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.

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.

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.

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.

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.

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.

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.

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%.

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.

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.

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.

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.

Individual and combined effects of fumonisin b1 and moniliformin on clinicopathological and cell-mediated immune response in Japanese quail.

A total of 390 one-day-old quail chicks (Coturnix coturnix japonica) were divided into 4 groups (3 replicates per treatment), viz. CX, FX, MX, and FM, containing 75, 105, 105, and 105 birds, respectively. Birds in the control group (CX) were fed quail mash alone, whereas birds in group FX were fed 200 ppm of fumonisin B(1) (FB(1)) from Fusarium verticillioides culture material; group MX was fed 100 ppm of moniliformin (M) from Fusarium fujikuroi culture material; and group FM was fed a combination of 200 ppm of FB(1) and 100 ppm of M. Diets were fed from d 1 to 35 to study clinical signs, growth response, serum biochemical changes, and cell-mediated immune response. Birds fed FB(1) (FX) showed ruffled feathers and poor growth. Birds in group MX appeared more stunted than those in group FX and exhibited signs of poor feathering and decreased feed and water intake. Clinical signs observed in group FM were more or less similar to those observed in groups FX and MX. Total mortality was 12.38, 7.62, and 20.95% for groups FX, MX, and FM, respectively. Mean BW in groups FX, MX, and FM were significantly lower than those in the control group (CX) at almost all intervals. Total serum proteins, albumin, cholesterol, aspartate transaminase, lactate dehydrogenase, and creatine kinase values were higher in all treatment groups compared with the control group. Cell-mediated immune response was more or less comparable in groups CX and MX, whereas the presence of FB(1) in the diet of groups FX and FM was found to be associated with a gradual increase in skin thickness, and the mononuclear inflammatory cell response was poor as compared with groups CX and MX throughout the study. Except for mortality (additive effect) and serum aspartate transaminase values (less than an additive effect up to 14 DPF), no additive or synergistic effects were observed for any of the other response variables measured in the current study, where all statistical differences were attributed to either one mycotoxin or the other.

Efficacy of turmeric (Curcuma longa), containing a known level of curcumin, and a hydrated sodium calcium aluminosilicate to ameliorate the adverse effects of aflatoxin in broiler chicks.

A 3-wk feeding study was conducted to evaluate the efficacy of turmeric (Curcuma longa) powder (TMP), containing a known level of curcumin, and a hydrated sodium calcium aluminosilicate (HSCAS; Improved Milbond-TX, IMTX, an adsorbent, Milwhite Inc., Houston, TX) to ameliorate the adverse effects of aflatoxin B(1) (AFB(1)) in broiler chicks. Four pen replicates of 5 chicks each were assigned to each of 7 dietary treatments, which included the basal diet not containing TMP, HSCAS, or AFB(1) (control); basal diet supplemented with 0.5% food grade TMP that contained 1.48% total curcuminoids (74 mg/kg); basal diet supplemented with 0.5% HSCAS; basal diet supplemented with 1.0 mg/kg AFB(1); basal diet supplemented with 0.5% TMP and 1.0 mg/kg AFB(1); basal diet supplemented with 0.5% HSCAS and 1.0 mg/kgAFB(1); and basal diet supplemented with 0.5% TMP, 0.5% HSCAS, and 1.0 mg/kg AFB(1). The addition of TMP to the AFB(1) diet significantly (P < 0.05) improved the weight gain of chicks, and the addition of HSCAS to the AFB(1) diet significantly (P < 0.05) improved feed intake and weight gain, and reduced relative liver weight. The addition of TMP or HSCAS and TMP with HSCAS ameliorated the adverse effects of AFB(1) on some of the serum chemistry parameters (total protein, albumin, cholesterol, calcium). Further, decreased antioxidant functions in terms of level of peroxides, superoxide dismutase activity, and total antioxidant concentration in liver homogenate due to AFB1 were also alleviated by the inclusion of TMP, HSCAS, or both. The reduction in the severity of hepatic microscopic lesions due to supplementation of the AFB(1) diet with TMP and HSCAS demonstrated the protective action of the antioxidant and adsorbent used in the present study.

Pathologic changes in extrahepatic organs and agglutinin response to Salmonella Gallinarum infection in Japanese quail fed Fusarium verticillioides culture material containing known levels of fumonisin B1.

Three hundred 1-day-old Japanese quail (Coturnix coturnix japonica) were divided into two groups of 150 each. One group was maintained on quail mash alone, whereas Fusarium verticillioides culture material (FCM) was added to quail mash in the second group from 5 days of age and supplied 150 mg FB1/kg mash. At day 21, each group was further subdivided into two groups, yielding four groups with 75 birds apiece, which served as the control (group CX), the Salmonella Gallinarum alone group (group CS), the FB1 alone group (group FX), and the group fed FB1 and infected with Salmonella Gallinarum (group FS). An oral challenge with Salmonella Gallinarum organisms (2 x 10(4) colony-forming units [cfu]/ml) was given to groups CS and FS at 21 days of age. Three quail each, were necropsied on day 21 (0 day interval) from groups CX and FX, whereas at subsequent intervals, i.e., 1, 2, 3, 5, 7, 10, 14, and 21 days postinfection (DPI), they were sacrificed from all four groups (CX, CS, FX, and FS) to study the agglutinin response to Salmonella Gallinarum and pathologic changes. The agglutinin titers to Salmonella Gallinarum in the combination group (FS) were generally lower when compared with those in group CS. A reduction in the size of spleen along with depletion of white pulp, thinning of cardiomyocytes, lymphoid cell depletion from bursal follicles, and renal tubular nephrosis were characteristic pathologic changes in group FX. In contrast, there was mild to severe enlargement of spleen accompanied by necrosis and reticuloendothelial cell hyperplasia, pericarditis, myocarditis, and focal interstitial nephritis in groups CS. Similar but more severe lesions were observed in the combination group (FS). In addition, the flabby texture of heart, hydropericardium, and ascites were mainly observed in group FS. It is concluded that continuous presence of fumonisins at 150 mg/kg diet increases the severity of Salmonella Gallinarum infection in young Japanese quail.

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.

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.

Widespread Occurrence of Ergot in Upper Midwestern U.S. Barley, 2005.

Ergot, caused by Claviceps purpurea (Fr.) Tul., occurs every year on cereals and grasses in North Dakota, but the occurrence on barley (Hordeum vugare L) is generally sporadic with a very low incidence of sclerotia. Disease surveys conducted during the 2005 growing season revealed an unusually widespread occurrence. This is of concern since barley production in North Dakota was estimated at 1.25 million metric tons, 27% of the total 2005 U.S. production. Barley samples (n = 304, ~0.50 kg) collected in all crop-reporting districts of North Dakota, northwestern Minnesota, and northeastern Montana, as part of an annual regional survey of barley crop quality (4), were examined for sclerotia. All barley samples were cleaned for dockage, and ergot (% [w/w]) was estimated on subsamples of ~100 g from a sample divider. Of all barley samples collected, 62% contained ergots. The regional average for ergot infested kernels was 0.06%, and samples ranged from <0.01 to 1.19%. Approximately 15% of all samples were in excess of 0.10% ergots and would have been downgraded to ergoty barley under the Official United States Standards for Grain. Occurrence of ergot was most common in northwestern Minnesota and the three eastern and north-central districts of North Dakota. Ergot was less frequent in the south-central and three western districts of North Dakota and was not detected in samples from northeastern Montana. Floret infection occurs during and up to 15 days after anthesis (2), and in the three eastern and north-central districts of North Dakota that occurred around the last week in June and first week in July. Between 22 June and 4 July, the North Dakota Agricultural Weather Network Stations in that region recorded average daily temperatures of 99% of the 30-year norm, but multiple precipitation events amounted to 227% of the 30-year norm. Rain splash and associated high relative humidity favor conidiation and spread of the fungus (1) and may have contributed to the high disease incidence. Average sclerotia weight for a sample ranged from <10 to 70 mg. However, large sclerotia (37 to 180 mg) often were removed by the no. 6 riddle of the dockage tester and were not counted in the ergot estimates as per U.S. Grading Standards. Samples containing 1.19, 0.81, 0.22, 0.14, 0.05, and 0.02% ergots were analyzed for ergopeptine alkaloids (3). These were found to contain 27.9, 25.4, 2.4, 1.1, 1.7, and 5.7 µg/g ergopeptine alkaloids, respectively. The average ratio of ergosine/ergotamine/ergocornine/ergocryptine/ergocristine was approximately 1:2:2:3:9. There also was widespread occurrence of the Fusarium mycotoxin deoxynivalenol (DON) on North Dakota barley in 2005. While there was no apparent relationship between the level of the DON and the amount of ergot in the samples (r = 0.042), more than 90% of samples with ergot had detectable levels (0.1 to 69 µg/g) of DON. While only DON is routinely measured in the crop survey (4), other tricothecenes and zearalenone have also been detected. This should be of concern to livestock producers and grain processors since the potential interactions of multiple mycotoxins are not well known. References: (1) G. M. Marshall. Ann. Appl. Biol. 48:19, 1960. (2) S. B. Puranik and D. E. Mathre. Phytopathology 61:1075, 1971. (3) G. E. Rottinghaus et al. J. Vet. Diag. Invest. 5:242 1993. (4) P. B. Schwarz et al. J. Am. Soc. Brew. Chem. 64:1, 2006.

Effects of feeding Fusarium verticillioides (formerly Fusarium moniliforme) culture material containing known levels of fumonisin B1 in Japanese quail (Coturnix coturnix japonica).

One hundred fifty 1-d-old quail chicks (Coturnix coturnix japonica) were divided into 2 groups. The 2 groups were designated as controls (CX) and fumonisin-fed birds (FX) with each containing 50 and 100 chicks, respectively. The birds in group CX were maintained on quail mash alone, whereas the birds in group FX were maintained on diets supplemented with 300 ppm of fumonisin B1 from Fusarium verticillioides (formerly Fusarium moniliforme) culture material from 1 d. Quail chicks in both groups were examined daily for clinical signs and mortality. Five randomly selected quail from each group were individually weighed on 0, 7, 14, 21, and 28 d post-feeding (DPF). After weighing, blood was collected from these birds at 7, 14, 21, and 28 DPF for hematological studies and at 14, 21, and 28 DPF for biochemical studies. Fumonisin B1-fed birds (FX) had ruffled feathers, reduced feed and water intake, poor body growth, and greenish mucus diarrhea with 59% mortality. Nearly 30% of the fumonisin B1-fed birds showed nervous signs during the 4-wk experimental period. From 7 DPF onward, BW in group FX were significantly lower than those in group CX. Fumonisin feeding significantly increased hemoglobin, packed cell volume, total erythrocyte count, and total leukocyte count. There was also a significant increase in aspartate transaminase and alanine transaminase in the fumonisin-fed group. Fumonisins significantly increased concentrations of total serum protein and albumin on 14 and 21 DPF, serum calcium and cholesterol levels from 14 DPF onward, and creatinine from 21 DPF onward. This study revealed that the addition of F. verticillioides culture material supplying a level of 300 ppm of FB1/kg of diet is highly toxic to quail chicks, resulting in heavy mortality, decreased growth rate, and significant alterations in hemato-biochemical parameters.

In vitro and in vivo assessment of humic acid as an aflatoxin binder in broiler chickens.

The in vitro affinity and adsorption capacity of a humic acid, oxihumate, for aflatoxin B1 (AFB1) was evaluated, utilizing Langmuir and Freundlich adsorption isotherms. Oxihumate showed a high in vitro affinity for AFB1. The Freundlich isotherm fitted the data better than the Langmuir isotherm, and binding capacities of 10.3, 7.4, and 11.9 mg of AFB1/g of oxihumate at pH 3, 5, and 7, respectively, were calculated. The in vivo efficacy of oxihumate as an aflatoxin binder in male broiler chickens exposed to aflatoxin-contaminated feed from 7 to 42 d of age was also assessed. The efficacy of oxihumate was compared with a commercially available product with a brewers dried yeast (BDY) and brewers fermentation solubles as main active ingredients. A total of 420 birds were assigned to 28 pens, with 15 birds per pen. The following treatments were applied: 1) 0 mg of AFB1 + 0 additives, 2) 1 mg of AFB1/kg of feed + 0 additives, 3) 1 mg of AFB1/kg of feed + 3.5 g of oxihumate/kg of feed, 4) 1 mg of AFB1/kg of feed + 3.5 g of BDY/kg of feed, 5) 2 mg of AFB1/kg of feed + 0 additives, 6) 2 mg of AFB1/kg of feed + 3.5 g of oxihumate/kg of feed, and 7) 2 mg of AFB1/kg of feed + 3.5 g of BDY/kg of feed. Each treatment consisted of 4 replicates. Oxihumate was effective in diminishing the adverse effects caused by aflatoxin on BW of broilers (P < 0.05). Oxihumate also showed protective effects against liver damage, stomach and heart enlargement, as well as some of the hematological and serum biochemical changes associated with aflatoxin toxicity (P < 0.05). Results indicated that oxihumate, but not BDY, could alleviate some of the toxic effects of aflatoxin in growing broilers. Oxihumate might, therefore, prove to be beneficial in the management of aflatoxin-contaminated feedstuffs for poultry when used in combination with other mycotoxin management practices. Additional studies are warranted to assess its efficacy under a wide variety of circumstances.