Exploring mycotoxin exposure in Parkinson’s disease: no plasma level differences but dietary correlations emerge

Aflatoxins (AFs), ochratoxin A (OTA), citrinin (CIT), fumonisin B1 (FB1), zearalenone (ZEN), and deoxynivalenol (DON) are mycotoxins that may contaminate diets, especially in low-income settings, with potentially severe health consequences. This study investigates the exposure of 439 pregnant women in rural Bangladesh to 35 mycotoxins and their corresponding health risks and links their exposure to certain foods and local stimulants. Overall, 447 first-morning urine samples were collected from pregnant women between July 2018 and November 2019. Mycotoxin biomarkers were quantified by DaS-HPLC–MS/MS. Urinary concentration of frequently occurring mycotoxins was used to estimate dietary mycotoxin exposure. Median regression analyses were performed to investigate the association between the consumption of certain foods and local stimulants, and urinary concentration of frequently occurring mycotoxins. Only in 17 of 447 urine samples (4%) were none of the investigated mycotoxins detected. Biomarkers for six major mycotoxins (AFs, CIT, DON, FB1, OTA, and ZEN) were detected in the urine samples. OTA (95%), CIT (61%), and DON (6%) were most frequently detected, with multiple mycotoxins co-occurring in 281/447 (63%) of urine samples. Under the lowest exposure scenario, dietary exposure to OTA, CIT, and DON was of public health concern in 95%, 16%, and 1% of the pregnant women, respectively. Consumption of specific foods and local stimulants—betel nut, betel leaf, and chewing tobacco—were associated with OTA, CIT, and DON urine levels. In conclusion, exposure to multiple mycotoxins during early pregnancy is widespread in this rural community and represents a potential health risk for mothers and their offspring.

Human biomonitoring of multiple mycotoxins in the Belgian population: Results of the BIOMYCO study

ABSTRACTContamination of grains with mycotoxins results in a dietary background exposure of the general population. In occupational settings such as during processing of raw materials as in milling, an additional mycotoxin exposure by inhalation is possible. Biomonitoring is an integrative approach to assess human exposure from various sources and by all routes. To investigate possible workplace exposure to mycotoxins, a pilot study was conducted that compared levels of urinary biomarkers in mill workers to those in a control group with dietary mycotoxin intake alone. Workers (n = 17) from three grain mills in North Rhine Westphalia, Germany, provided spot urines during shift; volunteers (n = 13, IfADo staff) with matched age structure served as control group. The mycotoxins selected for biomarker analysis were citrinin (CIT) deoxynivalenol (DON), ochratoxin A (OTA), and zearalenone (ZEN). Immunoaffinity columns (CIT, DON, ZEN) or liquid–liquid extraction (OTA) was employed for urine sample cleanup prior to targeted analysis by liquid chromatography with tandem mass spectrometry (LC-MS/MS) or by high-performance liquid chromatography (HPLC). In addition, mycotoxin metabolites that may be formed in the organism were analyzed, including deepoxy-deoxynivalenol (DOM-1), ochratoxin alpha (OTα), dihydrocitrinone (DH-CIT), and α- and β-zearalenol (α- and β-ZEL), as well as phase II metabolites that were hydrolyzed with β-glucuronidase/arylsulfatase prior to sample cleanup. All analyte concentrations were adjusted for creatinine (crea) content in the spot urine samples. Citrinin, DON, OTA, and ZEN were detected in nearly all urine samples from mill workers and controls. Interestingly, DH-CIT was found at higher mean levels than the parent compound (~0.14 and 0.045 µg/g crea, respectively), suggesting an effective metabolism of CIT in humans. Other metabolites DOM-1, OTα, and α- and β-ZEL were detected less frequently in urine. Deoxynivalenol was detected at the highest concentrations (mean ~6 µg/g crea), followed by OTA (mean ~0.08 µg/g crea); ZEN (mean ~0.03 µg/g crea) and its metabolites appeared in urine at lower levels. Mycotoxin biomarker levels in urine from mill workers and controls were not significantly different. From these results it is concluded that biomarker levels measured in urine samples from the two cohorts reflect mainly dietary mycotoxin exposure. An additional occupational (inhalational) exposure of mill workers, if any, is apparently low at the investigated workplaces.

Multiple mycotoxin exposure assessment through human biomonitoring in an esophageal cancer case-control study in the Arsi-Bale districts of Oromia region of Ethiopia

Delving into the possible role of Ochratoxin A in Parkinson's disease: a computational study spotlighting the potential mechanisms of action.

In this study, the modulation of key enzymes involved in epigenetic regulation was assessed in immortalized bovine macrophages (BoMacs) following in vitro exposure to the following Penicillium mycotoxins: citrinin (CIT), ochratoxin A (OTA), patulin (PAT), mycophenolic acid (MPA), penicillic acid (PA), or a combination of one of the above with OTA at the concentration that inhibits BoMac proliferation by 25% (IC25). Real-time PCR analysis of the genes coding DNA methyltransferases (DNMTs), histone demethylases (JMJD-3 and UTX), as well as the class-1 histone deacetylases (HDAC-1, -2, and -3) and histone acetylase (Bmi-1) was assessed following 6 h of mycotoxin exposure. A change in the expression of JMJD-3 as well as HDAC-3, MPA (p = 0.1) and PA (p = 0.08), by at least one of the treatments was observed at their respective IC25. The expression of JMJD-3 was significantly induced by PA, but synergistically suppressed by CIT + OTA. The combination of CIT + OTA also synergistically suppressed the expression of DNMT-3a and DNMT-3b. The combination of PAT + OTA reduced DNMT-3a expression, while PA + OTA reduced DNMT-3b expression. Lastly, MPA and PA slightly reduced HDAC-3 expression, while OTA in combination with CIT, PAT, MPA or PA synergistically suppressed HDAC-3 expression. The results of this study demonstrate that Penicillium mycotoxin exposure, specifically OTA and other mycotoxin combinations, can alter the expression of BoMac enzymes that are involved in epigenetic regulation. These findings suggest a potential novel regulatory mechanism by which mycotoxins can modulate macrophage function.

Biomonitoring studies can provide valuable insights into human mycotoxin exposure, especially when food contaminant data are scarce or unavailable as in Bangladesh. First biomonitoring data in Bangladeshi adults indicated exposure to the nephrotoxic mycotoxins ochratoxin A (OTA) and citrinin (CIT). This led us to conduct a follow-up study with analysis of urinary biomarkers for both CIT and OTA to investigate regional and seasonal influences on mycotoxin exposure in two Bangladeshi cohorts. In total, 164 urines were collected (n=69 in summer, n=95 in winter) from residents of a rural and an urban area, among which there were 62 participants enrolled in both sampling periods. Most urines had detectable biomarker levels (OTA, CIT and its metabolite dihydrocitrinone, HO-CIT), with more or less pronounced differences with regard to season and region. In both cohorts, OTA was found at a mean level of 0.06±0.10ng/mL urine (range 0.01-0.55ng/mL) in summer and a mean of 0.19±0.38ng/mL (range 0.01-1.75ng/mL) in winter season. A season difference was significant in the rural cohort, but not in the urban cohort, and slightly higher mean OTA levels in the rural compared to the urban cohort were only observed in winter urines. CIT biomarkers showed more pronounced variations, with a CIT mean of 0.10±0.17ng/mL (range 0.02-1.22ng/mL) and HO-CIT mean of 0.42±0.98ng/mL (range 0.02-5.39ng/mL) in summer, and CIT mean of 0.59±0.98ng/mL (range 0.05-5.03ng/mL) and HO-CIT mean of 3.18±8.49ng/mL (range 0.02-46.44ng/mL) in winter urines of both cohorts. In both seasons, total CIT biomarker concentrations were significantly higher in the rural cohort than in the urban cohort. A provisional daily intake for CIT was calculated and exceeded a preliminary value set by EFSA (0.2µg/kg/d) in 10 and 24% of participants in summer and winter, respectively. No significant correlations were found between urinary biomarker levels and intake of certain types of food, except for a positive trend for higher rice consumption. Our results in the Bangladeshi population indicate frequent co-exposure to nephrotoxic mycotoxin food contaminants that vary by season and region.

Hepatotoxic effect of ochratoxin A and citrinin, alone and in combination, and protective effect of vitamin E: In vitro study in HepG2 cell

Evaluating the human neurotoxicity and toxicological interactions impact of co-occurring regulated and emerging mycotoxins

Prenatal ochratoxin A exposure, birth outcomes and infant growth in rural Burkina Faso: A human biomonitoring sub-study from the MISAME-III trial

Determination of multiple mycotoxins in paired plasma and urine samples to assess human exposure in Nanjing, China

Citrinin (CTN) and ochratoxin A (OA) were fed alone and in combination to broilers from day of hatch until 3 weeks of age. Dietary concentrations of 300 micrograms CTN/g and 3.0 micrograms OA/g were used. Birds fed CTN had significantly (P less than or equal to 0.05) lower body weights than controls on days 14 and 21 and increased water consumption on days 7, 14, and 21. Birds fed OA had significantly lower body weights than controls on days 7, 14, and 21 and increased water consumption on day 14. Birds fed CTN and OA in combination had lower body weights than controls and increased water consumption during the experiment, but the alterations were intermediate in severity when compared with those in birds fed CTN or OA alone. Birds fed OA alone or combined with CTN had higher liver and kidney weights than controls, but birds fed CTN alone had only higher kidney weights. Birds fed both CTN and OA had concentrations of serum constituents similar to those in birds fed OA alone, except the levels of cholesterol and triglycerides were not significantly different from those in the controls. Histological evaluation of the kidney indicated no lesions in birds fed CTN alone, but birds fed OA, alone or in combination with CTN, had increased tubular casts and tubular hyperplasia compared with controls. These data suggest that there were no additive or synergistic toxic interactions when 300 micrograms CTN/g and 3.0 micrograms OA/g were fed simultaneously to broiler chicks for 3 weeks. However, the severe growth depression resulting from OA and the increased water consumption associated with CTN toxicosis were ameliorated when CTN and OA were fed in combination. These data may be useful in diagnosing field cases of mycotoxicosis where both CTN and OA are involved.

Mycotoxins are potent fungal toxins that frequently contaminate agricultural crops and foods. Mycotoxin exposure is frequently reported in humans, and children are known to be particularly at risk of exceeding safe levels of exposure. Urinary biomonitoring is used to assess overall dietary exposure to multiple mycotoxins. This study aims to quantify multi-mycotoxin exposure in UK children and to identify major food groups contributing to exposure. Four repeat urine samples were collected from 29 children (13 boys and 16 girls, aged 2.4–6.8 years), and food diaries were recorded to assess their exposure to eleven mycotoxins. Urine samples (n = 114) were hydrolysed with β-glucuronidase, enriched through immunoaffinity columns and analysed by LC-MS/MS for deoxynivalenol (DON), nivalenol (NIV), T-2/HT-2 toxins, zearalenone (ZEN), ochratoxin A (OTA) and aflatoxins. Food diaries were analysed using WinDiet software, and the daily intake of high-risk foods for mycotoxin contamination summarised. The most prevalent mycotoxins found in urine samples were DON (95.6% of all samples), OTA (88.6%), HT-2 toxin (53.5%), ZEN (48.2%) and NIV (26.3%). Intake of total cereal-based foods was strongly positively associated with urinary levels of DON and T-2/HT-2 and oat intake with urinary T-2/HT-2. Average daily mycotoxin excretion ranged from 12.10 µg/d (DON) to 0.03 µg/d (OTA), and co-exposure to three or more mycotoxins was found in 66% of samples. Comparing mycotoxin intake estimates to tolerable daily intakes (TDI) demonstrates frequent TDI exceedances (DON 34.2% of all samples, T-2/HT-2 14.9%, NIV 4.4% and ZEN 5.2%). OTA was frequently detected at low levels. When mean daily OTA intake was compared to the reference value for non-neoplastic lesions, the resulting Margin of Exposure (MoE) of 65 was narrow, indicating a health concern. In conclusion, this study demonstrates frequent exposure of UK children to multiple mycotoxins at levels high enough to pose a health concern if exposure is continuous.

Dietary exposure assessment and risk characterization of citrinin and ochratoxin A in Belgium

Total aflatoxin and ochratoxin A levels, dietary exposure and cancer risk assessment in dried fruits in Türkiye