Further survey of the occurrence of Fusarium toxins in wheat grown in southwest Germany.

A total of 53, 54, 57, 52 and 60 wheat samples for feed use were collected randomly after the 1989, 1990, 1991, 1992 and 1993 crops, respectively, from farms in an area of southwest Germany. Deoxynivalenol (DON), 3- and 15-acetyldeoxynivalenol (3-, 15-ADON), nivalenol (NIV), HT-2 toxin (HT-2), T-2 toxin (T-2), diacetoxyscirpenol (DAS), and fusarenon-X (FUS-X) were determined by gas chromatography, combined with mass selective detection (GC-MS), zearalenone (ZEA), alpha- and beta-zearalenol (alpha-beta-ZOL) were determined by HPLC. DON was the major toxin, with incidences at 77 to 93% and mean contents at 167 to 735 micrograms/kg. In contrast, incidences of ZEA, 3-ADON, NIV, HT-2, and T-2 were at 13 to 37%, 10 to 44%, 15 to 67%, 0 to 11%, and 0 to 12%, respectively, with mean contents in positive samples between 2 and 73 micrograms/kg, except for 948 micrograms/kg 3-ADON in samples from 1993. 15-ADON and FUS-X were assayed in samples from 1991, 1992 and 1993. 15-ADON was found in 0 to 11% of samples at mean levels < or = 17 mu/kg, DAS, alpha- and beta-ZOL, and FUS-X were not detected in any sample. Over the years, incidences and levels of toxins remained constant, decreased or increased, with most differences between years being slight and insignificant. The risk for livestock due to DON, HT-2 and ZEA was estimated based on maximum tolerated levels recommended for these toxins in some countries.     

Occurrence of mycotoxins in feed ingredients and complete feeds obtained from the Beijing region of China

Background

The current study was carried out to provide a reference for the control of mycotoxin contamination in feed ingredients and complete feeds for swine.

Methods

A total of 55 feed ingredients, including 14 corn, 13 wheat bran, 11 soybean meal and 17 dried distillers grains with solubles (DDGS) as well as 76 complete swine feeds including 7 creep feeds, 14 starter feeds, 14 grower feeds, 18 grower-finisher feeds, 10 gestating sow feeds, and 13 lactating sow feeds were randomly collected from 15 swine farms located in the Beijing region of China from July to August 2011. Immunoaffinity clean-up, using High-Performance Liquid Chromatography (HPLC) in combination with UV or Fluorescence Detection, was used for quantitative analysis of aflatoxin B₁ (AFB₁), deoxynivalenol (DON), zearalenone (ZEA) and ochratoxin A (OTA) in the ingredients and complete feeds.

Results

DON and ZEA were the most prevalent mycotoxins found. DON was detected at percentages of 93, 92, 54, 100 and 97% with a mean level of 1.01, 0.44, 0.05, 1.36 and 0.65 ppm in the samples of corn, wheat bran, soybean meal, DDGS and complete feeds, respectively. The detected percentages of ZEA were 100, 100, 54, 100 and 100 with mean levels of 109.1, 14.9, 9.2, 882.7 and 58.9 ppb in the same samples. In the wheat bran and soybean meal samples, the content of all four mycotoxins were below the maximum limits set by Chinese regulations while the percentage of samples that exceeded regulatory limits were 7, 57 and 7% for corn, and 7, 14 and 3% for the complete feeds for AFB₁, DON and OTA respectively. DDGS showed the most serious mycotoxin contamination and the percentage of samples that exceeded regulatory limits were 6, 88 and 41%, for AFB₁, DON and ZEA, respectively.

Conclusions

This paper is the first to present data on the natural occurrence of AFB₁, DON, ZEA and OTA in ingredients and complete feeds obtained from swine farms in China’s Beijing region. The data shows that feed ingredients and complete swine feeds obtained from these farms are most often contaminated with DON followed by contamination with AFB₁ and ZEA.     

脱氧雪腐镰刀菌烯醇与玉米赤霉烯酮联合暴露对体外培养鸡脾脏淋巴细胞内环境稳态的影响

本试验旨在研究脱氧雪腐镰刀菌烯醇(DON)与玉米赤霉烯酮与(ZEA)联合暴露对体外培养鸡脾脏淋巴细胞内环境稳态的影响。分别以0.012 50μg/mL DON+0.006 25μg/mL ZEA、0.050μg/mL DON+0.025μg/mL ZEA、0.2μg/mL DON+0.1μg/mL ZEA、0.8μg/mL DON+0.4μg/mL ZEA对体外培养鸡脾脏淋巴细胞进行联合暴露培养,48 h后测定细胞膜ATP酶(Ca~(2+)-ATP酶、Na~+/K~+-ATP酶)活性以及细胞内pH、Ca~(2+)水平和钙调蛋白(CaM)的mRNA表达水平。同时设不添加毒素的空白对照组。结果表明:添加毒素的各试验组间,细胞内Ca~(2+)水平、CaM mRNA表达水平随毒素浓度的升高而增加,且添加毒素的各试验组均显著或极显著高于空白对照组(P0.05或P0.01)。细胞内pH以及细胞膜Ca~(2+)-ATP酶与Na~+/K~+-ATP酶活性均随毒素浓度的升高而降低,且添加毒素的各试验组均显著或极显著低于空白对照组(P0.05或P0.01)。由此得出,DON、ZEA联合暴露导致体外培养鸡脾脏淋巴细胞内酸化、离子平衡失调等一系列细胞内环境稳态失衡,且呈剂量依赖性。     

Effects of ZEA,DON and Their Combined Effects on the Secretion of Cytokines in CTLL-2 Cells

To analyze the effects of ZEA or/and DON on the immunologic function,CTLL-2 cell was used as experimental materials. After exposing the CTLL-2 cell to different concentrations of ZEA (0,5,10,20 μg/mL),DON (0,0.5,1,2 μg/mL) and ZEA+DON (blank group,5 μg/mL ZEA,0.5 μg/mL DON,5 μg/mL ZEA+0.5 μg/mL DON) for 48 h,the concentration of GZMB,PFP,IFN-γ and TNF-α in CTLL-2 cell and supernatant fluid were detected by ELISA method. The result showed that DON and ZEA could reduce the concentration of PFP,GZMB and IFN-γ in the CTLL-2 cells and the culture supernatant fluids,and increase the concentration of TNF-α. The intoxicated group had significant or extremely significant difference compared to control group (P <0.05;P <0.01),which showed the dose effect. The group exposed to the combined of ZEA and DON showed an additive effect. The result indicated that ZEA,DON and the two combined could reduce the killing activity of immunological cell by affecting the secretion of immunological cytokines, and indirectly affect the negative feedback regulation of humoral immunity and cellular immunity,which led to the decline of immune function in animals.     

ZEA、DON及其联合染毒对CTLL-2细胞因子分泌的影响

为研究镰刀菌毒素玉米赤霉烯酮（ZEA）、脱氧雪腐镰刀菌烯醇（DON）及其联合作用对动物免疫功能的影响,试验以CTLL-2细胞（细胞毒性T淋巴细胞株）为材料,用不同浓度的ZEA （0、5、10、20 μg/mL）、DON （0、0.5、1、2 μg/mL）及联合（空白组、5 μg/mL ZEA、0.5 μg/mL DON、5 μg/mL ZEA＋0.5 μg/mL DON）处理CTLL-2细胞48 h,采用ELISA法检测了细胞内及培养上清液中颗粒酶B （GZMB）、穿孔素（PFP）、干扰素γ（IFN-γ）和肿瘤坏死因子α（TNF-α）等细胞因子的含量。结果显示,ZEA、DON能够降低CTLL-2细胞胞内及培养上清液中PFP、GZMB、IFN-γ的浓度,增加TNF-α浓度,染毒组与对照组相比均有显著或极显著差异（P＜0.05;P＜0.01）,且均呈剂量效应关系;ZEA、DON联合染毒表现为加性效应。结果表明,ZEA、DON及其联合作用可通过影响免疫细胞因子的分泌,降低免疫细胞杀伤活力,间接影响机体体液免疫和细胞免疫的负调节,从而导致动物机体免疫机能下降。     

Simultaneous detection of multiple mycotoxins in broiler feeds using a liquid chromatography tandem-mass spectrometry

Mycotoxins are secondary fungal metabolites that are typically present in grain and feed ingredients used for animal feeds. An analytical method using LC-ESI-MS/MS was developed to quantify nine mycotoxins, consisting of aflatoxin B₁ (AFB₁), AFB₂, AFG₁, AFG₂, T-2 toxin, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA) and ochratoxin A (OTA) in broiler feeds. In total, 100 samples of broiler feeds were collected from poultry farms in Central Thailand. The survey found that AFB₁ and ZEA were the most prevalent mycotoxins in the feed samples at percentages of 93% and 63%, respectively. The limit of detections (LODs) of investigated mycotoxins was 0.20–0.78 ng/g. AFB₂, DON, AFG₁, NIV and T-2 toxin were also detectable at low contamination levels with percentages of 20%, 9%, 7%, 5% and 1%, respectively, whereas OTA and AFG₂ were not detected in any of the feed samples. These results suggest that there is a very low level of risk of the exposure to mycotoxins in feeds obtained from broiler farms in Central Thailand.     

Absorption, metabolism and excretion of 3-acetyl DON in pigs.

The absorption, metabolism and excretion of 3-acetyldeoxynivalenol (3-aDON) in pigs were studied. Pigs with a faecal microflora known to be able to de-epoxidate trichothecenes were used in the experiment. The pigs were fed a commercial diet with 3-aDON added in a concentration of 2.5 mg/kg feed for 2.5 days. No traces of 3-aDON or its de-epoxide metabolite were found in plasma, urine or faeces. Deoxynivalenol (DON) was detected in plasma as soon as 20 min after start of the feeding. The maximum concentration of DON in plasma was reached after 3 h and decreased rapidly thereafter. Only low concentrations close to the detection limit were found in plasma 8 h after start of the feeding. A significant part of the DON in plasma was in a glucuronide-conjugated form (42 +/- 7%). No accumulation of DON occurred in plasma during the 60 h of exposure. The excretion of DON was mainly in urine (45 +/- 26% of the toxin ingested by the pigs) and only low amounts of metabolites of 3-aDON (2 +/- 0.4%) were recovered in faeces. De-epoxide DON constituted 52 +/- 15% of the total amount of 3-aDON-metabolites detected in faeces. The remaining part in faeces was DON. DON was still present in the urine and faeces at the end of the sampling period 48 h after the last exposure. The results show that no de-epoxides are found in plasma or urine in pigs after trichothecene exposure, even in pigs having a faecal microflora with a de-epoxidation activity. The acetylated form of the toxin is deacetylated in vivo. Furthermore, the experiment shows that the main part of DON is rapidly excreted and does not accumulate in plasma, but a minor part of the toxin is retained and slowly excreted from the pigs.     

Occurrence of mycotoxins in Polish animal feed in years 2006-2009

We performed a 4‐year survey (2006–2009, 1255 samples) of fungal secondary metabolites in feed material (cereal and corn grains) and feedstuffs (silages, mixed feeds). Five major mycotoxin groups were studied, including aflatoxins (AF), ochratoxin A (OTA), trichothecenes [deoxynivalenol (DON), nivalenol (NIV), T‐2 toxin, HT‐2 toxin], zearalenone (ZEA) and fumonisins (FUM). The metabolites were identified using HPLC methods with fluorescent, UV and MS/MS detection. Both immunoaffinity and SPE columns were used for sample preparation. In eleven samples, the concentration of several mycotoxins exceeded the recommended guidelines for feedstuffs. DON was detected at the highest concentration in the majority of analysed samples (cereal grains, silages and mixed feeds, maximum values ranged from 409 to 14 470 ng/g). Corn grains also contained other Fusarium toxins (FUM) at maximum levels ranging from 435 to 9409 ng/g. The highest average_positive concentration of the other trichothecenes (NIV, T‐2 and HT‐2 toxins) was <5.0–139 ng/g. ZEA was found at the highest concentration in corn grains and silages (maximum values ranging from 292 to 603 ng/g and 116 to 1150 ng/g, respectively). The highest average_positive concentration and the maximum level of OTA were detected in cereal grains (33.0 ng/g in 2009 and 760 ng/g in 2007, respectively). Less than 7% of the 557 samples were contaminated with AF at low levels (maximum of 0.61 ng/g). Our results support the need for further monitoring of mycotoxins in Polish feedstuffs and their components.     

黄曲霉毒素B1、玉米赤霉烯酮和呕吐毒素的累加细胞毒性研究

由于饲料中多种霉菌毒素并存的几率比较高，本研究以仔猪肠上皮细胞（IPEC-J2）为模型，研究黄曲霉毒素B1（AFB1）、玉米赤霉烯酮（ZEA）和呕吐毒素（DON）的叠加细胞毒性。细胞毒性试验选用AFB1、ZEA和DON三种毒素作为响应面Box-Behnke设计的三个因素，以AFB1：10、20、30 μg/L，ZEA：150、300、450 μg/L，DON：500、1000、1500 μg/L作为Box-Behnke设计的三个编码水平。利用响应面设计构建得到17组复合霉菌毒素组合，以其对IPEC-J2细胞活力的影响作为参考指标，得到对细胞损伤程度最高和最低的霉菌毒素添加比例。结果表明：经方程预测后，得到细胞活力最低（霉菌毒素毒性最高）的AFB1、ZEA和DON组合为30、150 μg/L和1500 μg/L，经测定细胞活力为32.32%|得到细胞活力最高（霉菌毒素毒性最低）的AFB1、ZEA和DON组合为10、150 μg/L和600 μg/L，经测定细胞活力为53.01%。该结果为多种霉菌毒素叠加毒性的研究提供了依据。 [关键词] IPEC-J2细胞|黄曲霉毒素B1|玉米赤霉烯酮|呕吐毒素|细胞毒性     

Effects of Fusarium toxin-contaminated wheat grain on nutrient turnover, microbial protein synthesis and metabolism of deoxynivalenol and zearalenone in the rumen of dairy cows

The aim of the present study was to examine the effects of feeding Fusarium toxin-contaminated wheat to dairy cows on nutrient utilization in the rumen and on duodenal flow of deoxynivalenol (DON), zearalenone (ZON) and their metabolites. Six dairy cows fitted with a large rumen cannula and a simple T-shaped cannula at the proximal duodenum was used in two experiments. The experiments included a control period in which the uncontaminated control wheat was fed and a period in which the control wheat was replaced by the Fusarium toxin-contaminated wheat (8.05 and 7.15 mg DON/kg and 0.26 and 0.1 mg ZON/kg in Expts 1 and 2 respectively). The wheat portion of the daily ration amounted to 50% on a dry matter (DM) basis and rations were completed with hay or grass silage. Five of the six cows were non-lactating and the total daily DM-intake ranged between 4 and 12 kg. The pH-values and the concentration of volatile fatty acids in ruminal fluid were not significantly influenced by feeding the contaminated wheat. In contrast, the postprandial ammonia concentration was consistently higher when the mycotoxin-contaminated wheat was fed. Moreover, the flow of microbial protein and utilizable protein at the duodenum were reduced at the same time. The concentrations of DON and ZON and of their metabolites in freeze-dried duodenal digesta were either not detectable or negligible during the control periods whereas distinct concentrations were measured during the periods where the contaminated wheat was fed. DON was nearly completely metabolized to de-epoxy-DON and the flow at the duodenum ranged between 4% and 28% of DON-intake. The ZON metabolites alpha-zearalenol (ZOL) and beta-ZOL were recovered at the duodenum beside the parent toxin ZON. Their recovery as a percentage of ZON-intake ranged between 43% and 132%. In conclusion, feeding of Fusarium toxin-contaminated wheat altered the ruminal protein utilization. The question of whether this effect was a result of the mycotoxin being present in the rumen or of Fusarium growth-related structural (cell wall) changes of the wheat grain needs to be clarified. The low recovery of DON at the duodenum would indicate either a nearly complete degradation of the molecule in the rumen or an absorption by the mucosa of the rumen, whereas the higher ZON recovery would suggest a lower degradation of the parent toxin in the rumen and/or recovery of some bile-originating entero-hepatic cycling ZON/metabolites.     

Deoxynivalenol-contaminated wheat in swine diets

Two studies were conducted using Fusarium graminearum-infected (scabby) wheat containing 6.8 ppm deoxynivalenol (DON), commonly called vomitoxin, substituted for normal wheat in starter pig diets to give varying levels of DON. After 3 wk on experimental treatments, one-half of the pigs in trial one were sacrificed to evaluate the effects of DON on heart, kidney, spleen and liver. Analyses for DON residues in these tissues were also performed. The remaining 16 pigs were placed on a conventional diet for 4 wk to evaluate effects of DON on subsequent animal performance. A different sample of scabby wheat containing 4.9 ppm of DON was substituted for sorghum grain in growing-finishing pig diets to give varying concentrations of DON. At the end of the 42-d feeding period, eight pigs were slaughtered to evaluate the effects of DON on selected tissues. Results of the three trials suggest that feed intake was reduced when DON concentrations in the swine diets neared or exceeded 1 ppm. No apparent signs of disease, including vomiting, were observed in experimental animals. Histological evaluation revealed no significant lesions or abnormalities related to DON ingestion in tissues examined. Traces of DON (8 to 28 ppb, wet weight) were found in kidney, liver, spleen and heart of starter pigs consuming the diets containing DON up to time of slaughter. No DON was found in tissues of growing-finishing pigs that were withdrawn from feed about 12 h before slaughter.     

Occurrence of some mycotoxins (aflatoxin, ochratoxin A, citrinin, zearalenone and vomitoxin) in various Egyptian feeds

Ninety five samples of various Egyptian feedstuffs were investigated for the aflatoxin B1, B2, G1, and G2 means thin layer chromatography (TLC). Out of these samples 44.2% were positive (maize, rice crack, rice germ, rice germ cake, rice bran, wheat bran, cotton seed, cotton seed cake, peanut, and mixed feed for broilers, egg production, calf fattening and milk production). High percentage (90.5%) of the positive samples were contaminated with less than 100 ppb total aflatoxins. Peanut from "Ismailia" showed the highest contamination-mean of 400 ppb aflatoxin B1. The contamination relationship between kernels and shell of the same pods of the peanut was 1:7. The lowest contamination-mean was 5 ppb B1 in soya bean samples. All samples of horse bean and fish meal were negative. Aflatoxin B1 was present alone so frequently (in 76.2% of the positive samples). The relationship between the concentrations of aflatoxins B2:G1:B1 was 1:2.3:22.4. 51 different samples of foods and feeds from various Egyptian regions were collected and investigated for the nephrotoxic mycotoxin ochratoxin A means TLC. Twelve samples (23.5%) from them were designated as positive samples. The positive samples belonged to white maize, wheat, wheat bran, beans, rice germ, rice germ cake, broilers feed, egg production feed, and milk production feed; whereas the yellow maize (hybrid), soya beans, wheat soya meal, rice crack, cotton seed, cotton seed cake, and fish meal samples were negative. The contamination range was from 4 ppb to 577 ppb with an average of 58.2 +/- 22.9 ppb. Half of the positive samples was contaminated with 10-100 ppb whereas 41.7% from the positive samples had less than 10 ppb and 8.3% only had more than 100 ppb. Citrinin is existing in Egyptian food and feedstuffs. Out of 52 different samples--from various Egyptian regions-15.4% were positive. These were rice bran, rice germ, maize (white), wheat bran, cotton seed cake and fish meal. The highest contamination was in fish meal (40-70 ppb) whereas the lowest was in wheat bran (3 ppb). Mean of the contamination level was 25.9 +/- 3.4 ppb with a range of 3-70 ppb. For the first time in the Egyptian foods and feeds will be informed about the presence of the mycotoxin zearalenone with a high concentration. From several Egyptian places, 64 samples were collected (4 samples for each food or feed stuff).(ABSTRACT TRUNCATED AT 400 WORDS)     

Molecular analysis of Clostridium difficile isolates recovered from horses with diarrhea

Clostridium difficile is an important cause of diarrhea in horses, causing sporadic and epidemic disease of varying severity. This study evaluated the molecular characteristics of 48 C. difficile isolates recovered from diarrheic horses admitted to a veterinary hospital by using PCR-ribotyping and toxin gene profile. Additionally, feces were tested for the presence of C. difficile toxin A/B via enzyme immunosorbant assay (EIA) in 38 horses. The toxin genes tcdA, tcdB and cdtB were present in 27 (56.25%), 35 (72.91%) and 2 (4.1%) strains, respectively. Eight isolates (16.6%) were A(-)B(+) variants. Thirteen of forty-eight isolates (27.0%) did not posses any toxin genes (A(-)B(-)CDT(-)). A positive EIA result was reported in 17 (44%) of the cases. There was no association between the presence of different ribotypes or strains and toxin gene(s) profiles and the clinical outcome.     

Deoxynivalenol and T-2 Toxin in Raw Feeds for Horses

In all, 72 samples of raw materials for equine feed were collected from farms located in different parts of northern Italy, and the presence of deoxynivalenol (DON) and T-2 toxin was evaluated using enzyme-linked immunosorbent assay. DON was detected in 38.9% of the samples tested, at levels ranging from 0.2 to 1.9 mg/kg. Maize was found to have the highest concentrations of DON, whereas barley was found to be the most commonly contaminated grain (73.3%). T-2 toxin was found in maize and rice bran at levels ranging from 12 to 102 μg/kg, with an overall incidence of 12.3% in the samples analyzed. In almost all the samples, T-2 toxin was found only in combination with DON. The occurrence of contamination observed in this survey, especially the presence of DON, is noteworthy. The levels detected are not very high, but even long-term exposure to low doses of these mycotoxins may represent a threat to horse health.     

Detection of lysergic acid in ruminal fluid, urine, and in endophyte-infected tall fescue using high-performance liquid chromatography.

Ergot alkaloids present in endophyte-infected tall fescue induce fescue toxicosis in livestock consuming the plant. The lysergic acid (LA) ring structure is a common moiety among the ergot alkaloids. Little is known about the bioavailability of LA because of limitations in available analytical protocols. Thus, a high-performance liquid chromatography procedure was developed to analyze biological matrices for LA. The biological matrices of interest were tall fescue straw and seed, and ruminant feces, urine, and ruminal fluid. Lysergic acid was added to each matrix at a high (150 ng/ml) or low (30 ng/ml) level. Using the high-level addition, the greatest recovery of LA was obtained from ruminal fluid, feces, and urine (P < 0.05), with an average 85.1% recovered. At the low level, a greater recovery of added LA was observed in the ruminal fluid, urine, and feces (82.1%; P < 0.05) than that in the other 2 matrices (62.6%). The limit of quantitation (LOQ) in ruminal fluid and urine was 5.5 and 18.4 ng/ml, respectively. Seed, straw, and feces had higher LOQ (24.2, 14.5, and 36.0 ng/g, respectively). Limit of detection (LOD) was 1.64, 10.80, 4.35, 5.52, and 7.26 ng/g for ruminal fluid, feces, urine, seed, and straw, respectively. To test the assay in vivo, samples of ruminal fluid and urine were collected from steers consuming a diet containing 400 ng of ergovaline/g and 30 ng of LA/g. All matrices sampled resulted in levels above the LOD and LOQ for the assay, indicating that this assay is sufficiently sensitive for use in assessing the bioavailability of LA.     

Usefulness of a commercially available enzyme immunoassay for Shiga-like toxins I and II as a presumptive test for the detection of Escherichia coli O157:H7 in cattle feces.

The performance of a commercially available enzyme immunoassay (EIA) for determining the presence of Shiga toxin I and II in human diarrheal stool samples was evaluated for use as a presumptive test for the presence of Escherichia coli O157:H7 in nondiarrheal bovine fecal samples collected from 10 Kansas cow-calf ranches. The prevalence of E. coli O157:H7 in 2,297 samples, as determined by selective bacterial culture, was 1.6%. The sample prevalence of non-E. coli O157:H7 Shiga toxin-producing bacteria, as detected by the Shiga toxin EIA, was 5.8%. Only 2 of 136 samples that tested positive with the Shiga toxin EIA were positive for E. coli O157:H7 by culture. Compared with bacterial culture, the sensitivity of the Shiga toxin EIA was 5.5% and the specificity was 94.1%. Agreement between the 2 tests, as measured by the kappa statistic, was poor (kappa = -0.002). Although the Shiga toxin EIA was not a good presumptive test for the determination of E. coli O157:H7 in bovine fecal samples because of its low sensitivity (5.5%), it might be a useful test for the detection of Shiga toxin producing non-E. coli O157:H7 organisms in bovine feces.     

14C]-aflatoxin B1 metabolism in lactating goats and rats

Four dairy goats in the second to third month of lactation were administered [14C]-Aflatoxin B1 ( [14C]-AFB1). Two goats were dosed intravenously (iv) with 130 muCi (182 muCi/mumol) and two were dosed orally with 196 muCi (256 muCi/mumol). Urine, milk and feces were collected for 120 h after [14C]-AFB1 administration. Recoveries (average of two animals) of 14C in urine, milk and feces were, respectively; 22.7, .97 and 65% (iv dose) and 30.9, 1.05 and 52.3% (oral dose). Aflatoxin M1 (AFM1) was found in milk in the highest concentration. Aflatoxin Q1 (AFQ1) and aflatoxicol (AFL) were found in trace quantities in milk of animals administered the oral dose. In general, AFM1 could account for the majority of dicholromethane-soluble 14C. Liver contained 7.3 and 4.9% of the dose after 120 h for the iv and oral doses respectively. Kidney, heart, lung and spleen all contained .1% or less of the dose at 120 h. Muscle contained .48% of the dose at 120 h from the goats administered [14C]-AFB1 orally. There was no detectable radioactivity in the fat of any goat at 120 h. Six lactating Sprague-Dawley rats with 12 nursing pups were used for comparison of ruminants and simple-stomached animals. Rats were administered 2 muCi of [14C]-AFB1 (125 muCi/mumol) iv (three animals) and orally (three animals). Mean recovery of 14C in urine, mammary plus milk and feces were, respectively; 9.5, 2.0 and 60.7% (iv) and 8.8, 2.6 and 65.0% (orally).(ABSTRACT TRUNCATED AT 250 WORDS)     

Plasma haptoglobin and immunoglobulins as diagnostic indicators of deoxynivalenol intoxication

This study aimed to discover potential biomarkers for dioxynivalenol (DON) intoxication. B6C3F1 male mice were orally exposed to 0.83, 2.5 and 7.5 mg/kg body weight (bw) DON for 8 days and the differential protein expressions in their blood plasma were determined by SELDI - Time-of-Flight/Mass Spectrometry (TOF/MS) and the immunoglobulins (Igs) G, A, M and E in the serum were investigated. 11.7 kDa protein was significantly highly expressed according to DON administration and this protein was purified by employing a methyl ceramic HyperD F column with using optimization buffer for adsorption and desorption. The purified protein was identified as a haptoglobin precursor by peptide mapping with using LC/Q-TOF/MS and MALDI-TOF/MS and this was confirmed by western blotting and ELISA. IgG and IgM in serum were decreased in a dose-dependent manner and IgA was decreased at 7.5 mg/kg bw DON administration, but the IgE level was not changed. To compare the expressions of haptoglobin and the Igs patterns between aflatoxin B1 (AFB1), zearalenone (ZEA) and DON intoxications, rats were orally administered with AFB1 1.0, ZEA 240 and DON 7.5 mg/kg bw for 8 days. Haptoglobin was increased only at DON 7.5 mg/kg bw, while it was slightly decreased at ZEA 240 mg/kg bw and it was not detected at all at AFB1 1.0 mg/kg bw. IgG and IgA were decreased by DON, but IgG, IgA, IgM and IgE were all increased by AFB1. No changes were observed by ZEA administration. These results show that plasma haptoglobin could be a diagnostic biomarker for DON intoxication when this is combined with examining the serum Igs.     

Effects of graded levels of Fusarium toxin contaminated maize in diets for female weaned piglets.

A dose response study was carried out with piglets to examine the effects of increasing amounts of Fusarium toxins in the diet on performance, clinical serum characteristics, organ weights and residues of zearalenone (ZON) and deoxynivalenol (DON) and their metabolites in body fluids and tissues. For this purpose, Fusarium toxin contaminated maize (1.2 mg ZON and 8.6 mg DON per kg maize) was incorporated into a maize based diet for piglets at 0, 6, 12.5, 25 and 50% at the expense of control maize. The experimental diets were tested on 100 female piglets allotted to 20 boxes (five animals per box) covering a body weight range of 12.4 +/- 2.2 kg to 32.5 +/- 5.6 kg. Voluntary feed intake and, consequently, body weight gain of the animals receiving the highest proportion of Fusarium toxin contaminated maize were significantly decreased while the feed conversion ratio was not affected by the treatment. The mean weight of the uterus related to the body weight of the animals of the same group was increased by almost 100% as compared to the control. For this group, significantly decreased values of total serum protein were determined, while the serum activity of the liver enzyme glutamate dehydrogenase and the serum concentration of the follicle stimulating hormone were decreased for all treatment groups receiving 6% contaminated maize or more in the diet. Serum concentrations of immuneglobulins were not consistently altered by the treatment. Corresponding to the dietary exposure, increasing concentrations of ZON and alpha-zearalenol were detected in the bile fluid, liver and in pooled urine samples. The metabolite beta-zearalenol was detected only in bile fluid. The total concentration of ZON plus its metabolites in bile fluid correlated well with the diet contamination (r = 0.844). DON was found in serum, bile fluid and pooled urine samples while de-epoxy-DON was detected only in urine. The serum concentration of DON correlated well with the respective toxin intake 3-4 h prior to slaughtering (r = 0.957). For all mentioned analyses of residues it has to be noted that toxin residues were detectable even if negligible concentrations were present in the diet.     

Effect of long-term feeding of graded levels of deoxynivalenol (DON) on growth performance,nutrient utilization,and organ health in finishing pigs and DON content in biological samples

The prevalence of deoxynivalenol (DON) is a concern for swine producers, and although there has been extensive research into the effects of DON in pigs, focus has been in young pigs and/or in short-term studies. The objective of the study was to determine the effect of long-term exposure to DON-contaminated diets in finisher pigs. A total of 200 pigs (76.6 ± 3.9 kg initial weight) were group housed (five pigs per pen; n = 10 pens/treatment) in a 6-wk trial. Pigs were fed a wheat-barley-soybean meal-based control (CONT) diet with no DON or the basal diet in which clean wheat was replaced by DON-contaminated wheat and wheat screenings to provide DON content of 1, 3, or 5 ppm (DON1, DON3, and DON5, respectively). Individual BW and pen feed intake were recorded weekly to calculate average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F). Blood was collected on days 0, 14, and 43 and analyzed for indicators of liver and kidney health. Nitrogen (N)-balance was conducted immediately following the growth performance period to determine the effect of DON on nutrient utilization. Blood and urine samples collected during N balance were analyzed for DON content. Feeding DON reduced (P < 0.05) ADFI and ADG from days 0 to 28 compared with CONT, after which there was no effect of diet on ADFI and ADG. The G:F was lower (P < 0.05) in DON5 fed pigs compared with all treatments during days 0 to 7; however, no treatment effects on G:F was observed from days 8 to 42. Nitrogen retention was lower (P < 0.05) in DON3 and DON5 compared with DON1-fed pigs. Nitrogen retention efficiency was higher (P < 0.05) in DON1 compared with DON3 and DON5 and protein deposition for DON1 pigs was higher (P < 0.05) than all treatments. There were no treatment effects on indicators of liver and kidney health. As dietary DON intake increased, concentration of DON in blood and urine increased. Overall, although there was an initial decrease in ADG and ADFI in pigs receiving diets containing >1 ppm DON, pig performance recovered after a period of time, whereas nutrient utilization continued to be affected after recovery of performance. Moreover, the lack of DON on G:F indicates that the negative effects of DON on growth performance are largely due to reduced feed intake. Overall, although pigs maybe capable of adapting to intake of DON-contaminated diets, their final body weight will be reduced when fed diets containing >1 ppm DON.