Ersilia Alexa

Assessment of inhibitory potential of essential oils on natural mycoflora and Fusarium mycotoxins production in wheat

BackgroundIn the last years essential oils from different plants were used in the prevention of fungi and mycotoxins accumulation in cereals. The most attractive aspect derived from using of essential oils as seed grains protectants is due to their non-toxicity. This study was focused on assessment the inhibitory effect of some essential oils: Melissa officinalis (O1), Salvia officinalis (O2), Coriandrum sativum (O3), Thymus vulgaris (O4) Mentha piperita (O5) and Cinnamomum zeylanicum (O6) against natural mycoflora and Fusarium mycotoxins production correlated with their antioxidants properties.ResultsAll essential oils showed inhibitory effect on fungal contamination of wheat seeds. This ability was dose-dependent. The highest inhibitory effect on Fusarium and Aspergillus fungi was recorded after 5 days of treatment. Fungi such as yeast (Pichia, Saccharomyces and Hyphopichia) were predominantly on seeds mycoflora after 22 days. Each treatment had a selective inhibitory effect on frequency of fungus genera. After 5 days of treatment the most fungicidal effect was recorder for O4, followed by O1. In terms of essential oils effect on mycotoxins development, the best control on fumonisins (FUMO) production was recorded for O6. The antioxidant properties of essential oils decreased in order: O4 > O1 > O6 > O5 > O2 > O3. Also, our data suggested that there is a significant negative correlation between antioxidant properties and seed contamination index (SCI), but there was not recorded a good correlation between antioxidant properties and FUMO content.ConclusionsBased on proven antifungal and antimycotoxin effects as well as their antioxidant properties, the essential oils could be recommended as natural preservatives for stored cereals. The highest inhibition of fungal growth was noted after 5 days of treatment and decreased after 22 days.

Physico-chemical comparison of betulinic acid, betulin and birch bark extract and in vitro investigation of their cytotoxic effects towards skin epidermoid carcinoma (A431), breast carcinoma (MCF7) and cervix adenocarcinoma (HeLa) cell lines

Betulin and betulinic acid are pentacyclic triterpenes present in the bark of the birch tree and other vegetal sources. Quantitatively, in birch bark betulin is more significant than betulinic acid; therefore, birch can be a large and feasible source of raw material for betulin extraction. Betulin can be used as extracted or, after chemical modification, as a starting compound for its acid, betulinic acid, with both substances possessing various interesting pharmacological properties. The purpose of this study is to analyse the betulin and betulinic acid content of a birch tree bark extract, as well as its cytotoxic activity. The extraction was done using a Soxhlet extractor and chloroform/dichlormethane/methanol (1 : 1 : 1) as solvent. The betulin and betulinic acid content of the extract was estimated using standards of pure betulin and betulinic acid, by thermal analysis as opposed to pure substance (thermogravimetric and differential thermal analysis). The extract and the main compounds were also analysed by NMR. The results indicated a high amount of betulin in the final extract (up to 50%), and an important quantity of betulinic acid: over 3%. The cytotoxic activity indicated a high proliferation inhibition for the birch tree extract but was still comparable with betulinic acid and betulin.

Nutritive quality of romanian hemp varieties (Cannabis sativa L.) with special focus on oil and metal contents of seeds

BackgroundThe study aims to determine the nutritional value of hemp seed expressed by the oil content and by the concentration of metals (Ca, Mg, K, Fe, Mn, Zn and Cd), for five varieties of monoecious and dioecious hemp seeds approved in Romania, comparative with the concentration of these metals in the soil.ResultsThe content of oil in hempseed registers a slight decrease in the production records of 2011, losses due to drought and low levels of precipitation during the growth period. The greatest loss is found in Diana monoecious variety (26.54-20.82%) followed by Zenit varieties (27.37-22.97%), Armanca (29.27-25.32%), Silvana (28.89-25.04%) and Denise (26.96-25.30%). Siccative hemp oil has a yellowish green color and an iodine index of 140–156 g I2/100 g oil. Hemp seed are rich in mineral based Ca (144–955 mg/100 g seed), Mg (237–694 mg/100 g seed), K (463–2821 mg/100 g seed), Fe (1133-2400 mg.kg-1), Mn (63–110 mg.kg-1) and Zn (42-94 mg.kg-1). For the soil the following macroelements concentrations were determined: Ca (2100–2520 mg.kg-1), Mg (320–376 mg.kg-1) and K (232–257 mg.kg-1). Mn (156–197 mg.kg-1) and Zn (54–67 mg.kg-1) remain within normal limits for Romania. The soils in the experience area contain large amounts of Fe (19000–20430 mg.kg-1). The presence of K in large quantities determines the accumulation of large quantities of Fe in the soil.ConclusionHempseed belonging to the five Romanian varieties are rich source of nutrients (Ca, Mg, K) and unsaturated oil easily digestible by the body, but the presence of Cd concentrations above the upper limit puts a question mark over the use of seeds in various food products. Hemp extracts easily certain metals from the soil. Significant amounts of Fe (1133–2400 mg.kg-1), Mn (63–110 mg.kg-1), Zn (42–94 mg.kg-1) and Cd (1.3-4.0 mg.kg-1) are found in hemp seeds. Hemp (Cannabis sativa L.) is included among plants suitable for phytoremediation of soil contaminated with cadmium, zinc and iron.

Tracking antioxidant properties and color changes in low-sugar bilberry jam as effect of processing, storage and pectin concentration

BackgroundRecently, an increased interest in the identification of valuable possibilities for preserving the antioxidant properties of products obtained by thermal processing of fruits rich in bioactive compounds can be noticed. In this regard, an extensive analysis is necessary in terms of thermal processed products behavior in relation to various factors. The purpose of the present study was to assess the effect which processing and storage at 20°C has on the antioxidant properties and color quality of low-sugar bilberry jam with different low-methoxyl pectin (LMP) concentrations.ResultsFor all measured parameters, it should be noted that thermal processing induced significant alterations reported to the values registered for fresh fruit. Most important losses due to thermal processing were recorded for total monomeric anthocyanins (TMA) (81-84%), followed by L-ascorbic acid (L-AsAc) content (53-58%), total phenolics (TP) content (42-51%) and FRAP (ferric reducing antioxidant power) values (36-47%). Moreover, depreciation of the investigated compounds occurred during storage at 20°C. Jam storage for 7 months resulted in severe losses in TMA content in the range 58-72% from the value recorded one day after processing. This coincided with marked increases in polymeric color percent of these products after 7 months of storage. Also, bilberry jam storage for 7 months resulted in a decrease in L-AsAc content of 40-53% from the value recorded one day after processing, 41-57% in TP content and 33-46% from the value recorded one day after processing for FRAP values. By decreasing of LMP concentration in the jam recipe from 1 to 0.3% there has been an increase in losses of investigated compounds.ConclusionOverall, the results indicated that bilberry jams can also represent a good source of antioxidant compounds, although compared to the fruit, important losses seem to occur. Practical application of this work is that this kind of information will be very useful in optimizing the jam processing technology and storage conditions, in order to improve the quality of these products.

The occurrence of mycotoxins in wheat from western Romania and histopathological impact as effect of feed intake

BackgroundThe goal of this study has been to evaluate the extent of mycotoxins contamination and their co-occurrence in wheat grain intended for animal feed. A total of 52 wheat samples were collected from the harvest of two consecutive years (2010, 2011) from two counties (Timis and Arad) located in Western Romania and the presence of ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZON) and fumonisins (FUMO) was determined by enzyme-linked immunosorbent assay (ELISA). In order to evaluate the toxicological impact of mycotoxins, naturally contaminated wheat samples were administered in rats feed for one month.ResultsThe mycotoxin with the highest incidence in wheat samples was DON due to agro-climatic conditions typical for the west part of Romania. DON was found in 73.08% of samples harvested in 2010 and the highest level of contamination was 3390 ppb. The incidence of DON in sample from 2011 was lower than those of 2010, with a frequency of occurrence of 19.23%. The occurrence of ZON was in the range 69.23–76.92%, with an average value of 187.74 ppb. The OTA content in wheat was below the maximum tolerable limit established by EU Commission regulation for feed legislation (250 ppb). For FUMO the lowest percentage of positive samples was registered (15.38% in wheat sample harvested in 2010 and 11.54% positive samples in 2011). With respect to the co-occurrence of Fusarium mycotoxins, the results proved that ZON was found as a co-contaminant together with DON, especially when climatic conditions for development of fungus are favorable (high air humidity). The differences recorded between investigated localities and their classifications according to the mycotoxin kind and year of harvest were carried out by principal components analysis (PCA). The histopathological and immunohistochemical evaluation performed by hematoxylin and eosin (HE) staining technique as well as by assessing the vascular endothelial growth factor (VEGF) revealed significant modification of kidney, liver and spleen cells in the case of DON and FUMO. In terms of toxicity induced by OTA and ZON it was highlighted mixed normal and necrotic areas in liver, while histological assessment revealed normal VEGF expression in kidneys.ConclusionsAlthough none of the analyzed samples exceeding the stipulated maximum limits for cereals used as feed, a high incidence of mycotoxins produced by Fusarium species have been recorded (DON and ZON) in wheat samples harvested in Western Romania. Also, histopathological evaluation revealed significant tissue lesions in liver and kidney of rats after one month of feeding with natural contaminated wheat.

Use of ATR-FTIR spectroscopy to detect the changes in extra virgin olive oil by adulteration with soybean oil and high temperature heat treatment

Abstract The structural changes induced in extra virgin olive oil (EVOO) by adulteration with soybean oil (SBO) and heat treatment at 185°C for 4 and 8 h were investigated using Attenuated Total Reflectance – Fourier Transform Infrared (ATR-FTIR) spectroscopy. Our results revealed that the band around 3006 cm–1 recorded shifts versus the percentage of adulterant. The changes in the absorbance at 3006 cm−1 (A3006) and in the ratio of the maximum heights of the bands at 3006 and 2925 cm−1 (A3006/A2925) were used to evaluate the EVOO adulteration. The regression analysis of A3006 and A3006/A2925 versus the percentage of adulterant was used to calculate the detection limits of adulteration. The time course of spectral changes showed that the oil heating caused notable modifications in the intensity of the absorption bands and induced no shifts in their exact position. The most relevant changes were reflected by conjugation and cis-trans isomerisation of double bonds, the formation of epoxides and widening of the band in the C=O region due to formation of secondary oxidation products. This study highlights that ATR-FTIR spectroscopy may be a promising means to differentiate among pure and adulterated oils and to study the thermooxidative processes in oils undergoing thermal stress. Graphical Abstract

Mycoflora and Ochratoxin A Control in Wheat Grain Using Natural Extracts Obtained from Wine Industry By-Products

The aim of this study was to evaluate the effect of some natural extracts obtained from grape pomace (GPE) and grape seeds (GSE) as compared to a synthetic food, antioxidant-butylated hydroxytoluene (BHT), in order to control fungal population and ochratoxin A (OTA) production in naturally contaminated wheat. The results showed that the addition of these extracts was efficient with OTA content decreasing. On treatment with these extracts the loss of OTA content after 14 days was in the range 7.8–28.3% relative to the control sample, but increased up to 26.48–37% after 28 days. The highest loss in OTA content was recorded for treatment with GPE at the 500 ppm level. Regarding the fungal development, the obtained results show that the total fungal populations were significantly reduced by using natural extracts. The most efficient extract was GPE. Both BHT and GPE inhibit the growth of Penicillium verrrucosum, for all doses, but less with Aspergillus genera. GPE affects the growth of other types of moulds such as Rhizopus microsporus, Fusarium graminearum, Alternaria infectoria and Cladosporium herbarum. Our data suggested that GPE and GSE are able to provide fungicidal and fungistatic protection and to control the OTA accumulation in wheat, at least in a similar manner to BHT.

Synergistic Antifungal, Allelopatic and Anti-Proliferative Potential of Salvia officinalis L., and Thymus vulgaris L. Essential Oils

The current study aimed to investigate the chemical composition and the synergistic potential of two essential oils (EOs), as obtained from Salvia officinalis L. (SEO), and Thymus vulgaris L. (TEO). The antifungal potential was tested in vitro against Fusarium graminearum (Fg 06_17), the herbicidal effect was studied using weed seeds of Amaranthus retroflexus (ARET), Chenopodium album (CALB), Echinochloa crus-galli (EGAL), but also wheat seeds (WS) of the Lovrin variety and tomato seeds Saint-Pierre of the variety. The GC-MS profile highlights that the mains compounds identified in SEO were: caryophyllene (25.364%), camphene (14.139%), eucalyptol (13.902%), and β-pinene (11.230%), while in TEO, the predominant phytochemicals were: γ-terpinene (68.415%) and p-thymol (24.721%). The results indicated that the tested EOs alone as well as in combination have allelopathic effect against investigated seeds, while the synergistic effect of TEO and SEO in terms of fungal growth was demonstrated at a level of 0.06%. Thyme and sage EOs exhibited in vitro anti-proliferative activity on two melanoma cell lines, namely A375 human melanoma and B164A5 mouse melanoma alone, as well as in combination. SEO was most effective in terms of decreasing the cell viability of murine and human melanoma cell lines when compared to TEO.

Phytochemical Screening and Biological Activity of Mentha × piperita L. and Lavandula angustifolia Mill. Extracts

The present study aimed to investigate the phytochemical composition of Mentha × piperita L. (MP) and Lavandula angustifolia Mill. (LA) extracts in terms of hydroxycinnamic acid (HCAs) content, in particular, caffeic (CA), p-cumaric (CU), ferulic (FE), and rosmarinic (RS) acids using LC-MS. Also, the in vitro antimicrobial effect against Staphylococcus aureus and the antiproliferative activity against two cancerous cell lines (A375 and MDA-MB-231) using the MTT assay were tested. The extracts were prepared using aromatic water which resulted from the extraction of oils from plants as extraction medium, with/without acid. The results showed that RS and FE represent the majority of HCAs compounds; the highest content of FE is found in LA (7.47 mg·g–1d.m.), and the maximum content of RS in MP (6.36 mg·g–1d.m.). Regarding the antimicrobial effect against Staphylococcus aureus, the two extracts showed a simulative role on the growth rate of Staphyloccocus aureus, but a slightly inhibitory effect (69.12%) can be attributed to the acidic environment. In terms of biological activity against MDA-MB-231 breast carcinoma cell line, and A375 human melanoma cell line, at the highest employed concentration, 150 μg·mL–1, the tested extracts present a weak antiproliferative effect.

Application of QuEChERS - High PerformanceLiquid Chromatography with PostcolumnFluorescence Derivatization (HPLC-FLD) methodto analyze Eprinomectin B1a residues froma pour-on conditioning in bovine edible tissues

Abstract A QuEChERS in house method for determining the marker residue of eprinomectin (eprinomectin B1a) by HPLC-FLD in bovine tissues and milk provided from treated animals was developed and applied. Briefly: all samples were extracted with acetonitrile using a dispersive SPE purification stage. The ascertained detection limits were 1 μg kg-1 and the quantification limits 2 μg kg-1. Recoveries on tissue samples fortified in the range of 10 μg kg-1 to 200 μg kg-1 were from 80.0% to 87.2%, with variation coefficients between 2.7% to 10.6%. The confirmation of residues in the purified extracts was made by LC-MS/MS after separation on an XTerra MS C18 (10 cm × 2.1 mm, 3.5 μm) column with a mobile phase of acetonitrile / formic acid 0.1% (97:3, v/v) at a flow rate of 0.2 mL min-1 and MRM monitoring of three characteristic ions (m/z 896.1, m/z 467.9 and m/z 329.9), resulting from the fragmentation of molecular ions [M-H]+ (m/z 914.6) of eprinomectin and the comparison of the abundance ratio of fragmented ions was obtained in the booth, sample and standard at comparative concentrations. In conclusion, this method has proven its advantage and versatility as a routine drug residues analysis method. Graphical Abstract