Individual and combined effects of deoxynivalenol (DON) with other Fusarium mycotoxins on rainbow trout (Oncorhynchus mykiss) growth performance and health.

This study assessed whether the toxicological effects of deoxynivalenol (DON) produced by Fusarium graminearum in rainbow trout (Oncorhynchus mykiss) are altered by the co-exposure to a mixture of toxins produced by Fusarium verticillioides (FUmix). This FUmix contained fusaric acid and fumonisin B1, B2 and B3. Four diets were formulated according to a 2 × 2 factorial design: CON-CON; CON-FUmix; DON-CON; and DON-FUmix. Diets with and without DON contained on average 2700 and 0 µg/kg feed, respectively. The sum of the analysed FUmix toxins was 12,700 and 100 µg/kg feed in the diets with and without FUmix, respectively. The experiment consisted of a 6-week restrictive feeding period immediately followed by a 2-week ad libitum feeding period. Growth performance measurements were taken per feeding period. Histopathological measurements in the liver and gastrointestinal tract (pyloric caeca, midgut and hindgut) were assessed at the end of week 1 and week 6 of the restrictive feeding period and at week 8, the last day of the ad libitum feeding period. During both restrictive and ad libitum feeding, the effects of FUmix and DON on growth performance were additive (no interaction effect; p > 0.05). During the restrictive feeding period, exposure to DON (p ≤ 0.001) and FUmix (p ≤ 0.01) inhibited growth and increased feed conversion ratio (FCR). During this period, DON exposure decreased the protein (p ≤ 0.001) and energy retention (p ≤ 0.05) in the trout. During the ad libitum feeding period, FUmix affected HSI (p ≤ 0.01), while DON exposure reduced feed intake (p ≤ 0.001) and growth (p ≤ 0.001) and increased FCR (p ≤ 0.01). In general, for both liver and intestinal tissue measurements, no interaction effects between DON and FUmix were observed. In the liver, histopathological analysis revealed mild alterations, increased necrosis score by DON (p ≤ 0.01), increased glycogen vacuolization by FUmix (p ≤ 0.05) and decreased percentage of pleomorphic nuclei by FUmix (p ≤ 0.01). DON had a minor impact on the intestinal histological measurements. Over time, some of the liver (glycogen vacuolization score, pleomorphic nuclei; p ≤ 0.01) and intestinal measurements (mucosal fold and enterocyte width; p ≤ 0.01) were aggravated in fish fed the FUmix contaminated diets, with the most severe alterations being noted at week 8. Overall, the co-exposure to FUmix and DON gave rise to additive effects but showed no synergistic or antagonistic effects for the combination of DON with other Fusarium mycotoxins.

Time- and Dose-Dependent Effects of Dietary Deoxynivalenol (DON) in Rainbow Trout (Oncorhynchus mykiss) at Organism and Tissue Level.

This study with juvenile rainbow trout evaluated the effects of dietary exposure to deoxynivalenol (DON) at industrially relevant doses (up to 1.6 mg/kg) on growth performance, the liver, and the gastrointestinal tract. Fifteen groups of 30 fish each were given one of five dietary treatments in triplicate: (1) control diet (CON; DON < 100 µg/kg feed), (2) naturally DON-contaminated diet (ND1) with a DON content of 700 µg/kg in the feed, (3) ND2 with a DON content of 1200 µg/kg feed, (4) a pure DON-contaminated diet (PD1) with 800 µg/kg of DON in the feed, and (5) PD2 with DON at a concentration of 1600 µg/kg in the feed. The feeding trial lasted eight weeks: six weeks of restrictive feeding followed by two weeks of ad libitum feeding. Exposure to DON during restrictive feeding for six weeks did not affect the growth performance of trout but did lead to a reduction in retained protein in fish fed with higher doses of DON in the ND2 and PD2 groups. During the two following weeks of ad libitum feeding, feed intake was similar among all groups, but body weight gain was lower in the ND2 and PD2 groups and feed efficiency was higher in PD2 (week 8). Histopathological assessment revealed liver damage, including altered nuclear characteristics and haemorrhages, in groups fed higher doses of natural DON (ND2) after just one week of restrictive feeding. Liver damage (necrosis and haemorrhage presence in ND2) was alleviated over time (week 6) but was again aggravated after ad libitum exposure (week 8). In contrast, gastrointestinal tract damage was generally mild with only a few histopathological alterations, and the absence of an inflammatory cytokine response was demonstrated by PCR at week 8. In conclusion, ad libitum dietary exposure of rainbow trout to either natural or pure DON resulted in reduced growth (dose-dependent), while restrictive exposure revealed time-dependent effects of natural DON in terms of liver damage.

The Occurrence of Mycotoxins in Raw Materials and Fish Feeds in Europe and the Potential Effects of Deoxynivalenol (DON) on the Health and Growth of Farmed Fish Species-A Review.

The first part of this study evaluates the occurrence of mycotoxin patterns in feedstuffs and fish feeds. Results were extrapolated from a large data pool derived from wheat (n = 857), corn (n = 725), soybean meal (n = 139) and fish feed (n = 44) samples in European countries and based on sample analyses by liquid chromatography/tandem mass spectrometry (LC-MS/MS) in the period between 2012-2019. Deoxynivalenol (DON) was readily present in corn (in 47% of the samples) > wheat (41%) > soybean meal (11%), and in aquafeeds (48%). Co-occurrence of mycotoxins was frequently observed in feedstuffs and aquafeed samples. For example, in corn, multi-mycotoxin occurrence was investigated by Spearman's correlations and odd ratios, and both showed co-occurrence of DON with its acetylated forms (3-AcDON, 15-AcDON) as well as with zearalenone (ZEN). The second part of this study summarizes the existing knowledge on the effects of DON on farmed fish species and evaluates the risk of DON exposure in fish, based on data from in vivo studies. A meta-analytical approach aimed to estimate to which extent DON affects feed intake and growth performance in fish. Corn was identified as the ingredient with the highest risk of contamination with DON and its acetylated forms, which often cannot be detected by commonly used rapid detection methods in feed mills. Periodical state-of-the-art mycotoxin analyses are essential to detect the full spectrum of mycotoxins in fish feeds aimed to prevent detrimental effects on farmed fish and subsequent economic losses for fish farmers. Because levels below the stated regulatory limits can reduce feed intake and growth performance, our results show that the risk of DON contamination is underestimated in the aquaculture industry.