-cineole, block excitability of rat sciatic nerve in vitro.

1. The effects of the essential oil of Croton nepetaefolius (EOCN) and its major constituent, 1,8-cineole, on the compound action potential (CAP) of nerve were investigated. 2. Experiments were performed in sciatic nerves dissected from Wistar rats, mounted in a moist chamber and stimulated at a frequency of 0.2 Hz, with electric pulses of 100 micros duration at 20-40 V. Evoked CAP were displayed on an oscilloscope and recorded on a computer. The CAP control parameters were as follows: peak-to-peak amplitude 8.1 +/- 0.6 mV (n = 15); conduction velocity 83.3 +/- 4.2 m/s (n = 15); chronaxie 58.0 +/- 6.8 msec (n = 6); and rheobase 2.8 +/- 0.1 V (n = 6). 3. Lower concentrations of EOCN (100 and 300 microg/mL) and 1,8-cineole (153 and 307 microg/mL; i.e. 1 and 2 mmol/L, respectively) had no significant effects on CAP control parameters throughout the entire recording period. However, at the end of 180 min exposure of the nerve to the drug, peak-to-peak amplitude was significantly (P < 0.05) reduced to 27.4 +/- 6.7 and 1.7 +/- 0.8% of control values by 500 and 1000 microg/mL EOCN, respectively (n = 6), and to 76.5 +/- 4.4, 70.0 +/- 3.9 and 14.8 +/- 4.1% of control values by 614, 920 and 1227 microg/mL (i.e. 4, 6 and 8 mmol/L) 1,8-cineole, respectively (n = 6). Regarding conduction velocity, at the end of the 180 min exposure period, this parameter was significantly reduced to 85.8 +/- 7.3 and 48.7 +/- 12.3% (n = 6) of control values by 500 and 1000 microg/mL EOCN, respectively, and to 86.4 +/- 4.5 and 76.1 +/- 5.2% (n = 6) by 920 and 1227 microg/mL 1,8-cineole, respectively. Chronaxie and rheobase were significantly increased by the higher concentrations of both EOCN and 1,8-cineole. 4. It is concluded that EOCN and its main constituent 1,8-cineole block nerve excitability in a concentration-dependent manner, an effect that was totally reversible with 1,8-cineole but not with EOCN. This suggests that other constituents of EOCN, in addition to 1,8-cineole, may contribute to the mediation of this effect of EOCN.

Ovicidal and larvicidal activity of Melia azedarach extracts on Haemonchus contortus.

Haemonchus contortus is responsible for severe economic losses in sheep and goat breeding in the Northeast of Brazil. However, the effectiveness of control is compromised due to anthelmintic resistance and misuse. In the search for natural anthelmintics, Melia azedarach L., a plant indigenous to India but now distributed throughout Brazil, was selected due to the reported anthelmintic properties of its seeds. The aim of this study was to evaluate the anthelmintic activity of the seed and leaf extracts of the Brazilian adapted plant and investigate the type of organic chemical compounds present in the most active extracts. The ovicidal and larvicidal activity of M. azedarach extracts on H. contortus was evaluated through egg hatching and larval development tests. Hexane and ethanol extracts of seeds and chloroform and ethanol extracts of leaves of M. azedarach were used in the tests. To perform the larval development test, feces of an animal free from parasites were mixed with third instar H. contortus larvae and extracts in several concentrations. The coprocultures were incubated for 7 days at 30 degrees C, then the larvae were recovered and counted. LC50 was calculated by probits using the SPSS 8.0 program. The seed ethanol extract was the most active on eggs (LC50=0.36mgmL(-1)) and the leaf ethanol extract showed the best inhibition of larval development (LC50=9.18mgmL(-1)). Phytochemical analysis of the most active extracts revealed the presence of condensed tannins, triterpenes and alkaloids.

Anthelmintic activity of Azadirachta indica A. Juss against sheep gastrointestinal nematodes.

Gastrointestinal nematode control has been performed through use of anthelmintics. However, the development of resistant populations has required research into new alternatives. There are popular reports about anti-parasitic activity of Azadirachta indica in animals and plants. The aim of this study was to evaluate the anthelmintic activity of A. indica after feeding sheep with the dried leaves. In this experiment, 40 sheep were allotted into four treatment groups. Group I received a treatment of A. indica dry leaves mixed in a concentrate at a rate of 0.1 g/kg dose for 3 months. Group II was treated with double the dose of Group I. Group III was treated with closantel (Diantel) at the manufacturer-recommended dose once at the beginning of the study and Group IV was not treated. To compare treatment effects, the following parameters were evaluated: egg count per gram of feces (EPG), worm burden, weight gain and haematocrit. EPG and worm burden results were statistically evaluated using the Kruskal-Wallis test. Haematocrit and live weight gain were submitted to analysis of variance (ANOVA) and the means evaluated by Tukey's test with 95% probability. None of the evaluated parameters of the treatment groups were statistically different when compared to the control group, demonstrating that, with the protocol used, A. indica has no anthelmintic effect.