Ppb Aflatoxin Research Articles

Developing statistical models of aflatoxin risk in peanuts using historical weather data

AbstractAflatoxin contamination in peanuts (Arachis hypogaea L.) is a significant public health risk. Aflatoxin is detected postharvest after inspection of loads associated with grading at peanut buying points, leaving growers and shellers in a precarious position. Stricter limits on aflatoxin contamination could restrict the United States access to international markets. Predicting aflatoxin risk remains challenging, but improved tools could help inform postharvest storage segregation decisions and alert industry stakeholders to seasonal threats. This study aimed to develop and evaluate multiple statistical models that estimate the regional status of peanut aflatoxin contamination based on preharvest weather conditions. Our approach expanded on an existing peanut aflatoxin model for which a new geographic area and time period were tested. Weather variables served as independent variables to predict the risk of aflatoxin as the proportion of samples with greater than 20 ppb and 4 ppb aflatoxin (PGT20 [the proportion of samples with greater than 20 ppb aflatoxin] and PGT4 [the proportion of samples with greater than 4 ppb aflatoxin], respectively) across 10 counties in Georgia for 2018–2022. Best‐performing models were developed through multiple linear stepwise regression explaining more than 72% and 41% of the variability in PGT20 and PGT4, respectively. Model performance further varied whether it was a year of low or high aflatoxin incidence, with temperature observed as a key influencing factor across best‐performing models. This study established an adaptive approach to monitoring and managing aflatoxin risk through statistical predictive modeling, with output targeting farmers, industry, regulators, and public health officials. Future model development will aim to improve interpretation and confidence with in‐season aflatoxin prediction and efficacy testing of this approach across space and time.

Aflatoxins in Wheat Grains: Detection and Detoxification through Chemical, Physical, and Biological Means.

Wheat (Triticum aestivum L.) is an essential food crop in terms of consumption as well as production. Aflatoxin exposure has a widespread public health impact in economically developing nations, so there is a need to establish preventive techniques for these high-risk populations. Pre-harvest and post-harvest practices are the two strategies used to control aflatoxin contamination, which include the use of genetically modified crops that show resistance against Aspergillus infection, the use of pesticides, changing the planting and harvesting time of crops, and physical, chemical, and biological methods. In this research, aflatoxin detection and quantification were performed in different wheat varieties to determine quantitative differences in comparison to the European Commission's limit of 4 ppb aflatoxins in wheat. TLC for qualitative and the ELISA kit method for quantitative analysis of aflatoxins were used. Out of 56 samples, 35 were found contaminated with aflatoxins, while the remaining 21 samples did not show any presence of aflatoxins. Out of the 35 contaminated samples, 20 samples showed aflatoxin contamination within the permissible limit, while the remaining 15 samples showed aflatoxin concentration beyond the permissible level, ranging from 0.49 to 20.56 ppb. After quantification, the nine highly contaminated wheat samples were detoxified using physical, chemical, and biological methods. The efficiency of these methods was assessed, and they showed a significant reduction in aflatoxins of 53-72%, 79-88%, and 80-88%, respectively. In conclusion, the difference in aflatoxin concentration in different wheat varieties could be due to genetic variations. Furthermore, biological treatment could be the method of choice for detoxification of aflatoxins in wheat as it greatly reduced the aflatoxin concentration with no harmful effect on the quality of the grains.

Biochemical, Histopathological, and Immunological Effects of Mannan and/or Chitosan Oligosaccharides in Broilers intoxicated with Aflatoxins

Food additive such as carboxy methyl chitosan (CMC) and mannan (MOS) may have detoxification effect on aflatoxins (AFs). The current study aimed to investigate the role of chitosan and/or mannan in reducing the toxicity of AFs in broiler chickens (cobb500), through improving biochemical, hematological, oxidative stress, and antioxidant parameters. Furthermore, we investigated the impact on histopathological parameters and immune gene expression analysis (caspase 3 and interleukin 1-β). Five equal groups of 50 broilers each were selected (n=10); (G1) given the basal diet, (G2) basal diet intoxicated with 100 ppb AFs kg-1diet; (G3) given as basal diet intoxicated with 100 ppb AFs kg-1diet and supplemented with MAN (1 g kg-1 diet); (G4) given basal diet intoxicated with 100 ppb AFs kg-1diet and supplemented with CMC (0.5 g kg-1 diet) ; and (G5) given basal diet intoxicated with 100 ppb AFs kg-1diet and supplemented with MAN (1 g kg-1 diet)and CMC (0.5 g kg-1 diet). The current study's findings showed that the administration of both MAN and CMC brought hematological parameters back to the level of control. Chicks treated with MAN and/or CMC after being intoxicated with aflatoxins showed a considerable improvement in both renal and hepatic indicators with a notable improvement in antioxidant indicators in the groups receiving CMC and MAN supplements after being exposed to aflatoxicosis. Additionally, chicks treated with MAN and/or COS after being intoxicated with aflatoxins showed a considerable improvement in the gene expression of Caspase-3 and IL-1β. Furthermore, supplemental MAN and/or CMC decreased aflatoxin accumulation in the tissues of the muscles, liver, and kidneys. The groups G3, G4, and G5 showed notable improvements in the hepatic parenchymal architecture in their liver sections. The results of this study confirmed the hypothesis that adding natural additives such mannan and chitosan might enhance the antioxidant capacity and vital parameters of broilers exposed to 100 ppb aflatoxins.

The protective effect of Lactopluse-keragen Longlife® in Mitigating the Adverse Effects of Aflatoxin in Broiler Chickens

This study aimed to investigate the potential effects of growth promoter, namely "chelated minerals," on aflatoxin-induced clinical signs, post-mortem lesions, growth performance, mortality rates, hematological, biochemical and histopathological alterations in broiler chicks. A total of 150 one-day-old Cobb broiler chicks were divided into three groups and subjected to different dietary treatments for 39 days. Control group fed on basal diet without any medication. Aflatoxin premedicated group received basal diets with 10 ppb aflatoxins. Group 3 was fed 10 ppb aflatoxins with Lactopluse-keragen Longlife® (1g/kg).Growth performance. Hematological parameters, serum biochemistry, and histopathological lesions were evaluated. The results revealed that aflatoxin supplementation had detrimental effects on growth performance, hematological parameters, serum biochemistry, and histopathological lesions in various body organs. It also resulted in increased mortality rates. However, the concurrent administration of Lactopluse-keragen Longlife® supplements successfully alleviated the hazardous effects of aflatoxin. This growth promoter improved growth rates, as demonstrated by weight gain in the treated groups. In conclusion, the concomitant oral administration of Lactopluse-keragen Longlife® proved to be an effective strategy for mitigating the adverse effects of aflatoxin in chickens. This intervention improved growth performance and ameliorated the detrimental effects of aflatoxin exposure. Continued research in this area is essential to enhance our understanding and develop optimal preventive measures against aflatoxin contamination in poultry production.

Nursery pigs fed with feed contaminated by aflatoxin B1 (Aspergillus flavus) and anti-mycotoxin blend: Pathogenesis and negative impact on animal health and weight gain

The present study aimed to evaluate whether a moderate dose of aflatoxin B1 in pigs' diet interferes with pigs' growth and health in the nursery phase and whether an anti-mycotoxin mixture minimizes the adverse effects of the toxin. One blend with Saccharomyces cerevisiae lysate, zeolite, silicon dioxide, propylene glycol, Carduus marianus extract, soy lecithin, and carbonate was used as an anti-mycotoxin. Four treatments, with six repetitions per treatment and three pigs/pen: Afla0-AntiMyc0 - negative control (without aflatoxin); Afla500-AntiMyc0 - positive control (500 ppb of aflatoxin); Afla0-AntiMyc1000 - 1000 mg/kg of anti-mycotoxin blend; Afla500-AntiMyc1000 - 500 ppb aflatoxin +1000 mg/kg of anti-mycotoxin blend. It was observed that pigs in the positive control (Afla500-AntiMyc0) had lower body weight and weight gain when compared to the other treatments during the experimental period. Also, pigs from Afla500-AntiMyc0 had lower feed intake between days 1–20 and 1 to 30 than Afla0-AntiMyc0. The pigs from Afla500-AntiMyc0 had higher levels of liver enzymes aspartate aminotransferase and alanine aminotransferase compared to other treatments. The pigs from Afla500-AntiMyc0 had higher villus height than the other treatments, while the folded size was smaller in this treatment. Crypts were deeper in the intestines of pigs in both treatments that consumed aflatoxin. In general, it is concluded that the intake of aflatoxin B1 by nursery pigs has negative impacts on the health and, consequently, the animals' growth performance; however, the addition of the contaminated feed with an anti-mycotoxin blend was able to protect the pigs, minimizing the adverse effects caused by the mycotoxin.

Effect of Different Levels of Aflatoxin B1 in Concentrated Livestock Diets on Some Characteristics of Rumen Fluid in Laboratory with Sodium Bentonite

This study was conducted in the nutrition laboratory of the Department of Animal Production in the College of Agricultural Engineering Sciences at the University of Baghdad, Al-Jadriya. The experiment lasted for 90 days to investigate the effects of adding sodium bentonite (SB) to a concentrated diet contaminated with aflatoxin B1 on some characteristics of rumen fluid in a laboratory setting. Four diets were prepared, each contaminated with different concentrations of aflatoxin B1 (0, 20, 40, 60 ppb), and four different percentages of sodium bentonite (0, 3, 5, 7%) were added to each ration. The results showed a significant increase in pH values among the different treatments, with the diet contaminated with 40 ppb aflatoxin B1 (T3) recording the highest acidity compared to the control treatment at 0% concentration. However, there were no significant differences in pH rates between the treatments. The pH rates of 7% sodium bentonite addition did not differ significantly from 5% and 0%, while 3% had the lowest pH rate. Regarding volatile fatty acids (VFA), there was a significant increase in their percentage with the increase in sodium bentonite concentrations. The second treatment contaminated with 20 ppb aflatoxin B1 recorded the highest VFA percentage among the treatments, while the fourth treatment (T4) at 0% concentration recorded the lowest VFA percentage. Significant differences in VFA percentages were observed among all the treatments, with the highest VFA percentage in T2, followed by T3 and T1, while T4 recorded the lowest percentage. The analysis of ammonia nitrogen values showed a significant increase among the different treatments, with T2 recording the highest value among the treatments, and T4 at 0% concentration recording the lowest ammonia nitrogen value. Significant differences in the average percentage of ammonia nitrogen were observed among all the experimental treatments, with the highest value in T2, followed by T1 and T3, while T4 recorded the lowest percentage. The total number of microorganisms showed a significant increase with the increase in sodium bentonite concentrations, with T2 at 7% concentration having the highest number of microorganisms, while T4 at 0% concentration recorded the lowest number. Significant differences in the average total number of microorganisms were observed among all the experimental treatments, with T2 having the highest number, followed by T1 and T3, while T4 had the lowest number.

Effect of Nigella sativa Seed Supplementation on Hematology, Acid-base Parameters, and Serum Biochemical Parameters in Nubian Goat Fed an Aflatoxin Contaminated Diet

This study aimed to investigate the effect of Nigella sativa (NS) seed supplementation on hematology, acid-base parameters, and serum biochemical parameters in Nubian goats fed an Aflatoxin-contaminated diet. In a completely randomized design, 20 growing male goats (aged 8-9 months; 11±0.5 kg) were allocated to five treatments (4 goats/treatment). The control group (G1) received a basal diet. The treatment groups received the same diet contaminated with 150 ppb Aflatoxin (G2), and other treatments received an Aflatoxin-contaminated diet supplemented with different levels of crushed NS seeds 2% (G3), 4% (G4), and 6% (G5). Blood samples were collected after 40 day feeding period to determine blood pH, glucose, hematological and biochemical parameters. Statistical analysis was performed to assess the significant differences among the treatments. Hemoglobin concentration (Hb), total erythrocytes count (TEC), mean corpuscular hemoglobin (MCH), serum total protein (TP), and globulins (GB) were significantly (P≤0.05) decreased by Aflatoxin-contaminated diet, whereas total leukocytes count (TLC) increased (P≤0.05). Supplementing NS seeds to an Aflatoxin-contaminated diet significantly (P≤0.05) increased Hb, TEC, TP, and GB. Lipid profile and serum liver enzymes were significantly (P≤0.05) increased by an Aflatoxin-contaminated diet. Supplementing NS seeds to an Aflatoxin-contaminated diet caused a decrease (P≤0.05) in lipid profile and serum liver enzymes. Supplementing NS seeds to an Aflatoxin-contaminated diet resulted in a good performance and improved physiological status, the superior effect to an Aflatoxin-contaminated diet supplemented with 6% NS seeds. The study recommended supplementing 6% NS seeds to goat diets to reduce suspected Aflatoxin contamination. Further investigations are needed to assess the protective effect of NS seeds in other animal species fed on Aflatoxin-contaminated diets.

Effects of Nigella Sativa Seed on Rumen Parameters, Digestibility, and Nitrogen Balance of Nubian Goat Fed Aflatoxin Contaminated Diet

Introduction: Animal health and productivity can be affected by Aflatoxin in feeds, which leads to reduced growth, decrease feed efficiency and potential contamination of animal-derived products. The current study investigated the influences of supplementing Nigella sativa (NS) seeds to an Aflatoxin-contaminated diet on rumen parameters, digestibility, and nitrogen balance of male Nubian goats. Materials and methods: A total number of 20 healthy male Nubian goat kids aged 8-9 months and an average weight of 11 ± 0.5 kg were randomly assigned to 5 equal groups. The control group (T1) received a basal diet. The second treatment group (T2) received the same diet contaminated with 150 ppb Aflatoxin and other treatments received an aflatoxin-contaminated diet supplemented with different levels of crushed NS seeds including 2% (T3), 4% (T4), and 6% (T5) of the total dry matter. Through using naturally infected groundnut cake, Aflatoxin was mixed with other ingredients of ration in different proportions to formulate treated diets. High-performance liquid chromatography was used to determine Aflatoxin concentration. Rumen liquor samples were collected after 40 days feeding period to evaluate rumen pH, volatile fatty acids (VFAs), ammonia nitrogen (NH3-N), rumen electrolytes, and protozoa count. Results: Rumen pH, ammonia nitrogen, ruminal sodium, and potassium were not significantly affected by the treatment groups. Rumen volatile fatty acids, protozoa count, nutrient digestibility, and nitrogen retention were significantly reduced by an Aflatoxin-contaminated diet (T2) compared to other groups. Supplementation of NS seeds to an Aflatoxin-contaminated diet caused a significant increase in VFAs, decreased protozoa count, and improved digestibility and nitrogen retention in all treated groups compared to T2 and T1 groups. Conclusion: Supplementing NS seeds to an Aflatoxin-contaminated diet positively affected rumen ecology and improved digestibility and nitrogen retention. The study recommended supplementing 6% of NS seeds to goat diets can reduce the negative effects of Aflatoxin in the diet.

A balancing act: Synthetic oversampling corrects skewed distribution of aflatoxin in maize and improves prediction of contaminated kernels

Current spectral models for detecting aflatoxin (AF) in maize kernels are limited by very low representation of kernels with AF levels above the models’ specified contamination thresholds. With the aim of improving prediction model sensitivity to AF-contaminated kernels, two methods were evaluated for artificially enriching the representation of kernels containing ≥150 parts per billion (ppb) AF in a large spectral dataset: humid incubation and synthetic oversampling. The addition of synthetic training samples improved prediction accuracy of kernels containing ≥150 ppb AF from 51% to 80%. Humid incubation contributed additional kernels with intermediate levels of AF contamination (primarily 5–75 ppb), but did not change the overall distribution of AF contamination. Feature importance distributions overlapped among models at 329–345 nm, 380–385.5 nm, 415–425 nm, 639–668 nm, and 1013.5–1060 nm. These spectral ranges could be applicable to the development of limited wavelength grain sorting devices for low-resource settings.

Mycotoxins associated with maize wastes treated with comprised capsule of Spirulina platensis biomass

Spirulina platensis has roles in biotechnological product advancement due to its safety for humans and animals. Analysis of S. platensis extract by high-performance liquid chromatography reflected the presence of 14 phenolic and flavonoid compounds with different concentrations such as ellagic acid (333 µg/g), gallic acid (294 µg/g), methyl gallate (147 µg/g), naringenin (144 µg/g), and chlorogenic acid (142 µg/g). The isolated Aspergillus flavus from silage of maize was tested to evaluate the effect of S. platensis extract on aflatoxins B1, B2, G1, and G2 production. The treated silage with S. platensis extract showed the presence of 37±0.33 ppb, 0.59±0.16 ppb, 0.26±0.22 ppb, and 0.21±0.18 ppb compared to un-treated silage that showed 3.14±0.15 ppb, 0.81±0.08 ppb, 0.46±0.05 ppb, and 0.26±0.23 ppb aflatoxins of B1, B2, G1, and G2. These results were shown on the 10th day of incubation. On the 15th day of incubation, the treated silage showed less mycotoxins than un-treated silage. At different incubation periods, glucosamine was estimated as a growth development biomarker. The content of glucosamine was inhibited as a result of the effect of S. platensis extract on fungus growth with 53.71%, 49.19%, 47.84%, 38.47%, and 35.72% inhibition on the 3rd, 6th, 9th, 12th, and 15th day.

Spectral kernel sorting based on high-risk visual features associated with mycotoxin contamination reduces aflatoxin and fumonisin contamination in maize from Ghana

Rapid single kernel analysis could enable physical sorting to remove mycotoxins from bulk grains. The purpose of this study was to use visual characteristics previously associated with aflatoxin and fumonisin contamination of maize kernels to calibrate a multi-spectral sorter and then sort mycotoxin contaminated lots. A total of 76 corn samples were collected from poultry farmers in the Dorma-Ahenkro area, Ghana. Paired 400-g subsamples were used for bulk analysis and single kernel sorting. Individual kernels were selected from contaminated samples by 2 levels of stratification: visible high-risk kernels (n = 1000) and visible low risk kernels (n = 1000). High-risk kernels had one or more of 3 features: fluorescence under UV (366 nm) light, mold, or brokenness. Kernels were used to calibrate a multi-spectral sorter (individual wavelengths from 470 to 1070 nm) to remove high-risk kernels. Then, kernel samples were sorted and the reject and accept streams individually ground and tested for aflatoxin and fumonisin contamination using ELISA. Bulk sample levels ranged between 0.78 and 67 ppb aflatoxin and <2.5 × 10−3 – 5.7 ppm fumonisin. Classification algorithms to reject visible high-risk spectra were 63% sensitive and 90% specific. After sorting, samples showed a significant aflatoxin reduction (p < 0.001, 73/76 samples reduced, mean reduction 31 ppb, range −9.7 – 67 ppb); all samples showed a significant fumonisin reduction (p < 0.001, mean reduction 1.9 ppm, range 9.3 × 10−2 – 6.1 ppm). From the accepted stream, 61/76 samples tested <15 ppb aflatoxin, significantly more than the 40/76 prior to sorting (p < 0.001); all accepted samples tested <2 ppm fumonisin concentration compared to only 2/76 prior to sorting. From sorting, average mass rejected was 12% (range 1.2%–36%), and that rejected mass contained an average of 46% of the total aflatoxin (range 4.3%–97%) and 88% of the total fumonisin (range 10%–84%). Visual characteristics associated with mycotoxin contamination can inform classification models which can enable sorting contaminated maize to reduce aflatoxin and fumonisin contamination.

Potential and challenge assessment of tannin extracts from black tea in male rabbits fed contaminated diet by Mycotoxins

Abstract The focus of this research was to investigate how tannin extract from black tea influenced the hematological and histopathological evaluations of male rabbits fed a mycotoxin-contaminated feed, including 17 ppb aflatoxin, 5 ppb ochratoxin and 2 ppb fumonisin. A total of 28 local male rabbits were allocated into four groups, the first of which was a control group. The second was fed a Mycotoxin-contaminated meal. The third was given a Mycotoxin feeding with tannin extracts (125mg/mL per head) administered orally, while the fourth was fed a Mycotoxin contaminated diet with tannin extracts (250 mg/mL per head) given orally. The findings revealed that tannin extracts seemed to have a significant positive impact on haematological results, particularly RBCs and WBCs, throughout the period; however, the 250 mg/mL dose showed no significant differences in Hb levels. Nonetheless, as compared to the Mycotoxin group, the histopathological sections of tannin extract demonstrated a reduction in the toxicity of the Mycotoxin diet on the liver and kidney tissues; conversely, the tissue sections of the control animals showed no abnormalities. In male rabbits fed contaminated Mycotoxin diets, tannin extract from black tea was found to have a favourable impact on hematological activities and vital organs such as the liver and kidney.

Efficacy of sodium bentonite (SB) to ameliorate adverse effects of aflatoxin on In Vitro rumen fermentation of wheat straw

The present experiment was conducted to study the effect of sodium bentonite (SB) in ameliorating the adverseeffects of aflatoxin on in vitro rumen fermentation of wheat straw. Five treatment groups, viz. T1: control (wheatstraw); T2: T1 + 300 ppb aflatoxin B1 (AFB1); T3: T2 + 0.33% SB; T4: T2 + 0.5% SB and T5: T2 + 1.0% SB wereprepared and incubated in vitro. Truly degradable dry matter (TDDM), truly degradable organic matter (TDOM),gas production (GP), microbial biomass production (MBP) and partitioning factor (PF) values in T1 were higherthan those of other treatment groups (T2 to T5). TDDM, TDOM, GP, MBP and PF values in T2 were lower thanthose of other treatment groups. The values of these parameters improved with increasing concentration of SB.Total volatile fatty acids (TVFA), acetate (A), propionate (P) and butyrate (B) values in T1 were higher than thoseof other treatment groups (T2 to T5). TVFA and A value between T2 and T3 were statistically similar. TVFA, A, Pand B values improved with increasing level of SB, however, these values were lower than that of control even athighest inclusion level of SB. A:P ratio among various treatments did not vary significantly. It was concluded thataflatoxin contamination of feed at 300 ppb level significantly affected the in vitro rumen fermentation in terms ofreduced truly degradable dry matter, truly degradable organic matter, gas production, microbial biomass production, partitioning factor and total volatile fatty acids concentration. Incorporation of sodium bentonite up to 1.0% level to the aflatoxin-contaminated wheat straw, partially ameliorated the adverse effects of aflatoxin on in vitro rumen fermentation parameters.

English

The performance of twenty four 6-8 week old apparently healthy male and female weanling rabbits of mixed breed fed diets treated with aflatoxin, which contained 5% sweet orange (Citrus sinensis) fruit peel meal was evaluated in an 8 week feeding trial. Sweet orange peels were collected from orange sellers, sun-dried and milled. Fungal strain of Aspergillus flavus was cultured and inoculated into groundnut cake to produce aflatoxin using solid state fermentation method. Treated groundnut cake was incubated for seven days with incremental incubation temperature from 20-25 &ordm;C The groundnut cake was autoclaved, milled and, aflatoxin extracted from 10 g sample of the milled cake with 50 ml chloroform, and its concentration quantified by Thin Layer Chromatography (TLC). Treated groundnut cake was included at 0, 50, 100 and 150 gram in grower rabbit diets to produce diets T1, T2, T3 and T4, having 0 ppb, 50 ppb, 100 ppb and 150 ppb aflatoxin, respectively. The rabbits were randomly allocated to four diets at the rate of six per diet, housed singly in rabbit hutches, fed and served water free choice. The result showed significant (P&lt;0.05) negative effect&nbsp; of diets on final live weight, weight gain, feed intake, feed conversion ratio, water consumption, protein intake and protein efficiency ratio as the dietary aflatoxin increased from 0 ppb to 150 ppb. Also, diets had significant (P&lt;0.05) negative effect on dressed weight and carcass length and, kidney. Total aflatoxin residue varied significantly (P&lt;0.05) from 0 &mu;g/kg - 2.76&mu;g/kg,&nbsp; 0 &mu;g/kg - 1.94&mu;g/kg and 0 &mu;g/kg - 0.85&mu;g/kg for liver, kidney and meat tissue, respectively as the dietary aflatoxin increased from 0 ppb to 150 ppb. Performance response of rabbits was affected negatively by aflatoxin, thereby showing the inability of 5% dietary inclusion of sweet orange (Citrus sinensis) peel meal to mitigate the adverse consequences of aflatoxin intake by rabbit. Key words: Growth responses, residual level, visceral organs, mitigate.

Long-term Feeding Effects of Calcium-bentonite Clay in Aflatoxin B1 Contaminated Diet on Egg Production, Egg Quality and Immune Status of Laying Hens

Background: This experiment was conducted to determine the effect of synthetic aflatoxin B1 and calcium bentonite clay as a mycotoxin binder in the laying hen diet on performance, egg quality and immune response to vaccination. Methods: A total of 144 Lohmann brown classic hens (22 weeks old) were divided into 4 groups of 6 replications with 6 hens each. This experimental design was completely randomized design and there were 4 experimental diets: (1) Basal diet (Positive control), (2) Basal diet +0.25% calcium bentonite clay, (3) Basal diet+200 ppb aflatoxin B1 (negative control) and (4) Basal diet+200 ppb aflatoxin B1 and +0.25% calcium bentonite clay. All hens were vaccinated with Newcastle (ND) and Infectious bronchitis (IB) vaccines during 24 weeks feeding trial. Result: At the end of the 24 weeks of feeding trial, dietary treatments did not influence the egg production, egg mass, feed conversion ratio (FCR) and IB titer of hens; however, aflatoxin B1 significantly (P less than 0.05) deceased final body weight and deceased the eggshell color score (P less than 0.05). Moreover, the ND titer was reduced by aflatoxin B1 at 8th and 12th weeks of trial (P=0.05 and less than 0.05; respectively). Thus, supplementation of 0.25% calcium bentonite clay in feed improved body weight, eggshell color and ND titer of laying hens similar to those fed the positive control diet.

Single kernel aflatoxin and fumonisin contamination distribution and spectral classification in commercial corn

Aflatoxin and fumonisin contamination distribution in corn is non-homogeneous. Therefore, bulk sample testing may not accurately represent the levels of contamination. Single kernel analysis could provide a solution to these problems and lead to remediation strategies such as sorting. Our study uses extensive single kernel aflatoxin (AF) and fumonisin (FM) measurements to (i) demonstrate skewness, calculate weighted sums of toxin contamination for a sample, and compare those values to bulk measurements, and (ii) improve single kernel classification algorithm performance. Corn kernels with natural contamination of aflatoxin and fumonisin (n = 864, from 9 bulk samples) were scanned individually twice for reflectance between the ultraviolet–visible–near infrared spectrum (304 nm–1086 nm), then ground and measured for aflatoxin and fumonisin using ELISA. Single kernel contamination distribution was non-homogeneous with 1.0% (n = 7) of kernels with ≥20 ppb aflatoxin (range 0 - 4.2×105 ppb), and 5.0% (n = 45) kernels with ≥2 ppm fumonisin (range 0 - 7.0×102 ppm). A single kernel weighted sum was calculated and compared to bulk measurements. Average difference in mycotoxin levels (AF = 0.0 log(ppb), FM = 0.0 log(ppm), weighted sum – measured bulk levels) calculated no systematic bias between the two methods, though with considerable range of −1.4 to 0.7 log(ppb) for AF and −0.6 to 0.8 log(ppm) for FM. Algorithms were trained on 70% of the kernels to classify aflatoxin (≥20ppb) and fumonisin (≥2ppm), while the remaining 30% of kernels were used for testing. For aflatoxin, the best performing algorithm was stochastic gradient boosting model with an accuracy of 0.83 (Sensitivity (Sn) = 0.75, Specificity (Sp) = 0.83), for both training and testing set. For fumonisin, the penalized discriminant analysis outperformed the rest of the algorithms, with a training accuracy of 0.89 (Sn = 0.87, Sp = 0.88), and testing accuracy of 0.86 (Sn = 0.78, Sp = 0.87). The present study improves the foundations for single kernel classification of aflatoxin and fumonisin in corn, and can be applied to high throughput screening. This study demonstrates the heterogeneous distribution of aflatoxin and fumonisin contamination at single kernel level, comparing bulk levels calculated from those data to traditional bulk tests, and utilizing a UV–Vis–NIR spectroscopy system to classify single corn kernels by aflatoxin and fumonisin level.

Interaction effects of sunflower oil and aflatoxin at graded levels in diet on performance, serum and tissue biochemical profile, organ weights and immuneresponse in broiler chicken.

The dietary supplementation of fat has great potential in countering the toxic effects of aflatoxin (AF) in chickens, but the issue was less researched upon. An experiment was conducted to evaluate the response of broiler chickens to graded levels of AF B1 (0, 150 and 300 ppb) and sunflower oil (SFO) (0, 1.5 and 3.0%) in the diet in a 3 × 3 factorial manner to understand their interaction effects. A total of 360 broiler chickens divided into 9 equal groups were fed the diets during 0 to 35 days of age, and their response was evaluated in terms of performance, serum biochemical profile, organ weights, liver fat content and bone mineralization. Sunflower oil at 1.5% in diet countered (P ≤ 0.01) the adverse effects of 150 ppb AF on body weight, whereas at 300 ppb AF, such a response was seen at the higher level (3%) of SFO. Aflatoxin decreased (P ≤ 0.01) feed intake by 4 and 11% at 150 and 300 ppb concentration, respectively at 35 days of age, which was increased (P ≤ 0.01) with each incremental level of SFO supplementation (by 3.0 and 8.8%, respectively at 1.5 and 3%, respectively). Serum protein concentration increased (P ≤ 0.01) by SFO supplementation only at the higher concentration (300 ppb) of AF (by 42.4%), whereas total cholesterol and triglyceride concentration, and immune response to SRBC inoculation increased (P ≤ 0.01) with SFO at either level of AF (by 16.8, 18.7 and 75.6% at 1.5% SFO and 33.1, 36.9 and 94.2% at 3.0% SFO, respectively at 35 days of age). Weights of the liver, giblets, kidneys and pancreas increased (P ≤ 0.01) by 23.2, 14.7, 34.2 and 16.9%, respectively, and thymus weight decreased (P ≤ 0.04) by 25.4% with 300 ppb AF, and SFO at 3% in diet countered the effect on weight of the liver and giblets. Fat deposition in the liver increased (P ≤ 0.01) as the concentration of AF increased in diet (by 9.4 and 17.3%, respectively at 150 and 300 ppb AF), which was significantly (P ≤ 0.05) countered by SFO at 3% in diet. Tibia bone Ca content increased by 2.4% (P ≤ 0.01) with SFO supplementation in AF-fed chickens. It is concluded that dietary SFO supplementation countered the adverse effects of AF in broiler chicks in a dose-dependent manner, and higher level of oil (3% in diet) was required at the higher concentration of AF (300 ppb) in diet.

Evaluation of the Impact of Skewness, Clustering, and Probe Sampling Plan on Aflatoxin Detection in Corn.

Probe sampling plans for aflatoxin in corn attempt to reliably estimate concentrations in bulk corn given complications like skewed contamination distribution and hotspots. To evaluate and improve sampling plans, three sampling strategies (simple random sampling, stratified random sampling, systematic sampling with U.S. GIPSA sampling schemes), three numbers of probes (5, 10, 100, the last a proxy for autosampling), four clustering levels (1, 10, 100, 1,000 kernels/cluster source), and six aflatoxin concentrations (5, 10, 20, 40, 80, 100 ppb) were assessed by Monte‐Carlo simulation. Aflatoxin distribution was approximated by PERT and Gamma distributions of experimental aflatoxin data for uncontaminated and naturally contaminated single kernels. The model was validated against published data repeatedly sampling 18 grain lots contaminated with 5.8–680 ppb aflatoxin. All empirical acceptance probabilities fell within the range of simulated acceptance probabilities. Sensitivity analysis with partial rank correlation coefficients found acceptance probability more sensitive to aflatoxin concentration (−0.87) and clustering level (0.28) than number of probes (−0.09) and sampling strategy (0.04). Comparison of operating characteristic curves indicate all sampling strategies have similar average performance at the 20 ppb threshold (0.8–3.5% absolute marginal change), but systematic sampling has larger variability at clustering levels above 100. Taking extra probes improves detection (1.8% increase in absolute marginal change) when aflatoxin is spatially clustered at 1,000 kernels/cluster, but not when contaminated grains are homogenously distributed. Therefore, taking many small samples, for example, autosampling, may increase sampling plan reliability. The simulation is provided as an R Shiny web app for stakeholder use evaluating grain sampling plans.

The effectiveness of Saccharomyces cerevisiae in inhibiting aflatoxin production and reducing the harmful effects of aflatoxin in ducklings

Sacharomyces cerevisiae isolates were obtained from baker's yeast, soil, fruit and identified with PCR. Twenty-seven isolates of S. cerevisiae were screened for capacity of inhibition of aflatoxin production of Aspergillus flavus (A. flavus) on coconut extract agar media (CEA). The results showed that the coculture method of S. cerevisiae isolates and aflatoxin producing A. flavus on CEA medium could be used for screening the strains that are able to antagonize aflatoxin-producing A. flavus. On ground corn medium, with the rate of 104 aflatoxin-producing A. flavus spores and 108 S. cerevisiae yeast cells/g, S. cerevisiae was able to reduce the amount of aflatoxin produced by A. flavus in corn. In an in vivo experiment, it was found that addition of 108 cells of S. cerevisiae to one kg of duck feed contaminated with 300 ppb aflatoxin from 1 to 10 days of age reduced adverse effects of aflatoxin on the liver and kidneys of ducks

Aflatoxicosis in fingerling common carp (Cyprinus carpio) and protective effect of rosemary and thyme powder: Growth performance and digestive status

Dietary aflatoxin B1 contamination is a demanding challenge for aquaculture industry. The present study was conducted to elucidate the effect of dietary aflatoxin B1 contamination on growth and digestive status of juvenile common carp. The effect of dietary inclusion of rosemary and thyme to prevent such effects was also investigated. Fingerlings were subjected to seven treatments including various dietary aflatoxin B1 contamination (0, 50, 100, 200, 300 and 400 ppb) and a group received diet contained 400 ppb aflatoxin B1 and 4% rosemary and thyme powder. The experiment lasted for 12 weeks. Results revealed that dietary aflatoxin B1 contamination did not affect growth performance. However, intestinal alkaline protease, lipase and amylase activity of fish were increased according to aflatoxin B1 diet levels. In addition, intestine tissue lesions including necrosis, immune cell infiltration and fibroplasia were manifested with increasing dietary aflatoxin B1 content. Digestive enzymes activity was ameliorated by dietary incorporation of the plant powder in those fish received diet with 400 ppb aflatoxin B1, but it did not prevent intestinal lesions. In conclusion, juvenile common carp showed no changes regarding various growth indices even at 400 ppb aflatoxin B1 level. Although, 50 ppb dietary aflatoxin B1 did not significantly affect growth or digestive enzymes activity of juveniles, it caused mild intestinal tissue deteriorative changes. Including the medicinal powder blend in diet with 400 ppb aflatoxin B1 could ameliorate the increased digestive enzymes activity; however, it did not prevent the intestinal tissue pathological changes. Understanding the exact mechanisms behind such protective effect requires further studies.