Feed additives containing sequestrant clay minerals and inactivated yeast reduce aflatoxin excretion in milk of dairy cows.

The objective was to evaluate the efficacy of 2 dietary mycotoxin sequestrants, Toxy-Nil (TN) or Unike Plus (UP), in reducing aflatoxin (AF) M1 concentrations in milk of dairy cows challenged with dietary AF. Thirty-two mid-lactation Holstein cows were blocked by parity, days in milk, and milk yield and were randomly assigned within block to receive one of the following treatments: (1) 2.8 mg of AF/cow per d (positive control, PC), (2) 2.8 mg of AF + 100 g of TN/cow per d, (3) 2.8 mg of AF + 100 g of UP/cow per d, or (4) no AF and no additives (negative control, NC). For 7 d, treatments, dispersed in 150 g of sweet feed carrier, were top-dressed twice daily by mixing into the top portion of the TMR at each feeding. After the experimental period, cows were fed the NC diet and clearance of AFM1 via milk was monitored for 7 d. Feed and water were available ad libitum throughout the trial. Treatments had no effect on feed intake, milk yield, milk composition, or milk somatic cell count. Relative intake of AF was similar among PC, TN, and UP, averaging 106.5, 107.6, and 102.5 ± 2.9 µg/kg of diet dry matter, respectively. Relative intake of mycotoxin sequestrants was similar between TN and UP, averaging 0.4 and 0.4 ± 0.1% of diet dry matter, respectively. Concentration and mass of AFM1 secreted in milk and in urine were similar between TN and UP, but were lower than PC; concentrations in milk averaged 0.2, 0.3, and 0.6 ± 0.1 µg/kg, respectively, and mass secreted in milk averaged 8.1, 9.8, and 20.5 ± 1.7 µg/d. Concentrations in urine averaged 6.9, 7.4, and 14.2 ± 1.5 µg/L, respectively, and mass secreted in urine averaged 225.7, 250.8, and 521.6 ± 53.1 µg/d. Likewise, concentration and mass of free AF excreted in feces were similar between TN and UP, but were lower than PC; concentrations averaged 7.7, 8.9, and 12.4 ± 0.6 µg/kg, respectively, and mass excreted averaged 57.8, 69.6, and 95.6 ± 4.8 µg/d. Transfer of AF from feed to AFM1 in milk was reduced by 63 and 52%, and in urine, by 57 and 52% for TN and UP, respectively. Transfer of AF from feed to free AF in feces was reduced by 38 and 26% for TN and UP, respectively. The clearance rate of AFM1 in milk did not differ among PC, TN, and UP (46.1, 66.5, and 50.0 ± 6.7%/d, respectively). Results indicate that dietary inclusion of 100 g of TN or UP significantly reduced AFM1 in milk of cows consuming TMR containing approximately 105 µg of AF/kg of diet dry matter. Results also suggest that both TN and UP reduced absorption of AF.

Carryover of aflatoxins from feed to lambari fish (Astyanax altiparanae) tissues.

The aim of this study was to verify the carryover of aflatoxin B1 from feed to lambari fish. Aflatoxins (AF) were incorporated into feed, checking the levels by HPLC. Treatments were: Control, feed without toxin; A, feed + 10 µg AFB1 kg-1; B, feed + 20 µg AFB1 kg-1; and C, feed + 50 µg AFB1 kg-1. Juveniles of lambari fish were placed in 12 aquariums at a density of 50 fish/m2. Fish were fed twice a day with extruded feed, at 5% of animal biomass. The unit sample was constituted by a pool of 10 fish. AFs B1, B2, G1, G2 and M1 were quantified by HPLC in fish muscle and liver after 30, 60, 90 and 120 days of experiment. There was accumulation of AFs is fish liver and muscle, mainly after 90 days. Fish from treatment C had higher levels of AFB1 in muscle when compared with the others, and AFB1 in muscle at 120 days was similar to the levels in feed. Therefore, when lambari fish is exposed on a daily and long-term basis to AFs in feed, the regulation limits for AFs in animal feed do not guarantee safety for consumers.

The efficacy of raw and concentrated bentonite clay in reducing the toxic effects of aflatoxin in broiler chicks.

The objective of this study was to evaluate the efficacy of two adsorbents, a raw bentonite clay (RC) and a concentrated bentonite clay (CC), in ameliorating the toxic effects of aflatoxin B1 (AFB1). Results of the in vitro study (pH 3.0) indicated the CC adsorbed more AFB1 than RC (93.39 mg/g vs. 79.30 mg/g) suggesting that CC may be more effective than RC in reducing the toxic effects of AFB1. One hundred and eighty day-old straight run broiler chicks were assigned to 6 replicate pens of 5 chicks each and assigned to 6 dietary treatments from hatch to day 21. Dietary treatments included: 1) basal diet (BD) containing no AFB1 or adsorbents; 2) BD plus 0.50% RC; 3) BD plus 0.50% CC; 4) BD plus 2.0 mg AFB1/kg; 5) BD plus 2.0 mg AFB1/kg plus 0.50% RC; and 6) BD plus 2.0 mg AFB1/kg plus 0.50% CC. Dietary AFB1 concentrations were confirmed by analysis and diets were screened for other mycotoxins prior to the start of the experiment. The addition of AFB1 to the feed reduced (P < 0.05) growth performance and increased (P < 0.05) relative liver weight (RLW) and kidney weight (RKW) of chicks fed AFB1 compared to control chicks on day 21. These changes were ameliorated (P < 0.05) by the addition of RC and CC to the AFB1 diet. Mild to moderate lesions of aflatoxicosis (2.25) were observed in chicks fed AFB1 alone on day 21. The addition of both RC and CC to the AFB1 diet decreased (P < 0.05) but did not prevent liver lesions (0.92 and 1.42, respectively). Results indicate that both RC and CC were effective in reducing the toxic effects of AFB1, however the cost of processing of CC would make the RC a more economical product for reducing the effects of AFB1 in young broiler chicks.

Efficacy of adsorbents (bentonite and diatomaceous earth) and turmeric (Curcuma longa) in alleviating the toxic effects of aflatoxin in chicks.

A study was conducted to determine the efficacy of bentonite clay (BC), diatomaceous earth (DE) and turmeric powder (TUM) in alleviating the toxic effects of aflatoxin B1 (AFB1). A total of 250 Ross-308 d-old male broiler chicks were assigned to 10 dietary treatments (5 replicates of 5 chicks) from hatch to d 21. Dietary treatments were: basal diet; basal diet plus AFB1 (2 mg) or BC (0.75%), or DE (0.75%), or TUM (200 mg/kg curcuminoids) and different combinations of AFB1, BC, DE and TUM. Feed intake (FI), body weight gain (BWG) and feed gain (FG) of the birds fed on BC or DE separately were not different from control birds. Birds fed on TUM only had similar FI and FG but lower BWG than control chicks. Aflatoxin B1 reduced FI, BWG and serum concentrations of glucose, albumin, total protein calcium, but increased FG and relative liver and kidney weights. Chicks fed on the combination of AFB1 and BC had similar FI and FG to control chicks. Chicks fed on the combination of DE and AFB1 had lower FI (23.1%) and BWG (28.6%) compared with control chicks. Chicks fed on the combination of TUM and AFB1 also had decreased FI (26.2 %) and BWG (31%) compared with control chicks. Chicks fed on the combination of AFB1, BC and TUM consumed significantly higher amounts of feed compared with chicks fed on only AF, but gained less when compared with control diet chicks. Chicks fed on the combination of AFB1, DE and TUM diet had poorer growth performance than those fed on AFB1 alone. None of the combination diets reduced the severity of liver lesions.

Efficacy of beer fermentation residue containing Saccharomyces cerevisiae cells for ameliorating aflatoxicosis in broilers.

This study aimed to determine the aflatoxin B1 (AFB1) binding capacity of a beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells, and the efficacy of BFR to ameliorate the toxic effects of AFB1 on performance, serum biochemistry, and histology of broilers. The BFR was collected from a microbrewery, and the yeast cells were counted, dried, and milled before it was used in the study. In vitro evaluation of the BFR was conducted using different concentrations of AFB1 (2.0, 4.0, 8.0, 16.0, and 32.0 µg AFB1/mL) and 100 mg/10 mL of BFR at pH 3.0 or 6.0. Two hundred 1-day-old male broilers (Ross 308) were assigned to chick batteries and allowed ad libitum access to feed and water. A completely randomized design was used with 5 replicate pens of 5 chicks assigned to each of 4 dietary treatments from hatch to 21 d, which included: 1) basal diet (BD), with no BFR or AFB1; 2) BD supplemented with 1% BFR; 3) BD supplemented with 2 mg AFB1/kg of feed; and 4) BD supplemented with 2 mg AFB1/kg feed and 1% BFR. Performance variables were determined weekly, while serum analyses were performed on d 14 and 21. At the end of the study, chicks were anesthetized with carbon dioxide, euthanized by cervical dislocation, and the kidney, liver, and bursa of Fabricius were removed for determination of relative weights, and for histological evaluation. In vitro assays showed that the higher the initial AFB1 concentration in solution, the greater the AFB1 amount adsorbed by BFR at both pHs tested. Feed intake, BW gain, and concentrations of albumin, total protein, and globulin increased (P < 0.05) in broilers fed BFR+AFB1 (Diet 4), when compared to the birds receiving only AFB1 (Diet 2). Although BFR was not able to reduce or prevent the effects of AFB1 on relative weights of kidneys and liver, it reduced the severity of histological changes in the liver and kidney caused by AFB1.

Occurrence of Polyphenols, Organic Acids, and Sugars among Diverse Elderberry Genotypes Grown in Three Missouri (USA) Locations.

Elderberry (Sambucus spp.) is an emerging horticultural crop used in a variety of foods, wines, and dietary supplements. A better understanding of the elderberry juice complex including its putative health-promoting compounds in relation to genetic and environmental parameters is needed. A multi-location planting of nine elderberry genotypes was established in 2008 at three geographically-diverse sites in Missouri, USA. Fruits were harvested from replicated plots 2009-2011, frozen, and later prepared for laboratory analysis. Polyphenols, organic acids, and sugars were quantified by HPLC and the results evaluated for response to genotype, site, and year. The American genotypes 'Ocoee' and 'Ozark' were consistently higher in chlorogenic acids compared to other genotypes, whereas 'Ocoee' was significantly higher in rutin than 'Ozark'. The European 'Marge' was significantly higher in isoquercitrin and other flavonoids compared to most North American genotypes. Significant differences in polyphenols were also detected among sites and production years. Malic, citric, and tartaric acids varied significantly among genotypes, sites, and years, whereas succinic, shikimic, and fumaric acids generally did not. Levels of lactic, acetic, and propionic acids were negligible in most samples. The American genotype 'Ocoee' was higher in citric and tartaric acids, while lower in malic acid. The sugars glucose and fructose also responded significantly to genotype, site, and year. 'Ocoee', 'Ozark', and 'Marge' perform very well in Missouri horticulturally and appear to have additional potential as cultivars based on their unique juice characteristics.

Assessment of dietary intake of fumonisin B1 in São Paulo, Brazil.

In this study, fumonisin B1 (FB1) consumption was assessed through determination of FB1 in corn meal, corn flour, corn flakes, polenta, canned corn and popcorn collected from homes of residents of Pirassununga, São Paulo, Brazil, and using a Food Frequency Questionnaire (FFQ) filled out by the residents. One hundred and twenty samples were collected from 39 residents on four separate occasions. FB1 was determined by high performance liquid chromatography using a validated method that uses SAX column clean-up. The highest levels of FB1 were found in corn meal at a mean concentration of 474.6 µg kg(-1). However, none of the samples tested for FB1 had levels above the tolerance limit established in Brazil. The mean probable daily intake (PDIM) of FB1 was 63.3 ng kg(-1)body weight day(-1), which is approximately 3% of the provisional maximum tolerable daily intake (PMTDI) recommended for fumonisins.

Lipid metabolism of commercial layers fed diets containing aflatoxin, fumonisin, and a binder.

Aflatoxins (AF) and fumonisins (FU) are a major problem faced by poultry farmers, leading to huge economic losses. This experiment was conducted to determine the effects of AF (1 mg/kg of feed) and FU (25 mg/kg of feed), singly or in combination, on the lipid metabolism in commercial layers and investigate the efficacy of a commercial binder (2 kg/t of feed) on reducing the toxic effects of these mycotoxins. A total of 168 Hisex Brown layer hens, 37 wk of age, were randomized into a 3 × 2 + 1 factorial arrangement (3 diets with no binder containing AF, FU, and AF+FU; 3 diets with binder containing AF, FU, and AF+FU; and a control diet with no mycotoxins and binders), totaling 7 treatments. The hens contaminated with AF showed the characteristic effects of aflatoxicosis, such as a yellow liver, resulting from the accumulation of liver fat, lower values of plasma very low-density lipoprotein and triglycerides, and higher relative weight of the kidneys and liver. Hepatotoxic and nephrotoxic effects of FU were not observed in this study. On the other hand, the FU caused a reduction in small intestine length and an increase in abdominal fat deposition. The glucan-based binder prevented some of the deleterious effects of these mycotoxins, particularly the effects of AF on hepatic lipid metabolism, kidney relative weight, and FU in the small intestine.

In vitro and in vivo efficacy of a hydrated sodium calcium aluminosilicate to bind and reduce aflatoxin residues in tissues of broiler chicks fed aflatoxin B1.

The aim of this study was to determine the binding capacity of a hydrated sodium calcium aluminosilicate (HSCAS) for aflatoxin B(1) (AFB(1)), and the efficacy of the HSCAS to reduce the concentrations of residual AFB(1) and its metabolites in the liver and kidney of broilers fed AFB(1). One hundred 1-d-old male broilers (Ross 708) were maintained in chick batteries and allowed ad libitum access to feed and water. A completely randomized design was used with 5 replicate pens of 5 chicks assigned to each of 4 dietary treatments from hatch to 21 d. Dietary treatments included the following: A) basal diet (BD), with no HSCAS or AFB(1), B) BD supplemented with 0.5% HSCAS only, C) BD supplemented with 2.5 mg of AFB(1)/kg of feed, and D) BD supplemented with 2.5 mg of AFB(1)/kg of feed and 0.5% HSCAS. On d 21, 5 chicks from each treatment were anesthetized with carbon dioxide, killed by cervical dislocation, and samples of liver and kidney were collected for analysis of AFB(1) residues. The percentage of AFB(1) bound for each concentration of adsorbent (100, 10, 1, 0.5, 0.25, and 0.05 mg/10 mL) was 100, 91.1, 81.8, 75.4, 40.1, and 8.8%, respectively. Concentrations of aflatoxin residues (AFB(1), aflatoxicol, aflatoxins B(2) and G(1)) were lower (P < 0.05) in livers and kidneys of birds fed AFB(1) plus HSCAS (diet D), when compared with birds fed AFB(1) alone (diet C). However, histopathology data from the in vivo study indicated that HSCAS did not prevent lesions associated with aflatoxicosis. The decrease in the bioavailability of AFB(1) caused by the HSCAS reduced aflatoxin residues in liver and kidney, but not enough to completely prevent the toxic effects of AFB(1) in broilers.

Pathology of ochratoxin A-induced nephrotoxicity in Japanese quail and its protection by sea buckthorn (Hippophae rhamnoides L.).

The present study was designed to study the protective effect of sea buckthorn (SBT) against renal damage induced by ochratoxin A (OTA) in Japanese quail. Day-old quail chicks were divided into six groups and fed a basal quail chick mash containing 2% SBT leaf powder (group SX), OTA at a dietary level of 3 ppm (group OX), 25 ppm L-beta-phenylalanine (Phe) plus 3 ppm OTA (group OP), 2% dietary level of SBT leaf powder plus 3 ppm OTA (group OS), SBT leaf extract at a level of 10%/L of drinking water plus 3 ppm OTA (group OSS), and a standard toxin-free feed (group CX, control) for 21 days. OTA at 3 ppm level in diet grossly revealed mild to moderate renal swelling in OX birds, and the severity was less in the case of OS, OSS, and OP birds. Microscopically, degenerative, necrotic, and inflammatory changes were observed in OX birds, but the changes were less severe in OS, OSS, and OP birds. Ultrastructural studies revealed remarkable and consistent changes in the proximal convoluted tubules (PCTs), with severe damage of mitochondria and endoplasmic reticulum in OX birds, whereas SBT-treated birds (groups OS, OSS) had mild changes in mitochondria. A moderate to marked increase in number of peroxisomes in the cytoplasm of PCTs was a consistent finding in the Phe- and SBT-treated groups kept on OTA in comparison to the group fed OTA alone. In conclusion, the inclusion of 2% SBT leaf powder in feed and SBT leaf extract in water provided partial protection against OTA-induced nephropathy in Japanese quail.

Toxic interaction between fumonisin B1 and moniliformin for cardiac lesions in Japanese quail.

This study examined the effects of fumonisin B1 (FB1) and moniliformin (M) on the heart of Japanese quail (Coturnix coturnix japonica). Three hundred and ninety day-old Japanese quail were randomly divided into four groups: 1) FB1 alone (FX), 2) M alone (MX), 3) FB1 and M (FM), and 4) chick mash alone (CX). We used three pen replicates of 35 quail per pen in groups FX, MX, and FM and three pen replicates of 25 quail per pen in group CX. Gross and microscopic changes in the heart were studied in nine birds (three birds per replicate) from each group at weekly intervals up to 28 days postfeeding (DPF). Ultrastructural changes were studied in the heart of three birds (one bird per replicate) from each group at 21 DPF. Thinning of the heart was the only significant gross lesion in group FX. In contrast, mild-to-severe cardiomegaly was a significant finding in groups MX and FM throughout the study. Microscopically, thinning of cardiomyocytes was evident at 7 DPF in group FX. In addition to the hypertrophy of cardiomyocytes evident as early as 7 DPF, myocardial karyomegaly, nuclear hyperchromasia, and myofibril disarray exhibiting a wavy pattern were more pronounced at 28 DPF in group MX. Similar but more severe lesions were observed in the FM combination group that included myocardial hemorrhages, vacuolar changes, hypertrophy of cardiomyocytes, focal myocarditis, and loss of myofibrils cross-striations. Via transmission electron microscopy, the maximum effect of FB1 toxicity was observed on mitochondria. In addition to an increase in the number of mitochondria, the mitochondria seemed invariably swollen and pleomorphic, although the outer membrane was intact, and the membrane cristae were usually distinct. Myofibrils seemed thinner, without much disruption in their architecture. Large numbers of vacuolar bodies of irregular size, both in the sarcoplasm and in between the myofibrils, were conspicuous in group FX. In contrast to group FX, the increase in number of mitochondria resulted in widespread separation of muscle fibers in group MX. In addition, the mitochondria were swollen and varied from round to oval to slightly elongated and occasionally forked, and vacuolation was rarely noticed in group MX. In the FM combination group, a significant increase in the number of mitochondria caused muscle fibers to look much thinner and assume a wavy pattern. We conclude that the effect of M on the heart is exaggerated in the presence of FB1. Although the overall interactive effect of FB1 and M was less than additive, the interactive effects between the two toxins for cardiac lesions were greater than additive to synergistic up to the second week, raising serious concerns on early age exposure to a combination of these two mycotoxins.

Use of different levels of ground endophyte-infected tall fescue seed during heat stress to separate characteristics of fescue toxicosis.

Studies of fescue toxicosis using whole seed diets show reduced feed intake and thermoregulatory ability, but much of the seed passes undigested through the animal. Cattle were fed ground tall fescue seed at different levels to potentially facilitate digestion and absorption of toxins and identify toxin sensitivity for major characteristics of the condition [i.e., hyperthermia, reduced feed intake (FI), reduced blood prolactin]. Steers (n = 18; 350 kg BW) were housed in the Brody Climatology Laboratory at thermoneutrality (TN; 19°C) and randomly assigned to daily diet treatments with either ground endophyte-infected [E+; low and high doses at 20 and 40 µg ergovaline/(kg BW/d), respectively] or endophyte-free [E-; control at 0 µg ergovaline/(kg BW/d)] tall fescue seed. After 12 d at TN, animals received 2 d of transition to heat stress (HS; 36°C daytime, 25°C nighttime) and maintained for 14 more days. Cattle were fed twice daily at 0800 and 1600 h, with water ad libitum. Feed intake was measured at 0700 h, with skin and rectal temperatures, and respiration rate at 0600, 1100, 1600, and 2100 h. Blood was sampled on selected days for prolactin and leptin determinations. Steers fed ground E+ diet decreased (P ≤ 0.0001) FI below controls at TN, with no dose effect. Maximum FI reduction with E+ treatment was 25% at TN, with an additional 46% decrease during HS (P ≤ 0.05). By the end of HS, E+ FI increased (P > 0.05) to that of E-, suggesting recovery. Prolactin was reduced (P ≤ 0.05) in high E+ cattle below controls at study end. Leptin blood concentrations were unaffected by E+ treatment (P > 0.05) but was reduced (P ≤ 0.05) by the end of HS. Pattern of rectal temperature response to HS showed a more rapid initial increase and decline for both E+ groups compared with controls (P ≤ 0.05). Skin temperature was the only variable that identified E+ dose differences. Although there were no treatment differences at TN, skin temperature was lower (P ≤ 0.05) for high E+ steers compared with controls during HS when air temperature was reduced each day. In general, FI was more responsive to E+ toxins than body temperature or blood prolactin, declining even at TN and exhibiting dynamic activity during HS. Although body temperature response to E+ toxins appears to stabilize during HS, this is misleading as rapid change in air temperature exposes effects on skin temperature.

Effects of melamine in young broiler chicks.

A study was conducted to determine the toxicity of melamine in young broilers fed graded levels of melamine. An additional objective was to determine melamine residual levels in selected tissues. One hundred and seventy-five 1-d-old male Ross broiler chicks were sorted to a randomized block design in stainless steel battery pens. Chicks were assigned to 7 dietary treatments containing 0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0% melamine. Each dietary treatment was fed to 5 replicate pens of 5 chicks for 21 d. Mortality increased quadratically (P<0.001) with increasing dietary concentrations of melamine. However, compared with controls, mortality was only higher (P<0.001) in birds fed≥2.5% melamine. Feed intake decreased linearly (P<0.001), whereas BW gain decreased quadratically (P<0.02) with increasing dietary concentrations of melamine. Compared with controls, both feed intake and BW gain were lower (P<0.001) only in birds fed≥1.0% melamine. Relative kidney weights increased linearly (P<0.001), whereas relative liver weights increased quadratically (P<0.05) with increasing dietary concentrations of melamine. Melamine residues in breast muscle and liver tissue increased linearly (P<0.001) with increasing dietary concentrations of melamine, whereas melamine residues in kidney and bile increased quadratically (P<0.02) with increasing dietary concentrations of melamine. Compared with controls, melamine concentrations in liver and kidney were higher (P<0.001) in birds fed all levels of melamine, whereas melamine concentrations in breast muscle and bile were only higher (P<0.001) in birds fed≥1.0% melamine. Serum albumin, total protein, globulin, and calcium increased quadratically (P<0.02) in birds as dietary melamine increased, whereas serum aspartate transaminase and gamma gluatamyltransferase increased linearly (P<0.01) with increasing levels of melamine in the diet. Renal histopathology revealed nonpolarizable melamine crystals in the collecting tubules and ducts of birds fed≥1.5% melamine. In summary, dietary melamine was toxic to broilers at concentrations≥1.0%.

Effects of dietary aflatoxin on the health and performance of growing barrows.

Aflatoxins, especially aflatoxin B1 (AFB1), can be greater in dried distillers grains with solubles (DDGS) because it can be concentrated during the ethanol production process. Increased use of DDGS in swine diets could potentially lead to an increased incidence of aflatoxicosis, a disease associated with decreased feed intake, reduced BW gain, and impaired liver function. The objective of this study was to determine the effects of AFB1 on the health, performance, and serum profile of growing barrows. Ninety Duroc × Yorkshire crossbred barrows were purchased (age = 35 ± 5 d; BW = 14.2 ± 3.0 kg), allocated to 9 pens with 10 pigs per pen, and randomly assigned to receive diets containing 0 µg/kg of AFB1 (CON), 250 µg/kg of AFB1 (LO), or 500 µg/kg of AFB1 (HI) for 7, 28, or 70 d in a 3 × 3 factorial arrangement of treatments. Feed intake was measured daily, and pigs were weighed and blood samples collected weekly. Serum was analyzed for concentrations of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (BILI), and blood urea nitrogen (BUN). Both ADFI and ADG were negatively affected (P ≤ 0.001) by AFB1 treatment. Average daily feed intake was less (P < 0.05) in HI barrows than in CON barrows from wk 5 to 10 and was less (P < 0.05) in LO barrows than in CON barrows in wk 5 and again from wk 8 to 10. Also, ADFI was less (P = 0.022) in HI barrows than LO barrows in wk 10. Decreased ADG (P < 0.05) was observed in HI barrows than in CON barrows in wk 8 and 10; no differences (P ≥ 0.665) in ADG were noted between CON and LO barrows. There was no effect (P ≥ 0.080) of AFB1 treatment on ALT or BILI concentrations. However, both AST and BUN were affected (P < 0.05) by AFB1 treatment. Serum AST was greater (P = 0.010) in LO barrows than CON barrows in wk 5, and serum BUN was greater (P = 0.004) in CON barrows than LO barrows in wk 3. Results from this study demonstrate that the performance and health of young growing barrows were affected by consumption of an AFB1-contaminated diet, especially when fed for a more extended period.

Genetic variation at bx1 controls DIMBOA content in maize.

The main hydroxamic acid in maize (Zea mays L.) is 2-4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA). DIMBOA confers resistance to leaf-feeding by several corn borers. Most genes involved in the DIMBOA metabolic pathway are located on the short arm of chromosome 4, and quantitative trait loci (QTLs) involved in maize resistance to leaf-feeding by corn borers have been localized to that region. However, the low resolution of QTL linkage mapping does not allow convincing proof that genetic variation at bx loci was responsible for the variability for resistance. This study addressed the following objectives: to determine the QTLs involved in DIMBOA synthesis across genetically divergent maize inbreds using eight RIL families from the nested association mapping population, to check the stability of QTLs for DIMBOA content across years by evaluating two of those RIL families in 2 years, and to test the involvement of bx1 by performing association mapping with a panel of 281 diverse inbred lines. QTLs were stable across different environments. A genetic model including eight markers explained approximately 34% of phenotypic variability across eight RIL families and the position of the largest QTL co-localizes with the majority of structural genes of the DIMBOA pathway. Candidate association analysis determined that sequence polymorphisms at bx1 greatly affects variation of DIMBOA content in a diverse panel of maize inbreds, but the specific causal polymorphism or polymorphisms responsible for the QTL detected in the region 4.01 were not identified. This result may be because the causal polymorphism(s) were not sequenced, identity is masked by linkage disequilibrium, adjustments for population structure reduce significance of causal polymorphisms or multiple causal polymorphisms affecting bx1 segregate among inbred lines.

Effects of turmeric (Curcuma longa) on the expression of hepatic genes associated with biotransformation, antioxidant, and immune systems in broiler chicks fed aflatoxin.

The objective of the present study was to evaluate the efficacy of curcumin, an antioxidant found in turmeric (Curcuma longa) powder (TMP), to ameliorate changes in gene expression in the livers of broiler chicks fed aflatoxin B(1) (AFB(1)). Four pen replicates of 5 chicks each were assigned to each of 4 dietary treatments, which included the following: A) basal diet containing no AFB(1) or TMP (control), B) basal diet supplemented with TMP (0.5%) that supplied 74 mg/kg of curcumin, C) basal diet supplemented with 1.0 mg of AFB(1)/kg of diet, and D) basal diet supplemented with TMP that supplied 74 mg/kg of curcumin and 1.0 mg of AFB(1)/kg of diet. Aflatoxin reduced (P < 0.05) feed intake and BW gain and increased (P < 0.05) relative liver weight. Addition of TMP to the AFB(1) diet ameliorated (P < 0.05) the negative effects of AFB(1) on growth performance and liver weight. At the end of the 3-wk treatment period, livers were collected (6 per treatment) to evaluate changes in the expression of genes involved in antioxidant function [catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST)], biotransformation [epoxide hydrolase (EH), cytochrome P450 1A1 and 2H1 (CYP1A1 and CYP2H1)], and the immune system [interleukins 6 and 2 (IL-6 and IL-2)]. Changes in gene expression were determined using the quantitative real-time PCR technique. There was no statistical difference in gene expression among the 4 treatment groups for CAT and IL-2 genes. Decreased expression of SOD, GST, and EH genes due to AFB(1) was alleviated by inclusion of TMP in the diet. Increased expression of IL-6, CYP1A1 and CYP2H1 genes due to AFB(1) was also alleviated by TMP. The current study demonstrates partial protective effects of TMP on changes in expression of antioxidant, biotransformation, and immune system genes in livers of chicks fed AFB(1). Practical application of the research is supplementation of TMP in diets to prevent or reduce the effects of aflatoxin in chicks fed aflatoxin-contaminated diets.

Efficacy of Solis, NovasilPlus, and MTB-100 to reduce aflatoxin M1 levels in milk of early to mid lactation dairy cows fed aflatoxin B1.

An experiment was conducted to determine the efficacy of 3 adsorbents, Solis (SO; Novus International Inc.), NovasilPlus (NOV; Engelhard Corp.), and MTB-100 (MTB; Alltech), in reducing aflatoxin (AF) M(1) concentrations in milk of dairy cows fed an AF-contaminated diet. Twelve early to mid lactation dairy cows averaging 163 d in milk were used in a 4 x 4 Latin square design with 3 replications. Cows were blocked by parity, body weight, and milk production and were provided ad libitum access to feed and water. Within each replicate, cows were randomly assigned to the 4 dietary treatments for 4 consecutive 7-d periods. Dietary treatments included AF [112 microg of AFB(1)/kg of diet dry matter (DM)]; AF + 0.56% SO; AF + 0.56% NOV; and AF + 0.56% MTB. Milk samples were collected on d 6 and 7 of each of the experimental periods. Feed intake, milk production, milk fat percentage, milk protein percentage, and linear somatic cell scores were not affected by dietary treatments and averaged 22.20 kg/d of DM, 33.87 kg/d, 3.78%, 2.95%, and 1.60, respectively, across all treatments. Transfer rates of AF from feed to milk averaged 2.65, 1.48, 1.42, and 2.52% for cows fed AF, AF + SO, AF + NOV, and AF + MTB, respectively. Daily AFM(1) excretion in milk averaged 66, 37, 35, and 63 microg/d for cows fed AF, AF + SO, AF + NOV, and AF + MTB, respectively. The addition of SO and NOV to the AF diet resulted in a significant reduction in milk AFM(1) concentrations (SO, 45%; NOV, 48%) and AFM(1) excretion (SO, 44%; NOV, 46%). In contrast, MTB was not effective in reducing milk AFM(1) concentrations (4%), AFM(1) excretion (5%), or AF transfer from feed to milk (2.52%). Results indicated that SO and NOV at 0.56% of the diet were effective in reducing milk AFM(1) concentrations in cows consuming a total mixed ration containing 112 microg of AFB(1)/kg of diet DM.

Effects of short-term heat stress on endophytic ergot alkaloid-induced alterations in rat hepatic gene expression.

Exposure to ergot alkaloids in endophyte-infected fescue (E+) is associated with impaired animal productivity, especially during heat stress, which is commonly referred to as fescue toxicosis. To elucidate the pathogenesis of this condition, the effects of short-term heat stress (HS) on hepatic gene expression in rats exposed to endophytic ergot alkaloids were evaluated. Rats implanted with telemetric transmitters to continuously measure core temperature were fed an E+ diet and maintained under thermoneutral (TN) conditions (21 degrees C) for 5 d, followed by TN or 31 degrees C (HS) conditions for 3 d. Feed intake (FI) and BW were monitored daily. The E+ and HS-induced alterations in hepatic genes were evaluated using DNA microarrays and PCR analyses. Hepatic antioxidant enzyme activities, as well as the incidence of apoptosis, were determined. As expected, intake of E+ reduced FI and BW from pretreatment levels under TN conditions, with greater reductions during short-term HS. Genes involved in gluconeogenesis and apoptosis were upregulated, whereas genes associated with oxidative phosphorylation, xenobiotic metabolism, antioxidative mechanisms, immune function, cellular proliferation, and chaperone activity were all downregulated with short-term HS. Hepatocytic apoptosis was increased and antioxidant enzyme activity decreased in the livers of rats exposed to HS. The hypothesized, exacerbating effects of HS on the direct, endophytic toxin-related and indirect, reduced caloric intake-associated alterations in hepatic gene expression were clearly demonstrated in rats and may help to elucidate the pathogenesis of fescue toxicosis in various animal species.

Toxicological and gene expression analysis of the impact of aflatoxin B1 on hepatic function of male broiler chicks.

The objective of this study was to determine the effects of dietary aflatoxin B1 (AFB1) on hepatic gene expression in male broiler chicks. Seventy-five 1-d-old male broiler chicks were assigned to 3 dietary treatments (5 replicates of 5 chicks each) from hatch to d 21. The diets contained 0, 1 and 2 mg of AFB1/kg of feed. Aflatoxin B1 reduced (P<0.05) feed intake, BW gain, serum total proteins, and serum Ca and P, but increased (P<0.01) liver weights in a dose-dependent manner. Microarray analysis was used to identify shifts in genetic expression associated with the affected physiological processes in chicks fed 0 and 2 mg of AFB1/kg of feed to identify potential targets for pharmacological/toxicological intervention. A loop design was used for microarray experiments with 3 technical and 4 biological replicates per treatment group. Ribonucleic acid was extracted from liver tissue, and its quality was determined using gel electrophoresis and spectrophotometry. High-quality RNA was purified from DNA contamination, reverse transcribed, and hybridized to an oligonucleotide microarray chip. Microarray data were analyzed using a 2-step ANOVA model and validated by quantitative real-time PCR of selected genes. Genes with false discovery rates less than 13% and fold change greater than 1.4 were considered differentially expressed. Compared with controls (0 mg of AFB1/kg), various genes associated with energy production and fatty acid metabolism (carnitine palmitoyl transferase), growth and development (insulin-like growth factor 1), antioxidant protection (glutathione S transferase), detoxification (epoxide hydrolase), coagulation (coagulation factors IX and X), and immune protection (interleukins) were downregulated, whereas genes associated with cell proliferation (ornithine decarboxylase) were upregulated in birds fed 2 mg of AFB1/kg. This study demonstrates that AFB1 exposure at a concentration of 2 mg/kg results in physiological responses associated with altered gene expression in chick livers.

Influence of toxic endophyte-infected fescue on sperm characteristics and endocrine factors of yearling Brahman-influenced bulls.

Sixteen (mean age = 1.1 +/- 0.1 yr; mean BW = 478 +/- 34 kg) Brahman-influenced bulls were used to determine the influence of fescue type on sperm characteristics and serum concentrations of prolactin, cortisol, and testosterone. Bulls were blocked by BW, scrotal circumference (SC), and pregrazing sperm characteristics and randomly assigned to graze toxic endophyte-infected (EI; 4 bulls/pasture; 2 pastures) or novel endophyte-infected (NE; 4 bulls/pasture; 2 pastures) tall fescue for 121 d. Semen was collected by electroejaculation, and SC was measured and blood samples collected monthly. Sperm were evaluated for motility and morphology with an integrated visual optical system. Overall mean concentration of prolactin was decreased more (P < 0.01) in EI bulls than NE bulls from May to August. Scrotal circumference was not affected by fescue type (P = 0.58); overall SC averaged 36.7 +/- 2.3 cm. Percentage of live sperm was not different (P = 0.24) between NE bulls (80%) than EI bulls (67%) in July and August. Bulls grazing NE fescue had more (P < 0.06) motile sperm than EI bulls in July and August. Percentages of progressive (57 vs. 38%, NE and EI, respectively; P < 0.06) and rapid (67 vs. 46%, NE and EI, respectively; P = 0.04) sperm were greater from bulls grazing NE than EI bulls in July and August. Average velocity of the smoothed sperm path and progressive velocity in a straight line from the beginning to the end of the sperm track were slower (P < 0.09) in EI bulls than NE bulls and were slower (P = 0.04) in August compared with July. Mean width of head oscillation as the sperm swims was less (P < 0.06) in August than July. Concentrations of cortisol and testosterone were not (P > 0.10) influenced by fescue type. Semen from bulls grazing EI had reduced motility and morphology than bulls grazing NE. Detrimental effects of toxic fescue may not be mediated by cortisol, testosterone, or both. Semen quality of bulls grazing toxic EI tall fescue was decreased with increased maximum ambient temperatures.