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a Universidade Regional do Cariri, URCA, Cel Ant^onio Luis, 1161, 63105-000, Pimenta, Crato, CE, Brazil b Universidade Federal do Paran a, UFPR, XV de Novembro, 1299, 80.060-000, Centro, Curitiba, PR, Brazil c Universidade Regional de Blumenau, FURB, Ant^onio da Veiga, 140, 89030-903, Itoupava Seca, Blumenau, SC, Brazil d Laborat orio de investiga¸c~ao em Gen etica e Hematologia Translacional, Instituto Gon¸calo Moniz (IGM), Funda¸c~ao Oswaldo Cruz (FIOCRUZ), Salvador, BA 40296- 710, Brazil e Centro Universit ario Doutor Le~ao Sampaio (Unile~ao), Av. Maria Leticia Leite Pereira s/n, Lagoa Seca, Juazeiro do Norte, Brazil

Background/Purpose The fractions were tested to investigate their chemical composition and antifungal potential in vitro. Methods/Results The results obtained through the chromatographic analysis allowed the identification of 28 compounds. Microbiological assays were carried out in order to determine the IC50, cell viability curve and inhibitory effect of the association of the fractions with Fluconazole (FCZ), against Candida spp. through the microdilution methodology. The assays for verification of change in morphology and Minimal Fungicidal Concentration were performed in solid media. The IC50 of the isolated and associated fractions were calculated to be between 5.10 and 926.56 μg/mL, when associated with FCZ the concentrations were reduced to 512 μg/mL and 1024 μg/mL, the IC50 value of the fractions associated with fluconazole varied between 1.38 and 925.56 μg/mL. Conclusions The fractions affect the morphological transition and have a relevant antifungal potential, as they caused fungal inhibition in isolated use and potentiated the action of FCZ.

(1) Background

The chemical composition of the Cynophalla flexuosa hydroethanolic extract (CFHEE) was identified and its antioxidant, antifungal and antipleomorphic activities against C. albicans and C. tropicalis strains were evaluated. Phytochemical prospecting evidenced polyphenolic and triterpenoid compounds while UPLC-MS analysis revealed the presence of Isopropyl/n-propyl-GLS; Methylpropyl-GLS/butyl-GLS; Methylbutyl-GLS; Quercetin O-di-hexoside; Quercetin-pentosyl-hexoside; Rutin; Quercetin O-glucoside; Kaempferol O-rhamnosyl-hexoside; Kaempferol O-pentosyl-hexoside and Lariciresinol hexoside. The CFHEE inhibited the effect of the DPPH● free radical, both when evaluated individually and in combination with the antifungal fluconazole. When associated with 256 μg/mL fluconazole, the extract, at concentrations from 128 μg/mL, reduced the effect of the DPPH● free radical with values ranging from 37.7% to 95.8%. The extract’s antifungal effect was considered clinically irrelevant and its combination with the antifungal triggered an antagonistic effect against the two strains, thus indicating the popular use of bravo beans in the form of teas or infusions should not be combined with the intake of the drug Fluconazole as this may lead to a reduction of its clinical effect. The extract, however, inhibited morphological changes (pleomorphism) in Candida species, preventing the development of hyphae. The CFHEE possesses an antioxidant effect and a potential pharmacological activity for the inhibition of one of the Candida spp. virulence factors.

Croton heliotropiifolius Kunth (Euphorbiaceae), whose occurrence has already been registered in the most varied Brazilian biomes, is commonly found in the Chapada do Araripe, Ceará. The species is traditionally used to treat fungal, parasitic, and degenerative diseases. This study investigated the chemical composition and pharmacological potential (antioxidant, antifungal, antiparasitic, and cytotoxic) of an aqueous extract obtained from the roots of C. heliotropiifolius. Following a qualitative phytochemical screening, the chemical constituents were identified by ultra-efficiency liquid chromatography coupled witha quadrupole/time-of-flight system (UPLC-QTOF). The antioxidant potential was verified by thin-layer chromatography (TLC). The direct and combined antifungal activity of the extract against opportunistic Candida strains was investigated using the microdilution method. The minimal fungicidal concentration (MFC) was determined by subculture, while the modulation of the morphological transition (fungal virulence) was evaluated by light microscopy. The in vitro antiparasitic activity was analyzed using epimastigotes of Trypanosoma cruzi and promastigotes of Leishmania braziliensis and Leishmania infantum, while cytotoxicity was determined in cultures of mouse fibroblasts. The phytochemical analysis identified the presence of acids, terpenes, flavonoids, lignans, and alkaloids. Among these constituents, the presence of polar and non-polar phenolic compounds with known antioxidant action was highlighted. While the extract showed clinically ineffective antifungal effects, it could enhance the effectiveness of fluconazole, in addition to inhibiting the morphological transition associated with increased virulence in Candida strains. Although the extract showed low cytotoxicity against fibroblasts, it also had weak antiparasitic effects. In conclusion, Croton heliotropiifolius is a source of natural products with antifungal and antioxidant potential.

The bioprospection of zootherapeutic products can be a source of new drugs and to the creation of new strategies of natural resources conservation and management of endangered species. This fact is supported by ethnobiological studies indicating that the usage of zootherapeutic products can be replaced by the use of natural products isolated from plants and domestic animals. The emergence of antibiotic-resistant bacteria has increased the need for research for new active principles. Ethnoveterinary studies in Brazil have shown that Sus scrofa domesticus fat is used for diseases associated with bacterial pathogens. The objective of this study was to identify the chemical composition and to evaluate the antibacterial activity of the fixed oil of Sus scrofa domesticus (OFSC) when used alone or associated with antibiotics. In the analysis of the oil composition, there were 4 constituents identified, with oleic acid being the major constituent. The OFSC did not present antibacterial activity when tested alone; however, it showed synergism in the modulating activity when associated with antibiotics Amikacin and Amoxicillin.

Antimicrobial resistance and the consequent inefficiency of antibiotics are the main problems faced by medicine. In view of this, numerous researches have been aimed at looking for new agents with antibacterial activity, among them natural products. Thus, this study aims to evaluate the antioxidant activity of aqueous and hydroalcoholic extracts of the leaves and roots of Spondias tuberosa by Thin Layer Chromatography (TLC), as well as to verify the antibacterial action of the extracts alone and in combination with commercial antibiotics to evaluate its potential in action of change of antibiotics. The extracts showed polar and nonpolar phenolic substances with antioxidant action. The Minimum Inhibitory Concentration (MIC) of the hydroalcoholic extracts of the leaves and roots was 1,024 μg/mL compared to the Staphylococcus aureus 25923 strain, whereas with the other strains, the products showed an MIC ≥ 2,048 µg/mL. The effect of combining extracts with amikacin, ampicillin and norfloxacin against the multidrug-resistant bacteria Escherichia coli 06, Staphylococcus aureus 10 and Pseudomonas aeruginosa 24 resulted in synergistic effects with aminoglycoside alone, achieving up to 75 % reduction in the MIC of the antibiotic. In view of the results obtained, it can be concluded that the extracts of S. tuberosa presented polar and nonpolar phenolic substances, in the antibacterial activity it can positively modify the effect of the aminoglycoside antibiotic against multi-resistant bacteria, but future studies are necessary to discover the mechanism of action of such an effect. La resistencia a los antimicrobianos y la consiguiente ineficacia de los antibióticos son los principales problemas a los que se enfrenta la medicina. Ante esto, numerosas investigaciones se han dirigido a buscar nuevos agentes con actividad antibacteriana, entre ellos productos naturales. Así, este estudio tiene como objetivo evaluar la actividad antioxidante de extractos acuosos e hidroalcohólicos de las hojas y raíces de Spondias tuberosa mediante Cromatografía de Capa Fina (CCF), así como verificar la acción antibacteriana de los extractos solos y en combinación con antibióticos comerciales para evaluar su potencial en la acción del cambio de antibióticos. Los extractos mostraron sustancias fenólicas polares y apolares con acción antioxidante. La Concentración Mínima Inhibitoria (CMI) de los extractos hidroalcohólicos de hojas y raíces fue de 1.024 μg / mL en comparación con la cepa Staphylococcus aureus 25923, mientras que con las otras cepas, los productos mostraron una CMI ≥ 2.048 µg / mL. El efecto de combinar extractos con amikacina, ampicilina y norfloxacina contra la bacteria multirresistente Escherichia coli 06, Staphylococcus aureus 10 y Pseudomonas aeruginosa 24 resultó en efectos sinérgicos con el aminoglucósido solo, logrando reducción de hasta un 75 % en la CMI del antibiótico. A la vista de los resultados obtenido, se puede concluir que los extractos de S. tuberosa presentó sustancias fenólicas polares y apolares, en el actividad antibacteriana puede modificar positivamente el efecto del antibiótico aminoglucósido contra bacterias multirresistentes, pero el futuro son necesarios estudios para descubrir el mecanismo de acción de tal efecto. A resistência antimicrobiana e a consequente ineficiência de antibióticos são os principais problemas enfrentados pela medicina. Diante disto, inúmeras pesquisas têm sido destinadas a procurar novos agentes com atividade antibacteriana, dentre eles os produtos naturais. Dessa forma, esse estudo tem como objetivo avaliar a atividade antioxidante dos extratos aquosos e hidroalcoólicos das folhas e raízes de S. tuberosa Arruda (umbu) por Cromatografia em Camada Delgada (CCD), bem como verificar a ação antibacteriana dos extratos isoladamente e em combinação com antibióticos comerciais para avaliar seu potencial na ação de modificação de antibióticos. Os extratos apresentaram substâncias fenólicas polares e apolares com ação antioxidante. A Concentração Inibitória Mínima (CIM) dos extratos hidroalcoólicos das folhas e raízes foi 1024 μg/mL frente a cepa Staphylococcus aureus 25923, já com as demais cepas os produtos demonstraram uma CIM ≥ 2.048 µg/mL. O efeito da combinação dos extratos com amicacina, ampicilina e norfloxacina frente às bactérias multirresistentes Escherichia coli 06, Staphylococcus aureus 10 e Pseudomonas aeruginosa 24 resultou em efeitos sinérgicos apenas com o aminoglicosídeo, alcançando até 75 % de redução da CIM do antibiótico. Diante dos resultados obtidos, pode-se concluir que os extratos de S. tuberosa apresentaram substâncias fenólicas polares e apolares, na atividade antibacteriana pode modificar positivamente o efeito do antibiótico aminoglicosídeo contra bactérias multirresistentes, mas estudos futuros é preciso para descobrir o mecanismo de ação de tal efeito.

Candida infections represent a threat to human health. Candida albicans is the main causative agent of invasive candidiasis, especially in immunosuppressed patients. The emergence of resistant strains has required the development of new therapeutic strategies. In this context, the use of liposomes as drug carrier systems is a promising alternative in drug development. Thus, considering the evidence demonstrating that sesquiterpene farnesol is a bioactive compound with antifungal properties, this study evaluated the activity farnesol-containing liposomes against different Candida strains. The IC50 of farnesol and its liposomal formulation was assessed in vitro using cultures of Candida albicans, Candida tropicalis, and Candida krusei. The Minimum Fungicidal Concentration (MFC) was established by subculture in solid medium. The occurrence of fungal dimorphism was analyzed using optical microscopy. The effects on antifungal resistance to fluconazole were assessed by evaluating the impact of combined therapy on the growth of Candida strains. The characterization of liposomes was carried out considering their vesicular size, polydispersion index, and zeta medium potential, in addition to electron microscopy analysis. Farnesol exerted an antifungal activity that might be associated with the inhibition of fungal dimorphism, especially in Candida albicans. The incorporation of farnesol into liposomes significantly increased its antifungal activity against C. albicans, C. tropicalis, and C. krusei. In addition, liposomal farnesol potentiated the action of fluconazole against C. albicans and C. tropicalis. On the other hand, the association of unconjugated farnesol with fluconazole resulted in antagonistic effects. In conclusion, farnesol-containing liposomes have the potential to be used in antifungal drug development. However, further research is required to investigate how the antifungal properties of farnesol are affected by the interaction with liposomes, contributing to the modulation of antifungal resistance to conventional drugs.

The increase in microorganisms with resistance to medications has caused a strong preoccupation within the medical and scientific community. Animal toxins studies, such as parotoid glandular secretions from amphibians, possesses a great potential in the development of drugs, such as antimicrobials, as these possess bioactive compounds. It was evaluated Rhinella jimi (Stevaux, 2002) glandular secretions against standard and multi-resistant bacterial strains; the effect of secretions combined with drugs; and determined the toxicity using two biologic in vivo models, and a in vitro model with mice livers. Standard strains were used for the determination of the Minimum Inhibitory Concentration (MIC), while for the modulatory activity of antibiotics, the clinical isolates Escherichia coli 06, Pseudomonas aeruginosa 03 and Staphylococcus aureus 10 were used. Modulatory activity was evaluated by the broth microdilution method with aminoglycosides and β-lactams as target antibiotics. The secretions in association with the antibiotics have a significant reduction in MIC, both the aminoglycosides and β-lactams. The toxicity and cytotoxicity results were lower than the values used in the modulation. R. jimi glandular secretions demonstrated clinically relevant results regarding the modulation of the tested antimicrobials.

The aim of this study was to identify and evaluate the chemical compositions and effects of the S. tuberosa leaf and root hydroalcoholic extracts (HELST and HERST) against different strains of Candida. Chemical analysis was performed by Ultra-Performance Liquid Chromatography Coupled to Quadrupole/Time of Flight System (UPLC-MS-ESI-QTOF). The Inhibitory Concentration of 50% of the growth (IC50) as well as the intrinsic and combined action of the extracts with the antifungal fluconazole (FCZ) were determined by the microdilution method while the minimum fungicidal concentrations (MFCs) and the effect on fungal morphological transitions were analyzed by subculture and in humid chambers, respectively. From the preliminary phytochemical analysis, the phenols and flavonoids were the most abundant. The intrinsic IC50 values for HELST ranged from 5716.3 to 7805.8 &micro, g/mL and from 6175.4 to 51070.9 &micro, g/mL for the HERST, whereas the combination of the extracts with fluconazole presented IC50 values from 2.65 to 278.41 &micro, g/mL. The MFC of the extracts, individually, for all the tested strains was &ge, 16384 &micro, g/mL. When fluconazole was combined with each extract, the MFC against CA URM 5974 was reduced (HELST: 2048 and HERST: 4096 &micro, g/mL). Synergism was observed against standard C. albicans (CA) and C. tropicalis (CT) strains and with the root extract against the CT isolate. The leaf extract inhibited the morphological transition of all strains while the root extract inhibited only CT strains.

The secular use of plants in popular medicine has emerged as a source for the discovery of new compounds capable of curing infections. Among microbial resistance to commercial drugs, species such as Piper diospyrifolium Kunth, which are used in popular therapy, are targets for pharmacological studies. With this in mind, antimicrobial experiments with the essential oil from the P. diospyrifolium (PDEO) species were performed and its constituents were elucidated. The oil compounds were identified by gas chromatography coupled to mass spectrometry (GC/MS). The broth microdilution method with colorimetric readings for bacterial tests (Escherichia coli and Staphylococcus aureus) and spectrophotometric readings for fungal tests (Candida albicans and Candida tropicalis), whose data were used to create a cell viability curve and calculate its IC50 against fungal cells, were used to determine the minimum inhibitory concentration of the oil and its combined action with commercial drugs. The oil's minimal fungicidal concentration and its action over fungal morphological transition were analyzed by subculture and microculture, respectively. Chemical analysis revealed Z-Carpacin, Pogostol and E-Caryophyllene as the most abundant compounds. Results from the intrinsic analysis were considered clinically irrelevant, however the oil presented a synergistic effect against multiresistant E. coli and S. aureus strains when associated with gentamicin, and against the standard and isolated C. tropicalis strains with fluconazole. A fungicidal effect was observed against the C. albicans isolate. Candida spp. hyphae inhibition was verified for all strains at the highest tested concentrations. The P. diospyrifolium essential oil presented a promising effect when associated with commercial drugs and against a fungal virulence factor. Thus, the oil presented active compounds which may help the development of new drugs, however, new studies are needed in order to clarify the oil's mechanism of action, as well as to identify its active constituents.

The family Piperaceae is known for presenting in its species flavoring, healing and antimicrobial properties among others. The objective of the present study was: to study the chemical profile of the essential oil of Piper rivinoides (EOPR); to analyze its anti-bacterial and antifungal potential, as well as to evaluate the antifungal and antibiotic-modifying capacity. The chemical constituents were identified by gas chromatography with flame ionization detector (GC-FID), allowing the identification of 7 constituents of a total of 86.99%. E-Isoelemicin was identified as the main constituent of petroleum (40.81%). Clinically relevant MIC results were obtained against fungi in which the inhibitory concentration remained

Fungal pathologies caused by the genus Candida have increased in recent years due to the involvement of immunosuppressed people and the advance of resistance mechanisms acquired by these microorganisms. Liposomes are nanovesicles with lipid bilayers in which they store compounds. α-Bisabolol is a sesquiterpene with proven biological activities, and in this work it was tested alone in liposomes and in association with Fluconazole in vitro to evaluate the antifungal potential, Fluconazole optimization, and virulence inhibitory effect in vitro. Antifungal assays were performed against standard strains of Candida albicans, Candida tropicalis, and Candida krusei by microdilution to identify the IC50 values and to obtain the cell viability. The Minimum Fungicidal Concentration (MFC) was performed by subculturing on the solid medium, and at their subinhibitory concentration (Matrix Concentration (MC): 16,384 µg/mL) (MC/16), the compounds, both isolated and liposomal, were associated with fluconazole in order to verify the inhibitory effect of this junction. Tests to ascertain changes in morphology were performed in microculture chambers according to MC concentrations. Liposomes were characterized from the vesicle size, polydispersity index, average Zeta potential, and scanning electron microscopy. The IC50 value of the liposomal bisabolol associated with fluconazole (FCZ) was 2.5 µg/mL against all strains tested, revealing a potentiating effect. Liposomal bisabolol was able to potentiate the effect of fluconazole against the CA and CT strains by reducing its concentration and completely inhibiting fungal growth. α-Bisabolol in liposomal form inhibited the morphological transition in all strains tested at a concentration of MC/8. The liposomes were homogeneous, with vesicles with diameters of 203.8 nm for the liposomal bisabolol and a surface charge potential of −34.2 mV, conferring stability to the nanosystem. Through scanning microscopy, the spherical shapes of the vesicles were observed.

Ethnopharmacological relevance The use of popular plants has guided pharmaceutical research aimed at combating pathogenic microorganisms. Psidium guajava L. is a plant of great versatility and it has been used both as food and as a therapeutic agent. Root, bark, leaves, fruits, flowers and seeds are used for medicinal purposes, especially in infusions and decoctions for oral and topical use . P. guajava is utilized in symptomatology treatment related to organ malfunction and of diseases caused by the action of pathogenic and/or opportunistic microorganisms. Many pharmacological studies have been conducted to scientifically assess its therapeutic potential. Aims of study The aim of the current study is to relate the popular use of this plant and its bioscientific assessment as a therapeutic agent in the treatment of diseases and symptoms caused by the action of protozoa, fungi, bacteria and viruses, and also evaluate the safety for the usage and the interaction with drugs. Materials and methods A bibliographic database the ethnobiology of Psidium guajava (2005–2015) and the pharmacological infections and parasitic diseases (2010–2015). Searches were done in scientific disclosure databases such as PubMed, Web of Science, and Scopus. Results P. guajava leaf extracts were scientifically investigated for the treatment of diseases caused by protozoa (leishmaniasis, malaria, giardiasis, amoebiasis and trichomoniasis), fungi (dermatosis, systemic and mucocutaneous diseases), bacteria (respiratory, mucocutaneous and gastrointestinal infections, cholera, gastritis and stomach ulcers, oral and periodontal infections, venereal diseases and urinary infections) and viruses (herpes, influenza, rotavirus disease and AIDS). The toxicity assays indicates the safet for usage. Conclusions Highlight and elucidate the therapeutic potential and versatility of P. guajava . They also justify using ethnobiology efficiency to guide pharmacological studies. Some limitations can be observed in this kind of study, as the lack for ethnobiological informations and the absence of some controls in the assays.

Introduction The survival and spreading of resistant bacterial strains has boosted research which focusses on discovering new antimicrobial agents derived from plant species. Several studies have demonstrated that substances present in plant extracts can modify the activity of antibiotics, increasing their efficacy. Species of the genus Psidium have been popularly used to treat bacterial infections. However, their modulatory effect on antibiotic activity remains to be elucidated. The aim of this study was to evaluate the modulatory effect of the hydroalcoholic extracts obtained from the leaves of Psidium guajava L. and Psidium brownianum Mart ex DC on bacterial proliferation, both separately and in combination with antibiotics. Methods The assays were performed using the microdilution method. The minimum inhibitory concentration (MIC) of the extracts and drugs were determined against standard and clinical isolates of Escherichia coli , Pseudomonas aeruginosa and Staphylococcus aureus . To verify the potentiation of the antibiotic activity, the MIC of the antibiotics were determined alone and in association with the extracts. Results The extracts of P. guajava and P. brownianum did not present clinically significant activity against the Gram negative bacteria evaluated, with MIC values against S. aureus of 256 and 512 μg/mL, respectively. However, when combined at sub-inhibitory concentrations with antibiotics, both extracts presented significantly synergistic effects. Conclusions Our results demonstrated the effectiveness of P. guajava and P. brownianum on modulating bacterial growth, suggesting that these natural products might be used in drug development in association with antibiotics, reducing bacterial resistance and thus, improving the treatment of bacterial infections.

The therapeutic combinations have been increasingly used against fungal resistance. Natural products have been evaluated in combination with pharmaceutical drugs in the search for new components able to work together in order to neutralize the multiple resistance mechanisms found in yeasts from the genus Candida. The aqueous and hydroethanolic extracts from Psidium brownianum Mart ex DC. and Psidium guajava L. species were evaluated for their potential to change the effect of commercial pharmaceutical drugs against Candida albicans and Candida tropicalis strains. The tests were performed according to the broth microdilution method. Plate readings were carried out by spectrophotometry, and the data generated the cell viability curve and IC50 of the extracts against the yeasts. A chemical analysis of all the extracts was performed for detection and characterization of the secondary metabolites. The total phenols were quantified in gallic acid eq/g of extract (GAE/g) and the phenolic composition of the extracts was determined by HPLC. Fluconazole and all extracts presented high Minimum Inhibitories Concentrations (MICs). However, when associated with the extracts at sub-inhibitory concentrations (MIC/16), fluconazole had its effect potentiated. A synergistic effect was observed in the combination of fluconazole with Psidium brownianum extracts against all Candida strains. However, for Psidium guajava extracts the synergistic effect was produced mainly against the Candida albicans LM77 and Candida tropicalis INCQS 400042 strains. The IC50 values of fluconazole ranged from 19.22 to 68.1 μg/mL when it was used alone, but from 2.2 to 45.4 μg/mL in the presence of the extracts. The qualitative chemical characterization demonstrated the presence of phenols, flavonoids and tannins among the secondary metabolites. The concentration of total phenols ranged from 49.25 to 80.77 GAE/g in the P. brownianum extracts and from 68.06 to 82.18 GAE/g in the P. guajava extracts. Our results indicated that both P. brownianum and P. guajava extracts are effective on potentiating the effect of fluconazole, and therefore, these plants have the potential for development of new effective drugs for treating fungal infections.

We assessed extracts from Psidium brownianum for antifungal activity and identified the phenolic phytocompounds. Minimum inhibitory concentration was determined by microdilution and IC50 was calculated. The minimun fungicidal concentration and the morphology of Candida were evaluated. Extracts analyzed by high-performance liquid chromatography demonstrated flavonoids and phenolic acids. The minimum inhibitory concentration was 8192 µg/mL and the IC50 varied between 1056 and 5128 µg/mL. Extracts showed fungistatic effect and altered the dimorphism of the strains, being the better result observed using the decoction, that affected the fungal dimorphism of the strain CA ATCC40006 at 4096 µg/mL.

(1) Background: Infections by Candida species represent a serious threat to the health of immunocompromised individuals. Evidence has indicated that nerolidol has significant antifungal properties. Nonetheless, its use is restricted due to a low water solubility and high photosensitivity. The incorporation into liposomes may represent an efficient alternative to improve the physicochemical and biopharmaceutical properties of this compound. The present study aimed to characterize the antifungal properties of liposomal nerolidol, alone or in combination with fluconazole. Of note, this is the first study reporting the antifungal activity of liposomal nerolidol and its potentiating effect in association with fluconazole. (2) Methods: The Inhibitory Concentration 50%-IC50 and minimum fungicide concentrations (MFC) of the substances against Candida albicans (CA), Candida tropicalis (CT), and Candida krusei (CK) were established by subculture in a solid medium. To evaluate the antifungal-enhancing effect, the MFC of fluconazole was determined in the presence or absence of subinhibitory concentrations of nerolidol (free or liposomal). The analysis of fungal dimorphism was performed through optical microscopy and the characterization of liposomes was carried out considering the vesicular size, polydispersion index, and zeta medium potential, in addition to a scanning electron microscopy analysis. (3) Results: The physicochemical characterization revealed that liposomes were obtained as homogenous populations of spherical vesicles. The data obtained in the present study indicate that nerolidol acts as an antifungal agent against Candida albicans and Candida tropicalis, in addition to potentiating (only in the liposomal form) the effect of fluconazole. However, the compound had little inhibitory effect on fungal dimorphism. (4) Conclusions: The incorporation of nerolidol into liposomes improved its antifungal-modulating properties.

The search for natural substances such as plant extracts with antimicrobial properties has considerably increased, given that biofilms constitute a barrier against antifungal therapy, where these can be formed on any surface, such as acrylic resin prosthesis. The objective of this study was to identify the chemical composition of the Persea americana Mill. leaf ethanol extract (EEFPa) using the UPLC-QTOF-MS/MS technique, to verify its antifungal activity through a sensitivity test according to the conditions described in the documents in M27-A3 (CLSI, 2008) and M60 (CLSI, 2017), to induce biofilm formation in acrylic resin discs and quantify their formation using tetrazolium salt reduction (MTT), as well as to treat these with the extract and fluconazole. Ten of the twelve compounds present in the extract were identified. In the sensitivity test the lowest minimum inhibitory concentration observed was 512 μg/mL, while fluconazole concentrations ranged from 64 to 1 μg/mL. During biofilm induction, all the isolates were able to form biofilms within 48 h. During biofilm treatment, the extract was less effective at biofilm reduction than Fluconazole. The EEFPa showed significant antifungal activity against some of the strains in this study, however the extract showed lower effect when compared to fluconazole against the biofilm formation.

The Piper mikanianum species were investigated by the antimicrobial potential and chemical composition. Chemical analysis was performed by gas chromatography coupled to mass spectrometry (GC/MS). The Minimum Inhibitory Concentration (MIC) as well as the 50% Inhibitory Concentration against Candida strains were determined by microdilution. The effect of the drug-oil combination was also evaluated to verify possible synergism. The Minimum Fungicidal Concentration (MFC) was evaluated by subculturing the microdilution in Petri dishes and the anti-pleomorphism potential of the oil was tested in humid chambers. Chemical analysis revealed safrol as the major compound. The results from the intrinsic activity evaluation of the oil did not reveal a clinical importance, however, it presented a synergistic effect when associated with gentamicin against the multidrug resistant E. coli strain and when associated with fluconazole against fungal strains. Moreover, the oil possessed a fungistatic effect. Total inhibition of filamentous structures occurred in both Candida species in the anti-virulence test. The P. mikanianum essential oil showed a potentiating activity of drugs for which resistance exists and an inhibitory effect of one of the main virulence factors of the Candida genus, morphological transition, which has been previously shown to be responsible for causing invasive infections in human tissues.

Ethnopharmacological relevance Northeast Brazilian ethnoveterinary studies associated with the medicinal use of zootherapies have shown that ruminants’ body fat such as sheep (Ovis aries), goats (Capra hircus) and cows (Bos taurus) are used in diseases affecting domestic animals. Aim of the study The objective of this study was to evaluate the antibacterial activity of the fixed oils from these ruminants in isolation and in association with antibiotics. Results Ovis aries (OFOA), Capra hircus (OFCH) and Bos taurus (OFBT) fixed oils were extracted using a Soxhlet apparatus with hexane as the solvent. Through the use of gas chromatography coupled to mass spectrometry (GC-MS) the methyl esters from the ruminants’ fixed oils were obtained and the fatty acids present in these oils were indirectly determined. The OFOA, OFCH and OFBT antibacterial and antibiotic modifying activities against standard and multi-resistant bacterial strains were carried out using the broth microdilution test. The fixed oils from these species did not present antibacterial activity when tested in isolation, obtaining Minimal Inhibitory Concentration (MICs) values ≥ 1024 μg/mL. However, when associated with antibiotics, OFBT and OFCH showed a synergistic activity for the Amicacin, Amoxicillin, Norfloxacin and Oxytetracycline antibiotics. Conclusion The OFOA promoted a synergistic action for the same antibiotics with the exception of Norfloxacin.

In the Northeast of Brazil, ethnoveterinary studies have shown that the body fat from Gallus gallus domesticus and Meleagris gallopavo are used for diseases that affect domestic animals. The objective of this study was to identify the chemical composition and to evaluate the antibacterial activity of the Gallus gallus domesticus (OFGG) and Meleagris gallopavo (OFMG) fixed oils in isolation and in association with antibiotics. The OFGG and OFMG from the poultry's body fat were extracted using hexane as a solvent in Soxhlet. Their composition was indirectly determined using fatty acid methyl esters. The OFGG and OFMG antibacterial and modulatory activities against standard and multi-resistant bacterial strains were performed through the broth microdilution test. In the OFGG chemical composition, 4 constituents were identified. The saturated fatty acid (AGS) and unsaturated fatty acid (AGI) percentages were 35.1% and 64.91% respectively, with linoleic acid being the major component. In the OFMG, 3 constituents were identified. The AGS percentage was 27.71% and 72.29% for AGI, with oleic acid as the most abundant component. The oils did not present antibacterial activity when tested in isolation, presenting Minimum Inhibitory Concentrations (MICs) > 512 μg/mL. However, when associated with antibiotics the OFGG showed synergistic activity with the antibiotics Amikacin, Amoxicillin, Norfloxacin and Oxytetracycline, while the OFMG promoted a synergistic action with the antibiotics Amikacin, Amoxicillin and Norfloxacin.

Psidium guajava L. is a plant widely used for food and in folk medicine all over the world. Studies have shown that guava leaves have antifungal properties. In this study, Flavonoid and Tannic fractions were tested to investigate their chemical composition and antifungal potential in vitro.21 compounds in the two fractions, presenting a higher content of phenolic compounds. The antifungal assays were performed against Candida albicans, Candida tropicalis and Candida krusei by microdilution to determine the IC50 and the cell viability curve. Minimal Fungicidal Concentration(MFC) and the inhibitory effects of the association of the fractions with Fluconazole, as well as the assays used to verify any morphological changes were performed in microculture chambers based on the concentrations from the microdilution. The IC50 of the isolated fractions and the fractions associated with each other were calculated, varying from 69.29 to 3444.62 μg/mL and the fractions associated with fluconazole varied from 925.56 to 1.57 μg/mL, it was clear that the association of the natural product with the antifungal presented a synergism. The fractions affected pleomorphism capacity and have a potential antifungal activity as they caused fungal inhibition in isolated use, potentiated the action of Fluconazole, reducing its concentration and impeding morphological transition, one of the virulence factors of the genus.

Context: Animals are used for the treatment of diseases caused by inflammatory processes, although few studies evaluate their potential for these purposes.Objectives: To evaluate the anti-inflammatory potential of zootherapeutic products derived from vertebrates used in Brazilian traditional medicine.Material and methods: The species analyzed were Tupinambis merianae, Iguana iguana, Crotalus durissus, Boa constrictor, and Euphractus sexcinctus. The methods used in anti-inflammatory assays were ear edema (topical) and paw (systemic).Results: With regard to topical anti-inflammatory activity, the fat from T. merianae, C. durissus, I. iguana, B. constrictor, and E. sexcinctus reduced inflammation, while for systemic anti-inflammatory activity, only the fat and the skin of C. durissus, the skin of I. iguana and the fat from B. constrictor reduced inflammation.Conclusions: Studies should be conducted to evaluate the mechanisms of action for each product that demonstrated anti-inflammatory activity as w...

In this study, the Piper cernuum Essential Oil (EOPC) was tested against bacterial and fungal strains using the microdilution method to evaluate its minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC), as well as its antibiotic modifying effect, in addition to observing the oils ability to cause fungal dimorphism and to generate an IC50 cell viability curve. The EO was obtained through hydrodistillation using a Clevenger-type apparatus with the dry leaves and analyzed by GC-FID, allowing the identification of 14 compounds, with the main compound being 4-epi-cis-dihydromarofuran (28.97%). An EOPC intrinsic activity was identified only against Candida albicans 4127 (IC50 56.851 μg/mL). Given the MIC results from the EOPC intrinsic activity against Staphylococcus aureus 10 at the concentration of 406 μg/mL, its association with fluconazole potentiated it’s antifungal action. The action of gentamicin was potentiated by the addition of the EOPC to Staphylococcus aureus 10 and Escherichia coli 06 strains, both with MIC

Background.Phrynops geoffroanusis a small turtle that inhabits lakes, rivers, and streams throughout South America. The body fat of this animal is used as a folk medicine in Brazil for treating illnesses such as sore throats, ear aches, mumps, rheumatism, and arthritis. The present study evaluated the antimicrobial activity of oil extracted fromPhrynops geoffroanus(OPG), determined its chemical composition, and discussed the implications of its use in traditional medicine. The OPG was obtained from the ventral region of this turtle using hexane as a solvent. The antimicrobial activity of OPG was tested against standard and multiresistance strains of bacteria and fungi and its composition was determined indirectly by analyzing the methyl esters of the component fatty acids. The OPG presented a clinically relevant antifungal activity againstCandida kruseiATCC 6258 (MIC 128 µg/mL). When the OPG was associated with the antibacterial and antifungal drugs, was observed a synergistic effect when associated the OPG with the gentamicin against the strainPseudomonas aeruginosa22. Our results indicated that OPG has clinically relevant antifungal activity againstC. krusei, and demonstrated synergetic antibacterial activity in combination with commercial antibiotics againstPseudomonas aeruginosa.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:42:28Z No. of bitstreams: 1 S1519-69842009000400027.pdf: 120656 bytes, checksum: 8c3a48fc99aacb9acc18e6ec10581cda (MD5) Made available in DSpace on 2013-08-22T18:42:28Z (GMT). No. of bitstreams: 1 S1519-69842009000400027.pdf: 120656 bytes, checksum: 8c3a48fc99aacb9acc18e6ec10581cda (MD5) Previous issue date: 2009-08-01 Made available in DSpace on 2013-09-30T19:29:45Z (GMT). No. of bitstreams: 2 S1519-69842009000400027.pdf: 120656 bytes, checksum: 8c3a48fc99aacb9acc18e6ec10581cda (MD5) S1519-69842009000400027.pdf.txt: 25784 bytes, checksum: c05d2af5add07efe26e097ac7da6b886 (MD5) Previous issue date: 2009-08-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:52:53Z No. of bitstreams: 2 S1519-69842009000400027.pdf: 120656 bytes, checksum: 8c3a48fc99aacb9acc18e6ec10581cda (MD5) S1519-69842009000400027.pdf.txt: 25784 bytes, checksum: c05d2af5add07efe26e097ac7da6b886 (MD5) Made available in DSpace on 2014-05-20T13:52:53Z (GMT). No. of bitstreams: 2 S1519-69842009000400027.pdf: 120656 bytes, checksum: 8c3a48fc99aacb9acc18e6ec10581cda (MD5) S1519-69842009000400027.pdf.txt: 25784 bytes, checksum: c05d2af5add07efe26e097ac7da6b886 (MD5) Previous issue date: 2009-08-01 Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Apresentamos dados sobre taxas de infecção pulmonar por parasitas nos lagartos Tropidurus hispidus e T. semitaeniatus que vivem simpatricamente na encosta da Chapada do Araripe, situada na região Nordeste do Brasil. Não encontramos infecção parasitária nos pulmões dos espécimes de T. semitaeniatus. Entretanto, foram encontradas duas espécies de parasitas nos hospedeiros T. hispidus, os pentastomídeos Raillietiella mottae e os nematódeos Rhabdias sp. A prevalência geral de infecção foi de 5%. A prevalência de infecção de R. mottae foi de 2,5% correspondendo a apenas um parasita por hospedeiro. A prevalência de infecção de Rhabdias sp. foi de 2,5% e a amplitude de infecção de 1-2 parasitas por hospedeiros. Esses resultados compreendem o primeiro registro de Rhabdias infectando lagartos da família Tropiduridae em toda a região Neotropical. Apresentamos também uma comparação com os parâmetros de infecção pulmonar por parasitas incluindo os estudos mais recentes no Brasil. We present data on pulmonary infection rates by parasites in the lizards Tropidurus hispidus Spix, 1825 and T. semitaeniatus (Spix, 1825) living sympatrically in the Chapada do Araripe mountain Range, northeastern Brazil. We found no parasite pulmonary infection in T. semitaeniatus. However, two pulmonary parasite species were found in the T. hispidus hosts, the pentastomid Raillietiella mottae Almeida, Freire and Lopes, 2008 and the nematode Rhabdias sp. Overall prevalence was 5%. Prevalence of R. mottae was 2.5% and corresponded to only one parasite on each infected host. Prevalence of Rhabdias sp. was 2.5% and the range of infection was 1-2 parasites per host. This represents the first record of Rhabdias infecting lizards of the family Tropiduridae in the Neotropical region. Furthermore, we present a comparison of parameters of infection by pulmonary parasites including some recent studies in Brazil. Universidade Regional do Cariri Centro de Ciências Biológicas e da Saúde Departamento de Química Biológica Universidade Federal da Paraíba Centro de Ciências Exatas e da Natureza Laboratório/Coleção de Herpetologia Universidade Estadual Paulista Laboratório de Parasitologia de Animais Silvestres Instituto de Biociências Universidade Estadual Paulista Laboratório de Parasitologia de Animais Silvestres Instituto de Biociências

Ethnozoological studies have shown that Spilotes pullatus Linn. (Colubridae: Ophidia), is associated with medicinal and magic-religious uses in Brazil.This study was designed to determine the chemical composition of the oil extracted from the body fat of S. pullatus and to test its antimicrobial properties, alone and in association with aminoglycosides, against fungi and bacterial strains in concentrations ranging between 1024 and 0.5 µg/mL.The snakes were collected in the Chapada do Araripe, county of Crato, Ceará State, Brazil. The oil was extracted in a Soxhlet apparatus using hexane. The methyl esters of the fatty acids present in the samples were identified using GC-MS. The antimicrobial and drug modulatory activities of oil were tested by microdilution against fungal and bacterial strains.The chemical composition of the fixed oils of S. pullatus identified 10 constituents representing 94.97% of the total sample. The percentages of saturated and unsaturated fatty acids were 33.59 and 61.38%, respectively, with the most abundant components being elaidic (37.26%). The oil did not demonstrate any antimicrobial or antifungal activity when tested alone, presenting MIC values ≥ 1024 µg/mL. However, when associated with antibiotics, it demonstrated synergistic effects with gentamicin against all the bacterial lineages assayed, and antagonistic effects with amikacin and neomycin against strains of Escherichia coli.Oil extracted from the body fat of S. pullatus did not demonstrate any inhibitory effects on bacterial or fungal activities, but was effective in modulating the effects of certain antibiotics.

Leptodactylus macrosternumandL. vastus(family: Leptodactylidae) are commonly encountered in the “Caatinga” biome in northern Brazil. The body fat ofL. vastusis used as a zootherapeutic for treating a number of human maladies. The aim of this work was to determine the chemical composition of the body fats ofL. macrosternumandL. vastusand to evaluate their antimicrobial activities as well as the ecological implications of their use in traditional folk medicine. Oils were extracted from body fat located in the ventral region ofL. macrosternum(OLM) andL. vastus(OLV) using hexane as a solvent. The fatty acids were identified by GC-MS. The antimicrobial activities of the oils, either alone or in combination with antibiotics and antifungal drugs, were tested on standard strains of microorganisms as well as on multiresistant strains ofEscherichia coliandStaphylococcus. OLM contained 40% saturated and 60% unsaturated fatty acids, while OLV contained 58.33% saturated and 41.67% unsaturated fatty acids. Our results indicated that both OLM and OLV demonstrated relevant antimicrobial activities (with MIC 256 μg/mL for both) againstPseudomonas aeruginosaandCandida krusei. However, no antimicrobial effects were observed when these oils were combined with antibiotics or antifungal drugs.

Amphibian skins have wide variety of biologically active compounds associated with the natural defenses of these animals.To study the in vitro anticancer activity of methanol extracts of the skin of Rhinella jimi Stevaux (Anura: Bufonidae).The extract was obtained by cold methanol extraction for 96 h using dried skins (295 mg). The methanol skin extract was dried under reduced pressure, giving a 5.5% yield. In order to test for growth-inhibitory activity, in vitro tests were performed with the following cancer cell lines using concentrations ranging between 0.25-250 µg/mL of the extract by 48 h: K562 (leukemia), MCF-7 (breast), NCI-ADR (breast with MDR phenotype), UACC-62 (melanoma), NCI460 (lung), PCO3 (prostate), HT-29 (colon), OVCAR (ovary), and 786-0 (kidney).The methanol extract of R. jimi produced a growth inhibition in a dose-dependent manner against the most of the assayed cell lines. In addition to the growth inhibition, the extract induced the cell death in the ovary and colon lines (EC₅₀ 0.125 and 0.2 µg/mL, respectively), demonstrating 100% of inhibition with 2.5 µg/mL. However, prostate and leukemia cell lines demonstrated less sensitivity, with EC₅₀ of 24 and 235 µg/mL, respectively. This is the first report about the anticancer activity by natural products from the skin of R. jimi.The methanol extracts of R. jimi significantly affected the growth of several cell lines, demonstrating that these compounds are a potential source of substances that could be utilized in cancer treatments.

Lizards of the genus Stenocercus Duméril and Bibron, 1837 are widely distributed throughout South America, being found both in lowlands and at high altitudes. Stenocercus squarrosus was recently described from, and is supposed to, be restricted to the region of Serra das Confusões National Park, Piauí State, Brazil. We here report the first record of this species in Ceará State, from a reasonably high altitudinal area in the municipality of Santana do Cariri, Ceará State, within the limits of the Chapada do Araripe Environmental Protection Area, and it is the first record of this genus for the state, which represents an increase of approximately 460km (airline) northeast of the previously known distribution of this species. Our finding reinforces the importance of conserving that area and indicates the necessity of additional inventories of the local fauna, especially in areas of Carrasco and Cerrado vegetation poorly sampled in Ceará. Extensão da distribuição de Stenocercus squarrosus Nogueira and Rodrigues, 2006 (Reptilia, Squamata, Tropiduridae) no nordeste do Brasil. Lagartos do gênero Stenocercus Duméril & Bibron, 1837 são amplamente distribuídos na América do Sul, podendo ocorrer tanto em terras baixas quanto em elevadas altitudes. Stenocercus squarrosus foi recentemente descrita para o estado do Piauí, na região do Parque Nacional Serra das Confusões, sendo até então considerada restrita. Nós relatamos aqui o primeiro registro de ocorrência dessa espécie no estado do Ceará, ocorrendo em uma área de altitude no município de Santana do Cariri, estado do Ceará, dentro dos limites da Área de Proteção Ambiental da Chapada do Araripe, sendo este o primeiro registro do gênero para o estado, ampliando a distribuição da espécie em cerca de 460 km em linha reta, sentido nordeste. Este registro reforça a importância de conservação da área, indicando a necessidade de mais estudos investigativos de fauna no local, sobretudo em áreas de Carrasco e Cerrado, as quais têm sido pobremente estudadas no Ceará.

Atractus ronnie was recently described from Serra de Baturité, a mountainous relictual forest enclave in the semiarid Caatinga, state of Ceará, northeastern Brazil. Here we report new records of A. ronnie in two other areas of relictual forests and provide additional data on pholidosic variation. The results presented herein reinforce the need for systematic inventory surveys in the relictual forests of Ceará, since the herpetofauna remains poorly known. Novos registros de Atractus ronnie (Reptilia, Serpentes, Colubridae) em florestas relictuais do estado do Ceará e comentários sobre dados merísticos e morfométricos. Atractus ronnie foi recentemente descrita para Serra de Baturité, uma área montanhosa de enclave de floresta relictual na Caatinga semi-árida, estado do Ceará, nordeste do Brasil. Apresentamos aqui novos registros de A. ronnie em duas novas áreas de floresta relictual e apresentamos dados adicionais de variação de sua folidose. Os resultados apresentados aqui reforçam a necessidade de inventários sistemáticos em florestas relictuais do Ceará, uma vez o conhecimento sobre a herpetofauna das florestas relictuais permanece pobremente conhecido.