Mycotoxins In Feed Research Articles

Mycotoxins in grains and feed - contamination and toxic effect in animals

Mycotoxins are secondary metabolites of moulds that represent a significant food safety issue and pose a risk to health and wellbeing of humans and animals, having a negative impact on economies all over the world. They can cause intoxication in animals and affect the less productive performance and nutritional value of animal feed. Also, mycotoxins may contaminate different types of food through directly and indirectly way and cause negative health effects in humans. To estimate the potential of their harmful effect it is necessary to know the terms of mycotoxins occurrence and frequency, toxicity and biotransformation in different animal species and humans, possibility of contamination prevention, and their inactivation or reduction in the case when the incidence is inevitable. The relatively small number of comparable data on the incidence of mycotoxins in feed from European countries show the necessity of systematic control and the application of validated analytical methods in their determination. This paper gives the overview of incidence of grains and feed contamination with most important and common mycotoxins and the evidenced toxic effects in animals.

Occurrence, Prevention and Limitation of Mycotoxins in Feeds

There has been a significant concern regarding the potential health risks for humans and animals via foods and feeds that are contaminated with different agents. Particularly, mycotoxin contamination is of great importance as it is widespread and unpreventable. In both foods and feeds, molds produce secondary metabolites called mycotoxins; these are produced generally after the fungi reach their maturity. Depending on the definition used, hundreds of fungal compounds are recognized as mycotoxins. However, the attention is mainly focused on aflatoxins, ochratoxins, fumunisins, and zearalenone which are considered the most important threats for human and animal health. Mycotoxin contamination causes a fundamental problem all over the world including developed countries. Additionally, the economic impact of mycotoxins is another global concern on the agricultural markets. These concerns are based on toxicological data, which show that naturally occurring levels of mycotoxins have adverse effects in farm and laboratory animals as well as humans. The diversity of mycotoxin structures induces various toxic effects. Owing to the significant health risks and economic impacts, considerable investigations are being performed to diminish their harmful effects and to prevent their formation. In order to limit their levels, much research has been focused on detecting the mycotoxins in contaminated food and feedstuffs. This review will focus on information about primary mycotoxins, their occurrence, related regulations, prevention and methods of detection within the light of the current literature.

Natural occurrence of emerging Fusarium mycotoxins in feed and fish from aquaculture.

A new analytical method for the simultaneous determination of enniatins (ENs) and beauvericin (BEA) in fish feed and fish tissues by liquid chromatography coupled to mass spectrometry with linear ion trap (LC-MS/MS-LIT) was developed. Results showed that the developed method is precise and sensitive. The presence of emerging Fusarium mycotoxins, ENs and BEA, was determined in samples of aquaculture fish and feed for farmed fish, showing that all feed samples analyzed were contaminated with mycotoxins, with 100% coexistence. In aquacultured fish samples, the highest incidence was found in edible muscle and liver. As for the exposure assessment calculated, it was found that average consumer intake was lower than tolerable daily intake (TDI) values for other Fusarium mycotoxins.

Evaluation of β-d-Glucan Biopolymer as a Novel Mycotoxin Binder for Fumonisin and Deoxynivalenol in Soybean Feed

The walls of yeast cells, which contain β-D-glucan biopolymers, have an active role in reducing mycotoxins in animal feed. This study aimed to evaluate the β-D-glucan biopolymers as a mycotoxin binder for fumonisin (FUM) and deoxynivalenol (DON) toxins as well as their effect on the nutritional value of soybean, which is considered one of the important feed row materials. The evaluation was carried out using some toxigenic Fusarium isolates (Fusarium solani, F. oxysporum, and F. verticillioides) in vitro and in vivo. The FUM and DON levels were determined by immune affinity column. The F. verticillioides was the most toxigenic, followed by F. oxysporum and lastly F. solani, while secretion of DON toxin was determined to be greater than FUM with all the tested fungi. The effectiveness of β-D-glucan biopolymers on FUM and DON absorption was greater than clay and calcium propionate. In vivo, treating soybean seeds with β-D-glucan biopolymers led to reduction in the level of FUM and DON toxins in seeds artificially inoculated by F. verticillioides. β-D-glucan treatment also has a low effect on nutritional components of the seeds compared to untreated ones. In conclusion, this study found a new approach to reduce Fusarium mycotoxins in feed to an allowable safe limit and at the same time maintaining the nutritional value of these materials.

Mycotoxin occurrence in feed and feed raw materials worldwide: long-term analysis with special focus on Europe and Asia

During an 8-year period, 17 316 samples of feed and feed raw materials from all over the world were analysed for contamination with aflatoxins, ochratoxin A, zearalenone, deoxynivalenol and fumonisins. Overall, 72% of the samples tested positive for at least one mycotoxin and 38% were found to be co-contaminated. Mycotoxin concentrations were generally low and the majority of the samples were compliant with the most stringent EU guidance values or maximum levels for mycotoxins in feed. However, in their present state these regulations do not address co-contamination and associated risks. Long-term trends are difficult to establish as strong yearly variations were observed regarding mycotoxin prevalence and contamination levels. In some cases unusual weather conditions can be linked with high observed mycotoxin loads. An exception to this rule is South-East Asia, where a steady increase of aflatoxin prevalence has been observed. The percentage of aflatoxin-positive samples in this region rose from 32% in 2005 to 71% in 2011.

Fungi and mycotoxins in silage: an overview

The present revision shows the early and current knowledge in the field of silage fungi and mycotoxins explaining the relevance of fungi and mycotoxins in silage. The problem does not end in animal disease or production losses as mycotoxins in feed can lead to the presence of their metabolic products in dairy products, which will be eventually affecting human health, mainly infants. Silage is green forage preserved by lactic fermentation under anaerobic conditions. This ecosystem maintains its quality and nutritional value depending on interactions among physical, chemical and biological agents. Forages used for ensilage are naturally in contact with yeasts and filamentous fungi, and the contamination often occurs in the field and can also occur during harvesting, transport, storage. Moreover, postharvest poor management can lead to a rapid spoilage. Studies on fungal contamination of dairy cattle feed have shown how corn silage influences the contamination degree of feed supplied to livestock. Increasing knowledge in this area will help elucidate the influence that this microbiota exerts on production and/or degradation of mycotoxins present in silage. Some of these fungi, although opportunist pathogens, are relevant epidemiologically and represent a high risk of contamination to farm workers who handle them improperly.

Influence of mycotoxin zearalenone on the swine reproductive failure

Reproductive failure in swine is often a difficult diagnostic problem. If diagnoses of infectious disease or management related problems are not obtained, feed quality and safety may be questioned. Mycotoxins are often present in swine feed in the amount that can have detrimental impact on production and reproduction. Problems are expressed only as alterations of the reproductive cycle, reduced feed intake, slow growth or impaired feed efficiency. In Serbia, generally speaking, high concentrations of mycotoxins were noticed, especially mycotoxin zearalenone. High presence of zearalenone in swine feed is probably due to climatic influence and should be monitored constantly. This paper includes field observations regarding the influence of moldy feed containing mycotoxin zearalenone on the occurrence of the reproductive failure in swine breeding categories (sows, gilts and boars). The material for this research was obtained from four swine farms where certain reproductive disorders and health problems in breeding animals were detected. Depending on the specificity of each evaluated case and available material, the applied research methods included: anamnestic and clinical evaluation, pathomorphological examination, standard laboratory testing for detection of aerobic and anaerobic bacteria, and microbiological feed testing, in order to examine the presence of fungi and mycotoxins by applying the method of thin layer chromatography. On the basis of the obtained results, it could be concluded that mycotoxin zearalenone was detected in all examined feed samples. The presence of mycotoxin in feed was directly related to the reproductive failures in the examined swine categories (vulvovaginitis, endometritis, rebreeding, infertility). Swine reproduction represents the base for intensive swine production. The presence of mycotoxins in swine feed have influence on the reproduction and health status of pigs and under certain conditions may significantly disturb the production process.

Simultaneous multiple mycotoxin quantification in feed samples using three isotopically labeled internal standards applied for isotopic dilution and data normalization through ultra‐performance liquid chromatography/electrospray ionization tandem mass spectrometry

Mycotoxins are typically present in grain and are also concentrated in distillers dried grains with solubles (DDGS), common feed ingredients for food animals. The diversity of mycotoxins and feed matrices has made the routine detection and quantification of mycotoxins in feed both complex and prohibitively expensive. Ultra-performance liquid chromatography/electrospray ionization triple quadrupole detection (UPLC/ESI-TQD) (tandem mass spectrometry, MS/MS) with (13) C-labeled isotopic dilution was used to analyze internal standard isotopologues of three mycotoxin molecules, as well as 29 other structurally differing mycotoxin molecules from four common feed matrices: corn, wheat, barley, or DDGS. Mycotoxins were extracted via a single-step procedure using a mixture of acetonitrile/water/formic acid. Labeled isotopologues were used as a surrogate to account for extraction quality and as internal standards for the evaluation of the feed matrix signal suppression/enhancement (SSE) contributed by each mycotoxin and by each matrix. The SSE was corrected by matrix-matched calibration with blank certified reference feed material. The limits of detection for individual mycotoxins in buffer ranged from 0.01 to 206.7 µg/mL but could increase by up to four times depending on the matrix effect. The accuracy and precision were enhanced by the use of isotopically labeled standards. The recoveries were somewhat negatively affected by the SSE contributed by each matrix. Each mycotoxin was successfully detected and assigned to one of four SSE categories: high (-66%), intermediate (-48%), low (-19%) signal suppression and signal enhancement (> +300%). An improved LC/MS method was validated, which offers a practical and economical means for large-scale detection and quantification of multiple mycotoxins in common animal-feed matrices, including DDGS.

Evaluation of the effect of mycotoxin binders in animal feed on the analytical performance of standardised methods for the determination of mycotoxins in feed

Recently, the use of substances that can suppress or reduce absorption, promote the excretion of mycotoxins or modify their mode of action in feed, so-called mycotoxin binders, has been officially allowed in the European Union as technological feed additives. The influence of the addition of mycotoxin binders to animal feed on the analytical performance of the official methods for the determination of mycotoxins was studied and the results are presented. Where possible standardised methods for analysis were applied. Samples of 20 commercial mycotoxin binders were collected from various companies. The following mycotoxins were included in the study: aflatoxin B1, deoxynivalenol, zearalenone, ochratoxin A, fumonisins B1 and B2, T-2 and HT-2 toxins. A binder (or binders combined in a group) was mixed with feed material containing the mycotoxin, and the feed material was analysed. For data evaluation, the mean values were compared by Student's t-test (an independent two-sample t-test with unequal sample sizes and equal variance). The repeatability standard deviation of each method was used as an estimate of method variability. No significant differences (p = 0.05) in mycotoxin levels between binder-free material and the material containing different binders were found. Further, the possible effects of binder addition in combination with processing (pelletising) on the amount of aflatoxin B1 determined in feed were studied. Three commercial mycotoxin binders containing hydrated sodium calcium aluminosilicate (HSCAS) as the main component were used in these experiments. Feed samples with and without mycotoxin binders were pelletised with and without steam treatment. After pelletising, materials were analysed for AFB1. Only the combination pelletising and a mixture of binders added at a total level of 1.2% had a significant effect (41% reduction) on the amount of AFB1 determined.

Porcine ear necrosis syndrome: A preliminary investigation of putative infectious agents in piglets and mycotoxins in feed

The aim of this study was to identify the causative factors of porcine ear necrosis syndrome (PENS) in 72 pigs, 5.5-10 weeks in age housed on nine farms. Biopsy samples of ear pinnae were collected from all piglets for bacteriology, histopathology and in situ hybridization for porcine circovirus type 2 (PCV2). At the same time, serum samples were taken for serological analysis and viral PCR, and feed was sampled for mycotoxin analysis. The initial lesion of PENS seemed to be a focal epidermal necrosis. Streptococci were isolated from 44 and staphylococci from 36 pinnae. PCV2 could not be detected by in situ hybridization or qPCR. Seven piglets were positive for porcine reproductive and respiratory syndrome virus, and one for Mycoplasma suis. One piglet had antibodies against Sarcoptes scabiei var. suis. No infectious agents were found in 15 samples. Positive virology and parasitology were often found alongside positive bacteriology. Deoxynivalenol, zearalenone and ergot alkaloids were detected in feed. The findings suggest that PENS is multifactorial in origin and that although infectious agents can be involved in the development of the syndrome they are not the exclusive triggering factor.

Mycotoxins in broilers: pathological alterations induced by aflatoxins and ochratoxins, diagnosis and determination, treatment and control of mycotoxicosis

Mycotoxins are secondary metabolites produced by fungi. Mycotoxins in feed are considered the second most serious issue in the poultry industry, following increased feed prices. There are different types of mycotoxins, including aflatoxins, ochratoxins, fumonisin, trichothecenes, zearalenone, patulin, but the most important are aflatoxins and ochratoxins. Ochratoxin is known as a nephrotoxin as it damages kidney tissues predominantly, whereas aflatoxin has more adverse effects upon the liver. Diagnosis and determination of toxins can be done in tissues and feed samples by using different techniques such as high performance liquid chromatography (HPLC), thin layer liquid chromatography (TLC), and fluorescence polarization immunoassay (FPIA). Treatment of mycotoxicosis can be done by decontamination or detoxification of feed by using mycotoxin binders, plant preparations (nutraceuticals) and vitamins. Mycotoxicosis can be controlled by using transgenic crops and improving harvesting management, storing feed in good environmental conditions and fresh feed for the birds. Hazard analysis critical control point (HACCP) should be referred to for the control of mycotoxins in the food chain.

Patented Techniques for Detoxification of Mycotoxins in Feeds and Food Matrices

Patented Techniques for Detoxification of Mycotoxins in Feeds and Food Matrices

Patented Techniques for Detoxification of Mycotoxins in Feeds and Food Matrices

Mycotoxins are found worldwide in cereal grains, animal feed and human food that are produced from contaminated grains. This presents food safety risks. A variety of methods has been developed to reduce the safety risks by removing mycotoxins from contaminated grain products, eliminate the bioavailability of mycotoxins in the gastrointestinal system, degrade mycotoxins in food and feed, reduce mycotoxins pre- and post-harvest, and prevent (mycotoxin involved) plant disease. These techniques include physical methods such as flotation, extraction and adsorption, chemical reactions such as alkalization, oxidation, heat and irradiation, as well as biological detoxifications using microorganisms, enzymes and resistant plants. Most of the recent patented techniques for detoxification of mycotoxins in feeds and food matrices are discussed for an improved understanding of the patent techniques and insights into detoxification of mycotoxins.

Fungi and Mycotoxins in Feed Intended for Sows at Different Reproductive Stages in Argentina

The aim of this study was to evaluate fungi and contamination levels of aflatoxin B1, ochratoxin A, fumonisin B1, and zearalenone in raw materials and finished feed intended for sows at different reproductive stages. Total fungi, Aspergillus, Penicillium, and Fusarium species occurrence, were examined. Aspergillus flavus, A. niger aggregate spp., and F. verticillioides were the prevalent species. Fungal counts exceeded the levels proposed as feed hygienic quality limits (1 × 104 colony forming units) at all reproductive stages. Aflatoxin B1, ochratoxin A, fumonisin B1, and zearalenone were detected by high-pressure liquid chromatography. Aflatoxin levels in 80% samples of finished sow feeds were over the permitted levels of 0.02 μg g−1 (mean 228.2 ± 95 μg Kg−1). Fumonisin B1 was detected in all tested raw materials at levels that varied from 50.3 to 1137.64 μg Kg−1 and finished feed samples at levels that ranged from 99.8 to 512.4 μg Kg−1. Aflatoxin B1, zearalenone, and ochratoxin A were not detected in raw materials. All finished feeds were negative for zearalenone contamination whereas all nonpregnant gilt samples were contaminated with low OTA levels (mean 0.259 ± 0.123). This fact requires periodic monitoring to prevent the occurrence of mycotoxicosis in animal production, to reduce the economic losses, and to minimize hazards to human health.

Occurrence of Mycotoxins in Feed as Analyzed by a Multi-Mycotoxin LC-MS/MS Method

Crops used for animal feed can be easily contaminated by fungi during growth, harvest, or storage, resulting in the occurrence of mycotoxins. Because animal feed plays an important role in the food safety chain, the European Commission has set maximum levels for aflatoxin B1 and recommended maximum levels for deoxynivalenol, zearalenone, ochratoxin A, and the sum of fumonisin B1 and B2. A multimycotoxin LC-MS/MS method was developed, validated according to Commission Decision 2002/657/EC and EN ISO 17025 accredited for the simultaneous detection of 23 mycotoxins (aflatoxin-B1, aflatoxin-B2, aflatoxin-G1, aflatoxin-G2, ochratoxin A, deoxynivalenol, zearalenone, fumonisin B1, fumonisin B2, fumonisin B3, T2-toxin, HT2-toxin, nivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, diacetoxyscirpenol, fusarenon-X, neosolaniol, altenuene, alternariol, alternariol methyl ether, roquefortine-C, and sterigmatocystin) in feed. The decision limits of the multimycotoxin method varied from 0.7 to 60.6 microg/kg. The apparent recovery and the results of the precision study fulfilled the performance criteria as set in Commission Decision 2002/657/EC. The analysis of three different feed matrices (sow feed, wheat, and maize) provided a good basis for the evaluation of the toxin exposure in animal production. In total, 67 samples out of 82 (82%) were contaminated; type B-trichothecenes and fumonisins occurred most often. The majority of the infected feed samples (75%) were contaminated with more than one type of mycotoxin.

Zearalenone, deoxynivalenol and aflatoxin B1 and their metabolites in pig urine as biomarkers for mycotoxin exposure

Methods to determine zearalenone (ZEA), deoxynivalenol (DON), aflatoxins (AF) and their metabolites in pig urine were developed as biomarkers for pig exposure to the mycotoxins in feed. Urine samples were incubated with β-glucuronidase to cleave conjugates, extracted and cleaned-up with solid phase and immunoaffinity columns, followed by HPLC with UV and fluorescence detection. Good recoveries (83-130%), low variation (2-10%), and low detection limits (0.3-9.9ng/ml) were obtained. The results of controlled AFB1 feeding trials found no difference in urine concentrations of AFB1 or AFM1 from pigs fed three different levels (127, 227, 327 µg/kg) of AFB1 in diets. The excretion of AFB1 and AFM1 in urine was on average 30% of the oral dose and the ratio AFB1 to AFM1 was around 23%. The analysis of 15 Vietnamese pig urine samples indicate a relatively high exposure of ZEA, DON and AF, which were found as toxin or metabolites in 47, 73, and 80% of the urine samples, respectively.

The clinical and pathomorphological diagnosis of mycotoxicosis in different swine categories

The issue of mycotoxins and mycotoxicosis in veterinary medicine is directly connected to the usage of mouldy and/or adversely stored grains (corn, wheat, barley) used in animal feed. In swine production, in our geographical region, the most common are mycotoxicosis caused by zearalenon (F-2 toxin), but aflatoxins, ochratoxin and trichothecenes can also be found. For the known mycotoxins of clinical importance, the response is usually subacute or chronic and the presenting clinical signs are often vague. Mostly the problems are expressed only as alterations of the reproductive cycle, reduced feed intake and slow growth. However, if we consider the clinical signs and pathomorphological picture of mycotoxicosis in different swine categories (breeding animals, suckling and weaned piglets, fatteners), the age dependent changes can be found. Some mycotoxins have hepatotoxic, nephrotoxic and immunosuppressive effects, which further complicate the clinical and pathomorphological picture and diagnosis of mycotoxicosis in swine. The material for this research included the samples provided from ten swine farms. In different swine categories health disorders, resembling to the problem with mycotoxins were detected. The applied research methods included clinical evaluation and pathomorphological examination and laboratory microbiological feed testing, in order to examine the presence of fungi and some mycotoxins (aflatoxins, zearalenon, ochratoxin A and trichothecenes). On the basis of the obtained results, it may be concluded that the most frequently detected mycotoxin in the examined feed samples was zearalenon. The presence of mycotoxin in feed was directly connected to the reproductive failures and diagnosed health disorders in the examined swine categories (vulvovaginitis, skin necrosis, pneumonia, gastroenteritis).

Mycotoxins in feedstuffs in Portugal: an overview

Mycotoxins are secondary metabolites produced by many genera of fungi in many commodities, under certain conditions. Mycotoxicological control of feed is a procedure that aims to protect human and animal health, avoiding the adverse effects of these undesirable substances. This component of the sanitary control of feed and food is essential to prevent the presence of those substances which can seriously affect the health of the animals. In Portugal, there is relatively few information related to the natural occurrence of mycotoxins in feed. In this context, the authors present results and data compilation concerning the occurrence of mycotoxins in raw materials and also feed for dairy cattle, swine, poultry, horses, fish, laboratory rats and pet; making a generic qualitative appreciation of the risks associated to the presence of mycotoxins in these feedstuffs. The mycotoxins studied: aflatoxins (AFs), ochratoxin A (OTA), fumonisin B1 and B2 (FB1, FB2) were analysed by High Performance Liquid Chromatoghraphy (HPLC). Deoxynivalenol (DON) and zearalenone (ZEN) were determined by Thin-Layer Chromatography (TLC). The results suggest that contaminations with these mycotoxins in feed are quite common, revealing the need for surveillance and monitoring programs for the prevention of the sanitary impacts of these "non desirable substances".

Safety of Improved Milbond-TX When Fed in Broiler Diets Limiting in Available Phosphorus or Containing Variable Levels of Metabolizable Energy

Improved Milbond-TX (IMTX), a hydrated sodium calcium aluminosilicate, was added to broiler diets at 0, 1, or 2% in 2 experiments. In experiment 1, three concentrations of available P, 0.22, 0.32, and 0.42%, were combined with the IMTX in isocaloric and isonitrogenous corn-soybean meal-based diets and fed to male broilers for 21 d. In experiment 2, the IMTX was added to isonitrogenous broiler diets calculated to contain 2,940, 3,020, or 3,100 kcal of ME/kg and also fed for 21 d. Deficient dietary P reduced feed intake and BW at 7, 14, and 21 d of age, but IMTX had no effect even when fed at 8 times the amount recommended by the manufacturer (0.25%) to ameliorate the toxic effects of feed mycotoxins. Similarly, increasing the energy level of the diet resulted in improved BW and feed conversion, but IMTX fed at 8 times the amount recommended by the manufacturer had no detrimental effect on chick performance. There was no interaction found between IMTX and energy or available P for any performance measure. Addition of IMTX to the diet above the recommended concentration of the manufacturer also had no detrimental effect on percentage of toe ash or excreta moisture.

Worldwide occurrence of mycotoxins in commodities, feeds and feed ingredients

Globalisation of the trade in agricultural commodities has contributed significantly to the discussion about potential hazards involved and has increased in particular the awareness of mycotoxins. Safety awareness in food and feed production has also risen due to the simple fact that methods for testing residues and undesirable substances have become noticeably more sophisticated and more available at all points of the supply chain. A 2-year survey program was initiated by feed additive producer Biomin ® in order to evaluate the incidence of mycotoxins in feed and feed raw materials in some of the major animal production regions. Fusarium mycotoxins tested were those known for their impact on feed industry and animal husbandry, namely deoxynivalenol (DON), T-2 toxin, zearalenone (ZON), fumonisins B1, B2, and B3. In addition, ochratoxin A and aflatoxin B1, which are not produced by Fusarium, were tested and are reported herein since interactions between Fusarium toxins and other non- Fusarium mycotoxins are possible. A total of 2753 analyses were performed on 1507 samples sourced from European and Mediterranean markets, and 6391 analyses were undertaken on 1291 samples originating from the Asian-Pacific region. More than half of materials sampled in Europe were contaminated at levels above the limit of quantification of methods applied, while one third of tests on Asian-Pacific sourced samples were positive. European samples had DON, ZON and T-2 toxin as major contaminants, materials from Asia and the Pacific tended to be contaminated with DON, ZON, fumonisins, and aflatoxins.