Chemical, antioxidant and cytotoxic profile of hydroalcoholic extracts of plants from Southern Brazil and their activity against pathogenic fungi isolated from dogs and cats with sensitivity and resistance to conventional antifungals.

We aimed to evaluate the chemical, antioxidant, cytotoxic, and antifungal activities of hydroalcoholic extracts of native plants from Southern Brazil: Schinus terebinthifolia (SCH), Persicaria hydropiperoides (PER), Eugenia uniflora (EUG) and Equisetum hyemale (EQU). Ethyl gallate, quercetin, and quinic acid were prevalent compounds identified by LC-MS. For total phenolic/flavonoid contents and the antioxidant potential against ABTS/DPPH radicals, the ascending order was EQU < PER < EUG < SCH. All extracts were low cytotoxic and kept a high Vero cell viability (>75%) at concentrations up to 12.5 mg/mL (MTT assay). By M38-A2/M27-A3 (CLSI) against 68 clinical isolates of animals and strains of Malassezia pachydermatis, Sporothrix brasiliensis, Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes, all extracts (MIC/MFC ≤3.13-100 mg/mL) were active, except EUG. SCH inhibited and killed S. brasiliensis (MIC/MFC50/90 3.12-12.5 mg/mL) and dermatophytes (MIC/MFC 6.25-25 mg/mL) resistant to ketoconazole and itraconazole. These findings support the promising use of the selected plant extracts as antifungal agents.

Multiresistant bacteria isolated from domestic and wild animals with skin lesions were susceptible to native plants from Southern Brazil.

We evaluated the chemical composition, toxicity, and antibacterial activity of Schinus terebinthifolia (SCH), Eugenia uniflora (EUG), Persicaria hydropiperoides (PER), Equisetum hyemale (EQU), Solidago chilensis (SOL), and Baccharis trimera (BRA). These plants were tested (7.5-0.01 mg/mL) against Gram-positive (G+; n = 32) and Gram-negative (G-; n = 26) isolates from animals (M07-A9, CLSI). Antibiogram (disk diffusion), chromatographic analysis (UPLC), and toxicity assay (HET-CAM) were also performed. A high incidence of resistance was noted, in which 18.4% (07/38) of G+ (Staphylococcus intermedius/Enterococcus faecium) and 17.7% (06/34) of G- (Pseudomonas aeruginosa/Escherichia coli/Proteus mirabilis) were multidrug-resistant. All bacteria were sensitive (MIC50) to SCH (both 3.75 mg/mL), EUG (3.75 mg/mL and 7.5 mg/mL, respectively) and PER (both 7.5 mg/mL). SCH/EUG/PER highlighted as antibacterial, probably due to the major compounds (ethyl gallate, quinic acid, quercetin). These extracts showed normal embryonic development (SCH/EUG: 7.5-0.94 mg/mL). These findings highlighted the promising use of native plants for therapeutic purposes.

Bioactive content of six passion fruit genotypes cultivated in southern Brazil

Passion fruit production in Brazil is concentrated in tropical regions, however, recently production has expanded to regions with temperate climate. Cultivar performance in different climate and soil can lead to variation in the contents of bioactive compounds in the juice and rind of the fruit. This study characterized the bioactive content of passion fruit rind and juice of six passion fruit genotypes ('Catarina Roxo', 'Catarina', 'Urussanga', 'BRS Gigante Amarelo', 'BRS Rubi do Cerrado', and 'BRS Sol do Cerrado') cultivated in southern Brazil. Phenolic, flavonoid, anthocyanin, carotenoid, and ascorbic acid contents and the antioxidant potential of each fruit portion from each genotype were evaluated. Fruit composition varied with genotype and evaluated tissue. Bioactive compound contents were greatest in the rind of 'Urussanga' and 'Catarina', whereas the juice of 'BRS Sol do Cerrado' and 'Catarina Roxo' had the greatest bioactive content. Given cultivar and tissue variability for bioactive content, postharvest use of passion fruit will determine the choice of cultivar to obtain a product with maximum bioactive compounds. Passion fruit genotypes evaluated in this study have shown to be potential sources of bioactive compounds. Fruit rind has the potential to be explored in the scientific and technological scope, due to its high amounts of phenolic compounds and antioxidant activity.

Susceptibility and resistance of Sporothrix brasiliensis to branded and compounded itraconazole formulations.

Itraconazole is the first drug of choice for the treatment of sporotrichosis and it is available at different concentrations for veterinary patients. However, therapeutic failure has been reported, limiting clinical treatment. This study evaluated the in vitro efficacy of brand-name and compounded itraconazole formulations against Sporothrix brasiliensis and estimated the itraconazole content in each tested formulation. Oral capsules were acquired from two brand-name products for human (H-IND) and veterinary (V-IND) uses, and three from compounding pharmacies in Pelotas, RS, for human (H-COMP1/H-COMP2) and veterinary (V-COMP) uses. Capsule purity was analyzed by liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS). Antifungal activity was determined against 29 Sporothrix brasiliensis by the M38-A2 guideline of CLSI. H-IND/H-COMP1/H-COMP2 had high efficacy against S. brasiliensis (approximately 70% of total isolated susceptible), V-COMP showed moderate efficacy (51.7%), and V-IND was the least effective formulation (37.9%). Thirty-four percent of the total isolates were resistant to all formulations. Furthermore, itraconazole content did not match the concentration indicated by the manufacturers, ranging from 387.70 to 7.81 µg/mg (H-COMP2 > V-COMP > H-IND > H-COMP1 > V-IND). Therefore, it is possible that the formulations showed different in vitro efficacy due to the difference in their itraconazole contents. Given the emergence of antifungal resistance for all formulations, the choice product to be used must follow susceptibility testing. Stringent quality control measures are recommended for product manufactures to assure drug content uniformity.

Hypolipidemic and anti-inflammatory properties of phenolic rich Butia odorata fruit extract: potential involvement of paraoxonase activity.

Aim: This study investigated the effects of polar Butia odorata fruit extract on metabolic, inflammatory, and oxidative stress parameters in rats submitted to a hyperlipidaemia condition induced by tyloxapol.Methods: Animals were divided into 3 groups: saline, saline plus tyloxapol, and B. odorata extract plus tyloxapol. Animals were treated for 15 days with a saline solution or B. odorata fruit extract and after hyperlipidaemia was induced by tyloxapol.Results: Treatment with B. odorata extract reduced serum triglyceride, total cholesterol, C-reactive protein, and adenosine deaminase and butyrylcholinesterase activities when compared to the tyloxapol group. HDL-cholesterol and paraoxonase 1 activity were higher in B. odorata extract treated animals when compared to tyloxapol-treated animals. No differences were observed in hepatic oxidative stress parameters. Phenolic compounds present in B. odorata fruit extract were identified and quantified by LC-MS/MS.Conclusion: These findings indicated that phenolic rich B. odorata extract has hypolipidemic and anti-inflammatory effects in hyperlipidemic rats.

Thermal and irradiation resistance of folic acid encapsulated in zein ultrafine fibers or nanocapsules produced by electrospinning and electrospraying.

The objectives of this study were to characterize zein fibers and capsules prepared by electrospinning and electrospraying techniques, respectively, and then use them to encapsulate folic acid. Folic acid containing fibers and capsules (0.5, 1.0, and 1.5%, w/v) were submitted to thermal treatment (100, 140, and 180 °C) and ultraviolet A light (UVA) irradiation to evaluate the resistance of folic acid. Zein fibers and capsules containing folic acid showed high encapsulation efficiency (>80%). Unencapsulated folic acid showed a reduction in folic acid content from 17.17 µg/mL to 5.44 µg/mL (approximately 70%) when exposed to 180 °C. Photodegradation of unencapsulated folic acid lowered its concentration from 17.17 µg/mL to 12.58 µg/mL (~26% of reduction), when exposed for 1 h to UVA irradiation. However, folic acid concentration when encapsulated in fibers (1.5%) was maintained or only slightly reduced from 9.73 µg/mg to 8.88 µg/mg after thermal treatment at 180 °C. The capsules containing 1.5% of folic acid also presented a slight reduction in folic acid concentration from 8.84 µg/mg to 7.88 µg/mg when exposed to 24 h of UVA irradiation. Zein fibers and capsules containing folic acid present promising characteristics for application in foods that require thermal processing or exposure to irradiation.

Selection of native bacterial starter culture in the production of fermented meat sausages: Application potential, safety aspects, and emerging technologies.

The development of standardized and safe food products with the typical characteristics of each region is highly desirable and can be obtained by using native starter cultures that influence the flavor, texture, and color of fermented foods. Therefore, scientists have been employing various techniques for screening and characterizing native bacteria (lactic acid bacteria and Gram-positive catalase-positive cocci) for application in fermented meat sausage. The present review outlines in vitro assays that evaluate the potential application and safety aspects of native isolates and introduces emerging omics technologies applied to the microbiology of fermented meat sausage. Results from current research are presented, and the strengths and limitations of each assay are provided, with references indicating where further details can be obtained. In choosing the most appropriate in vitro method, it is necessary to consider the available analytical infrastructure, the sensitivity and selectivity of the assay, the time it takes to get the results, the ease of the assay, and the costs involved.

Discrimination of genotype and geographical origin of black rice grown in Brazil by LC-MS analysis of phenolics.

Physicochemical properties, cooking time, and phenolics profile of two black rice genotypes grown at six different locations in Brazil were determined. The cultivar IAC 600 and the elite-line AE 153045 were used. The main growing locations for black rice were considered, as follows: Alegrete (ALG), Capão do Leão (CPL), Guaratinguetá (GUA), Roseira (ROS), Santa Vitória do Palmar (SVP), and Taubaté (TBT). Principal component analysis (PCA) and supervised partial least squares-discriminant analysis (PLS-DA) from liquid chromatography-mass spectrometry (LC-MS) data sets showed distinction among genotypes and locations. Quercetin-3-O-glucoside and vanillic acid were the most relevant compounds for discriminating genotypes. SVP location provided the most distinctive black rice, with greater total phenolics content. Characteristics of black rice from SVP location were associated to effects of latitude and wind conditions. Hesperetin, vanillic acid, quercetion-3-O-glucoside, and p-coumaric acid were the most relevant compounds for discriminating locations.

Changes in Phenolic Acid and Isoflavone Contents during Soybean Drying and Storage.

The changes in phenolic acid and isoflavone profile of soybean genotypes ( Nidera 5909 RR and BMX Força RR) dried at different temperatures and stored for 12 months were investigated. In both cultivars, there was a reduction of the germination capacity and an increase of fungal incidence with the increase of drying temperature and storage time. Multivariate analysis of phenolic acids allowed for the differentiation among treatments. Cultivar Nidera 5909 RR, dried at 110 °C, showed an interaction with characters of greater relevance for differentiation, being influenced by the increase of bound coumaric, and syringic, and free-hydroxybenzoic, syringic, and coumaric acids. Multivariate analysis of isoflavones showed a strong affinity of the aglycone isoflavones (genistein, glycitein, and daidzein) within the Nidera 5909 RR cultivar at all drying temperatures and with BMX Força RR cultivar at the highest temperatures. These results indicate that the release and interconversion of isoflavone malonyl-ß-glucosides and ß-glucosides into aglycone forms are simultaneous reactions during storage.

Polar Origanum vulgare (Lamiaceae) extracts with antifungal potential against Sporothrix brasiliensis.

Oregano (Origanum vulgare) has anti-Sporothrix spp. activity, including against strains that are resistant to antifungal drugs. As the studies are limited to the essential oil, the aim of this study was to evaluate the chemical, antioxidant and cytotoxic properties of polar oregano extracts and their anti-Sporothrix brasiliensis activity. Aerial plant parts were used in the preparation of 10 min (INF10) and 60 min (INF60) infusions, a decoction (DEC) and a hydroalcoholic extract (HAE). Six phenolic acids and four flavonoids were identified and quantified through liquid-chromatography (LC-MS). Extracts in increasing order of total phenolic and flavonoid contents were HAE 40 mg/ml for the other extracts. This is the first report of oregano extracts showing antifungal activity against S. brasiliensis. Its use in the treatment of sporotrichosis may be considered upon toxicity and safe-use conditions are tested.

Cooking quality properties and free and bound phenolics content of brown, black, and red rice grains stored at different temperatures for six months.

The changes in cooking quality and phenolic composition of whole black and red rice grains stored during six months at different temperatures were evaluated. Brown rice with known cooking quality properties and low phenolic levels was used for purposes comparison. All rice genotypes were stored at 13% moisture content at temperatures of 16, 24, 32, and 40°C. Cooking time, hardness, free and bound phenolics, anthocyanins, proanthocyanidins, and free radical scavenging capacity were analysed. The traditional rice with brown pericarp exhibited an increase in cooking time and free phenolics content, while rice with black pericarp exhibited a reduction in cooking time after six months of storage at the highest studied temperature of 40°C. There as increases in ferulic acid levels occurred as a function of storage temperature. Red pericarp rice grains showed decreased antioxidant capacity against ABTS radical for the soluble phenolic fraction with increased time and storage temperature.

Stability of bioactive compounds in butiá (Butia odorata) fruit pulp and nectar.

Butia odorata is a palm tree native to southern Brazil whose fruit (known as butiá) and leaves are used to make many food products and crafts. Butiá contain several biologically active compounds with potential health benefits. However, processing conditions can alter quality attributes including bioactive compound content. This study evaluated the stability of bioactive compounds in butiá pulp upon pasteurization, during 12months of frozen storage, and in butiá nectar after a 3-month storage period. Pulp pasteurization resulted in a reduction in phenolic, flavonoid, carotenoid, and ascorbic acid contents. After a 12-month frozen storage period, flavonoid, phenolic, and ascorbic acid contents decreased while carotenoid content remained unaltered. Carotenoid, ascorbic acid, and phenolic contents were unaffected by the 3-month storage of butiá nectar; however, flavonoid content and antioxidant potential were reduced. Despite bioactive compound degradation upon heat treatment and storage, butiá nectar remained rich in phenolics, especially (-)-epicatechin, rutin, and (+)-catechin.

Chemical composition and cytotoxicity of extracts of marjoram and rosemary and their activity against Sporothrix brasiliensis.

PURPOSE: Motivated by increasing reports of antifungal resistance in human and animal sporotrichosis, this study evaluated the chemical composition, cytotoxicity and anti-Sporothrix brasiliensis activity of extracts of marjoram (Origanum majorana) and rosemary (Rosmarinus officinalis). METHODOLOGY: Ten (INF10) and 60 min (INF60) infusions, a decoction and a hydroalcoholic extract (HAE, 70 %) were prepared from both plants (10 % w/v). The extract composition was analysed by liquid chromatography/mass spectrometry and the cytotoxicity was evaluated using a colorimetric assay in canine and feline kidney cells. Using a broth microdilution assay (CLSI M38-A2) adapted to the extracts, 30 Sporothrix brasiliensis isolates from dogs, cats and humans, and one Sporothrix schenckii were tested.Results/Key findings. The predominant phenolic compounds found in all extracts were 4-hydroxybenzoic acid, caffeic acid and chlorogenic acid. Luteolin was also one of the predominant compounds, but only in the HAE of marjoram. Extracts of marjoram maintained cell viability in concentrations up to 2.5 mg ml-1 for the feline cell line and up to 10 mg ml-1 for the canine cell line, whereas in rosemary, the cell viability for both kidney lines was maintained with concentrations up to 5 mg ml-1. The activity of rosemary extracts was low or absent. Among the marjoram extracts, HAE was highlighted and had fungistatic activity against Sporothrix brasiliensis (MIC5040 mg ml-1), including in all itraconazole-resistant isolates. S. schenckiisensu stricto was sensitive to marjoram extracts (MIC/MFC ≤5 mg ml-1), with the exception of INF10. CONCLUSION: These findings support the potential usefulness of the HAE of marjoram in the treatment of sporotrichosis.

Untargeted Metabolomic Analysis of Capsicum spp. by GC-MS.

INTRODUCTION: In order to conserve the biodiversity of Capsicum species and find genotypes with potential to be utilised commercially, Embrapa Clima Temperado maintains an active germplasm collection (AGC) that requires characterisation, enabling genotype selection and support for breeding programmes. OBJECTIVE: The objective of this study was to characterise pepper accessions from the Embrapa Clima Temperado AGC and differentiate species based on their metabolic profile using an untargeted metabolomics approach. MATERIAL AND METHODS: Cold (-20°C) methanol extraction residue of freeze-dried fruit samples was partitioned into water/methanol (A) and chloroform (B) fractions. The polar fraction (A) was derivatised and both fractions (A and B) were analysed by gas chromatography coupled to mass spectrometry (GC-MS). Data from each fraction was analysed using a multivariate principal component analysis (PCA) with XCMS software. RESULTS: Amino acids, sugars, organic acids, capsaicinoids, and hydrocarbons were identified. Outlying accessions including P116 (C. chinense), P46, and P76 (C. annuum) were observed in a PCA plot mainly due to their high sucrose and fructose contents. PCA also indicated a separation of P221 (C. annuum) and P200 (C. chinense), because of their high dihydrocapsaicin content. CONCLUSIONS: Although the metabolic profiling did not allow for grouping by species, it permitted the simultaneous identification and quantification of several compounds complementing and expanding the metabolic database of the studied Capsicum spp. in the AGC. Copyright © 2017 John Wiley & Sons, Ltd.

Mycotoxin and fungicide residues in wheat grains from fungicide-treated plants measured by a validated LC-MS method.

Wheat (Triticum aestivum) is an annual crop, cultivated in the winter and spring and susceptible to several pathogens, especially fungi, which are managed with fungicides. It is also one of the most consumed cereals, and can be contaminated by mycotoxins and fungicides. The objective of this study was to validate an analytical method by LC-MS for simultaneous determination of mycotoxins and fungicide residues in wheat grains susceptible to fusarium head blight treated with fungicides, and to evaluate the relationship between fungicide application and mycotoxin production. All parameters of the validated analytical method were within AOAC and ANVISA limits. Deoxynivalenol was the prevalent mycotoxin in wheat grain and epoxiconazole was the fungicide residue found in the highest concentration. All fungicidal treatments induced an increase in AFB2 production when compared to the control (without application). AFB1 and deoxynivalenol, on the contrary, were reduced in all fungicide treatments compared to the control.

Butia spp. (Arecaceae) LC-MS-Based Metabolomics for Species and Geographical Origin Discrimination.

The metabolic variability of fruit from Butia spp. (Arecaceae) genotypes from different geographical locations was characterized using untargeted metabolomics by liquid chromatography-mass spectrometry (LC-MS) followed by multivariate data analyses. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) from LC-MS data sets showed a clear distinction among Butia catarinensis, Butia odorata, Butia paraguayensis, and Butia yatay. The major metabolites that contributed to species discrimination were primary metabolites including sugars and organic acids and specialized metabolites such as tetrahydroxy-trans-stilbene and rutin. B. odorata fruit from Tapes, RS, Brazil, showed a high content of organic acids and flavonoids, whereas B. odorata fruits from Capão do Leão, RS, Brazil, showed a high sugar content. The results demonstrate that LC-ESI-qToF-MS-based metabolic profiling coupled with chemometric analysis can be used to discriminate among Butia species and between geographical origins of B. odorata and to identify primary and specialized metabolites responsible for the discrimination.

Preharvest UV-C radiation influences physiological, biochemical, and transcriptional changes in strawberry cv. Camarosa.

Ultraviolet C (UV-C) radiation is known for preventing fungal decay and enhancing phytochemical content in fruit when applied postharvest. However, limited knowledge is available regarding fruit responses to preharvest application of UV-C radiation. Thus, the effects of UV-C radiation on photosynthetic efficiency, dry matter accumulation and partitioning, fruit yield and decay, phytochemical content, and relative transcript accumulation of genes associated with these metabolic pathways were monitored in strawberry (Fragaria x ananassa Duch.) cv. Camarosa. A reduction in photosynthetic efficiency was followed by a decrease in light harvesting complex LhcIIb-1 mRNA accumulation as well as a decrease in yield per plant. Phenylalanine ammonia lyase activity, phenolic, anthocyanin, and L-ascorbic acid contents were higher in UV-C treated fruit. In addition, preharvest UV-C treatment reduced microorganism incidence in the greenhouse and on the fruit surface, increased the accumulation of ß-1,3-Gluc and PR-1 mRNA, and prevented fruit decay.

Bioactive and yield potential of jelly palms (Butia odorata Barb. Rodr.).

In this descriptive study, Butia odorata genotypes were evaluated for yield, fruit number, pulp yield, bioactive content (including phenolic compounds, carotenoid, anthocyanin, L-ascorbic acid, and fibre content), antioxidant potential, and phenotypic characteristics. Genotype 117 was the highest yielding, with an estimated fruit yield of 22,000 kg ha(-1) and pulp yield of 12,000 kg ha(-1). On the other hand, the lowest yielding genotype, accession 49, showed an estimated fruit yield of 8400 kg ha(-1). Jelly palm fruit were generally rich in phenolic content (280.50-398.50 mg 100(-1)g), carotenoid content (2.80-4.08 mg 100 g(-1)), and L-ascorbic acid content (34.63-63.84 mg 100g(-1)). While the highest yielding genotype was not the richest in bioactive content, the lowest yielding genotype showed the highest l-ascorbic acid content. Although fruit yield and phytochemical composition are desirable attributes in jelly palm fruit, none of the genotypes evaluated showed high levels of both. Therefore, fruit yield and bioactive phytochemical content appear to be inversely proportional.