Ochratoxin A (OTA) is a mycotoxin produced mostly by several species of Aspergillus and Penicillium. OTA is nephrotoxic in all animal species in which it has been tested and is cancerogenic in rodents. It is associated with Balkan endemic nephropathy. It is naturally present in many crop products such as cereals (barley, wheat, maize) and dried fruits, spices, coffee, wine, olives, and cocoa. The aim of this study was to assess the contamination of three Ivoirian spices with OTA (ginger, chili, and pepper) widely consumed by the population. A total of 90 spice samples (ginger: n = 30; chili: n = 30; pepper n = 30) was taken from various sales outlets of Abidjan. OTA was quantified using an HPLC apparatus coupled with a fluorimetric detector. The chili and ginger samples were contaminated with OTA at a mean concentration of 57.48 ± 174 and 0.12 ± 0.15μg/kg, respectively. No contamination of the pepper samples was detected. Eight (26.67%) of the chili samples exceeded the maximum limit of 15μg/kg established by European regulation. These results should serve as an alert on the risk to the consumer population of these products that are highly contaminated with OTA.

Using micro high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) a simple and fast method for the quantitative determination of 26 mycotoxins was developed. Sample preparation consists of a single extraction step and a dilute-and-shoot approach without further cleanup. With a total run time of 9 min and solvent consumption below 0.3 mL per chromatographic run, the presented method is cost-effective. All toxins regulated by the European Commission with maximum or guidance levels in grain products (fumonisins B1 and B2 (FB1 and FB2)); deoxynivalenol (DON); aflatoxins B1, G1, B2, and G2 (AFB1, AFG1, AFB2, and AFG2); ochratoxin A (OTA); T-2 and HT-2 toxins; and zearalenone (ZEN) can be quantified with this method. Furthermore, the enniatins B, B1, A, and A1 (EnB, EnB1, EnA, and EnA1); beauvericin (BEA); 3-acetyl-deoxynivalenol (3-AcDON); fusarin C (FusC); sterigmatocystin (STC); gliotoxin (GT); and the Alternaria toxins alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), tentoxin (TEN), and altertoxin I (ATX I) can also be quantified. For all regulated compounds, recoveries ranged between 76 and 120%. For all other toxins, the recovery was at least 51%. The method was applied for the analysis of 42 maize samples from field trials in South Africa.

Sterigmatocystin (STC) is a mycotoxin produced by several species of Aspergillus and other fungi. STC is a precursor of the carcinogen aflatoxin B1 and exerts carcinogenic, teratogenic, and mutagenic effects. A reliable GC-MS-based analytical method using on-column injection was developed and validated to determine STC content in grain. In this method, STC was extracted with acetonitrile (84%), and the filtered extract was diluted with phosphate buffer before immunoaffinity column (IAC) cleanup. After elution and N2 evaporation, the sample containing STC was dissolved in acetone and injected into the GC-MS system without derivatization. Separation of STC was carried out by a capillary column (0.25 mm i.d.×30 m, 0.25 μm) with a deactivated pre-column (0.53 mm i.d.×0.6 m). The matrix effect was investigated in maize, wheat, and rice, and an insignificant matrix effect was observed after IAC cleanup. The calibration curve was linear in the range of 20-300 pg (equivalent to 8-120 μg/kg in grain) with a coefficient of determination (R2) of 0.999, and the relative standard deviation of repeatability (RSDr) was less than 10 %. The limit of detection (LOD) of the method was 6 pg (equivalent to 2.4 μg/kg in grain), and the limit of quantification (LOQ) was 20 pg (equivalent to 8 μg/kg). The method described in this report is sensitive, reliable, and can be used to monitor STC contamination in grain.

This study describes the screening of soil bacteria for their capability to degrade zearalenone (ZEN), employing an enrichment technique in which ZEN is used as the sole carbon source. Two isolates that were able to degrade ZEN belonged to the genus Pseudomonas according to biochemical characterization and 16S rRNA gene sequence and were named as Pseudomonas alcaliphila TH-C1 and Pseudomonas plecoglossicida TH-L1, respectively. The results showed that the degradation rates of P. alcaliphila TH-C1 and P. plecoglossicida TH-L1 for ZEN (2 μg/ml) were 68 ± 0.85 % and 57 ± 0.73%, when incubated for 72 h at 30 °C in a rotary shaker (160 rpm) and detected by high-performance liquid chromatograms (HPLC). These results suggest that the two Pseudomonas strains are new bacterial resource for degrading ZEN and can provide a new approach for the detoxification of ZEN.

The suitability to assess zearalenone (ZEA) exposure in pigs of a commercial ELISA kit for ZEA analysis in urine was tested. A daily dose of 0, 5, 10, 20 and 40 μg synthetic ZEA per kilogram BW was administered via the feed to four gilts per dose group, and after 3 and after 7 days of ZEA intake, urine samples were assayed with the ELISA which has a relative cross-reactivity of 42% with α-zearalenol. The concentration of urinary ZEA equivalents (ZEA plus 42% of α-zearalenol present) did not differ between day 4 and day 8 (P = 0.50) within each dose group. The urinary ZEA equivalent/creatinine ratio was tightly correlated with ZEA intake (r = 0.95). The urinary ZEA equivalent/creatinine values at 0 and 40 μg/kg BW were distinctly different from those of the intermediate dose levels, whereas there was some overlapping of the individual values at the dose levels 5, 10 and 20 μg/kg BW. The urinary ZEA equivalent/creatinine ratio can be used as a biomarker for ZEA exposure in pigs provided that urine samples of several animals receiving the same diet are assayed, either separately or after pooling.

A total of 216 23-week-old laying hens from two different genetic backgrounds (half of the birds were Lohmann brown [LB] and [LSL] hens, respectively) and 24 adult roosters were assigned to a feeding trial to study the effect of increasing concentrations of deoxynivalenol (DON) in the diet (0, 5, 10mg/kg) on the reproductive performance of hens and roosters, and the health of the newly hatched chicks. Hatchability was adversely affected by the presence of DON in LB hens' diet, while the hatchability of the LSL chicks was significantly higher than LB chicks. An interaction effect between DON in the hens' diet and the breed was noticed on fertility, as the fertility was decreased in the eggs of LB hens receiving 10mg/kg DON in their diet and increased in the eggs of LSL hens fed 10mg/kg DON. Moreover, spleen relative weight was significantly decreased in the chicks hatched from eggs of hens fed contaminated diets, while gizzard relative weight was significantly decreased in LB chicks with 10mg/kg DON in their diet compared with the control group. On the other hand, the chicks' haematology and organ histopathology were not affected by the dietary treatment. Additionally, the presence of DON in the roosters' diet had no effect on fertility (the percentage of fertile eggs of all laid eggs). Consequently, the current results indicate a negative impact of DON in LB hens' diet on fertility and hatchability, indicating that the breed of the hens seems to be an additional factor influencing the effect of DON on reproductive performance of the laying hens.

The formation of guttation droplets is a long-known property of various fungi. However, their composition, biological function and metabolism in fungi have hardly attracted deeper research interest. The highly toxic mould Stachybotrys (S.) chartarum chemotype S is supposed to play-amongst other factors such as endotoxins and microbial volatile organic compounds (MVOCs)-an important role in indoor air toxicity, mainly after water damage. The way of toxins becoming airborne and leading to exposure via inhalation, however, is still under discussion. We hypothesised that guttation may be a factor for exudation of toxins into the environment. Therefore, selected isolates (n = 15) of our own culture collection of Stachybotrys spp. (S. chartarum chemotype S, S. chartarum chemotype A, S. chlorohalonta) originating from various habitats were cultivated on malt extract agar for 3weeks. All strains but one produced different amounts of guttation droplets, which were collected quantitatively and subjected to various independent analytical techniques like ELISA, effect-based bioassay (MTT cell culture test) and tandem mass spectrometry (LC-MS/MS). Actually, the toxigenic isolates (n = 5) produced highly toxic guttation droplets, which was confirmed by all methods. The concentration of macrocyclic trichothecenes, such as satratoxin G and H, ranged between the LOD and 7,160ng/ml exudate and 280 and 4,610ng/ml as determined by LC-MS/MS, respectively. According to our knowledge, the ability of S. chartarum to produce toxic exudates is reported for the first time, which possibly plays an important role regarding its toxic potential in indoor environments.

Deoxynivalenol (DON, vomitoxin) is a trichothecene mycotoxin which can be considered to be an indicator of Fusarium mycotoxin contamination in grain, feed and food. Recent studies have described the presence of glucose conjugated DON, which is a product of plant metabolism, but there is a lack of information available on DON conjugation by fungi. The aim of the current study was, therefore, to investigate the ability of fungi to metabolize DON into hydrolysable conjugated DON. Alternaria alternata (54028 NRRL) and Rhizopus microsporus var. rhizopodiformis (54029 NRRL) were found to be capable of metabolizing DON into hydrolysable conjugated DON. This ranged from 13-23% conjugation of DON in potato dextrose agar media and from 11-36% in corn-based media. There was, however, considerable variation between fungal strains in the ability to conjugate DON as only a slight increase in hydrolysable conjugated DON (1-6%) was observed when incubating with A. oryzae (5509 NRRL). A. oryzae (5509 NRRL) was also shown to degrade DON (up to 92%) over 21days of incubation on corn-based media. The current study shows that conjugation of DON can be achieved through fungal metabolism in addition to being a product of plant metabolism.

Zearalenone (ZEA) mainly contaminates grains such as corn and wheat, causing damage to livestock through ingestion of contaminated feed. Recently, various clays have been added to the feed to adsorb mycotoxins and to prevent mycotoxicosis of animals fed contaminated feeds. However the adsorption mechanism of the mycotoxin to clay is not well understood. In this study, a method to analyze the level of adsorption of ZEA to clay was developed using Japanese acid clay. Changes to the amount of the clay, concentration of ZEA, shaking time, and other parameters were evaluated to determine their influence on adsorption. The adsorption isotherms were also developed. Under conditions that mimic the gastrointestinal tract of swine, 100% of ZEA was adsorbed to clay at a pH equivalent to the stomach, while the level of desorption under intestinal basic conditions was 1.8%. Thus Japanese acid clay has a high ability to absorb ZEA with very little desorption under gastrointestinal conditions of the swine. Isothermal analysis suggests that the Japanese acid clay is potentially highly efficacious as a ZEA adsorbent.

Samples of stored maize from villages located in five different agroecological zones (southern lowlands, northern lowlands, Senqu river valley, foothills and mountains) of Lesotho were collected in 2009/10 and 2010/11 and assessed for contamination with toxigenic fungi. The water activity of all samples collected during the two seasons was <0.70. The total fungal populations of the maize from different regions in the two seasons was not significantly different (p > 0.05). Fusarium verticillioides, F. proliferatum and F. subglutinans predominated in different regions in both seasons based on molecular analyses. In the 2009/10 season, the isolates of these species all produced FB1, while in the 2010/11 season, very few produced FB1. A. flavus isolates (2009/10) were recovered from mountains and Senqu river valley samples while the 2010/11 isolates were predominantly from the foothills and northern lowlands. The mountain isolates of Aspergillus section Flavi produced the highest levels of AFB1 (20mgkg(-1)). Aspergillus parasiticus was only isolated from the foothills, Senqu river valley and southern lowlands samples, and the AFB1 levels produced ranged from 'none detected' to 3.5mgkg(-1). The Aspergillus ochraceous isolates were least frequently encountered in both seasons. In the 2009/10 season, the isolates from the northern lowlands produced ochratoxin A (OTA) in culture. No isolates of A. niger from different regions in both seasons produced any OTA. Multi-mycotoxin analyses of the maize samples were done for a range of mycotoxins. At least one sample from each region in both seasons was FB1-positive. FB1 levels for 2010/11 samples (7-936μgkg(-1)) were higher than in the 2009/10 season (2-3μgkg(-1)). In both seasons, the mountains registered the highest levels of FB1. Deoxynivalenol (DON) was recovered from all the samples analysed, with the highest mean contamination of 1,469μgkg(-1) in samples from the northern lowlands. Moniliformin (MON) was detected from all agroecological zones in the two seasons (5-320μgkg(-1) in 2009/10; 15-1,205μgkg(-1) in 2010/11). Emerging toxins such as fusaproliferin (FUS) and beauvericin (BEA) were also detected. OTA was not detected in any of the samples analysed. Only one 2009/10 sample in the Senqu river valley was positive for AFB1. This is the first report on toxigenic fungi and multi-mycotoxin contamination of maize samples from subsistence farmers' stores in different agroecological zones of Lesotho.