Carry-over of deoxynivalenol into eggs of laying hens — Preliminary results

Carry-over of deoxynivalenol (DON) into eggs was investigated within the scope of a 16-week experiment with laying hens, in which the birds were fed a maize-based diet containing DON at 11.9 mg/kg dry matter. Eggs were collected during weeks 2, 4, 8, and 16. DON and its metabolite deepoxy-DON were analysed separately in freeze-dried yolk and albumen. Yolk was extracted with water and the extract was purified using an immunoaffinity column (IAC). Albumen was extracted with acetonitrile-water and the extract was pre-cleaned before applying an IAC. All albumen and some yolk samples were incubated with β-glucuronidase prior to extraction. DON and de-epoxy-DON were determined by high performance liquid chromatography (HPLC) with diode array detection (DAD). The detection limits of both toxins were 20 ng/g and 15 ng/g in freezedried yolk and albumen, respectively, corresponding to approximately 10 ng/g and 2 ng/g in fresh samples. The recovery of DON/de-epoxy-DON in spiked samples (50–200 ng/g) was 87/83% (yolk) and 87/77% (albumen) with coefficients of variation of 4–15%. Neither DON nor de-epoxy-DON were detected in any of the samples. In order to achieve lower detection limits, the methods are currently optimized. However, these preliminary results indicate that eggs do not contribute significantly to the dietary DON intake of the consumer. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germany, May 17–19, 2004     

Serum cation profile of broilers at various stages of exposure to deoxynivalenol

The present experiment was carried out to investigate if levels of serum cations in broilers are modulated differently at various stages of exposure to deoxynivalenol (DON). Male broiler chicks at 7 days of age were fed a basal diet (0.27 mg of DON; 0.01 mg of zearalenone/kg), or either a low DON diet (1.68 mg of DON; 0.15 mg of zearalenone/kg) or a high DON diet (12.21 mg of DON; 1.09 mg of zearalenone/kg) produced using extracts from Fusarium graminearum cultures. Blood samples from the birds were collected during weeks 2, 4, and 5 of exposure. The high DON diet resulted in lower serum calcium levels compared to the basal diet at all the 3 sampling stages, while the low DON diet resulted in lower serum calcium levels only during weeks 2 and 5. Serum potassium levels were reduced under both the DON diets during weeks 2 and 5, while no diet-associated changes were found for serum levels of magnesium, sodium, and zinc. Under the present experimental conditions, the serum levels of calcium were consistently modulated in the broilers exposed to the DON-contaminated diets. The modulation of serum levels of potassium was, however, dependent upon the stage of exposure to DON.     

Dietary deoxynivalenol does not affect mineral element accumulation in breast and thigh muscles of broiler chicken

Deoxynivalenol (DON), a well-known contaminant of feed, can have negative effects on gut permeability and function in poultry, which then could affect major and trace element content of the broilers’ breast and thigh muscles, and ultimately reduce meat quality. To study this hypothesis, DON-contaminated diet was fed to broiler chicks. Two groups of birds were housed in metabolic cages with free access to water and feed, with or without DON (10 mg/kg). After 5 weeks, birds were dissected and samples of the breast and thigh muscles, feed and droppings were analysed for five macro (Ca, K, Mg, Na, and P) and ten micro elements (Al, Cr, Cu, Fe, Mn, Li, Mo, Ni, Pb, Rb, and Zn) by inductively coupled plasma optical emission spectrometry (ICP-OES) or inductively coupled plasma mass spectrometer (ICP-MS) methods. In both groups, increased (p?<?0.05) concentrations of Ca Na, Fe, Mn, and Zn were found in thigh muscles compared with the breast, whereas the concentrations of Mg, P, and Rb were higher in the breast muscles. DON had no effect on the elemental contents of the broilers’ breast and thigh muscles. In conclusion, DON at a level of 10 mg/kg feed to broiler chicken over of 5 weeks did not alter the macro or micro element composition in muscle meat.     

Humic substances failed to prevent the systemic absorption of deoxynivalenol (DON) and its adverse effects on piglets

The aim of this study was to examine the effects of a control diet (CON, 0.25?mg DON/kg diet) or a Fusarium toxin-contaminated diet (FUS, 4.49?mg DON/kg diet) without and with humic substances (HS) (CON-HS and FUS-HS, 0.23 and 4.56?mg DON/kg diet, respectively) on piglets during a 5-week growth trial starting after weaning (6.7?±?0.9?kg live weight, n?=?20/group). Feed intake was significantly reduced by feeding the FUS containing diets by approximately 21% compared with the CON diet irrespective of HS supplementation. The decrease in live weight gain paralleled the feed intake depression and amounted to approximately 26%. Feeding the FUS diet was clearly reflected by the DON levels in blood. While only traces of DON with median concentrations of 3?ng/ml (2?C5?ng/ml) and 2?ng/ml (0?C3?ng/ml) were detected in piglets fed the CON and CON-HS diets, respectively, significantly higher levels of 22.5?ng/ml (7?C30?ng/ml) and 23.5?ng/ml (15?C32?ng/ml) were found in piglets fed the FUS and FUS-HS diet, respectively. The urinary excretion of DON and its metabolite de-epoxy-DON as percentage of DON intake was not significantly influenced by HS supplementation and amounted to 24.1 and 20.2% for groups FUS and FUS-HS, respectively. In conclusion, the tested HS preparation cannot be recommended as a DON inactivating feed supplement for pigs.     

Urinary deoxynivalenol (DON) and zearalenone (ZEA) as biomarkers of DON and ZEA exposure of pigs

Four diets contaminated with 1.1 to 5.0 mg/kg deoxynivalenol (DON) and 0.4 to 2.4 mg/kg zearalenone (ZEA) were fed to four groups of six growing Large White pigs. Urine samples were collected after 3 to 4 days and again after 6 to 7 days on the diets. On each sampling day, half of the animals were sampled in the morning, after an 8-h fast, and the other half were sampled in the afternoon, after 7 h of ad libitum access to feed. The urinary concentrations of DON, DON-glucuronide, DON-3-sulphate, de-epoxy-DON, as well as of ZEA, ZEA-14-glucuronide, α-zearalenol and α-zearalenol-14-glucuronide, analysed using LC-MS/MS, were used to calculate urinary DON and ZEA equivalent concentrations (DONe and ZEAe). The urinary concentration of DONe (P?<?0.001), but not of ZEAe (P?=?0.31), was lower in the fasted than that in the fed animals. The urinary DONe/creatinine and ZEAe/creatinine ratios were highly correlated with DON and ZEA intake per kg body weight the day preceding sampling (r?=?0.76 and 0.77; P?<?0.001). The correlations between DON intake during the 7 h preceding urine sampling in the afternoon and urinary DONe/creatinine ratio (r?=?0.88) as well as between mean ZEA intake during 3 days preceding urine sampling and urinary ZEAe/creatinine ratio (r?=?0.84) were even higher, reflecting the plasma elimination half-time of several hours for DON and of more than 3 days for ZEA. ZEAe analysed in enzymatically hydrolysed urine using an ELISA kit was highly correlated with the LC-MS/MS data (r?=?0.94). The urinary DONe and ZEAe to creatinine ratios, analysed in pooled urine samples of several pigs fed the same diet, can be used to estimate their exposure to DON and ZEA.     

Effect of Dietary Supplementation with Copper Sulfate or Tribasic Copper Chloride on the Growth Performance, Liver Copper Concentrations of Broilers Fed in Floor Pens, and Stabilities of Vitamin E and Phytase in Feeds

An experiment was conducted using a total of 840, 1-day-old, Arbor Acres commercial male broilers to compare copper (Cu) sulfate and tribasic Cu chloride (TBCC, Cu₂(OH)₃Cl) as sources of supplemental Cu for broilers fed in floor pens. Chicks were randomly allotted to one of seven treatments for six replicate pens of 20 birds each, and were fed a basal corn–soybean meal diet (10.20 mg/kg Cu) supplemented with 0, 100, 150, or 200 mg/kg Cu from either Cu sulfate or TBCC for 21 days. Chicks fed 200 mg/kg Cu as TBCC had a higher (P?<?0.05) average daily gain (ADG) than those consuming other diets. Liver Cu contents of broilers fed diets supplemented with TBCC were numerically lower (P?>?0.05) than those of broilers fed diets supplemented with Cu sulfate. The vitamin E contents and the phytase activities in the feed fortified with TBCC were higher (P?<?0.01) and numerically higher (P?>?0.05) compared with those in the feeds fortified with Cu sulfate stored at room temperature, respectively. The vitamin E contents in liver and plasma of broilers fed diets supplemented with TBCC were higher (P?<?0.05) than those of birds fed diets supplemented with Cu sulfate. This result indicates that TBCC is more effective than Cu sulfate in improving the growth of broilers fed in floor pens, and it is chemically less active than Cu sulfate in promoting the undesirable oxidation of vitamin E in feeds.     

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.     

On the effects of the <Emphasis Type="Italic">Fusarium</Emphasis> toxin deoxynivalenol (DON) administered per os or intraperitoneal infusion to sows during days 63 to 70 of gestation

Six pregnant sows of 180.6 ± 5.6 kg were fed either a Fusarium-contaminated (4.42 mg DON and 48.3 μg ZON per kg, DON per os, n = 3) or a control diet (0.15 mg DON and 5 μg ZON/kg) in the period of days 63 and 70 of gestation. On day 63 of gestation, sows fed the control diet were implanted with an intraperitoneal osmotic minipump (delivery rate of 10 μL/h, for 7 days) containing 50 mg pure (98%) DON in 2 ml 50% DMSO (DON ip, n = 3). Frequent plasma samples were taken to estimate the kinetics after oral and ip DON exposure. The intended continuous delivery of DON by the intraperitoneal minipump could not be shown, as there was a plasma peak (C_max) of 4.2–6.4 ng DON/mL either immediately (sow IP-2+3) or 2.5 h (sow IP-1) after implantation of the pump followed by a one-exponential decline with a mean half-time (t_1/2) of 1.75–4.0 h and only negligible DON plasma concentrations after 12 h. Therefore, the DON ip exposure has to be regarded as one single dose 1 week before termination of experiment. The DON per os sows showed a mean basis level (after achieving a steady state) of DON plasma concentration of about 6–8 ng/mL, as also indicated by the plasma DON concentration at the termination of the experiment. On day 70, caesarean section was carried out, the fetuses were killed immediately after birth, and samples of plasma, urine, and bile were taken to analyze the concentration of DON and its metabolite de-epoxy-DON. At necropsy there were no macroscopic lesions observed in any organ of either sows or piglets. Histopathological evaluation of sows liver and spleen revealed no alterations. The proliferation rate of peripheral blood mononuclear cells (PBMC) with or without stimulation was not affected by the kind of DON treatment. The exposure of pregnant sows at mid-gestation (days 63–70, period of organogenesis) to a Fusarium toxin-contaminated diet (4.42 mg DON and 0.048 mg ZON per kg) or pure DON via intraperitoneal osmotic minipump did not cause adverse effects on health, fertility, maintenance of pregnancy, and performance of sows and their fetuses. However, DON was detected in fetus plasma, indicating that this toxin can pass the placental barrier and may cause changes in the proportion of white blood cells (lower monocyte and neutrophil and higher lymphocyte proportion in DON per os fetuses).     

Effects of low to moderate levels of deoxynivalenol on feed and water intake,weight gain,and slaughtering traits of broiler chickens

The aim of the study was to evaluate the effects of low to moderate oral exposure to the Fusarium toxin deoxynivalenol (DON; derived from culture material) on performance, water intake, and carcass parameters of broilers during early and late developmental phases. A total of 160 Ross 308 broilers were randomly allocated to four different feeding groups (n = 40/group) including 0 (control), 2.5, 5, and 10 mg DON/kg wheat-soybean meal-based feed. Three consecutive replicates of the experiment were performed. Half of the broilers were slaughtered in week 3 of the trial whereas the other half were slaughtered in week 5. Dry matter intake (DMI) and water intake (WI) were recorded on a daily basis and the body weight (BW) and BW gain (BWG) were determined weekly. The following carcass traits were recorded and calculated in absolute and relative data: dressed carcass weight, breast muscle weight, leg weight, and liver weight. Data showed that BW (P < 0.001), BWG (P = 0.005), and DMI (P < 0.001) were reduced by DON-feeding during the entire feeding period. The ratio of DMI to body weight gain (DMI/BWG) was not affected by the treatment. However, the ratio of water to DMI (WI/DMI) increased in DON-treated birds (P = 0.021). Contrast analysis showed that DON tendentially reduced slaughter weight (P = 0.082) and decreased leg yield (P = 0.037) in DON-fed chickens in week 5 of the experiment. Liver organ weight decreased in the 3-week-old DON-fed broilers compared to that in the control-fed birds (P = 0.037). In conclusion, the study suggests that DMI and BW were negatively affected under the experimental conditions at DON levels lower than the current guidance value in the European Union of 5 mg/kg feed. The study also indicates that broilers fed on low to moderate level DON-contaminated diets showed increased WI/DMI ratio which might have negative influence on wet litter syndrome.     

Lipopolysaccharides (LPS) modulate the metabolism of deoxynivalenol (DON) in the pig

Pigs might be exposed to lipopolysaccharides (LPS) and deoxynivalenol (DON) at the same time, and both toxins are thought to interactively affect the intestinal barrier, the innate immune system, and the xenobiotics metabolism. Hence, we aimed at examining the single and combined effects of both toxins on nutrient digestibility and DON metabolism. For this purpose, barrows (26?±?4 kg) were fed restrictedly either a control diet (CON) or a diet contaminated with 3.1 mg DON/kg (DON) for 37 days. At day 37 of the experiment, pigs were infused intravenously for 60 min either with 100 μg DON/kg body weight (BW) (CON-DON), 7.5 μg LPS/kg BW (CON-LPS, DON-LPS) or a combination of both substances (CON-DON?+?LPS), or physiological saline (CON-CON, DON-CON). Blood samples were collected frequently until 3.25 h before the pigs were sacrificed for bile, liver, and kidney collection. The apparent digestibility of N-free extractives was significantly increased by 1 % when the DON-contaminated diet was fed. The total DON content in blood was significantly higher in endotoxemic pigs (34.8 ng/mL; CON-DON?+?LPS) when compared to the pigs infused with DON alone (18.8 ng/mL; CON-DON) while bile concentrations were not influenced by LPS. DON residue levels in liver and kidney closely reflected the treatment effects as described for blood. In contrast to DON infusion, the LPS challenge resulted in a significantly lower total DON concentration (13.2 vs. 7.5 ng/mL in groups DON-CON and DON-LPS, respectively) when the pigs were exposed to DON through the diet. The conjugation degree for DON in blood and bile was not influenced by treatments. In conclusion, endotoxemic pigs are characterized by higher DON residue levels in blood, liver, and kidney, probably by a compromised elimination.     

Effects of oral exposure to sodium sulphite-treated deoxynivalenol (DON)-contaminated maize on performance and plasma concentrations of toxins and metabolites in piglets

The objective of the present study was to demonstrate the efficiency of the decontamination process applied to deoxynivalenol (DON)-contaminated maize by sodium sulphite (Na₂SO₃) treatment in vivo. Additionally, in vitro characterisation of the toxicity of the DON sulphonates (DONS 1, 2 and 3 denote structurally different forms), the resulting DON metabolites, on peripheral blood mononuclear cells (PBMC) should substantiate the inactivation of DON. In a piglet experiment, both DON-contaminated maize and -uncontaminated control maize either untreated (DON?, CON?) or Na₂SO₃-treated (DON+, CON+) were mixed into feed and fed for 42 d starting from weaning. The results showed that feed intake and daily weight gain of animals fed DON? were significantly lower compared to animals fed CON? and CON+, whereas group DON+ reached the control level or even exceeded it. The feed-to-gain ratio was unaffected (p = 0.45). Furthermore, DON concentrations in plasma markedly reflected the diets’ DON concentrations. These were < 0.1, < 0.1, 5.4 and 0.8 mg/kg feed for CON?, CON+, DON? and DON+, and amounted to 0.3, 0.4, 33.0 and 9.3 ng/ml in plasma, respectively. Whereas DONS 2 and 3 were detected in the DON+ diet, only DONS 2 was recovered in plasma. Regarding the toxicity of DONS, no or much lower toxicity was found compared to DON. DONS 1 and Na₂SO₃ did not affect the viability of PBMC. At 32.71μM DONS2 the viability was reduced by 50% and thus this compound was less toxic than DON by a factor of 73. Consequently, wet preservation of maize with Na₂SO₃ was an effective tool to avoid the adverse effects of DON on performance of piglets.     

Effects of Feed Contaminant Deoxynivalenol on Plasma Cytokines and mRNA Expression of Immune Genes in the Intestine of Broiler Chickens

An experiment was conducted to investigate the individual and combined effects of dietary deoxynivalenol (DON) and a microbial feed additive on plasma cytokine level and on the expression of immune relevant genes in jejunal tissues of broilers. A total of 40 broiler chicks were obtained from a commercial hatchery and divided randomly into four groups (10 birds per group). Birds were reared in battery cages from one day old for 5 weeks. The dietary groups were 1) control birds fed basal diet; 2) DON group fed basal diet contaminated with 10 mg DON/ kg feed; 3) DON + Mycofix group fed basal diet contaminated with 10 mg DON/ kg feed and supplemented with a commercial feed additive, Mycofix® Select (MS) (2.5 kg/ton of feed); 4) Mycofix group fed basal diet supplemented with MS (2.5 kg/ton of feed). At 35 days, the plasma levels of tumor necrosis factor alpha (TNF-α) and interleukin 8 (IL-8) were quantified by ELISA test kits. Furthermore, the mRNA expression of TNF-α, IL-8, IL-1β, interferon gamma (IFNγ), transforming growth factor beta receptor I (TGFBR1) and nuclear factor kappa-light-chain-enhancer of activated B cells 1 (NF-κβ1) in jejunum were quantified by qRT-PCR. The results showed that the plasma TNF-α decreased in response to DON, while in combination with MS, the effect of DON was reduced. DON down-regulated the relative gene expression of IL-1β, TGFBR1 and IFN-γ, and addition of MS to the DON contaminated diet compensates these effects on IL-1β, TGFBR1 but not for IFN-γ. Furthermore, supplementation of MS to either DON contaminated or control diet up-regulated the mRNA expression of NF-κβ1. In conclusion, DON has the potential to provoke and modulate immunological reactions of broilers and subsequently could increase their susceptibility to disease. The additive seemed to have almost as much of an effect as DON, albeit on different genes.     

On the toxicokinetics and the metabolism of deoxynivalenol (don) in the pig

Eleven castrated male pigs weighing 88.1?±?3.9?kg on average were adapted to a diet containing DON (4.2?mg DON/kg) over a period of 7 days. Feed was given restrictively with 1.1?kg per meal (two meals per day). On the day of measurement, all pigs were slaughtered at different time intervals following the morning meal containing DON (1, 2, 3, 4, 5, 6, 8, 15, 18 and 24?h after feeding), with the exception of one pig which was slaughtered unfed. DON and de-epoxy-DON were analysed in serum and digesta from consecutive segments of the digestive tract (stomach, small intestine divided into three parts of a similar length, caecum, colon, rectum). DON was rapidly and nearly completely absorbed while passing through the stomach and the proximal small intestine. Maximum serum concentration appeared 4.1?h after the DON-containing meal and half of the systemically absorbed DON was eliminated after 5.8?h. De-epoxy-DON appeared in increasing proportions from the distal small intestine and reached approximately 80% of the sum of DON plus de-epoxy-DON in faeces collected from the rectum. It was concluded that de-epoxydation of DON, which primarily occurs in the hindgut, probably does not contribute much to a detoxification in the pig.     

Metabolic changes in Brycon cephalus (Teleostei,Characidae) during post-feeding and fasting

Metabolic changes during the transition from post-feeding to fasting were studied in Brycon cephalus, an omnivorous teleost from the Amazon Basin in Brazil. Body weight and somatic indices (liver and digestive tract), glycogen and glucose content in liver and muscle, as well as plasma glucose, free fatty acids (FFA), insulin and glucagon levels of B. cephalus, were measured at 0, 12, 24, 48, 72, 120, 168 and 336 h after the last feeding. At time 0 h (the moment of food administration, 09.00 h) plasma levels of insulin and glucagon were already high, and relatively high values were maintained until 24 h post-feeding. Glycemia was 6.42+/-0.82 mM immediately after food ingestion and 7.53+/-1.12 mM at 12 h. Simultaneously, a postprandial replenishment of liver and muscle glycogen reserves was observed. Subsequently, a sharp decrease of plasma insulin occurred, from 7.19+/-0.83 ng/ml at 24 h of fasting to 5.27+/-0.58 ng/ml at 48 h. This decrease coincided with the drop in liver glucose and liver glycogen, which reached the lowest value at 72 h of fasting (328.56+/-192.13 and 70.33+/-14.13 micromol/g, respectively). Liver glucose increased after 120 h and reached a peak 168 h post-feeding, which suggests that hepatic gluconeogenesis is occurring. Plasma FFA levels were low after 120 and 168 h and increased again at 336 h of fasting. During the transition from post-feeding to fast condition in B. cephalus, the balance between circulating insulin and glucagon quickly adjust its metabolism to the ingestion or deprivation of food.     

Effects of graded levels of Fusarium toxin contaminated wheat in diets for fattening pigs on growth performance, nutrient digestibility, deoxynivalenol balance and clinical serum characteristics

A dose response study was carried out with pigs in order to examine the effects of increasing dietary deoxynivalenol (DON)-concentrations on performance, clinical serum characteristics, nutrient digestibility and DON-metabolism. For this purpose, wheat contaminated naturally with Fusarium toxins was incorporated into pig diets at increasing proportions to give calculated dietary DON-concentrations of 0, 2.3 and 4.6 mg/kg during the starter period of phase 1 (14 d) of the experiment, and 0/0, 1.2/1.4, 2.3/3.7 mg/kg starter/grower diet during phase 3 (56 d) of the experiment. Each diet was tested on 16 pigs of both sexes with an initial average live weight of approximately 28 kg. A recovery phase (phase 2, 21 d) was intercalated between phase 1 and 3 of the growth experiment where all groups were fed with the uncontaminated control diet since some pigs exposed to the highest dietary DON-concentration during phase 1 nearly completely refused the offered feed. Affected pigs completely recovered during this phase. In phase 3, when diets with lower DON-concentrations were fed, no differences in performance could be detected. Serum clinical characteristics (enzymes indicating liver damage, total protein, immunoglobulins) did not respond to increasing DON-concentration in the diets. DON-concentration in serum increased in a dose-response-related manner as dietary DON-concentration increased. However, this parameter was not or only weakly correlated to any of the examined performance parameters or serum characteristics. Also, nutrient digestibility of the diets and N-retention were not affected by treatments with the exception of crude fat digestibility which was not consistently influenced. Concentration of DON and its metabolite de-epoxy-DON increased in urine with increasing dietary DON-concentration in a strongly linearly related fashion. The proportion of the excretion of de-epoxy-DON of the total urinary excretion of DON plus de-epoxy-DON rose linearly up to approximately 4%. Total recovery of DON plus de-epoxy-DON as percentage of DON-intake varied between 45 and 57% and was not influenced by dietary DON-concentration. Only a very small fraction of approximately 0.1% of ingested DON was recovered in faeces.     

On the toxicokinetics and the metabolism of deoxynivalenol (DON) in the pig

Eleven castrated male pigs weighing 88.1 +/- 3.9 kg on average were adapted to a diet containing DON (4.2 mg DON/kg) over a period of 7 days. Feed was given restrictively with 1.1 kg per meal (two meals per day). On the day of measurement, all pigs were slaughtered at different time intervals following the morning meal containing DON (1, 2, 3, 4, 5, 6, 8, 15, 18 and 24 h after feeding), with the exception of one pig which was slaughtered unfed. DON and de-epoxy-DON were analysed in serum and digesta from consecutive segments of the digestive tract (stomach, small intestine divided into three parts of a similar length, caecum, colon, rectum). DON was rapidly and nearly completely absorbed while passing through the stomach and the proximal small intestine. Maximum serum concentration appeared 4.1 h after the DON-containing meal and half of the systemically absorbed DON was eliminated after 5.8 h. De-epoxy-DON appeared in increasing proportions from the distal small intestine and reached approximately 80% of the sum of DON plus de-epoxy-DON in faeces collected from the rectum. It was concluded that de-epoxydation of DON, which primarily occurs in the hindgut, probably does not contribute much to a detoxification in the pig.     

Role of Selenium from Different Sources in Prevention of Pulmonary Arterial Hypertension Syndrome in Broiler Chickens

Pulmonary arterial hypertension (PAH) syndrome in broilers is associated with hypoxia, which prevails at high altitude. Oxidative stress is the pathogenic mechanism underlying PAH. Because selenium is key element in the structure of antioxidant enzymes, we evaluated pulmonary hypertensive responses in broiler chickens fed with diets supplemented with organic or nano-selenium. One hundred forty-four broilers (starting at 5 days old) were fed with (i) control group: birds received a standard diet; (ii) nano-selenium group: birds were fed with basal diet supplemented with nano-selenium at 0.3 mg/kg; and (iii) organic selenium group: birds received basal diet supplemented with organic selenium at 0.3 mg/kg. We assessed growth performance, carcass characteristics, antioxidant variables, blood parameters, and small intestine morphology. Although Se supplementation did not affect growth performance, carcass traits, and organ weight (P > 0.05), the right to total ventricular weight ratio (RV:TV), malondialdehyde concentration in the liver, and heterophil to lymphocyte ratio were significantly lower in the nano-selenium group relative to the control (P < 0.05). Chickens that received nano-selenium also elicited significantly higher antibody titers after 24 h of an injection of sheep red blood cells (P < 0.05). Nano-selenium supplementation also significantly increased villus height, absorptive surface area, and lamina propria thickness relative to the control (P < 0.05) in different segments of the small intestine. In contrast, organic selenium supplement improved intestinal morphometry only in the jejunum. We conclude that dietary supplementation of 0.30 mg/kg nano-selenium could prevent right ventricular hypertrophy as reflected by reduced RV:TV, reduced levels of lipid peroxidation in the liver, and improved gut function.     

Effects of deoxynivalenol (DON), zearalenone (ZEN), and related metabolites on equine peripheral blood mononuclear cells (PBMC) in vitro and background occurrence of these toxins in horses

Both deoxynivalenol (DON), zearalenone (ZEN), and their metabolites are known to modulate immune cells in various species whereby viability and proliferation are influenced. Such effects were rarely examined in horses. Therefore, one aim of the present study was to titrate the inhibitory concentrations of DON, 3-acetyl-DON (3AcDON), de-epoxy-DON (DOM-1), ZEN, and α- and β-zearalenol (ZEL) at which viability and proliferation of equine PBMC were reduced by 50 % (IC₅₀) and 10 % (IC₁₀) in vitro. For evaluation of practical relevance of the in vitro findings, a further aim was to screen horses for the background occurrence of DON, ZEN, and their metabolites in systemic circulation and to relate toxin residues both to the inhibitory toxin concentrations and to hematological and clinical-chemical characteristics.The IC₅₀ (μM) for DON, 3AcDON, β-ZEL, α-ZEL, and ZEN were determined at 3.09, 25.90, 75.44, 97.44, and 98.15 in unstimulated cells, respectively, while in proliferating cells, the corresponding IC₅₀ values were 0.73, 6.89, 45.16, 75.96, and 82.51. Neither viability nor proliferation was influenced by DOM-1 up to a concentration of 100 μM.The in vivo screening (N?=?49) revealed the occurrence of ZEN (N?=?24), α-ZEL (N?=?3), β-ZEL (N?=?37), DON, and DOM-1 (N?=?2). The detected concentrations were much lower than the corresponding IC₅₀ while the IC₁₀ of DON and β-ZEL for proliferating PBMC corresponded to approximately 26 and 35 ng/mL which might be relevant when contaminated diets are fed.Clinical-chemical and hematological traits were not related to mycotoxin residue levels excepting blood urea nitrogen which was positively correlated to the sum of β-ZEL, α-ZEL, and ZEN concentration. Whether this reflects simply the feeding history of the horses or renal failures giving rise to a prolonged half-life of the toxins needs to be clarified further.     

Effects of deoxynivalenol and zearalenone on oxidative stress and blood phagocytic activity in broilers

Effects of dietary contamination with various levels of deoxynivalenol (DON) and zearalenone (ZEA) were investigated on Ross 308 hybrid broilers of both sexes. After hatching, all chickens were fed an identical control diet for two weeks. Then chickens of Group 1 received a diet contaminated with DON and ZEA, both being 3.4 mg · kg^?1, while Group 2 received DON and ZEA at 8.2 and 8.3 mg · kg^?1, respectively. The diet of the control group contained background levels of mycotoxins. Samples of blood and tissues were collected after two weeks. Intake of both contaminated diets resulted in a significantly decreased activity of glutathione peroxidase (GPx) and increased level of malondialdehyde (MDA) in liver tissue, while in kidneys the concentration of MDA was significantly increased only in Group 1. On the other hand, activities of blood GPx and plasma γ-glutamyltransferase (GGT) were elevated in Group 2 only. Activities of thioredoxin reductase in liver and GPx in duodenal mucosa tissues, superoxide dismutase (SOD) in erythrocytes as well as levels of MDA in duodenal mucosa and α-tocopherol in plasma were not affected by dietary mycotoxins. Blood phagocytic activity was significantly depressed in Group 1 and 2. These results demonstrate that diets contaminated with DON and ZEA at medium levels are already able to induce oxidative stress and compromise the blood phagocytic activity in fattening chickens.     

The Effects of Ziprasidone, Clozapine and Haloperidol on Lipid Peroxidation in Human Plasma (in vitro): Comparison

Oxidative injury in schizophrenia can be caused by the disease itself and probably by antipsychotics treatment. The aim of the study was to establish whether there is a difference between ziprasidone, clozapine and haloperidol effect on lipid peroxidation in human plasma, measured by the level of thiobarbituric acid reactive substances (TBARS). The samples of plasma from healthy subjects were incubated with the drugs (1 and 24 h) and compared with control samples. The levels of TBARS were measured spectrophotometrically, according to the Rice-Evans method. The multifactorial variance analysis ANOVA II test showed that the differences in TBARS levels significantly depended on the studied drugs (ziprasidone 40 ng/ml, haloperidol 4 ng/ml and clozapine 350 ng/ml) (F = 3.248 p = 0.047) and (ziprasidone 139 ng/ml, haloperidol 20 ng/ml and clozapine 420 ng/ml) (F = 2.248, p = 2.9 × 10^?5). Statistically increased levels of TBARS after 24 h incubation of plasma with ziprasidone 139 ng/ml and haloperidol 20 ng/ml (p < 0.001, p < 0.05 respectively) in comparison with control samples were observed. Clozapine did not significantly (p > 0.05) increase TBARS level in plasma in comparison with control samples. The results obtained in the study showed that ziprasidone and haloperidol contrary to clozapine induced a significant increase in plasma lipid peroxidation.