Chronic Effects of Fusarium Mycotoxins in Rations with or without Increased Concentrate Proportion on the Insulin Sensitivity in Lactating Dairy Cows.

The objective of this study was to investigate the effect of long-term exposure to a Fusarium toxin deoxynivalenol (DON, 5 mg/kg DM) on the energy metabolism in lactating cows fed diets with different amounts of concentrate. In Period 1 27 German Holstein cows were assigned to two groups and fed a control or mycotoxin-contaminated diet with 50% concentrate for 11 weeks. In Period 2 each group was further divided and fed either a diet containing 30% or 60% concentrate for 16 weeks. Blood samples were collected in week 0, 4, 8, 15, 21, and 27 for calculation of the Revised Quantitative Insulin Sensitivity Check Index and biopsy samples of skeletal muscle and the liver in w 0, 15, and 27 for analysis by real-time RT-qPCR. The DON-fed groups presented lower insulin sensitivities than controls at week 27. Concomitantly, muscular mRNA expression of insulin receptors and hepatic mRNA expression of glucose transporter 2 and key enzymes for gluconeogenesis and fatty acid metabolism were lower in DON-fed cows compared to the control. The study revealed no consistent evidence that DON effects were modified by dietary concentrate levels. In conclusion, long-term dietary DON intake appears to have mild effects on energy metabolism in lactating dairy cows.

Zearalenone (ZEN) metabolism and residue concentrations in physiological specimens of dairy cows exposed long-term to ZEN-contaminated diets differing in concentrate feed proportions.

A long-term feeding experiment with dairy cows was performed to investigate the effects of feeding a Fusarium toxin contaminated (FUS) and a background-contaminated control (CON) ration with a mean concentrate feed proportion of 50% during the first 11 weeks after parturition (Groups FUS-50, CON-50, Period 1), and with concentrate feed proportions of 30% or 60% during the remaining 17 weeks (Groups CON-30, CON-60, FUS-30 and FUS-60, Period 2), on zearalenone (ZEN) residue levels in blood serum, milk, urine and bile. ZEN, α-zearalenol (α-ZEL) and ß-zearalenol (ß-ZEL), zearalanone (ZAL), α-zearalanol (α-ZAL) and ß-zearalanol (ß-ZAL) were determined by HPLC with fluorescence detection. The ZEN concentrations of the rations fed to Groups CON-50, FUS-50 (Period 1), CON-30, CON-60, FUS-30 and FUS-60 (Period 2) amounted to 53.1, 112.7, 35.0, 24.4, 73.8 and 72.5 µg/kg dry matter, respectively. The concentrations of ZEN, α-ZEL, ß-ZEL, ZAN, α-ZAL and ß-ZAL in serum, urine and milk were lower than 1, 1, 4, 100, 50 and 200 ng/g, respectively, while ZEN, α-ZEL and ß-ZEL were detected in bile. Their levels changed with oral ZEN exposure in the course of the experiment and in a similar direction with concentrate feed proportion (Period 2 only). Thus the proportions of the individual ß-ZEL, α-ZEL and ZEN concentrations of their sum varied only in narrow ranges of 68-76%, 6-13% and 12-20%, respectively. Interestingly, the bile concentrations of ß-ZEL, α-ZEL and ZEN of Groups CON-60 and FUS-60 amounted to only approximately 50%, 45% and 62%, respectively, of those of Groups CON-30 and FUS-30 despite a similar or even lower ZEN exposure. The results indicate that conversion of ZEN to its detectable metabolites was not changed by different dietary concentrate feed proportions while their absolute levels were decreased. These findings might suggest concentrate feed proportion-dependent and rumen fermentation-mediated alterations in ZEN/metabolite degradation, and/or liver associated alterations in bile formation and turnover.

Effects of deoxynivalenol (DON) and related compounds on bovine peripheral blood mononuclear cells (PBMC) in vitro and in vivo.

The Fusarium toxin deoxynivalenol (DON) often co-occurs along with the acetylated derivatives 3-acetyl-DON and 15-acetyl-DON in diets for ruminants. De-epoxy-DON is formed by rumen micro-organisms, while the acetylated DON derivatives might also undergo ruminal metabolism with de-epoxy-DON as an end product. However, despite the fact that de-epoxy-DON is the predominant substance finally absorbed, a complete degradation of the mother compounds can not be assumed for all feeding and metabolic situations of the cow, and thus raising the question of their possible post-absorptive effects. Hence, the aim of the study was to examine the effects of all four compounds on the concanavalin A stimulated proliferation of bovine peripheral blood mononuclear cells (PBMC) using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) as indicator in vitro and ex vivo. Among the DON-related compounds, DON and 15-acetyl-DON resulted in a similar IC50 (i.e. the concentration where the proliferation was inhibited by 50%) of 0.5 µM, whereas 3-acetyl-DON was less toxic (IC50 = 2.6 µM), while actually no IC50 could be estimated for de-epoxy-DON which was characterized by a maximum inhibition of approximately 24% at the highest tested in vitro concentration of 18.29 µM. For the in vivo experiment, 14 Holstein cows were used and fed either an uncontaminated control diet (CON) or a diet contaminated with Fusarium toxins, with DON being the predominating toxin for 18 weeks when blood was collected for PBMC isolation and subsequent proliferation/viability assay. The complete diets for the CON and FUS group contained 0.4 and 4.6 mg DON/kg DM, respectively, at that time. Exposure of dairy cows to the FUS diet resulted in maximum serum de-epoxy-DON levels of 52 ng/ml (0.19 µM), while levels of the unmetabolized DON reached maximum levels of 9 ng/ml (0.03 µM). The PBMC of these cows were slightly less viable, by approximately 18% (p = 0.057), while stimulation capability was not decreased at the same time. Although de-epoxy-DON was characterized by the lowest in vitro toxicity among the tested DON-related compounds, there appeared to be a lower viability of the PBMC isolated from cows fed the FUS diet, which had nearly exclusively de-epoxy DON in serum beside slight traces of unmetabolized DON. Thus, the factors responsible for these apparent discrepancies need to be clarified.

No carry over of unmetabolised deoxynivalenol in milk of dairy cows fed high concentrate proportions.

To examine the carry over of deoxynivalenol (DON) and its metabolite de-epoxy DON (DOM-1) in milk, lactating German Holstein cows (n = 13) were fed an isoenergetic total mixed ration in Period 1 with 50% concentrates and 5.3 mg DON/kg dry matter (DM) over 11 wk and were compared with control cows (n = 14). In Period 2 (18 wk), an elevated concentrate proportion was compared to a low concentrate ration by dividing the cows into four Groups (n = 8): Control-30 (30% concentrates), Myco-30 (30% concentrates, 4.4 mg DON/kg DM), Control-60 (60% concentrates) and Myco-60 (60% concentrates, 4.6 mg DON/kg DM). Taken both periods together, no unmetabolised DON was detected in milk samples using the HPLC-UV method. DOM-1 concentrations ranged between below the LOD and 3.2 microg/kg milk in mycotoxin fed cows, while control cows did not excrete any measurable amounts of DOM-1. Regarding the concentrate effects, the carry over of DON as DOM-1 in milk was negligible (between 0.0001 and 0.0011) but significantly higher in Group Myco-30 than in Group Myco-60. This effect may result from an altered bioavailability of DON from maize silage which made up a higher proportion of the daily ration.

Ruminal fermentation patterns and parameters of the acid base metabolism in the urine as influenced by the proportion of concentrate in the ration of dairy cows with and without Fusarium toxin-contaminated triticale.

Feeding a total mixed ration with 50% concentrate and a mean deoxynivalenol (DON) concentration of 5.3 mg/kg DM to 13 German Holstein cows in early lactation (Myco group) resulted in alterations in the ruminal fermentation patterns (lower molar percentage of acetate and isobutyrate, higher molar percentage of valerate) compared to the 14 control cows (Period 1, 11 weeks). In the Myco group, significantly lower ruminal pH value occurred in weeks 4 and 8 and lower minimum pH values critical for developing subacute ruminal acidosis were detected. Accordingly, the net acid base excretion in the urine and the base-to-acid ratio were lower (significant in week 8 only). These effects probably resulted from a higher dry matter intake and are not related to the presence of Fusarium toxin. In Period 2, the same 27 cows plus five additional cows were divided into four groups over 18 weeks. The effects of an elevated concentrate proportion of 60% were tested with and without Fusarium toxin (Control-60, 0.4 mg DON/kg DM and Myco-60, 4.6 mg DON/kg DM) and compared to two groups fed 30% concentrate (Control-30, 0.6 mg DON/kg DM and Myco-30, 4.4 mg DON/kg DM). As expected, a high concentrate proportion significantly affected the ruminal fermentation patterns. Net acid base excretion and base-to-acid ratio did not reveal a distinct concentrate effect and the ruminal pH values were on a high level in all groups when ruminal fluids were taken after 3-5 hours of feed restriction (median between 6.8 and 7.2). Additional effects were observed on the profile of short chain fatty acids in the presence of Fusarium toxin at both concentrate levels. This indicates a switch in the microbial community due to direct mycotoxin effects and/or indirect effects of the Fusarium infection related alterations in the physico-chemical properties of the infected cereal on ruminal microbes.

On the effects of the concentrate proportion of dairy cow rations in the presence and absence of a fusarium toxin-contaminated triticale on cow performance.

The aim of the present study was to investigate the effects of a deoxynivalenol (DON) contaminated ration with a concentrate proportion of 50%, on the performance of dairy cows (Period 1), and to examine the effects when the concentrate proportion was elevated to 60% compared to a ration with 30% concentrates (Period 2). In Period 1, 13 lactating German Holstein cows (Myco group, on average 29 days in milk) were fed the experimental diet (on average 5.3 mg DON/kg DM) as total mixed ration over 11 weeks, while another 14 cows (on average 33 days in milk) received a control diet. Both rations contained 50% concentrates (on DM basis). In Period 2 (18 weeks), the same 27 cows plus five additional cows were divided into four groups: Control-30 (30% concentrates), Myco-30 (30% concentrates, 4.4 mg DON/kg DM), Control-60 (60% concentrates), Myco-60 (60% concentrates, 4.6 mg DON/kg DM). The overall performance level was characterised by a mean daily DM intake of 17.9 kg and a mean daily milk production of 26.7 kg fat corrected milk (FCM) in Period 1 and 17.3 kg DM intake and 24.5 kg FCM in Period 2, respectively. In both periods cows fed the Fusarium toxin-contaminated diets consumed more DM (in Period 2 only significant for group Myco-30) resulting from stimulating effects on the ingesta passage rate of the natural contaminated Fusarium-infected triticale. In Period 1, cows fed the Fusarium toxin-contaminated diet had a significantly higher milk yield, milk urea and somatic cell count, whereas milk fat and protein concentration and fat-to-protein-ratio (FPR) were significantly lower. In Period 2, on a low concentrate level, FCM was significantly higher in group Myco-30. On a high concentrate level, group Myco-60 produced significantly more milk, but milk fat and protein concentration, FPR and milk urea were significantly lower. A concentrate proportion of 60% had a depressing effect on milk fat concentration but was significantly more pronounced in the presence of Fusarium toxin-contaminated and Fusarium damaged-triticale.