Dietary supplementation with Lippia sidoides essential oil improves organ integrity but the specific activity of antioxidant enzymes is dose-dependent in Danio rerio.

The nutritional quality of food can affect the health of animals. This study examined the effects of dietary supplementation with Lippia sidoides essential oil (LSEO) on the physiology of Danio rerio. Four hundred fourty-eight fish were divided into 28 tanks and subjected to different dietary treatments: a control group with no supplementation, a group with grain alcohol supplementation and five groups with LSEO at concentrations of 0.25%, 0.50%, 0.75%, 1.00% and 1.25%. After 15 days, histological and enzymatic analyses were conducted. The 0.25% LSEO group exhibited lower glutathione peroxidase and catalase activity compared to the 1.00% group. Additionally, fish in the 0.25% LSEO group showed improved liver, kidney and splenic integrity indices. These findings support the inclusion of 0.25% LSEO in the diet of D. rerio, suggesting potential benefits for fish physiology and encouraging further research on phytotherapeutics in fish diets.

Efficacy of essential oil from ginger (Zingiber officinale) for anesthesia and transport sedation of pacu (Piaractus mesopotamicus).

This study evaluated the anesthetic and sedative effects of the essential oil of Zingiber officinale (EOZO) on juvenile pacu (Piaractus mesopotamicus). Experiment 1 evaluated concentrations of 0, 50, 100, 200 and 400 mg L-1 EOZO for times of induction and recovery from anesthesia. Furthermore, hematological responses and residual components of EOZO in plasma were determined immediately after anesthesia. Experiment 2 evaluated the effect of 0, 10, 20 and 30 mg L-1 EOZO on water quality, blood variables and residual components of EOZO in plasma and tissues (muscle and liver) immediately after 2 h of transport. Survival was 100%. The three main compounds of EOZO [zingiberene (32.27%), ß-sesquiphellandrene (18.42%) and ß-bisabolene (13.93%)] were observed in animal plasma and tissues (muscle and liver) after anesthesia and transport, demonstrating a direct linear effect among the evaluated concentrations. The concentration of 200 mg L-1 EOZO promoted surgical anesthesia of pacu and prevented an increase in monocyte and neutrophil levels, yet did not alter other hematological parameters. The use of 30 mg L-1 EOZO has a sedative effect on juvenile pacu, thereby reducing oxygen consumption during transport. Furthermore, the use of 30 mg L-1 EOZO in transport water prevented an increase in hemoglobin and hematocrit, with minimal influences on other blood variables.

Prospecting of essential oils in combination with florfenicol against motile Aeromonas isolated from tambaqui (Colossoma macropomum).

Natural products with antimicrobial activity and their association with synthetic antimicrobials are a sustainable option in fish farming. The objective of this study was to determine antimicrobial activity, antibiofilm potential and synergism of five essential oils (EOs) with florfenicol against motile Aeromonas isolated from Amazonian Colossoma macropomum. As their major constituent, the EOs of the species of Aloysia triphylla, Croton cajucara (red and white morphotype), Cymbopongo citratus and Lippia gracilis present ß-pinene (22.1%), germacrene D (11.5%), linalool (23%), geranial (45.7%) and carvacrol (42.2%), respectively. The EOs of L. gracilis and C. citratus showed the best antimicrobial activities against the Aeromonas strains (5 mg mL-1). All EOs interfered with biofilm formation and consolidated biofilm. The EOs of A. triphylla, C. citratus and L. gracilis showed a synergistic effect with florfenicol, reducing the amount of the chemical into the water systems while treatment.

Ovicidal effect of essential oils of Lippia alba, Lippia sidoides and Lippia gracilis on the acanthocephalan Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae).

Acanthocephalosis in tambaqui Colossoma macropomum culture is a limiting factor for the production of the species, being the most recurrent, which makes it necessary to develop strategies to control parasitosis even in the early stages of parasite development. This study aimed to elucidate the morphological characteristics of eggs of the acanthocephalan Neoechinorhynchus buttnerae and the ovicidal effect of the essential oils of Lippia alba, Lippia sidoides, and Lippia gracilis. The parasites were collected from hosts naturally infected with N. buttnerae, and the eggs obtained from the female 'parasites' abdominal cavity were added to 12-well plates containing the essential oils at different dilutions. After the eggs (N = 300) were separated and measured (length and width) with the aid of an optical microscope coupled to a digital program, the viability of the treated eggs was monitored 24 h after treatments, L. gracilis essential oil was 100% effective at the highest concentration, whereas L. sidoides essential oil showed over 50% efficacy at 2 mg/ml, reaching 100% efficacy at all higher concentrations. Lippia. alba essential oil did not show satisfactory efficacy in preventing N. buttnerae egg hatching.

Blood biochemical parameters and melanomacrophage centers in Nile tilapia fed essential oils of clove basil and ginger.

Nile tilapia (Oreochromis niloticus) is the most farmed freshwater fish in the world, however, disease outbreaks are the main cause of losses in production. Due to this, there is an increasing interest in natural products for enhancing disease resistance, without causing physiological impairment. The aim of this study was to verify the effect of the essential oils of clove basil (Ocimum gratissimum) and ginger (Zingiber officinale) on physiological and immunological parameters of nile tilapia. After 35 and 55 days of supplementation (0.5%, 1.0% and 1.5%), blood collection was performed for determination of metabolites (glucose, cholesterol, triglycerides, serum total protein and immunoglobulins) and lysozyme activity. Spleen tissue was also collected at 55 days for analyzing melanomacrophages centers. At 35 days, cholesterol showed significant reduction in the treatments 0.5% and 1.5 ginger and 1.5% clove basil. The serum triglycerides showed significant reduction in these same treatments, in addition to 1.0% clove basil. At 55 days all the supplemented groups showed significant reduction for both parameters. The lysozyme activity at 35 days was significantly higher in fish supplemented with 1.0% ginger and showed no difference among the treatments at 55 days. Regarding the melanomacrophage centers, at 35 days increased number in fish supplemented with 0.5% clove basil was observed but with no significant difference in the total percent area of spleen occupied by centers. The levels of glucose, total serum protein and immunoglobulins did not show significant differences in both sampling times. The addition of essential oils showed to be useful for improving physiological status of the fish, without excessive activation of defense mechanisms.

In vivo antimalarial activity and mechanisms of action of 4-nerolidylcatechol derivatives.

4-Nerolidylcatechol (1) is an abundant antiplasmodial metabolite that is isolated from Piper peltatum roots. O-Acylation or O-alkylation of compound 1 provides derivatives exhibiting improved stability and significant in vitro antiplasmodial activity. The aim of this work was to study the in vitro inhibition of hemozoin formation, inhibition of isoprenoid biosynthesis in Plasmodium falciparum cultures, and in vivo antimalarial activity of several 4-nerolidylcatechol derivatives. 1,2-O,O-Diacetyl-4-nerolidylcatechol (2) inhibited in vitro hemozoin formation by up to 50%. In metabolic labeling studies using [1-(n)-(3)H]geranylgeranyl pyrophosphate, diester 2: significantly inhibited the biosynthesis of isoprenoid metabolites ubiquinone 8, menaquinone 4, and dolichol 12 in cultures of P. falciparum 3D7. Similarly, 2-O-benzyl-4-nerolidylcatechol (3) significantly inhibited the biosynthesis of dolichol 12. P. falciparum in vitro protein synthesis was not affected by compounds 2 or 3. At oral doses of 50 mg per kg of body weight per day, compound 2 suppressed Plasmodium berghei NK65 in infected BALB/c mice by 44%. This in vivo result for derivative 2 represents marked improvement over that obtained previously for natural product 1. Compound 2 was not detected in mouse blood 1 h after oral ingestion or in mixtures with mouse blood/blood plasma in vitro. However, it was detected after in vitro contact with human blood or blood plasma. Derivatives of 4-nerolidylcatechol exhibit parasite-specific modes of action, such as inhibition of isoprenoid biosynthesis and inhibition of hemozoin formation, and they therefore merit further investigation for their antimalarial potential.

Lippia sidoides essential oil at concentration of 0.25% provided improvements in microbiota and intestine integrity of Danio rerio.

The study evaluated the effects of dietary supplementation with Lippia sidoides essential oil on the microbiota and intestinal morphology of Danio rerio. For this, 448 fish were randomly distributed in 28 tanks divided into a control group fed a commercial diet without supplementation, a group fed a commercial diet containing grain alcohol and five groups fed a commercial diet containing essential oil of L. sidoides (LSEO) at concentrations of 0.25%, 0.50%, 0.75%, 1.00% and 1.25%. After the period of dietary supplementation, biological materials were collected for microbiological and histological analyses. There were no significant differences regarding the microbiological count between the groups. Diversity of the microbiome was higher in 0.25% group than in control group. LSEO inhibited the growth of potentially pathogenic bacteria. Fish fed LSEO0.25% showed greater intestinal histomorphometric indices. The inclusion of LSEO at 0.25% in the diet of D. rerio provided improvements in fish microbiota and intestine integrity.

O estudo avaliou os efeitos da suplementação dietária com óleo essencial de Lippia sidoides sobre a microbiota e a morfologia intestinal de Danio rerio. Para isso, 448 peixes foram distribuídos aleatoriamente em 28 tanques divididos em grupo controle alimentado com dieta comercial sem suplementação, grupo alimentado com dieta comercial contendo álcool de cereais e cinco grupos alimentados com dieta comercial contendo óleo essencial de L. sidoides (LSEO) em concentrações de 0,25%, 0,50%, 0,75%, 1,00% e 1,25%. Após o período de suplementação alimentar, foram coletadas amostras biológicas para análises microbiológicas e histológicas. Não houve diferenças significativas quanto à contagem microbiológica entre os grupos. A diversidade do microbioma foi maior no grupo de 0,25% do que no grupo controle. LSEO inibiu o crescimento de bactérias potencialmente patogênicas. Peixes alimentados com LSEO 0,25% apresentaram maiores índices histomorfométricos intestinais. A inclusão de LSEO a 0,25% na dieta de D. rerio proporcionou melhorias na microbiota e na integridade intestinal dos peixes.

Isolation and Identification of Pigment-Producing Endophytic Fungi from the Amazonian Species Fridericia chica.

Pigments of fungal origin have aroused increasing interest in the food dye and cosmetic industries since the global demand for natural dyes has grown. Endophytic microorganisms are a source of bioactive compounds, and Amazonian plant species can harbor fungi with a wide range of biotechnological applications. Popularly known in Brazil as crajiru, Fridericia chica is a medicinal plant that produces a red pigment. In this study, a total of 121 fungi were isolated in potato dextrose agar from three plants. We identified nine pigment-producing endophytic fungi isolated from branches and leaves of F. chica. The isolates that showed pigment production in solid media were molecularly identified via multilocus analysis as Aspergillus welwitschiae, A. sydowii, Curvularia sp., Diaporthe cerradensis (two strains), Hypoxylon investiens, Neoscytalidium sp. (two strains) and Penicillium rubens. These isolates were subjected to submerged fermentation in two culture media to obtain metabolic extracts. The extracts obtained were analyzed in terms of their absorbance between 400 and 700 nm. The pigmented extract produced by H. investiens in medium containing yeast extract showed maximum absorbance in the red absorption range (UA700 = 0.550) and significant antioxidant and antimicrobial activity. This isolate can thus be considered a new source of extracellular pigment.

Immunomodulatory and Anticancer Effects of Fridericia chica Extract-Loaded Nanocapsules in Myeloid Leukemia.

Nanocapsules provide selective delivery and increase the bioavailability of bioactive compounds. In this study, we examined the anticancer and immunomodulatory potential of Fridericia chica (crajiru) extract encapsulated in nanocapsules targeting myeloid leukemias. Nanocapsules containing crajiru (nanocapsules-CRJ) were prepared via interfacial polymer deposition and solvent displacement. Size and polydispersity were measured by dynamic light scattering. Biological assays were performed on leukemia cell lines HL60 and K562 and on non-cancerous Vero cells and human PBMC. The anticancer activity was evaluated using cytotoxicity and clonogenic assays, while the immunomodulatory activity was evaluated by measuring the levels of pro- and anti-inflammatory cytokines in PBMC supernatants treated with concentrations of nanocapsules-CRJ. Nanocapsules-CRJ exhibited significant cytotoxic activity against HL60 and K562 cells at concentrations ranging from 0.75 to 50 µg/mL, with the greatest reductions in cell viability observed at 50 µg/mL (p < 0.001 for HL60; p < 0.01 for K562), while not affecting non-cancerous Vero cells and human PBMCs. At concentrations of 25 µg/mL and 50 µg/mL, nanocapsules-CRJ reduced the formation of HL60 and K562 colonies by more than 90% (p < 0.0001). Additionally, at a concentration of 12 µg/mL, nanocapsules-CRJ induced the production of the cytokines IL-6 (p = 0.0002), IL-10 (p = 0.0005), IL-12 (p = 0.001), and TNF-α (p = 0.005), indicating their immunomodulatory potential. These findings suggest that nanocapsules-CRJ hold promise as a potential therapeutic agent with both cytotoxic and immunomodulatory properties.

Red sacaca essential oil-loaded nanostructured lipid carriers optimized by factorial design: cytotoxicity and cellular reactive oxygen species levels.

Amazonian flora includes several species with the potential to develop pharmaceutical and biotechnological products. The essential oils from Amazonian species possess some biological properties, such as antioxidant, antibacterial, and cytotoxic activities. The essential oil of red sacaca (RSO), Croton cajucara Benth., contains metabolites characterized by antioxidant and anti-inflammatory activities. Nanostructured lipid carriers (NLC) are an advantageous alternative for the effective delivery of drugs because they can solubilize lipophilic actives and reduce their cytotoxicity. This study aimed to optimize the synthesis of RSO-loaded nanostructured lipid carriers (NLC-RSO) using a 23 factorial design and investigate their antioxidant and cytotoxic effects. The red sacaca essential oil (RSO) metabolite profile was characterized using gas chromatography coupled with a mass spectrometer (GC-MS), identifying 33 metabolites, with linalool and 7-hydroxy-calamenene as the major ones, as reported in the literature. The optimized NLC-RSO formulation had a particle size less than 100 nm and a polydispersity index lower than 0.25. After characterizing NLC-RSO using Fourier-transform infrared spectroscopy, powder X-ray diffraction, zeta potential, moisture content, and wettability, in vitro cytotoxicity were performed in A549 and BEAS-2B cell lines using the resazurin metabolism assay. The data indicated a lower IC50 for RSO than for NLC-RSOs in both cell lines. Furthermore, low cytotoxicity of blank nanoparticles (blank NP) and medium chain triglycerides-loaded nanostructured lipid carriers (NLC-MCT) towards both pulmonary cell lines was noted. At a concentration of 50-100 µg/mL, free RSO exhibited higher cytotoxicity than NLC-RSO, demonstrating the protective effect of this lipid carrier in reducing cytotoxicity during metabolite delivery. Similarly, free RSO showed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging than NLC-RSO, also indicating this protective effect. The 2',7'-dichlorofluorescein diacetate (DCFH-DA) intracellular reactive oxygen species (ROS) level assay did not show differences between the treatments at higher but non-cytotoxic dosages. Taken together, our results suggest that NLC-RSOs are potential RSO delivery systems for applications related to cancer treatment.

In vitro susceptibility of Plasmodium falciparum Welch field isolates to infusions prepared from Artemisia annua L. cultivated in the Brazilian Amazon.

Artemisinin is the active antimalarial compound obtained from the leaves of Artemisia annua L. Artemisinin, and its semi-synthetic derivatives, are the main drugs used to treat multi-drug-resistant Plasmodium falciparum (one of the human malaria parasite species). The in vitro susceptibility of P. falciparum K1 and 3d7 strains and field isolates from the state of Amazonas, Brazil, to A. annua infusions (5 g dry leaves in 1 L of boiling water) and the drug standards chloroquine, quinine and artemisinin were evaluated. The A. annua used was cultivated in three Amazon ecosystems (várzea, terra preta de índio and terra firme) and in the city of Paulínia, state of São Paulo, Brazil. Artemisinin levels in the A. annua leaves used were 0.90-1.13% (m/m). The concentration of artemisinin in the infusions was 40-46 mg/L. Field P. falciparum isolates were resistant to chloroquine and sensitive to quinine and artemisinin. The average 50% inhibition concentration values for A. annua infusions against field isolates were 0.11-0.14 µL/mL (these infusions exhibited artemisinin concentrations of 4.7-5.6 ng/mL) and were active in vitro against P. falciparum due to their artemisinin concentration. No synergistic effect was observed for artemisinin in the infusions.

Stability and antioxidant activity of semi-synthetic derivatives of 4-nerolidylcatechol.

4-nerolidylcatechol (4-NC) is an unstable natural product that exhibits important antioxidant, anti-inflammatory and other properties. It is readily obtainable on a multi-gram scale through straightforward solvent extraction of the roots of cultivated Piper peltatum or P. umbellatum, followed by column chromatography on the resulting extract. Semi-synthetic derivatives of 4-NC with one or two substituent groups (methyl, acetyl, benzyl, benzoyl) on the O atoms have been introduced that have increased stability compared to 4-NC and significant in vitro inhibitory activity against the human malaria parasite Plasmodium falciparum. Antioxidant and anti-inflammatory properties may be important for the antiplasmodial mode of action of 4-NC derivatives. Thus, we decided to investigate the antioxidant properties, cytotoxicity and stability of 4-NC derivatives as a means to explore the potential utility of these compounds. 4-NC showed high antioxidant activity in the DPPH and ABTS assays and in 3T3-L1 cells (mouse embryonic fibroblast), however 4-NC was more cytotoxic (IC50 = 31.4 µM) and more unstable than its derivatives and lost more than 80% of its antioxidant activity upon storage in solution at -20 °C for 30 days. DMSO solutions of mono-O-substituted derivatives of 4-NC exhibited antioxidant activity and radical scavenging activity in the DPPH and ABTS assays that was comparable to that of BHA and BHT. In the cell-based antioxidant model, most DMSO solutions of derivatives of 4-NC were less active on day 1 than 4-NC, quercetin and BHA and more active antioxidants than BHT. After storage for 30 days at -20 °C, DMSO solutions of most of the derivatives of 4-NC were more stable and exhibited more antioxidant activity than 4-NC, quercetin and BHA and exhibited comparable antioxidant activity to BHT. These findings point to the potential of derivatives of 4-NC as antioxidant compounds.

Therapeutic Potential of Leaves from Fridericia chica (Bonpl.) L. G. Lohmann: Botanical Aspects, Phytochemical and Biological, Anti-Inflammatory, Antioxidant and Healing Action.

Plants of the species Fridericia chica (Bonpl.) L. G. Lohmann (Bignoniaceae), which are widely distributed in Brazil and named crajiru in the state of Amazonas, are known in folk medicine as a traditional medicine in the form of a tea for the treatment of intestinal colic, diarrhea, and anemia, among other diseases. The chemical analysis of extracts of the leaves has identified phenolic compounds, a class of secondary metabolites that provide defense for plants and benefits to the health of humans. Several studies have shown the therapeutic efficacy of F. chica extracts, with antitumor, antiviral, wound healing, anti-inflammatory, and antioxidant activities being among the therapeutic applications already proven. The healing action of F. chica leaf extract has been demonstrated in several experimental models, and shows the ability to favor the proliferation of fibroblasts, which is essential for tissue repair. The anti-inflammatory activity of F. chica has been clearly demonstrated by several authors, who suggest that it is related to the presence of 3-deoxyanthocyanidins, which is capable of inhibiting pro-inflammatory pathways such as the kappa B (NF-kB) nuclear transcription factor pathway. Another important effect attributed to this species is the antioxidant effect, attributed to phenolic compounds interrupting chain reactions caused by free radicals and donating hydrogen atoms or electrons. In conclusion, the species Fridericia chica has great therapeutic potential, which is detailed in this paper with the objective of encouraging new research and promoting the sum of efforts for the inclusion of herbal medicines in health systems around the world.

In vivo and in vitro antimalarial activity of 4-nerolidylcatechol.

4-Nerolidylcatechol (4-NC) isolated from Piper peltatum L. (Piperaceae) was evaluated for in vitro antiplasmodial activity against Plasmodium falciparum (cultures of both standard CQR (K1) and CQS (3D7) strains and two Amazonian field isolates) and for in vivo antimalarial activity using the Plasmodium berghei-murine model. 4-NC exhibits significant in vitro and moderate in vivo antiplasmodial activity. 4-NC administered orally and subcutaneously at doses of 200, 400 and 600 mg/kg/day suppressed the growth of P. berghei by up to 63% after four daily treatments (days 1-4). Also, 4-NC exhibited important in vitro antiplasmodial activity against both standard and field P. falciparum strains in which 50% inhibition of parasite growth (IC(50) ) was produced at concentrations of 0.05-2.11 µg/mL and depended upon the parasite strain. Interestingly, healthy (non-infected) mice that received 4-NC orally presented (denatured) blood plasma which exhibited significant in vitro activity against P. falciparum. This is evidence that mouse metabolism allows 4-NC or active metabolites to enter the blood. Further chemical and pharmacological studies are necessary to confirm the potential of 4-NC as a new antimalarial prototype.

Piper peltatum: biomass and 4-nerolidylcatechol production.

Piper peltatum L. is used for the treatment of inflammation, malaria, and other ailments. 4-Nerolidylcatechol (4-NC) is a valuable natural product that has important anti-inflammatory, antimalarial, and antioxidant properties. 4-NC is a component of P. peltatum and P. umbellatum extracts, which are used in cosmetics. The aim of this work was to evaluate the production of plant biomass and the production of 4-NC in roots of cultivated P. peltatum over a full life cycle. Seedlings were produced in a greenhouse and then transplanted. The weight of dry plant parts (leaves, stems, roots, and inflorescences); numbers of stems, leaves, and inflorescences; and the leaf-to-stem ratio were evaluated at intervals of 60 days after transplanting (DAT). Extracts were prepared using 1:1 ethanol-chloroform and an ultrasound bath. Roots, leaves, and inflorescences contained 4-NC according to TLC photodensitometry analysis. Quantification of 4-NC in root extracts was performed using HPLC-DAD analysis. Per-hectare production of 4-NC by roots was estimated based on quantitative HPLC analysis and biomass data. Optimal per-hectare yields of 4-NC were obtained by harvesting roots between 350 and 400 DAT. In this period, the average yield was 27 kg 4-NC per hectare. Importantly, at the time of maximal overall production of root biomass (470 DAT), there was a decrease in the production of 4-NC (23.8 kg/ha), probably due to the onset of senescence.

Isolation of 4-nerolidylcatechol from leaves of Piper peltatum L., and evaluation of larvicidal activity in mosquito vectors, with emphasis on Aedes aegypti (Diptera: Culicidae)

ABSTRACT Synthetic chemical insecticides are widely used in population control of pests. Aedes aegypti is the primary vector of dengue, Zika, chikungunya and yellow fever to humans, and has proven resistance to chemical insecticides. As an alternative vector control method, the ethanolic extract of the leaves of Piper peltatum L. (Piperaceae) showed larvicidal activity against Ae. aegypti. Despite the wide medicinal use of this plant, the biological activity of its isolated constituents remains unexplored. In this sense, we isolated, identified and evaluated the larvicidal activity of 4-nerolidylcatechol (4-NC) from P. peltatum against Ae. aegypti, Culex quinquefasciatus and Anopheles darlingi, focusing on the larvicidal, adulticidal and genotoxic potential of 4-NC on Ae. aegypti. Larvae were captured in the city of Manaus, Amazonas state, Brazil. 4-NC was isolated from the extract of the leaves of P. peltatum via silica gel chromatography. This was identified using nuclear magnetic resonance spectroscopy and tested in Artemia franciscana (6.25, 12.5, 25, 50, 100 and 200 µg/mL). In the toxicity bioassay, Ae. aegypti larvae were exposed to 30, 50, 70, 90, and 110 µg/mL of 4-NC, while Cx. quinquefasciatus and An. darlingi were exposed to 6.25, 12.5, 25, 50 and 100 µg/mL. Ae. aegypti larvae were also subjected to 40 and 60 µg/mL of 4-NC (genotoxicity bioassay), and adult females to 62.5 to 1,000 µg/mL (adulticidal bioassay). The results of the 4-NC toxicity assays showed that there was 100% mortality in larvae of Ar. franciscana at the concentration of 200 µg/mL, with an LC50 of 8.0 μg/mL. In the larvae of Ae. aegypti, mortality was 100%, with an LC50 of 62 μg/mL. In larvae of Cx. quinquefasciatus, 97% mortality occurred, with an LC50 of 52.3 μg/mL, and in An. darlingi larvae there was an 83% mortality rate, with an LC50 of 55.8 μg/mL. In adults of Ae. aegypti, however, there was no adulticidal activity. In the larvae of Ae. aegypti, the genotoxic effect of 4-NC (40 and 60 µg/mL) showed significant frequency (p < 0.05) of cellular abnormalities (micronuclei, budding and nuclear bridges) of interphasic nuclei of neuroblasts and oocytes in relation to the negative control. This result may be associated with a decrease in oviposition of females, which was observed in two generations. We can confirm that 4-NC has larvicidal activity against Ae. aegypti, Cx. quinquefasciatus and An. darlingi. Although it does not present adulticidal activity in Ae. aegypti, it reduced the oviposition of females. Therefore, 4-NC seems to be a strong candidate for the development of an alternative method for the control of these mosquitoes in the immature phase.

Antiparasitic activity, histopathology and physiology of Colossoma macropomum (tambaqui) exposed to the essential oil of Lippia sidoides (Verbenaceae).

In vivo and in vitro antiparasitic activity of the essential oil of Lippia sidoides and blood and histological alterations were assessed in Colossoma macropomum (tambaqui). Essential oil concentrations of 10, 20, 40, 80, 160 and 320mg/L were assayed in vitro against monogenoideans Anacanthorus spathulatus, Notozothecium janauachensis and Mymarothecium boegeri from fish gills. Lippia sidoides essential oil concentrations of 320 and 160mg/L were 100% effective against monogenoideans in 10min and 1h of exposure, respectively. However, the effectiveness of 100% concentrations of 80mg/L and 40mg/L occurred in 3 and 6h, respectively. In the in vivo tests, juvenile fish were submitted to 60min of baths with 10mg/L and 15min of baths with 20mg/L of the essential oil of L. sidoides. These therapeutic baths were not efficient against Ichthyophthirius multifiliis, and monogenoideans present in the gills of C. macropomum. In addition, 10 and 20mg/L of the essential oil of L. sidoides caused an anesthetic effect on the fish and did not influence total glucose and protein plasma levels; however, it decreased the number of total erythrocytes in fish exposed to the higher concentration of this essential oil. Severe alterations and irreversible damage were observed in the fish gills just after L. sidoides essential oil baths and after 24h of recovery. The most recurrent lesions found were hyperplasia and fusion of the lamellar epithelium, vasodilation, detachment of the gill epithelium and lamellar aneurism, epithelial breakdown with hemorrhage, congestion, edema and necrosis, proliferation of the mucous cells and chloride cells and lamellar hypertrophy. Therefore, since the essential oil of L. sidoides has in vitro antiparasitic activity and low concentrations of it have shown toxic effects, the bioactive potential of its main chemical components should be investigated, as well as more efficient forms of its administration in therapeutic baths in order to eliminate fish parasites.

Physicochemical characterization and cosmetic applications of Passiflora nitida Kunth leaf extract

Abstract Passiflora nitida Kunth, an Amazonian Passiflora species, is little studied, although the specie's high biological potential. Herein the plant's pharmacognostic characterization, extract production, antioxidant potential evaluation, and application of this extract in cosmetic products is reported. The physical chemical parameters analyzed were particle size by sieve analysis, loss through drying, extractive yield, total ash content, laser granulometry, specific surface area and pore diameter (SBET), differential scanning calorimetry, thermogravimetry (TG), and wave dispersive X-Ray fluorescence (WDXRF). Total phenol/flavonoid content, LC-MS/MS analysis, DPPH and ABTS antioxidant radical assays, cytotoxicity, melanin, and tyrosinase inhibition in melanocytes test provided evidence to determine the content of the major constituent. P. nitida dry extract provided a fine powder with mesopores determined by SBET, with the TG curve showing five stages of mass loss. The antioxidant potential ranged between 23.5-31.5 mg∙mL-1 and tyrosinase inhibition between 400-654 µg∙mL-1. The species presented an antimelanogenic effect and an inhibitory activity of cellular tyrosinase (26.6%) at 25 µg/mL. The LC-MS/MS analysis of the spray-dried extract displayed the main and minor phenolic compounds constituting this sample. The results indicate that P. nitida extract has promising features for the development of cosmetic formulations

Efficacy of 11 Brazilian essential oils on lethality of the cattle tick Rhipicephalus (Boophilus) microplus.

Herbal extracts have been investigated as an alternative for parasite control, aiming to slow the development of resistance and to obtain low-cost biodegradable parasiticides. The goal of this study was to evaluate the efficacy, in vitro, of 11 essential oils from Brazil on reproductive efficiency and lethality of the cattle tick Rhipicephalus (Boophilus) microplus. The effects of oils extracted from Curcuma longa, Zingiber officinale, Lippia alba, Lippia gracilis, Lippia origanoides, Lippia sidoides, Mentha arvensis, Mentha piperita, Croton cajucara (white and red), and Croton sacaquinha on ticks were investigated by the Immersion Test with Engorged Females (ITEF) and the modified Larval Packet Test (LPT). Distilled water and 2% Tween 80 were used as control treatments. Chemical analysis of the oils was done with gas chromatography coupled with mass spectrometry. Analysis of the in vitro tests using Probit (SAS program) allowed the calculation of lethal concentrations (LCs). Lower reproductive efficiency indexes and higher efficacy percentages in the ITEF were obtained with the oils extracted from C. longa (24 and 71%, respectively) and M. arvensis oils (27 and 73%, respectively). Lower LC50 was reached with C. longa (10.24 mg/mL), L. alba (10.78 mg/mL), M. arvensis (22.31 mg/mL), L. sidoides (27.67 mg/mL), and C. sacaquinha (29.88 mg/mL) oils. In the LPT, species from Zingiberaceae and Verbenaceae families caused 100% lethality at 25 mg/mL, except for L. sidoides. The most effective oils were from C. longa, L. gracilis, L. origanoides, L. alba, and Z. officinale. The LC50 and LC90 were, respectively: 0.54 and 1.80 mg/mL, 3.21 and 7.03 mg/mL, 3.10 and 8.44 mg/mL, 5.85 and 11.14 mg/mL, and 7.75 and 13.62 mg/mL. The efficacy was directly related to the major components in each essential oil, and the oils derived from Croton genus presented the worst performance, suggesting the absence of synergistic effect among its compounds. Since C. longa, containing 62% turmerone, was the one most efficient against ticks, this compound may be potentially used for tick control, but further research is needed, especially to assess toxicity of these compounds to the host. These new studies, together with the results presented here, may provide a strong rationale for designing pre-clinical and clinical studies with these agents.

Piper aduncum against Haemonchus contortus isolates: cross resistance and the research of natural bioactive compounds.

The anthelminthic activity of the essential oil (EO) of Piper aduncum L. was tested in vitro on eggs and larvae of resistant (Embrapa2010) and susceptible (McMaster) isolates of Haemonchus contortus. The EO was obtained by steam distillation and its components identified by chromatography. EO concentrations of 12.5 to 0.02 mg/mL were used in the egg hatch test (EHT) and concentrations of 3.12 to 0.01 mg/mL in the larval development test (LDT). Inhibition concentrations (IC) were determined by the SAS Probit procedure, and significant differences assessed by ANOVA followed by Tukey's test. In the EHT, the IC50 for the susceptible isolate was 5.72 mg/mL. In the LDT, the IC50 and IC90 were, respectively, 0.10 mg/mL and 0.34 mg/mL for the susceptible isolate, and 0.22 mg/mL and 0.51 mg/mL for the resistant isolate. The EO (dillapiole 76.2%) was highly efficacious on phase L1. Due to the higher ICs obtained for the resistant isolate, it was raised the hypothesis that dillapiole may have a mechanism of action that resembles those of other anthelmintic compounds. We further review and discuss studies, especially those conducted in Brazil, that quantified the major constituents of P. aduncum-derived EO.