LC-MS Analysis and Antifungal Activity of Turnera subulata Sm.

Fungi of the Candida genus are responsible for invasive candidiasis, which affects people all over the world and has high mortality rates. This is due to their virulence factors, which give them great resistance and pathogenicity. In addition, the emergence of multidrug-resistant strains makes it difficult to treat these infections. In this way, natural products have emerged as an alternative to standard drugs, where plants known for their medicinal properties such as Turnera subulata become attractive to research. The present work aimed to analyze the ethanol extract of Turnera subulata leaves against standard strains of Candida albicans, Candida krusei and Candida tropicalis using broth microdilution techniques. The identification of the compounds in T. subulata leaves by LC-MS revealed the presence of a wide variety of substances such as carboxylic acids and terpenes, with flavonoids and fatty acids being more evident. The antifungal assays showed that the extract was not able to inhibit the growth of the tested strains at concentrations with a clinical relevance. However, at higher concentrations, it was able to inhibit the fungal dimorphism of C. albicans and C. tropicalis. It is possible that the T. subulata extract has potential as an inhibitor of fungal virulence factors without affecting the cell viability. Further research should be carried out in order to assess its inhibitory potential for other fungal virulence factors.

Single and combined toxicity of amino-functionalized polystyrene nanoparticles with potassium dichromate and copper sulfate on brine shrimp Artemia franciscana larvae.

The increasing use and disposal of plastics has become a persistent problem in the marine environment, calling for studies that refer to realistic scenarios to understand their effects on biota. Particularly, the understanding about the effects of co-exposure with nanoplastic particles and metals on aquatic organisms is still limited. The present work aimed to investigate the acute toxicity of amino-functionalized polystyrene nanoparticles (PS-NH2; 50 nm) as proxy for nanoplastics on brine shrimp Artemia franciscana larvae under different culture conditions and at different stages of development, as well as the combined effect with two reference toxicants - potassium dichromate (K2Cr2O7) and copper sulfate (CuSO4). Nauplii (instar II or III larval stages) were exposed to different concentrations of PS-NH2 (0.005 to 5 µg mL-1) for up to 48 h, with or without agitation in order to mimic a more realistic environmental scenario. Larval mobility and PS-NH2 accumulation were monitored under microscopy. PS-NH2 alone showed toxicity only at the highest concentration tested (5 µg mL-1) regardless the incubation method used (61.2 + 3.1% and 65.0 + 4.5% with and without agitation, respectively). Moreover, instar III stage was the most sensitive to PS-NH2 exposure (38.2% immobility in 24 h of exposure; 5 µg mL-1). Evidence of PS-NH2 retention in the gastrointestinal tract in a concentration- and time-dependent manner was also obtained. Mixtures of PS-NH2 (0.005 and 5 µg mL-1) with different concentrations of K2Cr2O7 increased the immobilization rate of the larvae after 48 h of exposure, when compared to the K2Cr2O7 alone. Similar results were observed for CuSO4 in the co-exposure conditions at different concentrations. However, exposing nauplii to a mixture of PS-NH2 (0.005 µg mL-1) and CuSO4 decreased immobilization rate, in comparison to the group exposed to CuSO4 alone. The present work highlights the potential risk posed by nanoplastics to zooplanktonic species through their interaction with other toxicants.

Chemical identification and antimicrobial potential of essential oil of Piper rivinoides kunth (BETIS-WHITE).

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 <256 µg/mL, as for Candida albicans 4127 (217.6 µg/mL). The association of EOPR with an antifungal showed a high synergistic affinity against the strains of C. tropicalis 40042 and 4262. We concluded that no intrinsic EOPR activity was observed at any concentrations tested against bacteria. However, EOPR associated with Gentamicin acted synergistically against S. aureus 10 and Escherichia coli 06, but with Erythromycin there was a synergistic effect against Escherichia coli 06, and antagonism with norfloxacin.

Chemical composition and anti-Candida potencial of the extracts of Tarenaya spinosa (Jacq.) Raf. (Cleomaceae).

Phytochemical prospecting was performed by HPLC-DAD. The Inhibitory Concentration of 50% of mortality the microorganisms (IC50) was determined and a cell viability curve was obtained. Minimum Fungicidal Concentration (MFC) was determined by subculture in Sabourad Dextrose Agar. The effect of the combination extract/fluconazole was verified by microdilution, with the extracts in subinhibitory concentrations (MFC/16). Caffeic acid was the major compound of both extracts, representing 6.08% in the aqueous extract and 7.62% in the ethanolic extract. The extracts showed a fungistatic effect (MFC ≥ 16,384 µg/mL). The IC50 results demonstrated that the combination of the extracts with fluconazole were more significant than the products tested alone, with values from 4.9 to 34.8 µg/mL for the ethanolic extract/fluconazole and 5 to 84.7 µg/mL for the aqueous extract/fluconazole. The potentiating effect of fluconazole action was observed against C. albicans and C. tropicalis. In C. krusei the aqueous extract had an antagonistic effect.

Chemical composition, antifungal activity and potential anti-virulence evaluation of the Eugenia uniflora essential oil against Candida spp.

The development of fungal resistance to antifungal drugs has been worsening over the years and as a result research on new antifungal agents derived from plants has intensified. Eugenia uniflora L. (pitanga) has been studied for its various biological actions. In this study the chemical composition and antifungal effects of the E. uniflora essential oil (EULEO) were investigated against Candida albicans (CA), Candida krusei (CK) and Candida tropicalis (CT) standard strains. The essential oil obtained through hydro-distillation was analyzed by gas chromatography coupled to mass spectrometry (GC-MS). To determine the IC50 of the oil, the cellular viability curve and the inhibitory effects were measured by means of the oil's association with Fluconazole in a broth microdilution assay with spectrophotometric readings. The Minimum Fungicidal Concentration (MFC) was determined by solid medium subculture with the aid of a guide plate while the assays used to verify morphological changes emerging from the action of the fractions were performed in microculture chambers at concentrations based on the microdilution. Two major oil constituents stand out from the chemical analysis: selina-1,3,7(11)-trien-8-one (36.37%) and selina-1,3,7(11)-trien-8-one epoxide (27.32%). The concentration that reduced microorganismal growth was ≥8,192 µg/mL while the IC50 varied, this being between 1892.47 and 12491.80 µg/mL (oil), 10.07 - 80.78 µg/mL (fluconazole) and 18.53 - 295.60 µg/mL (fluconazole + oil). The combined activity (fluconazole + oil) resulted in indifference and antagonism. A MFC of the oil in association with fluconazole was recorded at the concentration of 8,192 µg/mL against CA and CK. The oil caused the inhibition of CA and CT morphological transition. In view of the results obtained, additional research is needed to elucidate the activity of the E. uniflora oil over genetic and biochemical processes regarding its effect on Candida spp. virulence.