Browsing by Author "Visintin, L."
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- Early-life exposure to MYCOtoxins and its impact on health – a case studyPublication . Alvito, P.; Assunção, R.; Bastos-Amador, P.; de Boevre, M.; Duarte, E.; Martins, C.; Serrenho, i.; Silva, I.; Visintin, L.; Ferreira, M.Considering the potential impact on health and the scarce data available regarding early-life exposure to mycotoxins, earlyMYCO project (early-life exposure to MYCOtoxins and its impact on health) proposed to answer key questions: are pregnant women and infants until six months exposed to mycotoxins? Is this exposure a health threat? Does this early-life exposure influence the intestinal immune system development? Which is the burden derived from the exposure to mycotoxins? The earlyMYCO pilot study enrolled 19 pairs of mother and children, with a loss to follow-up ranging between 11% and 47% for different moments of observation. The mycotoxins’ biomarkers detected were AFB1, OTA, DON and bZEL in urine samples (mother and children), and AFB1, aZEL, FB1, FB2 and FB3 in breast milk samples. Food consumption data revealed that foods consumed more frequently during the week were bread, dairy products, non-alcoholic drinks (tea and coffee), animal products (meat and fish) and pasta. Regarding infants, 22% were fed with infant formula and 78% were exclusively breastfed. Considering the exposure levels, a low risk of mothers’ exposure to the main mycotoxins analyzed is expected, since urine samples did not reveal detectable levels of these compounds; however, infants’ urine samples presented a DON mean value of 14.8 ng/mL (corresponding to 148.0 μg/kg bw/day through reverse dosimetry), which could represent a risk for this population group. Notably, maternal exposure to AFB1 promoted an increase of overall T cell population, while it also resulted in a selective reduction of cytokine-producing innate lymphoid cells group 2 (ILC2) population in intestine of the progeny. These alterations were associated with decreased expression of Reg3b and Reg3g by the intestinal mucosa of progeny. Thus, these results indicate that maternal exposure to mycotoxins impacts the development of offspring intestinal immune system. An in vitro approach using intestinal cell lines Caco-2 and Caco-2/HT29-MTX models exposed to AFB1 during 24h, confirmed the deleterious effects of AFB1 on intestinal membrane integrity and its effect on mucus layer. To assess the impact of AFB1 on early-life microbiota, faeces from litters of AFB1 treated female mice and controls were assessed by metagenomics. Although the overall diversity (Shannon diversity index) of the microbiome wasn’t affected between groups, the microbiome composition varied between AFB1 and control faecal samples (Bray–Curtis dissimilarity index). In particular, some beneficial species were diminished in the litters from AFB1 treated females. Results emphasized the need for assessing the prenatal and lactation exposure to mycotoxins.
- Human biomonitoring of multiple mycotoxins in biological samples of Portuguese ‘earlyMYCO‘ mother-and-child pairsPublication . Visintin, L.; García-Nicolás, M.; Dhondt, E.; Martins, C.; Assunção, R.; Serrano, D.; Alvito, P.; Vidigal, C.; Almeida, E.; Nunes, C.; De Saeger, S.; De Boevre, M.Early-life exposure to hazardous compounds is an emerging research field that urgently needs to be investigated, as an attempt to contribute to exposome research. Mycotoxins are naturally-occurring food contaminants and recent studies reported that Portuguese children (< 3 years) as well as the adult population are exposed to multiple mycotoxins through food consumption constituting a potential health threat. Considering the above, a project designated by earlyMYCO (Early-life exposure to MYCOtoxins and its impact on health) is being developed to assess the risk of early-life exposure to mycotoxins in biological samples. The earlyMYCO project included a mother-and-child longitudinal study, with three moments of observation, starting from pregnancy (24-28 weeks) and going through the child’ first months of life (1-6 months). This study was conducted in the Primary Health Care Group Lisboa Central (ACES) and aimed to determine multiple mycotoxins and its biomarkers in breast milk and urine samples from mother-and-child pairs. The sample preparation procedure for breast milk samples (n=9) consisted of a solid-phase extraction (SPE) purification followed by a liquid-liquid extraction (LLE); urine samples (n=50) were prepared by applying the QuEChERS technique. Liquid chromatography coupled to ESI+ Xevo TQ-S mass spectrometry (UHPLC-ESI-MS/MS) was used to determine multiple mycotoxin biomarkers of exposure in a targeted approach, namely aflatoxins B1, G1, B2, G2 and M1 (AFB1, AFG1, AFB2, AFG2, AFM1), ochratoxin A and alpha (OTA, OT-α), deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEN), alpha and beta zearalenol (α-ZEL, β-ZEL), fumonisins B1, B2, B3 and hydrolyzed B1 (FB1, FB2, FB3, HFB1), sterigmatocystin (STE), and aflatoxicol (AFL) using a previously optimized and validated method. All the breast milk samples analysed contained multiple mycotoxins to whom the children were exposed to. Unfortunately, due to covid-19 the number of samples was restricted. All the breastmilk samples resulted to be positive for FB1 and FB2 (exception for one sample) with concentrations between 0.16 and 34.76 ng/mL and 0.14 and 11.90 ng/mL, respectively; 5 samples resulted positive also for FB3 (1.09-4.97 ng/mL). DON was detected in one sample as well as NIV with concentrations of 0.75 and 3.26 ng/mL, respectively. Moreover, AFB1 and AFG1 were detected respectively in three samples (0.36-0.64 ng/mL) and one sample (below the LOD), while in two samples OTA was present below the LOD. Besides the targeted approach on urine samples, an untargeted LC-HRMS approach was launched on both sample matrices.