Review of investigations regarding
mycotoxin control in poultry
throughout the years

We review the key aspects of mycotoxin control and its negative effects on poultry farming with Dr. Milad Manafi.

Milad Manafi

Department of Animal Science, Faculty of Agricultural Sciences,
Malayer University, Malayer, Iran.

Generally, different poultry species require an adequate supply of carbohydrates, proteins, fats/oils, vitamins, minerals, and water (Manafi et al., 2011).

However, poor raw material management and storage and high carbon and moisture rates in feedlots may lead to a final diet contaminated with fungi and mycotoxins (Döll y Dänicke, 2004).

Mycotoxins are recognized as secondary toxic metabolites mainly produced by various toxigenic fungal species of the Aspergillus, Fusarium, and Penicillium (Egbontan et al., 2017).

The mycotoxin type, level and frequency of exposure (acute or chronic), body mass index, gender, concomitant health issues, and possible synergistic effects of other chemicals affect the manifestation of the disease (Manafi et al., 2009).

In poultry, the significance of these mycotoxins’ effects depends on their presence in food and feed above the regulatory limits.

Different symptoms in poultry and other animals have been well documented by many scientists:

  • Weight loss, anorexia and impaired feed conversion competency
  • Immunosuppression and failed response to vaccination
  • Low fertility
  • Drop in egg production and high chances of egg blood spots
  • Kidney enlargement
  • Pale fatty liver and hepatitis
  • Gizzard erosions and oral lesions
  • Increased incidence of leg malformations
  • Increased death rates and visceral hemorrhages
  • Reduced villus height

(Alexandros and Jean, 2002; Döll et al., 2004; Jaynes et al., 2007; Marin et al., 2013; Mishra et al., 2013; Manafi et al., 2014; Manafi et al., 2015; Fowler et al., 2015; Manafi et al., 2016; Ji et al., 2016; Manafi, 2018; Manafi et al., 2019, Raj et al., 2021).

Furthermore, there is a high chance of mycotoxin carry-over (Figure 1) into edible byproducts obtained from poultry fed with contaminated feed. This phenomenon can lead to cancer, as some mycotoxins are recognized for their carcinogenicity by the World Health Organization’s (WHO) and International Agency for Research on Cancer (IARC) (Ganesan et al., 2021).

Figure 1. Schematic representation of Ochratoxin A (OTA) carry-over from feed to various parts of the chicken (adapted from Ganesan et al., 2021).


Commonly, mycotoxin analysis methods include:

  • Enzyme-linked immunosorbent assays (ELISA)
  • High-performance liquid chromatography (HPLC)
  • LC-MS/MS

The ELISA is a quick test with reliable accuracy in simple substrates, such as raw feed ingredients.

On the other hand, HPLC and LC-MS/MS-based analyses are relatively more accurate than rapid test kits and used for evaluating mycotoxin removers, especially in complex substrates, such as formulated feeds (Niderkorn et al., 2007)

Most of the mycotoxins are liposoluble compounds that can easily be absorbed from the site of exposure (gastrointestinal or respiratory tract) into the circulatory system, reaching the liver where they are metabolized by the microsomal system into active or detoxified metabolites and distributed throughout the organism (Haschek et al., 2002).

Through natural cellular processes of transcription and translation, mutations may manifest or even exacerbate the deregulation of cell growth (Manafi et al., 2012)


Increased efforts are being undertaken by scientists to develop cost-effective and safe products to achieve decontamination and remediation of different mycotoxins (preventive or curative) in feedstuffs (Manafi et al., 2019).

Preferably, the level of aflatoxins in feed should be zero. In any case, certain regulatory organizations have set their threshold as the maximum level for poultry.

Currently, the most effective way of protecting animals against mycotoxicosis is the inclusion of adsorbents in the feed (Ditta et al., 2018)

The ability of mycotoxin binders to remove toxins is determined by using in vitro and in vivo assays with their specific pros and cons.

For in vivo studies, to obtain consistent results specific bioassays should be conducted.

In contrast, in vitro experiments are relatively easy to perform, and they can shorten the time and cost of experimentation.

Keeping in mind that the ultimate goal of an in vitro study is to replace in vivo experiments in practice, the conditions of in vitro experiments should be tightly controlled and well-designed to obtain accurate research records (Hahn et al., 2015).

On this basis, Physical and Organic methods to control mycotoxin in poultry feed are detailed hereunder with the emphasis on the previous carried out research experiments.

Physical methods

The extraction of mycotoxins from feedstuff is an effective tool, as these secondary metabolites are highly soluble in organic solvents. The utilization of mycotoxin-binding adsorbents is the most applied physical method for protecting animals against mycotoxicosis.

Clays usually consist of two or more mineral-oxide layers and some of their particles can absorb moisture and expand. However, their efficiency depends on the chemical structure of the adsorbent and the mycotoxin, as it is important to ensure that the adsorbents do not remove essential nutrients from the diet (Manafi et al., 2012).

Zeolites, activated carbons, and hydrated sodium calcium aluminosilicate are among the most important clays used for controlling mycotoxicosis. Although these methods are comparatively expensive, their efficiency is partially efficient.


Zeolites are crystalline aluminosilicate compounds.

An improvement in 29-41% in body weight gain was reported in broilers exposed to 3.50 ppm of aflatoxin (AF) through dietary supplementation of commercial zeolite (Duarte y Smith, 2005).

Many other scientists have tried to ameliorate the adverse effects of mycotoxins in different animals using zeolite (Daković et al., 2005; Dhanasekaran et al., 2011; Rajendran et al., 2020; Raj et al., 2021).

Hydrated Sodium Calcium Aluminosilicate (HSCAS)

HSCAS, a phyllosilicate derived from natural zeolite, is perhaps the most extensively investigated mycotoxin adsorbent.

Phillips et al. (1988) showed that HSCAS has a high affinity for AFB1 after screening 38 different adsorbents that were representative of the major chemical class of aluminas, silicas, and aluminosilicates.

There is a wide range of efficacy observed in different studies incorporating HSCAS into the diets to reduce the toxicity of AF, OTA, and T-2 toxin in poultry (Huff et al., 1992; Jindal et al., 1993; Kubena et al., 1998; Raju y Devegowda, 2000; Huwing et al, 2001; Girish y Devegowda, 2004; Duarte y Smith, 2005; Khatoon et al., 2018; Wei et al., 2019).

Activated carbon

Activated carbon is an insoluble powder formed through the pyrolysis of different kinds of organic materials and is quite effective for adsorbing OTA.

Different studies have reported an improvement in the bodyweight broilers following the inclusion of activated charcoal in diets containing different mycotoxins (Ramos y Hernández, 1997; Solfrizzo et al., 2001; Huwing et al., 2001; Duarte y Smith, 2005; Mgbeahuruike et al., 2018).


Bentonite is a mineral clay with the unique characteristic of swelling to several times its original volume when placed in water. Due to their montmorillonite content, bentonites form thixotropic gels as a result of their ion exchange capabilities.

The bentonite forms a complex with the toxin, preventing the absorption of mycotoxins, such as aflatoxins, across the intestinal epithelium (Duarte y Smith, 2005).

Clay materials can bind to molecules of certain sizes and configurations and have been used effectively to decrease the effects of aflatoxin-contaminated diets in poultry. In this regard, there are plenty of publications reporting mycotoxin decontamination in poultry (Unsworth et al., 1989; Smith y Ross, 1991; Hagler et al., 1992; Santurio et al., 1999; Rosa et al., 2001; Vieira, 2003; Eralsan et al., 2005; Eraslan et al., 2006; Murugesan et al., 2015; Mgbeahuruike et al., 2018).

Organic methods

In this category of mycotoxin control tools, the application of herbal-antioxidant agents, vitamins, algae, enzymes, nutritional manipulation, and biological methods will be reviewed.


The application of some plant-derived extracts, such as turmeric (Curcuma longa) garlic (Allium sativum) and asafetida (Ferula asafetida), have been shown to counteract aflatoxicosis in poultry through their antioxidant activity by reducing the level of free radicals (Manafi et al., 2018).

The body’s antioxidant system mainly involves reducing agents (tocopherol, ascorbic acid, glutathione, carotenoids), peroxidases (glutathione peroxidase, catalase), enzymes (peptidases, proteases, vitamin A), and superoxide dismutase (Renzulli et al., 2004).

The most common functional chemical groups with radical scavenging properties are hydroxyl (phenolics), sulfhydryl (cysteine, glutathione), and amino groups (uric acid, spermine) (Lee et al., 2001).

There are several studies on the use of diverse herbal extracts against mycotoxicosis on different animals that could partially alleviate some negative effects:

  • Increased peroxides
  • Reduced antioxidant enzyme activities
  • Inhibited protein/DNA synthesis
  • Suppressed chromosomal aberration
  • Inhibited cytochrome P450 bioactivation of AFB1
  • Reduced antioxidant biomarkers (glutathione peroxidase and superoxide dismutase)
  • Inhibited AFB1 mutagenicity and increased formation AFB1-DNA adducts

(Iqbal et al., 1983; Nyandieka et al.,1990; Surai; 2001; Weiss, 2002; Gowda and Ledoux, 2008; Gowda et al., 2008; Dalleau et al., 2008; Ruan et al., 2019).


There are reports of regeneration of α-tocopherol on reaction with other reducing agents like glutathione, urate (Kagan y Tyurina, 1998) and ascorbate (May et al., 1998).

Chow (2001) stated that α-tocopherol is the most biologically active form, quickly scavenging peroxy radicals by forming a stable tocopheroxy radical and acting as a biological modifier.

Vitamin E pretreatment significantly lowered AF-induced lipid peroxidation (Verma y Nair, 2004).

Among vitamins, ascorbate (vitamin C) is very important due to its ability to scavenge superoxide, hydrogen peroxide, hydrogen radicals, hypochlorous acid, and singlet oxygen (Chow, 2001).

Riboflavin also has a protective action against AFB1 induced DNA damage in rats (Gowda y Ledoux, 2008).

Muchos otros estudios han demostrado los efectos de diferentes vitaminas en animales cuando son expuestos a micotoxinas (Nyandieka et al., 1990; Coelho, 1996; Hoehler y Marquardt, 1996; Diaz y Smith, 2005; Kabak, 2009; Wayne, 2012; Murugesan et al., 2015).

Algae (Spirulina platensis)

The nutritional value of some algae like Spirulina platensis is extremely high.

Spirulina is rich in amino acids, vitamins, gamma-linoleic acid, sugars, and trace elements (EFSA FEEDAP Panel, 2016).

It has also been reported that Spirulina platensis is effective against aflatoxicosis by Raju et al., (2004); Abdel-Wahhab and Aly (2005); Dal Bosco et al., (2008); Manafi et al., (2009); Manafi, (2011); Manafi et al., (2012) and Park et al., (2018).


Enzyme degradation reagents are used for the biodegradation of the toxic chemical structure of the mycotoxins into non-toxic metabolites by using microorganisms and their metabolites or specifically extracted components.

They are believed to break the functional atomic group of the mycotoxin molecule, thereby rendering it non-toxic (Kabak et al., 2006).

Some enzymes, such as carboxyesterase present in the microsomal fraction of the liver, esterase, and epoxidase are being tested for their practical applicability in field conditions (Pasteiner, 1997).

The demonstrated efficacy of using enzymes against mycotoxicosis is welldocumented by a wide range of researchers (Vekiru et al., 2010; Ji et al., 2016; Tso et al., 2019; Fruhauf et al., 2019).

Nutritional manipulations

An increase in the dietary protein levels and supplementation of L-phenylalanine has been revealed to be effective against aflatoxicosis and ochratoxicosis. Additionally, increasing the supplementation of riboflavin, pyridoxine, folic acid and choline showed a protective effect against aflatoxicosis (Ehrich et al., 1986).

Antioxidants, such as β-napthoflavone, vitamin C, and vitamin E offer protection against AF-induced genotoxicity in most in vitro studies (Johri et al., 1990).

Devegowda et al., (1998); Krska et al., (2008); Bryden, (2012), y Marin et al., (2013) have reported that additional supplementation of poultry diets with micronutrients supplementation can partially alleviate the adverse effects of mycotoxins.

Biological methods

A rapid explosion in the feed industry has opened up new possibilities through the degradation of mycotoxins by microorganisms (Alexandros y Jean, 2002).

Several yeasts, moulds, and bacterial strains possess the ability to destroy or transform mycotoxins successfully (Edlayne et al., 2009).

In this category, bacterial, protozoan, fungal and yeast degradations will be reviewed hereunder.


Several bacterial species have shown the ability to degrade AF (Kong et al., 2012).

Specifically, some acid-producing bacteria’s such as Lactobacillus plantarum and Lactobacillus acidophilus were found to detoxify AF, OTA, T-2 toxin, and zearalenone (ZEN) in different studies (Linderfelser y Ceigler, 1970; Bata y Lásztity, 1999; Turbic et al, 2002; Lahtinen et al., 2004; Kusumaningtyas et al., 2006; Niderkorn et al., 2007; Wu et al., 2009; Kolossova y Stroka, 2009; Rawal y Roger, 2010; Ma et al., 2012; Mishra et al., 2013; Fan et al., 2013 y Fan et al., 2015).


Protozoa are important agents for mycotoxin biodegradation in the rumen (Upadhaya et al., 2010).

Most of the studies on the subject indicate that mycotoxin degradation was achieved primarily by ruminal protozoa (Jouany et al., 2009). Besides Hussein y Brasel (2001) reported that up to 90% of T-2 toxin degradation was achieved by rumen protozoa.

Tetrahymena pyriformis, at a dose rate of 22x106 cells, detoxified AFB1 by converting it into its hydroxyl products to an extent of 5% in 24 hours and 67% in 48 hours (Robertson et al., 1970).

Intact rumen fluid containing various protozoa was reported to metabolize T-2 toxin and OTA, while no effect on AF was noted (Kiessling et al., 1984). However, the fact that protozoa cannot be cultured in vitro limits further understanding of their ability to degrade mycotoxins (Chaucheyras-Durand y Ossa, 2014; Newbold et al., 2015).

Studies involving culture-independent approaches based on analysis of particular genes and genomes potentially associated with mycotoxin degradation may help improve our understanding of the role of protozoa in the process (Garai et al., 2021).


An intracellular substance was found to be responsible for A. flavus and A. parasiticus to degrade the formed toxins in a culture when their mycelium was subjected to fragmentation.

Peroxidase enzymes produced by fungal mycelium, which can catalyze hydrogen peroxide into free radicals, react with mycotoxins (Dvorak, 1989; Alexandros y Jean, 2002; Edlayne et al., 2009).


The advent of biotechnology in the last decade has opened a new pathway for tackling the problem of mycotoxicosis by using yeast extracts.

Yeast is a rich source of numerous vitamins and certain species and strains can detoxify mycotoxins through their degradation (Biernasiak et al., 2006).

Mannanoligosaccharide (MOS) derived from the cell wall of Saccharomyces cerevisiae appears to have a high affinity for a wide range of mycotoxins (Biernasiak et al., 2006).

It is believed that the glucomannan matrix of MOS preparation traps the mycotoxins in an irreversible way (Afzali y Devegowda, 1998).

Besides, it is assumed that a small portion of modified-MOS might be taken up in the small intestine by M-cells, causing B-cell activation and subsequent activation of T-cells and macrophages, leading to an overall increase in the immune status of the birds (Savage et al., 1996).

The mycotoxin binding ability of MOS has been demonstrated in various in vitro and in vivo trials (Stanley et al., 1993; Devegowda et al., 1996; Afzali y Devegowda, 1998; Devegowda et al., 1998; Raju y Devegowda, 2000; Swamy y otros, 2004; Yegani y otros, 2006; Awaad y otros, 2011; Fowler y otros, 2015; Farooqui y otros, 2019; Arif y otros, 2020).




There is a wide range of commercially available mycotoxin binders and antifungal agents that have varying potency effects at reducing the presence or eliminating the toxicity of mycotoxins in poultry feeds.

The development of a successful commercial mycotoxin binder calls for the incorporation of the best of these active ingredients at the required concentration to ensure an overall reduction of the harmful effects of the presence of mycotoxins in poultry diets.


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