Measurement of urinary concentrations of the mycotoxins zearalenone and sterigmatocystin as biomarkers of exposure in mares.

Mycotoxins may affect animal health, including reproduction. Little is known about the clinical relevance of exposure of horses to contaminated feed. This study aimed at (i) monitoring the levels of the mycotoxins zearalenone (ZEN), with its metabolites α- and ß-zearalenol (α- and ß-ZOL), and sterigmatocystin (STC) in urine samples from thoroughbred mares in Japan and (ii) relating these findings to the potential effects on reproductive efficacy of breeding mares. Sixty-three urine samples of breeding mares from 59 breeding farms were used. Urine samples and reproductive records were collected from each mare when it was presented to the stallion station. Urinary concentrations of ZEN, α- and ß-ZOL, and STC were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). ZEN, α- and ß-ZOL were measurable in the urine of all examined mares, indicating the prevalence of ZEN in equine feeds. In seven of the 63 samples, STC was also detected at levels ranging from 1.3 to 18.0 pg/mg creatinine. No significant correlation between the concentrations of mycotoxins and pregnancy status was observed. In conclusion, measurement of mycotoxins in urine samples is a useful non-invasive method for monitoring the systemic exposure of mares to multiple mycotoxins.

Gas Chromatography-Mass Spectrometry for Metabolite Profiling of Japanese Black Cattle Naturally Contaminated with Zearalenone and Sterigmatocystin.

The objective of this study was to evaluate the metabolic profile of cattle fed with or without zearalenone (ZEN) and sterigmatocystin (STC)-contaminated diets using a gas chromatography-mass spectrometry metabolomics approach. Urinary samples were collected from individual animals (n = 6 per herd) from fattening female Japanese Black (JB) cattle herds (23 months old, 550-600 kg). Herd 1 had persistently high urinary ZEN and STC concentrations due to the presence of contaminated rice straw. Herd 2, the second female JB fattening herd (23 months old, 550-600 kg), received the same dietary feed as Herd 1, with non-contaminated rice straw. Urine samples were collected from Herd 1, two weeks after the contaminated rice straw was replaced with uncontaminated rice straw (Herd 1N). Identified metabolites were subjected to principal component analysis (PCA) and ANOVA. The PCA revealed that the effects on cattle metabolites depended on ZEN and STC concentrations. The contamination of cattle feed with multiple mycotoxins may alter systemic metabolic processes, including metabolites associated with ATP generation, amino acids, glycine-conjugates, organic acids, and purine bases. The results obtained from Herd 1N indicate that a two-week remedy period was not sufficient to improve the levels of urinary metabolites, suggesting that chronic contamination with mycotoxins may have long-term harmful effects on the systemic metabolism of cattle.

Measurement of sterigmatocystin concentrations in urine for monitoring the contamination of cattle feed.

This study aimed (1) at determining the levels of the fungal toxin sterigmatocystin (STC) in the feed and urine of cattle and (2) at evaluating the effects of supplementing the feed with a mycotoxin adsorbent (MA) on STC concentrations in urine. Two herds of female Japanese Black cattle were used in this study. The cattle in each herd were fed a standard ration containing rice straw from different sources and a standard concentrate; two groups of cattle from each herd (n = six per group) received the commercial MA, mixed with the concentrate or given as top-dressing, whereas a third group received no supplement and served as control. Urine and feed samples were collected at various time points throughout the experiment. STC concentrations were measured using liquid chromatography-tandem mass spectrometry (LC-TMS). STC concentrations in straw were higher in Herd 1 (range 0.15-0.24 mg/kg DM) than in Herd 2 (range <0.01-0.06 mg/kg DM). In Herd 1, STC concentrations in urine significantly declined 2 weeks after replacing the contaminated feed, whereas MA supplementation had no effect. In conclusion, mycotoxins in urine samples are useful biological markers for monitoring the systemic exposure of cattle to multiple mycotoxins, as well as evaluating the effectiveness of interventions.

[High levels of sterigmatocystin in patients with chronic liver diseases]. / Detectarea sterigmatocistinei la pacientii cu boala ii hepatica cronica.

UNLABELLED: Sterigmatocystin (STC) is a wide spread mycotoxin produced by Aspergillus fungi, with hepatotoxic and carcinogenetic proprieties. OBJECTIVES: To determine the STC concentration in blood and urine from patient with liver cirrhosis (LC) and hepatocellular carcinoma (HCC), with correlation with liver function parameters. MATERIAL AND METHODS: The study enrolled 166 patients divided in three groups: control--55 patients (27M, 28F); LC--58 patients (31M, 27F); HCC--53 patients (26M, 27F). 20 ml of blood and 50 ml of urine were collected from each patient and liver enzymes and alfa-fetoprotein (AFP) were measured. STC was determined by high performance liquid chromatography, with concomitant detection in ultraviolet and fluorescence. RESULTS: STC was detected in 26.2% of samples, more frequently in LC and HCC groups (p < 0.001). STC mean values were 0.014 ng/ml and 0.005 ng/ml in blood, respective urine of controls, rising to 0.626 ng/ml (p = 0.003) respective 1.053 ng/ml (p = 0.049) in LC and 2.02 ng/ml in blood (p < 0.0001) and 9.39 ng/ml in urine (p = 0.003) in patients with HCC. There is a perfect correlation between serum and urinary levels of STC in controls (r = 1), that become weak in patients with LC (r = 0.48) and insignificant in HCC (r = 0.15). AFP values were significantly correlated with STC concentration in patient with HCC, in both blood (r = 0.31) and urine (r = 0.84). CONCLUSIONS: STC values in patients with LC and HCC were significantly higher compared to controls. Strong positive correlation of STC with AFP in patients with liver cancer suggested a possible role of this mycotoxin in pathogenesis of the disease.

[High blood and urine concentration of hepatotoxic mycotoxins (aflatoxin B1, sterigmatocystin) in patients with liver cirrhosis]. / Detectarea micotoxinelor hepatotoxice la pacienti cu ciroza Hepatica.

UNLABELLED: Aflatoxins and sterigmatocystin are potent carcinogens, certainly involved in pathogenesis of liver cancer. AIM: To evaluate the risk of mycotoxin intake and to determine the presence of aflatoxin B1 (AFB1) and sterigmatocystin (STC) in patients with liver cirrhosis. MATERIAL AND METHOD: The study included 92 patients (33 controls, 59 liver cirrhosis) that completed a food frequency questionnaire (FFQ). Blood and urine samples were collected and mycotoxins determined by high performance liquid chromatography. RESULTS: 18.18% samples in controls and 72.88% in cirrhosis group presented detectable levels of mycotoxins. The mean values of AFB1 in blood were 0.7 ng/mL in controls and 1.67 ng/mL in test group (p = 0.11); STC presented 60 times higher levels in second group (p < 0.01). AFB1 presented a mean level of 1.2 ng/mL in urine of test group (not detected in controls); STC presented 256 time higher concentration in urine of cirrhotic patients, with a perfect correlation between blood and urine levels in control (r=1) and no correlation in test group (r = 0.05). There were no correlations between mycotoxin, liver enzymes, alpha-fetoprotein and mycotoxin intake risk estimated by FFQ. CONCLUSION: Most of the patients presented detectable levels of mycotoxins, significantly increased in cases with liver cirrhosis, probable due to a specific metabolic pattern.