The Presence of Molds and Their Secondary Metabolites in Purple Coneflower-Based Dietary Supplements (Echinacea purpurea (L.) Moench).

Purple coneflower (Echinacea purpurea (L.) Moench) is a plant in the family Asteraceae, mainly grown in North America. Echinacea purpurea has been used in conventional medicine. The plant has immuno-stimulating and antibacterial properties, but neither mold contamination nor a mycotoxin presence have been evaluated. Our goal is to determine the degree to which molds and mycotoxins contaminate dietary supplements based on purple coneflower distributed on the Polish market. We analyzed 21 samples divided into four groups: sachets (n = 5), dry raw material (n = 3), capsules (n = 9), and tablets (n = 4). The mycological analysis of dietary supplements shows that the average number of molds is 1012 cfu/g, and the most common molds are Aspergillus spp., Phoma spp. and Eurotium spp. The mycotoxins most common in the samples are ZEN (18/21), DON (5/21) and T-2 toxin (3/21).

Dietary Supplements Based on Red Yeast Rice-A Source of Citrinin?

Citrinin (CIT) is secondary metabolite of filamentous molds. This mycotoxin has nephrotoxic, hepatotoxic, embryocidal, and fetotoxic properties. It is also produced by several species of the three genera Penicillium spp., Aspergillus spp., and Monascus spp., which are used to make red yeast rice (RYR). The material for this study consisted of 15 dietary supplements containing an extract of fermented red rice, available on the Polish market. Samples were extracted using a MeOH-H2O mixture, cleaned-up with an immunoaffinity CitriTest HPLC column, and quantified by HPLC-FLD. None of the analyzed samples contained CIT above the established limit of detection (LOD). Studies on the presence of toxic metabolites in red yeast rice show the importance of regulating this product and of clear information on the label regarding the standardized amounts of monacolin.

Concentrations of zearalenone and its metabolites in female wild boars from woodlands and farmlands.

As a valuable food base and safe shelter, large agricultural fields create favourable living conditions for wild boars for most of the year. The occurrence of mould in these fields, causing hormonal disturbances, may lead to a lower age of sexual maturity and a prolonged breeding season, and therefore, the population growth in farmlands can be markedly higher than that in extensive woodlands. This study was initiated because of reproductive cycle disturbances in wild boar populations, which were presumably linked with habitat and food types, especially rotten maize. To determine if the major sources of zearalenone (ZEN) in female wild boars are maize cobs infected with mycotoxins, we compared the concentrations of ZEN and its metabolites in organs, tissues and body fluids of wild boars hunted in extensive maize fields and in extensive woodlands where no large maize fields were located within a distance of several km. Samples of blood, bile, liver, kidneys, muscles, urine, stomach and colon contents as well as ovaries from young female wild boars (40-60 kg) were collected in 2011-2014 and prepared for liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The results showed that ZEN was present in most of the samples, from both farmlands and woodlands, but its concentration within individual types of the analysed body fluids or tissues differed depending mainly on habitat type. In nearly all the analysed cases, higher concentrations of ZEN and its derivatives were detected in the samples collected from wild boars in farmlands, suggesting that wild boars living and feeding in extensive maize fields take in markedly higher amounts of mycoestrogens.

Ochratoxin A and citrinin in green coffee and dietary supplements with green coffee extract.

The aim of this study was to determine the degree of mold contamination and mycotoxin levels in commercially available green coffee products and dietary supplements with green coffee extract. The study included 34 samples from green coffee products: raw beans (n = 16), ground coffee (n = 15) and instant coffee (n = 3), as well as 22 samples from dietary supplements in form of capsules (n = 19), tablets (n = 2) and sachets (n = 1). Total mold count was determined with spread-plate method. Anamorphic mold were identified based on their microscopic morphology and the type of sporulation. Concentrations of mycotoxins, ochratoxin A and citrinin, were quantified by means of HPLC-fluorescence detection. Molds, typically Aspergillus spp. and Penicillium spp., were found in 94% of green coffee beans, 100% of ground and instant coffee samples, and 55% of dietary supplement samples. None of the samples contained detectable levels of citrinin. Ochratoxin A (0.4 ng/g) was detected in only one sample of raw green coffee beans, but in up to 40% and 67% of ground and instant coffee samples, respectively. Mean concentrations of ochratoxin A in ground and instant coffee samples were 3.28 ng/g and 4.09 ng/g, respectively, and maximum concentrations amounted to 6.65 ng/g and 7.44 ng/g, respectively. Ochratoxin A (mean concentration 9.60 ng/g, maximum level 31.4 ng/g) was also detected in up to 58% of the supplement capsules, but in none of tablets and sachets.

Cytotoxic effect of filtering respiratory protective devices from the waste sorting industry: is in vitro toxicology useful for risk characterization?

The use of Filtering Respiratory Protective Devices (FRPD) is mandatory in Portugal to protect workers from the waste industry of harmful exposures. Deleterious health effects of exposure to bioburden via inhalation and/or ingestion include respiratory symptoms and nephrotoxicity. Between January and February 2019, 118 FRPD samples were collected in one waste sorting industry and characterized regarding microbial contamination and cytotoxicity, defined as cell metabolic activity, through the MTT colorimetric assay (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide). Cytotoxic effect was classified according to percentage of extinction values with respect to the control group, as follows: absent (≥90); low (80%-90%, +); medium (60%-79%, ++); and high (below 60%, +++). For 113 samples the MTT assay revealed a cytotoxic effect in A549 cells, of which 81 presented high cytotoxicity. In SK cells, a cytotoxic effect was observed in 56 samples, of which five displayed a high cytotoxic effect. Several moderate (p < 0.05) to strong (p < 0.01) correlations were found between higher bacterial and fungal counts both in interior layers (fungi and bacteria) and in exhalation valves (fungi) of FRPD samples and reduced cell metabolic activity of SK cells. On the basis of the obtained results for the cytotoxic effect of FRPD samples on two different cells lines, it was determined that A549 cells exhibited a cytotoxic effect for a higher number of FRPD, whereas the SK cells model correlated better with the other assessed parameters, namely, bacterial and fungal counts and conditions of FRPD use. Although the results are not conclusive on the most appropriate cell line to assess FRPD cytotoxicity, they reinforce the importance of in vitro toxicology in exposure assessments to determine the cytotoxicity of mixtures of contaminants, for better risk characterization and selection of appropriate risk management measures.

Mycotoxin Contamination Concerns of Herbs and Medicinal Plants.

Plants and medicinal herbs that are available on the market do not always meet quality and safety standards. One particular concern is the risk of contamination with mycotoxins. Aflatoxins and ochratoxin A are the most frequently described mycotoxins in herbal products and have repeatedly been reported to occur at concentrations which exceed regulatory levels set by the European Union (EU). Possible solutions include enforcing existing limits, and for the new materials, establishing tighter limits and mandate the growth of medicinal plants in EU member countries under more strict conditions.

Advantageous Extraction, Cleanup, and UHPLC-MS/MS Detection of Patulin Mycotoxin in Dietary Supplements and Herbal Blends Containing Hawberry from Crataegus spp.

Patulin (PAT) is a highly genotoxic mycotoxin still found as the common contaminant of various kinds of spoiled fruits and related commodities which are often endorsed as the health-enhancing products. Thus, a fast and convenient liquid-solid extraction followed by a solid-phase cleanup with the MycoSep®228 AflaPat multifunctional column was used for the highly efficient isolation of PAT with an average recovery of 112.7% from commercial dietary supplements and herbal blends formulated with dried hawberry. Analysis of the PAT content was carried out using gradient elution with a Synergi Polar C18 column (150 × 2 mm, 4 µm) and UHPLC system equipped with a mass spectrometer. PAT was detected in all (n=14) commercial single-component dietary supplements formulated with dried hawberry belonging to Crataegus monogyna and/or Crataegus laevigata. Similarly, PAT was detected in 67% of the studied multicomponent commercial herbal blends (n=6) that contained-in addition to hawberry-different amounts of apple, chokeberry, elderberry, hibiscus, or mallow. Moreover, the PAT content was determined in the hawberry collected from the mature wild hawthorn trees belonging to three botanical species, Crataegus monogyna Jacq., Crataegus laevigata (Poiret) DC, and Crataegus rhipidophylla Gand, growing in the recreational forest areas and in the law-protected state national forest park in Poland. In conclusion, to prevent PAT accumulation and reduce the health risk of consumers in globalizing markets, the implementation of improved cultivation/processing practices of hawthorn trees and hawberry as well as increased analytical control related to the presence of PAT in dietary supplements and herbal blends formulated with fresh, dried, or frozen hawberry should be urgently recommended.