Preventive and Therapeutic Methods against the Toxic Effects of Mycotoxins – a review

Published on: 8/12/2016
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Ingredients used in animal feeds and their contamination with undesirable substances, such as mycotoxins, are fundamentally important both in terms of the quality of animal products and the potential human health impacts associated with the animal-based food production chain. Feed ingredients contaminated with mycotoxins may have a wide range of toxicological effects on animals. Therefore, mycotoxin contamination of feed ingredients constituting complete feed products represents an important potential hazard in farm animal production. This review summarises the potential effects of some preventive methods used during the storage of cereal grains as well as of nutritive (e.g. antioxidants, amino acids, fats) or non-nutritive compounds (e.g. pharmacological substances, carbon- or silicabased polymers) and detoxifying enzymes recommended for use against the toxic effects of different mycotoxins.


Mycotoxins pose a global problem and are estimated to affect as much as 25 percent of the world’s crop production each year (Lawlor and Lynch, 2005). The recognition of mycotoxins as a potential risk to the feed industry has made it necessary to adopt a suitable prevention and control strategy (FAO, 2001). Chronic exposure to mycotoxins may significantly alter the productivity of farm animals through direct exposure to mycotoxin-contaminated feed; however, mycotoxins may also be present in foods of animal origin, and thus pose a high risk to the consumer (JECFA, 2001).


The high prevalence of mycotoxins in animal feeds in the mild climatic zones of Europe and North America results in considerable economic losses. These toxins affect the health and productivity of all age groups of farm animals.


Prevention of the infection of plants with moulds and mycotoxin production during the storage and processing of feedstuffs

A variety of preventive methods have been proposed against the infection of plants with moulds, e.g. Fusarium sp., at field level. These include crop rotation, tillage, soil fertilisers, planting date, plant protection or breeding resistant plant varieties. Humidity and temperature control during storage would be useful methods of prevention (Jouany, 2007). Disinfection of feed delivery trucks, storage bins and feed conveyors with sodium hypochlorite (Paster et al., 1985), as well as sorting, washing, dehulling or thermal treatment of contaminated grains before storage or in the feed mill can also be used for the prevention of mould infection (Jouany, 2007). Addition of mould inhibitors, such as propionic acid, during storage (Ghosh et al., 1996) have been proposed as alternative preventive methods against moulds and mycotoxins. For the prevention of mycotoxin production in silages and roughages, the inoculation of these feeds with lactic acid producing bacteria having antifungal activity has been proposed for use against some mycotoxin-producing moulds including Aspergillus, Penicillium and Fusarium species (Magnusson et al., 2003). These lactic acid bacteria (e.g. Lactobacillus coryniformis, L. plantarum and Pediococcus pentosaceus) produce phenyl-lactic acid, 4-hydroxy-phenyl-lactic acid (Lavermicocca et al., 2000), some proteinaceous antifungal compounds (Magnusson and Schnürer, 2001) and cyclic dipeptides (Ström et al., 2002). Methods proposed for the detoxification of mycotoxin-containing feedstuffs include physical procedures (such as sorting, washing, dehulling, heat treatment, milling and irradiation) or chemical approaches including liquid extraction with organic solvents or water-based solutions of calcium chloride or sodium bicarbonate (Binder, 2007; Jouany, 2007), but large-scale and practical methods are not available at present.


Preventive nutritive methods

Various nutritional strategies have been proposed to alleviate the adverse effects of mycotoxins. For instance, in their pioneering work Smith et al. (1971) found that increased levels of crude protein (20–30%) in the diet of chickens alleviated the growth-depressing effect of aflatoxin, but such an effect was not found when ochratoxin A was used (Gibson et al., 1989). Of the amino acids, dietary supplementation of methionine or phenylalanine improved the health status of chickens fed diets contaminated with ochratoxin A (Gibson et al., 1990). The addition of dietary nucleotides (10 mg kg–1) was effective against DNA fragmentation but ineffective against the oxidative stress caused by deoxynivalenol or T-2 toxin in chicken spleen leukocytes (Frankic et al., 2006). These results underline the possible beneficial effect of nucleotides on the immune system of chicken during mycotoxin intoxication. Positive effects, such as lower mortality and higher growth rate, exerted by the addition of different lipids including olive oil or safflower oil as unsaturated fatty acid sources, were demonstrated in aflatoxicosis (Smith et al., 1971). Supplementation of diets with thiamine possibly ameliorates the toxicity of Fusarium mycotoxins because Fusarium moulds contain, in addition to mycotoxins, an anti-thiamine factor (Nagaraj et al., 1994).



Most of the mycotoxins, e.g. aflatoxin B1 (AFB1), T-2 toxin and ochratoxin (Balogh et al., 2007; Pál et al., 2009), provoke oxygen free radical formation. For this reason, the addition of natural (vitamins E and C, selenium, carotenoids, L-carnitine and melatonin) and also synthetic antioxidants (butylated hydroxytoluene, lipoic acid) is potentially efficacious because of the ability of these compounds to act as superoxide anion scavengers (Rogers, 2003; Citil et al., 2005; Surai, 2006; Dvorska et al., 2007). For instance, carotenoids (e.g. β-apo-8’-carotenal) were shown to inhibit aflatoxin B1-induced liver pre-neoplastic foci, DNA damage, carcinogenicity and genotoxicity in rats through the modulation of aflatoxin metabolism towards increased detoxification to less genotoxic products like aflatoxin M1 (Gradelet et al., 1998). Vitamin E supplementation was found to improve the utilisation of vitamin E, which is impaired by T-2 toxin (Weber et al., 2007). Some natural feed components (phenolic compounds, coumarin, chlorophyll and its derivatives, fructose) and also medicinal herbs and plant extracts have been found to act as an effective nutritional prevention against mycotoxicosis, mainly against aflatoxins (Galvano et al., 2001). The chemopreventive properties of natural or synthetic antioxidants are partially associated with their effect on the induction of antioxidant and phase II detoxifying enzymes including glutathione peroxidases or glutathione S-transferases. This effect has been proven in vivo by the supra-nutritional addition of methionine in aflatoxicosis (Veltmann et al., 1983) possibly through its action on the de novo biosynthesis of glutathione (Németh et al., 2004). Combined antioxidant treatment (CoQ10, L-carnitine, Zn, Mg, N-acetylcysteine, vitamin C, vitamin E, selenium and tamoxifen) was proven to be effective against the apoptosis-inducing action of ochratoxin A in mice (Atroshi et al., 2000). However, there are only few data about the positive effect of antioxidants on some important mycotoxins, such as fumonisin B1, T-2 toxin, zearalenone and citrinin, and no such studies have been done on recently discovered toxins such as beauvericin, fusaproliferin, moniliformin and fusaric acid (Atroshi et al., 2002). Chlorophyllin, a water-soluble derivative of chlorophyll, contains approximately 34% chlorophyll and 66% salt. Chlorophyllin was first proposed for use against mycotoxicosis as a chemoprotective agent because of its antioxidant properties (Atroshi et al., 2002). Its in vivo effect was demonstrated in rainbow trout in aflatoxin B1 toxicosis. Addition of chlorophyllin to the feed at a dose level of 4 g kg–1 significantly reduced the AFB1-DNA adduct formation (Breinholt et al., 1995; Breinholt et al., 1999).


Preventive methods with non-nutritive feed additives

Preventing or limiting the absorption of mycotoxins from the intestine using non-nutritive feed additives, commonly referred to as mycotoxin adsorbents or sequestrates, is the most widely accepted method (Huwig et al., 2001). The use of mycotoxin adsorbents as feed additives is one of the most prominent approaches to reduce the risk of mycotoxicoses in farm animals and to minimise the carry-over of mycotoxins from contaminated feeds into foods of animal origin (Sabater-Vilar et al., 2007). However, it must be pointed out that these additives are not authorised in Europe, but a new functional group has been established by the European Commission (286/2009/EC) within the main group of technological additives: ‘substances for additional reduction of contamination of mycotoxins: substances that can suppress or reduce the absorption or promote excretion of mycotoxins’ (EU, 2009). The use of binding agents or sequestrates, which can adsorb the mycotoxin molecules by means of ion exchange and thereby preclude their absorption from the gut, has gained considerable attention in recent times. These binders have small particles and a large surface area but a very narrow range of mycotoxin sequestrating ability. Effective binding ability mostly requires functional polar groups of the mycotoxin molecule, which means that most of the mycotoxins, except aflatoxins, have a low rate of binding to most of sequestrates (Devegowda et al., 1998a). Consequently, the reduction and/or prevention of human exposure require different practical and effective methods to detoxify mycotoxins or reduce/inhibit their absorption from the gastrointestinal tract of animals. However, some sequestrates also bind certain feed components, reduce their absorption and consequently impair the production traits.


Activated charcoal

The mycotoxin-sequestering capacity of activated charcoal depends on the specific surface area which is different (500 to 3500 m2 g–1) in lignin-based and in super-activated charcoal (Ramos et al., 1996). Based on that fact, the in vitro sequestrating ability of activated charcoal can be utilised for reducing the absorption of several fusariotoxins, such as deoxynivalenol and nivalenol (Avantaggaito et al., 2004) and fumonisins (Avantaggaito et al., 2005). However, activated charcoal is effective only when used in extremely high doses (5–20 g kg–1 feed), but sometimes it was found ineffective in animals even against aflatoxin toxicity (Bonna et al., 1991). The positive effect of activated charcoal was proved in vivo against aflatoxin B1 toxicity in chickens (Dalvi and Ademoyero, 1984), goats (Hatch et al., 1982), lactating dairy cows (Galvano et al., 1996) and mink (Bonna et al., 1991). Other authors found that activated charcoal was ineffective in reducing the toxicity of aflatoxin B1 in chickens (Kubena et al., 1990b) or turkeys (Edrington et al., 1996) or in decreasing the rate of excretion of aflatoxin M1 through the milk in dairy cows (Diaz et al., 2004). The positive effect of activated charcoal was proved in vivo against T-2 toxin toxicity in rats (Bratich et al., 1990) and also in fattening pigs (Poppenga et al., 1987). Activated charcoal was found to be ineffective against ochratoxin A toxicity in chickens (Rotter et al., 1989) and against fumonisin B1 toxicity in rats (Solfrizzo et al., 2001).


Silica-based polymers

Two subclasses of silica-based polymers or clays have importance as potential mycotoxin-sequestrating agents: the phyllosilicate subclass (groups of montmorrilonite/ smectite, kaolinite and illite) and the lectosilicate subclass (group of zeolites). These clay additives have been used to pelletise feeds and improve their flow characteristics, and they are not efficient against most of the mycotoxins, with the exception of aflatoxins (Masimango et al., 1979). The main problem with silica-based polymers is that they are required to be in the feed at a high inclusion level (5 to 20 g kg–1 feed). Furthermore, it must be noted that clays absorb several micronutrients, such as trace elements (Ward et al., 1991). For instance, clays were reported to lower serum chloride concentration in broiler chickens (Scheideler, 1993) and significantly reduced the apparent absorption of magnesium, manganese and zinc in sheep (Chestnut et al., 1992). Contrary to the effects seen in animals, in humans there was no interference of clays with micronutrients (vitamins and minerals) even when clays were used at a high dose (1.5 g day–1 of montmorrilonite; Afriyie-Gyawu et al., 2008). Natural clays are sometimes contaminated with heavy metals (e.g. lead, cadmium) or dioxins. For that reason, the US Food and Drug Administration (FDA) stated that ‘The use of sodium aluminosilicate and hydrated calcium sodium aluminosilicate as binders for mycotoxins is not considered to be generally recognised as safe’ (FDA, 1999). Additionally, clays accumulate in the manure and may have detrimental effects on the soils and pastures after the manure has been spread onto the field.



Montmorrilonite is the main constituent of bentonite which can be classified into calcium, magnesium, potassium and sodium bentonites. Bentonites have ion-exchange capabilities and have been widely used as mycotoxin sequestering agents. According to some recent in vitro data the mycotoxin-, particularly aflatoxin B1-, sequestrating capacity of montmorrilonites depends on their charge. Low-charge montmorrilonites, such as bentonite and sepiolite, retained more AFB1 than their high-charge counterparts, such as montmorrilonites (Jaynes et al., 2007). Montmorrilonite was found to be an effective absorbent in vivo against the genotoxicity caused by sterigmatocystin (0.5 mg kg–1 b.w.) in the Nile tilapia (Abdel-Wahhab et al., 2005). Its effect was also proved in vivo against aflatoxin B1 toxicity in chickens (Santurio et al., 1999), fattening pigs (Lindemann et al., 1993), freshwater fishes (Shehata et al., 2003) and rats (Abdel-Wahhab et al., 2002), and montmorrilonite also reduced the aflatoxin content of cow’s milk (Diaz et al., 1997). The effect of bentonite was proved against T-2 toxicosis in rats (Carson and Smith, 1983a), but only at an extremely high dose of application. Among the other phyllosilicates, some purified and chemically modified polymers have also been investigated as mycotoxin sequestrates and have been found effective even at a low inclusion level against the oestrogenic effect of zearalenone in prepubertal gilts (Malone et al., 2007).



Zeolites are a group of about 45 naturally occurring minerals, all of them alumino-silicates. They have a capacity to hold positive cations and also larger molecules and are normally used as technological additives (anti-caking agents). Synthetic aluminosilicate (Zeolite A) is approved as a feed additive in the category of technological additives in the European Union (E554) as a binding agent. Zeolites are sensitive to pH, and below pH 4.0 they are partly hydrolysed and their crystal structure is destroyed (Cook et al., 1982). The positive effect of zeolite was proved in vivo against aflatoxin B1 toxicity in chicken (Scheideler, 1993) and Japanese quail (Parlat et al., 1999), but it was also found that the addition of zeolite to feed did not protect against acute liver damage in acute aflatoxicosis of chicken (Sova et al., 1991). Among the different zeolites, clinoptilolite was found to be effective against aflatoxicosis in chicken (Harvey et al., 1993) but not in pregnant rats (Mayura et al., 1998), and it was also ineffective against cyclopiazonic acid toxicosis in chicken (Dwyer et al., 1997).


Hydrated sodium calcium aluminosilicate (HSCAS)

HSCAS is an artificially modified natural zeolite and it is possibly the most widely studied mycotoxin- sequestering agent among the mineral clays. However, the FDA stated that HSCAS as a binder for mycotoxins is not considered to be generally recognised as safe (FDA, 1999). Ramos and Hernandez (1997) reviewed more than 20 publications, which demonstrated the in vivo capacity of HSCAS as an aflatoxin B1 sequestering agent in chickens, turkeys, weaned pigs, dairy cattle, lambs, goats and mink. The ability of HSCAS to bind T-2 toxin was found to be moderate (Fazekas et al., 2000) or very low (Garcia et al., 2003), but HSCAS has been shown to be a highly effective binder of ochratoxin A in vivo in chicken (Garcia et al., 2003). However, other researchers found no significant effects of HSCAS on ochratoxin (Huff et al., 1992) and T-2 and diacetoxyscirpenol toxicosis in chicken (Kubena et al., 1990a), zearalenone toxicosis in mink (Bursian et al., 1992) and deoxynivalenol toxicosis in pigs (Dänicke et al., 2004). HSCAS was also ineffective against in vivo fescue toxicosis in sheep (Chestnut et al., 1992).



Cholestyramine is an anion-exchange resin utilised in human medicine for absorbing bile acids in the gastrointestinal tract. Cholestyramine was found to lower ochratoxin A content of the blood plasma of rats in vivo, to diminish the biochemical alterations developing as an effect of ochratoxicosis (Madhyastha et al., 1992) and to enhance the faecal excretion of ochratoxin (Kerkadi et al., 1998); however, it is not available for the feed industry.


Carbon-based polymers

Plant fibres

The mycotoxin-sequestering capacity of different high-fibre feedstuffs, such as hays (e.g. alfalfa hay) or straws (e.g. wheat straw), has been known for a long time, but there are mainly practical experiences, e.g. in equine nutrition, without scientific assessment. The positive effect of alfalfa fibre was first proved against zearalenone in rats and pigs (Smith, 1980; Stangroom and Smith, 1984) and also against T-2 toxicosis in rats (Carson and Smith, 1983b). However, it should also be mentioned that, besides its positive effects, alfalfa fibre is a potential source of Fusarium contamination, and its high inclusion rates (15–25%) required in the diet may cause digestive-physiological disturbances. Micronised wheat fibre has recently been found effective in decreasing the accumulation of ochratoxin A in rat tissues (liver and kidney). When used at an inclusion level of 20 g kg–1, it significantly increased the excretion of ochratoxin A via the faeces (Aoudia et al., 2008).


Mannan oligosaccharides

The other groups of fibre components are the cell wall components of the yeast Saccharomyces cerevisiae, the mannan oligosaccharides or their esterified form with β-D-glucan (esterified glucomannan), which showed considerable binding ability for several mycotoxins in vivo (Devegowda et al., 1998b). The sequestrating capacity of yeast cell wall for aflatoxin B1 was demonstrated in chicken (Stanley et al., 1993; Karaman et al., 2005; Devegowda and Murthy, 2005). Later the positive effect of partially purified yeast cell wall polymer was proved in chicken using aflatoxin B1, ochratoxin A and T-2 toxin (Raju and Devegowda, 2000; Denli and Okan, 2002; Basmacioglu et al., 2005). The mycotoxin-sequestering capacity of esterified glucomannan was also demonstrated for zearalenone (Yiannikouris et al., 2006), ochratoxin A (Raju and Devegowda, 2002) and ergot mycotoxins (Dvorska, 2005), for aurofusarin (a secondary metabolite of Fusarium graminearum) in Japanese quail (Dvorska et al., 2003) and for T-2 toxin in chicken (Dvorska et al., 2007). Positive effects of esterified glucomannan on the production parameters of chickens were also demonstrated if the birds were fed naturally contaminated grains containing low levels of aflatoxin, ochratoxin, zearalenone and T-2 toxin (Aravind et al., 2003). Esterified glucomannan was also found to be effective against natural contamination of the feed with Fusarium toxins in weaned piglets (Swamy et al., 2003), pigs (Swamy et al., 2002) and pregnant gilts (Diaz-Llano and Smith, 2006). The negative effects of feed naturally contaminated with Fusarium mycotoxins (DON, 15-acetyl-DON, fusaric acid and zearalenone at levels of 15.0, 0.8, 9.7 and 2.0 mg kg–1 feed, respectively) on the feed intake of horses were also diminished by the use of esterified glucomannan, but the differences as compared to non-contaminated feed were not significant. However, esterified glucomannan mitigated liver damage as proven by the significantly reduced increase of γ-glutamyltransferase activity (Raymond et al., 2003). The toxic effects of deoxynivalenol were not prevented by the addition of yeast cell wall polymer in weaned piglets (Dänicke et al., 2007). However, feeding a diet naturally contaminated with Fusarium mycotoxins (DON, 15-acetyl-DON, fusaric acid and zearalenone at 5.5, 0.5, 26.8 and 0.4 mg kg–1 feed, respectively) and supplemented with glucomannan polymer (1 g kg–1 feed) decreased the reduction of dopamine concentrations induced by Fusarium mycotoxins in the hypothalamus of fattening pigs (Swamy et al., 2002), without exerting a positive effect on the production traits. Glucomannans did not show a positive effect against the significant reduction of the rate of hepatic fractional protein synthesis in laying hens fed grains naturally contaminated with Fusarium mycotoxins (Chowdhury and Smith, 2005), and at the dose level of 2 g kg–1 feed they did not prevent the detrimental effects of feeding cereals naturally contaminated with deoxynivalenol, 15-acetyldeoxynivalenol, zearalenone and fusaric acid on feed intake and nutrient digestibility in dogs (Leung et al., 2007).


Humic acid and derivatives

Among the other carbon polymers, a high quality humic acid derivative called oxyhumate has also been reported to have mycotoxin-sequestrating capacity and recommended for use against aflatoxin toxicosis based on in vivo studies in chickens (Van Rensburg et al., 2006).


Animal waste

There are some suggestions (Gittings and Drakley, 2002) and experimental data (Abdelhamid et al., 2004) regarding the aflatoxin B1 binding capacity of egg-shell waste from egg processing and hatchery when used at a dose of 5 to 20 g kg–1 in the complete feed in a freshwater fish, the Nile tilapia.


Biotransformation of mycotoxins

Most of the adsorbents have only limited ability to control the detrimental effects of most of the mycotoxins. Biotransformation represents a different prospective way of mycotoxin control, which is based on findings that mycotoxins can be detoxified by the use of safe strains of soil and rumen bacteria, rumen protozoa or yeasts, or their purified enzymes (Bata and Lásztity, 1999; Heidler and Schatzmayr, 2003; Schatzmayr et al., 2006). Trichothecene mycotoxins, which contain an epoxide ring, lose their toxicity after de-epoxidation in the rumen (Swanson et al., 1987), with the exception of neosolaniol or T-2 toxin, which are degraded to other toxic metabolites, HT-2 toxin and T-2 triol (Westlake et al., 1989). Components of the intestinal microflora of pigs (mainly Eubacterium spp.) degrade nivalenol (NIV) and deoxynivalenol (DON) to their corresponding de-epoxy metabolites (Kollarczik et al., 1994; Binder et al., 2000). The above-mentioned detoxifying effect of epoxidases was proved in vivo in chicken in the case of T-2 toxicosis (Hofstetter et al., 2005). Ochratoxin A has an isocoumarin moiety linked by an amide bond to l-β-phenylalanine, which is hydrolysed by some enzymes of bacterial, protozoal or filamentous fungal origin (Abrunhosa and Venâncio, 2007), such as carboxypeptidase A (Pitout, 1969), lipase (Stander et al., 2000) and some proteases (Abrunhosa et al., 2006). A novel yeast strain, Trichosporon mycotoxinivorans, has been shown to be capable of degrading ochratoxin A and zearalenone (Molnar et al., 2004), and a novel metalloenzyme has been isolated from Aspergillus niger, which hydrolyses ochratoxin A to non-toxic ochratoxin α (Abrunhosa and Venâncio, 2007), which supports its use as novel feed additive against mycotoxin-related toxicosis. A recently isolated bacterial strain (BBSH 797) was found to degrade some mycotoxins of the trichothecene group. It transforms deoxynivalenol into its metabolite DOM-1, the non-toxic deepoxide of DON, scirpentriol was transformed into its non-toxic metabolite deepoxy-scirpentriol, while T-2 triol was degraded into its non-toxic deepoxy form and into T-2 tetraol, which was then further metabolised to deepoxy T-2 tetraol (Fuchs et al., 2002). Recently, another possible biotransformation of zearalenone has been found in the case of the mycoparasitic fungus Gliocladium roseum which synthesises a zearalenone-specific lactonase catalysing the hydrolysis of zearalenone, followed by spontaneous decarboxylation. The gene encoding the zearalenone lactonase (zes2) has been identified, and may be used for genetic transformation in the future (Utermark and Karlovsky, 2007).


Therapeutic methods

There have been some proposals to use pharmacological methods (e.g. steroidal anti-inflammatory agents, antihistaminic agents and opioid antagonists, multiple drug treatments and other therapeutic agents such as ‘cytoprotectives’, antibiotics, subcutaneous or intravenous fluids) to reduce the symptoms of mycotoxicosis, but only moderate effects were obtained (Ryu et al., 1987; Fricke et al., 1989). Among other drugs, N-acetylcysteine, which has been used safely in mammals as an antidote of several toxic and carcinogenic agents, also proved effective against the performance decrease, liver and renal damage and biochemical alterations induced by aflatoxin B1 in broiler chickens (Valdivia et al., 2001). The metabolism and/or elimination of absorbed mycotoxins occur mainly through the microsomal xenobiotic transforming enzyme system of the liver (cytochrome P450 superfamily), and thus the activation of that system using phenobarbital has also been proposed as a potentially useful method against aflatoxin B1 toxicosis (Chen et al., 1982).



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Paper first published in : Acta Veterinaria Hungarica 58 (1), pp. 1–17 (2010)
DOI: 10.1556/AVet.58.2010.1.1

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