Modified use of a commercial ELISA kit for deoxynivalenol determination in rice and corn silage

The analysis of deoxynivalenol (DON) in silage samples using enzyme-linked immunosorbent assay (ELISA) often leads to an overestimation. To better analyze DON in rice and corn silages using a commercially available ELISA kit, a cleanup method using a MultiSep #226 column was developed. As a result, overestimation of DON by the influence of specific cross-reaction with acetyldeoxynivalenol (AcDON) was confirmed. In samples where AcDON was not detected by liquid chromatography with mass spectrometry (LC-MS), no samples showed a significant difference (P?<?0.05) in DON amounts between ELISA with cleanup and LC-MS analysis. For the recovery study, blank silage was spiked with 0.5 or 1.0 mg/kg DON. The mean recoveries of DON determined by ELISA with cleanup and LC-MS analysis were 112 and 96 %, respectively, and the relative standard deviation for the repeatability (RSD_r) were 8.2 and 9.8 %, respectively. No samples showed a significant difference (P?<?0.05) in DON concentration determined by either ELISA or LC-MS analysis. A collaborative study to validate this rapid method was carried out using four samples, two rice and two corn silage, by 10 participating laboratories. Each sample was analyzed using blind duplicates. The mean values of DON detected were 1.5–2.3 mg/kg, RSD_r and the relative standard deviation for the reproducibility (RSD_R) were 4.1–12.7 and 7.6–23.4 %, respectively, and the HorRat values were 0.5–1.6. Therefore, the overestimation of DON by the influence of nonspecific cross-reaction with sample matrix was reduced by the cleanup method using a MultiSep #226 column, and analysis of DON in silage was improved. This use of this method for estimation of DON contamination in silage allows rapid detection at the place of use that is likely to result in improved animal health.     

Fusarium species and their mycotoxins in infected corn in Italy

Surveys of corn (infected plants and commercial kernels) forFusarium species and their mycotoxins were carried out on samples collected all over Italy and from some European and mediterranean countries.Investigations on samples of corn stalk and ear rot standing in the field, mainly collected in southern Italy, proved to be contaminated with zearalenone (ZON), zearalenols (ZOL), and deoxynivalenol (DON). TheFusarium species most frequently isolated, and their recorded toxigenic capability (in parentheses), were:F. moniliforme;F. culmorum (ZON, ZOL, DON, 3AcDON);F. equiseti (ZON, ZOL); andF. proliferatum (MF). Along with these species,F. graminearum group 2 (ZON, DON and/or 3AcDON or 15AcDON);F. chlamydosporum;F. acuminatum (type-A trichothecene derivatives); andF. semitectum were often found to be associated.F. heterosporum (ZON, ZOL);F. solani;F. crookwellense (ZON, ZOL, FUS, NIV);F. oxysporum (MF);F. avenaceum (MF);F. sporotrichioides (T-2 toxin and derivatives); andF. poae (DAS, MAS) were occasionally isolated.     

Survey and risk assessment of the mycotoxins deoxynivalenol,zearalenone, fumonisins,ochratoxin A,and aflatoxins in commercial dry dog food

The aim of the present study was to investigate the occurrence of mycotoxins in commercial dog food, as a basis to estimate the risk of adverse effects. Seventy-six dry dog food samples from 27 producers were purchased from retail shops, supermarkets, and specialized pet food shops in Vienna, Austria. The frequency and levels of deoxynivalenol (DON), zearalenone (ZEA), fumonisins (FUM), ochratoxin A (OTA). and aflatoxins (AF) in dry dog food were determined. Mycotoxin analysis were performed by commercial enzyme-linked immunosorbent assay (ELISA) test kits. Confirmatory analyses were done for DON, ZEA, and FUM by high performance liquid chromatography (HPLC) after extract clean-up with immunoaffinity columns. The correlations between ELISA and HPLC results for DON and ZEA were acceptable and indicated that ELISA could be a simple, low cost, and sensitive screening tool for mycotoxins detection, contributing to quality and safety of pet food. DON was the mycotoxin most frequently found (83% positives; median 308 μg/kg, maximum 1,390 μg/kg). ZEA (47% positives, median 51 μg/kg and maximum 298 μg/kg) and FUM (42% positives, median 122 μg/kg and maximum 568 μg/kg) were also frequently detected in dog food. OTA was less frequently found (5%, median 3.6 μg/kg, maximum 4.7 μg/kg. AF were not detected (<0.5 μg/kg) in any sample. The results show that dry dog food marketed in Vienna are frequently contaminated with mycotoxins (DON > ZEA > FUM > OTA) in low concentrations, but do not contain AF. The high frequency of Fusarium toxins DON, ZEA, and FUM indicates the need for intensive control measures to prevent mycotoxins in dog foods. The mycotoxin levels found in dry dog food are considered as safe in aspects of acute mycotoxicoses. However, repeated and long-time exposure of dogs to low levels of mycotoxins may pose a health risk.     

Fusarium Head Blight Reactions and Accumulation of Deoxynivalenol (DON) and Some of its Derivatives in Kernels of Wheat, Triticale and Rye

Fifty-three commercially grown cultivars and germplasm lines of winter triticale (n = 18), wheat (n = 13), and rye (n = 5) and spring triticale (n = 8), wheat (n = 7) and rye (n = 2) were inoculated at mid anthesis with a spore suspension consisting of a mixture of Fusarium culmorum, Fusarium avenaceum and Fusarium graminearum isolates of known toxinogenic activity. Reactions to Fusarium head blight were measured as disease severity, reductions of kernel number/head, kernel weight/head and 1000 kernel weight, number of Fusarium-damaged kernels and kernel content of deoxynivalenol (DON) and its acetyl-derivatives 3-AcDON, 15-AcDON, and moniliformin. None of the cereal genotypes was completely resistant to Fusarium head blight. Wheat suffered from the largest kernel weight reductions, and accumulated the largest amounts of deoxynivalenol (up to 39.5 mg/kg) and 3AcDON (up to 6.0 mg/kg) in kernels. Deoxynivalenol was not detected in grain samples of winter rye cv. Dańkowskie Z?ote, and spring rye cv. Ludowe. 15-AcDON was only detected in genotypes of triticale, and 3AcDON only in a few genotypes of winter wheat and rye. Moniliformin was detected at low concentrations (up to 0.092 mg/kg) in kernels of some genotypes selected for the mycotoxin analysis. A moderately strong Pearson correlation was found between head blight severity parameters and the accumulation of deoxynivalenol and its derivatives in grain of the cereal genotypes studied. Fusarium head blight severity parameters were correlated with the percentage of Fusarium-damaged kernels and reductions of yield components. However, some head blight-susceptible genotypes realized their potential yields, but accumulated high levels of mycotoxins in kernels. Both Fusarium head blight resistant and susceptible genotypes of the three cereal species accumulated deoxynivalenol in kernels. This finding suggests that the system regulating deoxynivalenol accumulation may be independent of Fusarium head blight reaction.     

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     

Influence of nitrogen fertilization on deoxynivalenol contamination of winter wheat — experimental field trials and evaluation of analytical methods

Experimental field trials were carried out to study the influence of N-fertilization on deoxynivalenol (DON) contamination of winter wheat. Within four years of investigation, no definite effect of mineral N-input at dosages varying between 0 and 240 kg N/ha could be observed on DON concentration in wheat grain. The main factors affecting DON contamination of wheat were theFusarium infection pressure, the weather conditions and the susceptibility of the wheat varieties againstFusarium head blight. DON was analyzed with enzyme-linked immunosorbent assay (ELISA) and, for comparison, some of the positive samples were additionally analyzed with high performance liquid chromatography (HPLC). There was a good correlation between the ELISA and the HPLC results for DON concentration in wheat.     

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.     

Zearalenone, deoxynivalenol and fumonisins in mixed-feed for laying hens

Fusarium toxins are secondary metabolites produced byfungi of these genera in many commodities under certain conditions. A study was carried out to investigate the co-occurrence of zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FB₁ and FB₂) in 52 samples of mixed-feed for poultry contaminated withFusarium verticillioides. The zearalenone and deoxynivalenol were checked using immunoaffinity column and the extraction of fumonisin was performed by strong anion exchange (SAX) solid phase column. Detection and quantification were determined by high performance liquid chromatography (HPLC). The limit of detection was 5 μg/kg for ZEN, 100 μg/kg for DON and 50 and 100 μg/kg for FB₁ and FB₂ respectively.Fusarium toxins were detected in 20 samples. Sixteen samples were positive for ZEN (30.7%) presenting levels that ranged from 7.4 μg/kg to 61.4 μg/kg (mean=27.0 μg/kg). 13.5% of the samples presented contaminations of DON, with levels ranging from 100.0 μg/kg to 253 μg/kg (mean=l18.07 μg/kg). FB₁ was detected in 19.2% of samples, with levels ranging from 50.0 μg/kg to 110.0 μg/kg (mean=73.6 μg/kg). FB2 was not detected in any sample. In positive samples simultaneously contamination with two or three mycotoxins were detected in 9 of them (17.3%).     

Simple method for isolation of 4-deoxynivalenol from rice inoculated with Fusarium graminearum.

A new method for preparative isolation of 4-deoxynivalenol (DON) is presented. This method avoids the loss of material during purification on silica gel by column chromatography. DON and 3-acetyldeoxynivalenol in crude extracts of rice inoculated with Fusarium graminearum were converted to triacetyldeoxynivalenol; the acetylated product was easier to purify by silica gel chromatography than DON is. After hydrolysis and further purification on a charcoal-alumina column, the 71% pure DON was recovered in yields as high as 450 mg of DON per kg of rice. Subsequent separation on a Sephadex LH20 column yielded DON that was greater than 90% pure.     

Simultaneous occurrence of mycotoxins in human food commodities from Cameroon

Eighty-two samples of dried food commodities from Cameroon were screened and quantified for different mycotoxins, including fumonisin B₁ (FB₁), zearalenone (ZEA), deoxynivalenol (DON), aflatoxin (AF) and ochratoxin A (OTA), by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), respectively. The percentage of positive samples was as follows: FB₁ 41%, AF 51%, ZEA 57%, DON 65% and OTA 3%. High FB₁ contents were found in maize, averaging 3,684 μg/kg (range: 37-24,225 μg/kg), whereas the highest average ZEA level was found in peanuts (70 μg/kg), followed by maize (69 μg/kg), rice (67 μg/kg) and beans (48 μg/kg) with no ZEA was detected in soybeans. DON contents were low, ranging from 13 to 273 μg/kg, and for AF the average content was 2.6 μg/kg with peanuts and maize as principal substrates. The incidence of OTA was low, with a mean level of 6.4 μg/kg recorded. The majority (79%) of samples contained more than one mycotoxin and the most frequent co-occurrence found was FB₁ + ZEA + DON, detected in 21% of samples (mainly maize) analysed. Co-contamination with FB₁ + ZEA + DON + AF was found in 11% of the samples. Although a large proportion of samples had fairly low levels of individual mycotoxins, this should be of concern as the co-occurrence of mycotoxins may generate additive or synergistic effect in humans, especially if the respective commodities are consumed almost on a daily basis.     

Determination of mycotoxins in bovine milk by liquid chromatography tandem mass spectrometry

Liquid chromatographic/tandem mass spectrometric methods using pneumatically assisted electrospray ionisation (LC-ESI-MS/MS) was developed for determination of 18 mycotoxins and metabolites-ochratoxin A, zearalenone, alpha-zearalenol, beta-zearalenol, alpha-zearalanol (zeranol), beta-zearalanol (taleranol), fumonisin B1, fumonisin B2, T-2 toxin, HT-2 toxin, T-2 triol, diacetoxyscirpenol (DAS), 15-monoacetoxyscirpenol (MAS), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON), deepoxy-deoxynivalenol (DOM-1) and aflatoxin M1--in milk. The mycotoxins were extracted and cleaned up simultaneously. Extraction and removal of lipophilic compounds was performed at pH 2 using a two-phase mixture of acetonitrile and hexane. The acetonitrile concentration of the aqueous phase was reduced and the pH was adjusted to 8.5 before clean up by solid phase extraction (SPE) on Oasis HLB. The toxins DON, DOM-1, 3-AcDON, 15-AcDON, ochratoxin A, zearalenone, alpha-zearalenol, beta-zearalenol, alpha-zearalanol and beta-zearalanol were detected in negative ion mode after separation on a Hypersil ENV analytical column, while the toxins T-2 toxin, HT-2 toxin, T-2 triol, DAS, MAS, fumonisin B1, fumonisin B2 and aflatoxin M1 were detected in positive ion mode after separation on a Luna C18 column. Two transition products were monitored for each compound. The extraction and SPE conditions were optimised to obtain maximum recovery and minimum signal suppression/enhancement. The detection capabilities related to the transition products of lowest abundance were in the range 0.020-0.15 microg/l. The mean true recoveries were in the range 76-108% at levels of 0.2-10 microg/l.     

Mycotoxins in cereal grain (part 15). Distribution of deoxynivalenol in naturally contaminated wheat kernels

The analysis of deoxynivalenol (DON) in naturally infected wheat samples, after having been separated into four fractions through laboratory sieves, showed very low levels of DON in the fraction of largest kernels >2.8 mm (0 up to 1 mg/kg). The highest concentration of DON was found in fractions 2.2 to 2.5 mm and <2.2mm with up to 14mg/kg and 15mg/kg DON, respectively. In two samples (fractions <2.2mm) nivalenol was detected in concentrations up to 1,4mg/kg.     

Decontamination ofFusarium mycotoxins,nivalenol, deoxynivalenol,and zearalenone,in barley by the polishing process

Korean dehusked and unhusked barley naturally contaminated withFusarium mycotoxins were polished using a Satake Grain Testing Mill. The pearled barley and bran fractions with different degrees of polishing were analyzed for nivalenol (NIV) and deoxynivalenol (DON) by gas chromatography with an electron capture detector, and for zearalenone (ZEN) by high-performance liquid chromatography with a fluorescence detector. NIV was detected in all the pearled barley fractions, but DON and ZEN were not detected in ≥27 % pearled barley fractions from dehusked barley and ≥36% pearled barley fractions from unhusked barley. However, for all degrees of polishing, NIV, DON, and ZEN were detected in bran fractions. The levels of NIV, DON, and ZEN in the bran fractions increased several fold over the original barley. Polishing was effective in removing DON and ZEN from the naturally contaminated barley, but not NIV.     

Aerobic and anaerobic de-epoxydation of mycotoxin deoxynivalenol by bacteria originating from agricultural soil

One hundred and fifty soil samples collected from different crop fields in southern Ontario, Canada were screened to obtain microorganisms capable of transforming deoxynivalenol (DON) to de-epoxy DON (dE-DON). Microbial DON to dE-DON transformation (i.e. de-epoxydation) was monitored by using liquid chromatography-ultraviolet-mass spectrometry (LC-UV–MS). The effects of growth substrates, temperature, pH, incubation time and aerobic versus anaerobic conditions on the ability of the microbes to de-epoxydize DON were evaluated. A mixed microbial culture from one composite soil sample showed 100% DON to dE-DON biotransformation in mineral salts broth (MSB) after 144 h of incubation. Treatments of the culture with selective antibiotics followed an elevated temperature (50°C) for 1.5 h considerably reduced the microbial diversity. Partial 16S-rRNA gene sequence analysis of the bacteria in the enriched culture indicated the presence of at least six bacterial genera, namely Serratia, Clostridium, Citrobacter, Enterococcus, Stenotrophomonas and Streptomyces. The enriched culture completely de-epoxydized DON after 60 h of incubation. Bacterial de-epoxydation of DON occurred at pH 6.0–7.5, and a wide array of temperatures (12–40°C). The culture showed rapid de-epoxydation activity under aerobic conditions compared to anaerobic conditions. This is the first report on microbial DON to dE-DON transformation under aerobic conditions and moderate temperatures. The culture could be used to detoxify DON contaminated feed and might be a potential source for gene(s) for DON de-epoxydation.     

Effects of deoxynivalenol (DON) on growth performance, nutrient digestibility and DON metabolism in pigs

Wheat infected naturally withFusarium, contaminated mainly with 18.53 mg DON per kg, was added to a total constant wheat proportion of 400g/kg diet. Control and DON contaminated feed was fed for 11 weeks underad libitum and restrictive feeding conditions to 48 pigs of both sexes, which were randomly divided into 4 groups. Effects on performance (live weight range between 26 and 100kg), duration of feed intake and blood parameters were investigated. Parallel to this study, a balance study was carried out to examine the effects on nutrient digestibility and DON metabolism. The group fed the DON contaminated rationad libitum consumed 15% less feed and gained 14% less live weight compared to thead libitum control group, while the feed to gain ratio was unaffected. Under restrictive feeding conditions (DON and control) pigs exhibited 33%, 25% and 10% lower feed consumption, live weight gain and feed to gain ratio, respectively, than the control group fedad libitum. Metabolizability of energy, digestibility of organic matter, crude protein, crude fat and N-retention were significantly increased by 4, 3, 6, 11 and 10%, respectively, in the DON group of the restrictively fed pigs. In average up to 43.2% of the ingested DON, as the parent toxin in both groups, was eliminated with the urine and up to 3.0% with faeces. DON fed animals needed more time to consume the restrictive ration than the control group. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germany, May 17–19, 2004. Financial support Deutsone Forschungsgemeingschaft     

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).     

Detoxification of Fusarium-contaminated maize with sodium sulphite – in vivo efficacy with special emphasis on mycotoxin residues and piglet health

A feeding experiment with piglets was performed to examine the efficacy of a wet preservation of Fusarium (FUS)-contaminated maize with sodium sulphite (SoS) based on deoxynivalenol (DON) and zearalenone (ZEN) residue levels in urine, bile and liquor and health traits of piglets. For this purpose, 80 castrated male piglets (7.57 ± 0.92 kg BW) were assigned to four treatment groups: CON? (control diet, with 0.09 mg DON and <0.01 mg ZEN/kg diet), CON+ (diet CON?, wet-preserved with 5 g SoS/kg maize; containing 0.05 mg DON and <0.01 mg ZEN/kg diet), FUS? (diet with mycotoxin-contaminated maize; containing 5.36 mg DON and 0.29 mg ZEN/kg diet), and FUS+ (diet FUS?, wet-preserved with 5 g SoS/kg maize; resulting in 0.83 mg DON and 0.27 mg ZEN/kg diet). After 42 d, 40 piglets (n = 10 per group) were sampled. A clear reduction of DON levels by approximately 75% was detected in all specimens of pigs fed diet FUS+. ZEN was detected in all urine, bile and liquor samples, while their metabolites were only detectable in urine and bile. Additionally, their concentrations were not influenced by SoS treatment. Among the health-related traits, feeding of FUS diets increased the total counts of leukocytes and segmented neutrophil granulocytes irrespective of SoS treatment. SoS treatment increased the total blood protein content slightly with a similar numerical trend in albumin concentration. These effects occurred at an obviously lower level in FUS-fed groups. Moreover, SoS treatment recovered the reduction of NO production induced by feeding diet FUS? indicating an effect on the redox level. As this effect only occurred in group FUS+, it is obviously related to the adverse effects of the Fusarium toxins. In conclusion, treatment of FUS-contaminated maize with SoS decreased the inner exposure with DON as indicated by the lower DON levels in various piglet specimens. However, health-related traits did not consistently reflect this decreased exposure.     

Progression of mycotoxin and nutrient concentrations in wheat after inoculation with Fusarium culmorum

The objective of this study was to follow the mycotoxin formation and changes in nutrient composition of wheat (cv. Ritmo) artificially inoculated with Fusarium culmorum. From anthesis until harvest, samples were taken once a week from the inoculated and control plots. The investigations were focused on monitoring the progression of the contamination of the wheat kernels with deoxynivalenol (DON) and zearalenone (ZON). Both the uncontaminated control kernels and the contaminated kernels were examined also for the presence of zearalenone-4-beta-D-glucopyranoside and several trichothecenes at harvest. Furthermore, the impact of the Fusarium inoculation on some nutrients as starch, crude protein, amino acid composition, crude ash, non starch polysaccharides (NSP) as well as viscosity and thousand seed weight (TSW) was examined. Also proteolytic and amylolytic activity as well as the NSP-degrading enzyme activities of inoculated and control samples were analysed at the time of harvest. DON was detected in higher concentrations and in earlier stages, while ZON was found later and in smaller amounts. On average 7.79?mg/kg DM of DON and 100?μg/kg DM of ZON were found in the inoculated kernels at the time of harvest. Neither in the contaminated nor in the control samples glucose conjugates of ZON (Zearalenone-4-beta-D-glucopyranoside) were detected. Moreover, the infection with Fusarium culmorum had pronounced effects on some quality parameters. The crude protein content of the inoculated kernels showed significantly higher values over the whole period compared to the control kernels. The protein content of the inoculated kernels amounted 13.9% DM at harvest, while only a concentration of 12.5% DM was detected in the control samples. Similarly, in almost all stages of development the crude ash content of inoculated samples was higher than in control samples. These distinct differences in kernel composition resulted possibly from the changes of the thousand seed weight. In the present work the grain harvested from the control plots showed a significantly higher TSW (24.2?g) as compared to their inoculated counterparts (15.5?g). Despite lower extract viscosity of inoculated samples at time of harvest, the content of soluble NSP of inoculated plots was higher than in control samples at the same time. Moreover, inoculation resulted in markedly increased activities of protease, amylase and several NSP-degrading enzyme activities. This would suggest that the cell wall penetrating properties of the fungus itself and/or that the fungus induced alterations of the metabolic activity of the embryo or other constituents of the wheat kernel could be responsible.     

The effects of Rhodiola rosea extract on 5-HT level, cell proliferation and quantity of neurons at cerebral hippocampus of depressive rats

The purpose of this study was to investigate the effects of Rhodiola rosea extract and depression on the serotonin (5-HT) level, cell proliferation and quantity of neurons at cerebral hippocampus of depressive rats induced by Chronic Mild Stress (CMS). Seventy male Sprague-Dawley rats were divided into seven groups (10 per group): normal control group, untreated depressive rat model group, negative control group, positive control group, low dosage Rhodiola rosea extract (1.5 g/kg) group, medium dosage Rhodiola rosea extract (3 g/kg) group and high dosage Rhodiola rosea extract (6 g/kg) group. After the depressive rats induced by CMS had received Rhodiola rosea extract for 3 weeks, the 5-HT levels at cerebral hippocampus were detected by high performance liquid chromatography. Bromodeoxyuridine (BrdU) was injected in vivo to label the proliferating cells at hippocampus, and morphometry was used to count the hippocampal neurons. The results showed that the 5-HT level of the three experimental groups had recovered to normal status. The immunohistochemistry of hippocampus BrdU positive cells had returned to the normal level in the group of depressive rats with low dosage Rhodiola rosea extract. In conclusion the results demonstrated that Rhodiola rosea extract could improve 5-HT level in hippocampus in depressive rats, and low dosage Rhodiola rosea could induce neural stem cell proliferation at hippocampus to return to normal level, repairing the injured neurons at hippocampus.