# Mycotoxins hazard: the European view

Philippe Pinton, Chloé Terciolo, Delphine Payros, Isabelle P Oswald

► To cite this version:

Philippe Pinton, Chloé Terciolo, Delphine Payros, Isabelle P Oswald. Mycotoxins hazard: the European view. Current Opinion in Food Science, 2025, 63, pp.101306. 10.1016/j.cofs.2025.101306. hal-05178405

HAL Id: hal-05178405

https://hal.inrae.fr/hal-05178405v1

Submitted on 23 Jul 2025

HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers.

L'archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d'enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés.

<figure><img src="image_1.png" /></figure>

Distributed under a Creative Commons Attribution 4.0 International License
<figure><img src="image_2.png" /></figure>

Review

# Mycotoxins hazard: the European view

Philippe Pinton, Chloe Terciolo*, Delphine Payros# and Isabelle P Oswald

<figure><img src="image_3.png" /></figure>

Food and feed are likely to be contaminated by mycotoxins, toxic secondary metabolites produced by molds mainly from *Aspergillus*, *Fusarium*, and *Penicillium* genera. To characterize the hazard of these contaminants, published data on their toxicokinetics and toxicity are scrutinized to determine the human health-based guidance values and to identify reference points (RPs) for animal category and species. This review describes how these values are constructed and summarizes the human tolerable daily intake (TDI) for the *Fusarium* toxins, deoxynivalenol, nivalenol, T-2 and HT-2 toxins, zearalenone, and fumonisins but also for ochratoxin A, and ergot alkaloids. For carcinogenic mycotoxins such as aflatoxins, it is not possible to establish TDI, and the risk for human health is determined by the margin of exposure between the benchmark dose's lower confidence limit and the exposure. This review also details, when they could be determined, the RPs for the same mycotoxins for ruminants (dairy cow, heifer, beef cattle, steer, sheep, goats), poultry (chicken, duck, turkey), porcine, solipeds, fish, rabbits, cats, and dogs.

It also addresses the difficulties and challenges involved in establishing TDIs and/or RPs with regard to (i) the lack of data (e.g. on emerging mycotoxins) and their quality, (ii) the availability of species- or sex-specific data, (iii) the consideration of clinical cases or a new, more sensitive endpoints, and (iv) the establishment of grouped TDIs/RPs and the consideration of possible interactions between mycotoxins.

## Address

Toxalim (Research Centre in Food Toxicology), Université de Toulouse, INRAE, ENVT, INP-Purpan, UPS, Toulouse, France

Corresponding author: Pinton, Philippe (Philippe.pinton@inrae.fr)

*Present address: Nantes Université, Univ Angers, INSERM, CNRS, Immunology and New Concepts in ImmunoTherapy, INCIT, UMR 1302/EMR6001, Nantes, France

# Present address: National Vet School of Toulouse (ENVT), INRAE, UMR 1225, IHAP, Université de Toulouse, 31300 Toulouse, France

**Current Opinion in Food Science 2025, 63:101306**

This review comes from a themed issue on **Food Toxicology**

Edited by **Paula Alvito**

For complete overview of the section, please refer to the article collection, "Food Toxicology 2024"

Available online 3 April 2025

https://doi.org/10.1016/j.cofs.2025.101306

2214-7993/© 2025 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

## Introduction

Food and feed supply chains may be colonized by filamentous fungi, which can result in moldy taste, reduced palatability, and nutritional loss. Additionally, these fungi can produce toxic secondary metabolites, known as mycotoxins. These toxins, primarily produced by mold belonging to the genera *Aspergillus*, *Fusarium*, and *Penicillium*, are the most prevalent natural contaminants in food and feed [1,2].

Cereals and dried fruit are the preferred substrates for toxigenic fungi, and contamination can occur in the field or during storage. It is influenced by a number of factors, including environmental conditions, crop varieties, agricultural practices, harvesting, and storage. Climate change may increase mycotoxin production by affecting these factors [3,4].

All over the world, more than 1000 mycotoxins have been identified, yet less than 20 are regulated. The chronic toxicity of mycotoxins is highly variable, with some, such as aflatoxins (AFs), being hepatotoxic and genotoxic carcinogens, while others have estrogenic, immunotoxic, nephrotoxic, or neurotoxic properties.

Numerous modifications of the 'native' forms of mycotoxins have been described. They may mainly result from biological or chemical transformations occurring by metabolization in animals or in plants [5]. While the effect of 'major' mycotoxins is beginning to be fairly well characterized, this is not true for 'emerging' mycotoxins. The latter are neither routinely determined nor regulated by law. Examples include enniatins, beauvericin, versicolorin, *Alternaria*, and NX toxins [2,3].

This review presents the hazards of main mycotoxins and the health-based guidance values (HBGVs) or reference points (RPs) established at the European level by the European Food Safety Authority (EFSA) for human but also for the different farm and companion animal. It especially focuses on toxins from *Fusarium* species deoxynivalenol (DON), nivalenol (NIV), T-2 and HT-2 toxins, zearalenone (ZEN), and fumonisins (FBs), but also on ochratoxin A (OTA), AFs, and ergot alkaloids (EAs).

## Determination of health-based guidance values and reference point

In order to characterize the mycotoxin hazard, health agencies conducted a comprehensive and meticulous
examination of the available published data on tox-
icokinetics (TK) and toxicity obtained *in vitro* or *in vivo*
[6]. This allows for the determination of the no-ob-
served-adverse-effect level (NOAEL), defined as the
highest dose with no significant adverse effects, and the
lowest-observed-adverse-effect level (LOAEL), defined
as the minimal dose with observed adverse effects. A
novel approach is to use of dose modeling to identify the
dose that produces a predetermined change in response
rate. This is referred to as the benchmark dose (BMD),
with a lower confidence limit (BMDL) [7]. Based on
these values, it is possible to establish HBGVs or, when
this is not feasible, to identify RPs and to compare them
with dietary exposure estimates in order to ascertain the
level of risk.

Because mycotoxins are naturally occurring con-
taminants, HBGVs for human chronic exposure to non-
carcinogenic toxins are characterized by tolerable daily
intakes (TDIs). The TDI represents the amount of a
substance that can be ingested on a daily basis per body
weight over a lifetime without appreciable health risk.
This value is derived from NOAEL or BMDL, with
uncertainty factors of at least 100 applied, to account for
interspecies and intraspecies differences.

In the case of genotoxic carcinogenic mycotoxins (e.g. AFB₁), it is not possible to establish a safety threshold and the ALARA (As Low As Reasonably Achievable) approach is used. However, the risk manager considers that a margin of exposure (MOE) of less than 10 000 between the BMDL and the exposure is of concern for human health [8].

The paucity of information available for farm and com-
panion animals allows only the establishment of RPs.
These are based on the NOAEL or on the LOAEL
divided by a factor of 3. Since the RPs are obtained for
the species considered, no safety factor is added.

Hazard assessment of the selected
mycotoxins

Deoxynivalenol

DON is a type B trichothecene (TCT), produced by *Fusarium graminearum* and *F. culmorum*. It contaminates cereals under temperate climates, during growing in the field [9]. DON co-occurs with several modified forms, especially 3- or 15-acetyl deoxynivalenol (3-Ac-DON or 15-Ac-DON) and the plants-derived DON-3-glucoside (DON-3Glc) [5].

At the molecular level, DON targets the peptidyl trans-
ferase center of the ribosome, blocking protein synthesis
and inhibiting RNA and DNA synthesis [10]. Resulting
activation of ribosome-associated mitogen-activated pro-
tein kinases provokes ribotoxic stress, apoptosis, decrease

in cell survival, or pro-inflammatory cytokine production.
Induced reactive oxygen species production impairs the
antioxidant defense system, increases lipid peroxidation,
and DNA and proteins damages [11,12].

Chronic exposure to DON causes intestinal damages, immunotoxicity, impaired reproduction, and development resulting from maternal toxicity, emesis, anorexia, and growth retardation. These later effects involve pro-inflammatory cytokines and satiety hormones [13]. Since 3- and 15-AcDON can be deacetylated and DON-3G de-glycosylated in the intestine, they are considered to have the same *in vivo* toxicity [14].

In human, a group TDI was established for the sum of DON, 3-Ac-DON, 15-Ac-DON, and DON-3Glc. This HBGV was based on reduction in body weight observed in mice. RPs were also identified for dairy cow, heifer, beef cattle, steer, sheep, goats, pig, laying hen, duck, broiler chicken, turkey, and fish (Table 1).

Nivalenol

NIV is another type B TCT produced by several *Fusarium* species and contaminating mainly cereals. As other TCT, it binds to ribosomes, leading to the inhibition of protein synthesis and mitogen-activated protein kinases activation. Less data exist for NIV compared to DON, but exposure to this mycotoxin has been described to induce immunotoxicity, hematotoxicity, as well as developmental and reproductive toxicity [15].

The modified form NIV-3Glc, potentially representing 50% of NIV in cereals, was assumed to be hydrolyzed to NIV in the intestine and included in the group TDI [16].

For human, this group TDI was based on the decreased white blood cells observed in rat. LOAELs were identified for pigs and poultry allowing the determination of RPs for NIV [17], while toxicity data were lacking for ruminant, rabbit, fish, and pet (Table 1).

T-2 and HT-2 toxins

T-2 and HT-2 toxins are type A TCT, mainly produced by *Fusarium* fungi under moist cool conditions and contaminate cereals. Multiple modified forms of T-2 and HT-2 have been described. These forms, such as neo-solaniol (NEO), T-2-triol, and T-2-tetraol can contribute up to 40% of the total T-2 and HT-2 concentration in cereals and foods and were thus included in the group TDI for human [18].

After absorption, T-2 toxin is rapidly hydroxylated, de-
epoxidized, glucuronidized, or acetylated, and HT-2
toxin is one of its main metabolites. T-2 toxin, like other
TCTs, causes ribotoxic stress by inhibiting protein and
<table><caption>**Table 1**

Human TDI and animal RP determined by EFSA for trichothecenes.</caption><thead><tr><th>Group of mycotoxins</th><th>Species</th><th>TDI or RP</th><th>Endpoint [reference]</th></tr></thead><tbody><tr><td>Grouped DON, 3-Ac-DON and 15-Ac-DON and DON-3Glc</td><td>Human</td><td>1 µg/kg bw/day</td><td>Reduced bodyweight gain in mice [14]</td></tr><tr><td rowspan="10">Grouped DON, 3-Ac-DON and 15-Ac-DON and DON-3Glc</td><td>Dairy cow</td><td>5 mg/kg feed</td><td rowspan="10">Reduced feed intake; reduced body weight gain [14]</td></tr><tr><td>Heifer</td><td>10 mg/kg feed</td></tr><tr><td>Beef cattle, steer</td><td>18 mg/kg feed</td></tr><tr><td>Sheep</td><td>16 mg/kg feed</td></tr><tr><td>Goats</td><td>5 mg/kg feed</td></tr><tr><td>Pig</td><td>0.7 mg/kg feed</td></tr><tr><td>Laying hen</td><td>5 mg/kg feed</td></tr><tr><td>Duck</td><td>7 mg/kg feed</td></tr><tr><td>Broiler chicken</td><td>0.6 mg/kg feed</td></tr><tr><td>Turkey</td><td>0.6 mg/kg feed</td><td>Decreased intestinal villus height, histological damages [43]</td></tr><tr><td>Fish</td><td>0.6 mg/kg feed</td><td>Reduced feed intake, body weight gain; lipid peroxidation; histopathological changes [14]</td></tr><tr><td rowspan="5">Grouped NIV and NIV-3Glc</td><td>Rabbit</td><td>4 mg/kg feed</td><td>Feed intake, body weight gain, and relative organ weights [14]</td></tr><tr><td>Cat</td><td>6 mg/kg feed</td><td>Reduced body weight gain [14]</td></tr><tr><td>Dog</td><td>4 mg/kg feed</td><td>Reduced body weight gain [14]</td></tr><tr><td>Horse</td><td>3.5 mg/kg feed</td><td>Reduced feed intake [38]</td></tr><tr><td>Human</td><td>1.2 µg/kg bw</td><td>Immuno- and hemato-toxicity in rats [16]</td></tr><tr><td rowspan="5">Grouped NIV and NIV-3Glc</td><td>Pig</td><td>0.8 mg/kg feed<br>32 µg/kg b.w./day</td><td>Hematological alteration; reduced proliferation of lymphocytes from blood, spleen, or thymus [17]</td></tr><tr><td>Broiler chicken</td><td>1 mg/kg feed<br>120 µg/kg b.w./day</td><td>Alteration of the gizzard [17]</td></tr><tr><td>Laying hen</td><td>0.3 mg/kg feed<br>15.9 µg/kg<br>b.w./day</td><td>Histological alteration of the kidney [17]</td></tr><tr><td>Human</td><td>0.02 µg/kg bw</td><td>Reduction in total leucocyte count in rat [18]</td></tr><tr><td>Cow</td><td>0.02 mg/kg feed<br>3.3 µg/kg b.w./day</td><td>Rumen acidosis [22]</td></tr><tr><td rowspan="10">Grouped T-2 and HT-2 and its modified forms<sup>a</sup></td><td>Lactating sheep</td><td>0.01 mg/kg feed<br>0.33 µg/kg<br>b.w./day</td><td>Rumen acidosis [22]</td></tr><tr><td>Goats</td><td>0.02 mg/kg feed<br>3.3 µg/kg b.w./day</td><td>Rumen acidosis [22]</td></tr><tr><td>Calve, lamb</td><td>100 µg/kg<br>b.w./day</td><td>Gastrointestinal lesions and alteration of serum proteins and hematology [21]</td></tr><tr><td>Pig</td><td>0.167 mg/kg feed<br>9.6 µg/kg b.w./day</td><td>Immunological or hematological effects [21]</td></tr><tr><td>Broiler chicken</td><td>0.167 mg/kg feed<br>13.3 µg/kg<br>b.w./day</td><td>Mucosal oral damage [21]</td></tr><tr><td>Laying hen</td><td>0.66 mg/kg feed<br>40 µg/kg b.w./day</td><td>Infertility, reduced egg production [21]</td></tr><tr><td>Duck</td><td>0.06 mg/kg feed<br>13.3 µg/kg<br>b.w./day</td><td>Reduced body weight gain [21]</td></tr><tr><td>Turkey</td><td>0.16 mg/kg feed<br>16 µg/kg b.w./day</td><td>Mucosal oral damage [21]</td></tr><tr><td>Fish</td><td>0.63 mg/kg feed<br>13 µg/kg b.w./day</td><td>Reduced feed intake and growth; alteration of hematocrit [21]</td></tr><tr><td>Rabbit</td><td>100 µg/kg<br>b.w./day</td><td>Alteration of drug-metabolizing enzymes; microsomal oxidative damages [21]</td></tr></tbody></table>

In italics RPs established from LOAELs.

<footnote>a T-2 HT-2 modified forms (and their relative potency factor): 19-HO-T-2 (1), NEO (0.3), 19-HO-HT-2 (0.3), T-2-triol (0.1), T-2-tetraol (0.1), T-2-3-Glc (1), T-2-3-diGlc (1), T-2-3-Sulf (1), T-2-3-GlcA (1), 3-Ac-T-2 (1), 3-Fer-T-2 (1), HT-2-3-Glc (1), HT-2-diGlc (1), HT-2-GlcA (1), HT-2-MalGlc (1), NEO-Glc (0.3), T-2-triol-Glc (0.1), T-2-tetraol-Glc (0.1).</footnote>
RNA/DNA synthesis. It leads to apoptosis, necrosis, lipid peroxidation, and cell membrane damage [19].

T-2 and HT-2 toxins cause weight loss, emesis, diarrhea, lethargy, immunosuppression, hemorrhage, and liver or stomach lesions in animals [20]. For humans, a group TDI for T-2 and HT-2 and its modified forms was established based on a pivotal rat study showing leukocyte count reduction [18] (Table 1). The modified forms being less toxic than T-2 and HT-2, relative potency factor, less than 1, were used for this group TDI [18].

Data on animals are scarce. However, for T-2 and HT-2, RP could be identified for ruminants (cow, sheep, and goats), poultry (broiler chicken, laying hen, duck, and turkey), fish, and rabbit (Table 1). No RP was identified for cat, dog, and horse [21,22].

## Zearalenone

ZEN is also produced by *F. graminearum* and *F. culmorum*. ZEN and its modified forms are predominantly found in cereal crops, cereal grains, and by-products [23]. Because it shares structural similarities with 17β-estradiol, it competes at the estrogen receptor and triggers a hyperestrogenic response. It acts as potential endocrine disruptor by altering the production of steroid sex hormones, affecting reproductive function. In animals, reproductive disorders have been observed, and prolonged exposure also leads to liver lesions, immunotoxicity, nephrotoxicity, and lipid peroxidation. Because of their ability to metabolize this toxin, sensitivity varies between species, with pigs being particularly sensitive [24].

Many modified forms of ZEN have been described, for which EFSA has established relative potency factors ranging from 0.2 for β-Zearalenol (β-ZEL) to 60 for α-Zearalenol (α-ZEL) [23].

In human, a group TDI of 0.25 µg/kg bw/day for ZEN and its modified forms was set based on estrogenic effects on pigs [25]. The RP established for animals (sheep, pig, poultry, fish, dog) reflects their different metabolism and susceptibility to ZEN. No RP were established for cattle, duck, goats, horse, rabbit, mink, and cat. Despite the lack of toxicology data for modified forms of ZEN in animal species, the same relative potency factor than the one determined for human were used (Table 2).

## Fumonisins

FBs are produced by *F. verticilioides* and *F. proliferatum* in maize and *Aspergillus niger* in grapes. This family includes over 30 members, with FB₁, FB₂, FB₃, and FB₄ being the most common and FB₁ the most abundant. Due to their biochemical similarity to ceramide, sphinganine (Sa), and sphingosine (So), FBs inhibit ceramide

synthases, disrupting sphingolipid metabolism. This affects cellular signaling, membrane function, growth, and differentiation and can lead to cell injury and apoptosis. The Sa/So ratio is a biomarker of FBs [26]. FBs can strongly interact with food or feed matrix macroconstituents by noncovalent binding, resulting in the so-called hidden forms. These forms are assumed to account for 60% of the dietary concentration of FBs and should therefore be considered in the exposure by multiplying the FBs concentration by a factor of 1.6 [27].

In humans, FBs are associated with neural tube defect, growth impairment but also esophageal and liver cancer. They are classified as ‘possibly carcinogenic to humans’ (group 2B) by the International Agency for Research on Cancer (IARC). In animals, FBs cause nephrotoxicity, hepatotoxicity, immunotoxicity, and intestinal barrier disturbances and provoke leukoencephalomalacia in horses and pulmonary edema in pigs [9].

In human, a group TDI (FB₁+FB₂+FB₃+FB₄) was derived from the liver toxicity observed in mice [27]. Different adverse effects were selected for animal species [28]; they were used to determine group RPs for FB₁+FB₂+FB₃ in cattle, pig, poultry, fish, rabbit, and horse (Table 2).

## Ochratoxins

OTA, the most widespread mycotoxin of ochratoxin (OT) family, is produced by various *Aspergillus* and *Penicillium* species under temperate and tropical climates. It contaminates wheat, grapes, coffee, cocoa, but also meat and cheese. Derivatives such as OTB, OTα, and OTC are produced in smaller quantities and are less toxic than OTA. Due to its similarity to phenylalanine, it inhibits phenylalanyl-tRNA synthetase, leading to decreased protein synthesis, mitochondrial respiration, cellular ATP depletion, disrupted calcium homeostasis, lipid peroxidation, and oxidative damage. OTA induces genetic mutations, DNA breaks, and chromosomal damage [29]. The carcinogenic mechanisms are still unclear, but OTA may cause DNA damage through oxidative stress [30].

OTA is linked to kidney diseases in humans and animals and classified as ‘possibly carcinogenic to humans’ (group 2B) by IARC. Thus, EFSA preferred to use a MOE. For neoplastic effects, it was based on renal tumor (adenoma and carcinoma) in rat (BMDL₁₀ = 14.5 µg/kg bw/day, and an MOE of 10 000). For non-neoplastic effects, it was based on kidney lesions observed in pigs (BMDL₁₀ = 4.73 µg/kg bw/day and an MOE of 200). RP were also identified for pig, chicken for fattening, hen, fish, and rabbits (Table 3). No RPs were identified for ruminant, soliped, and other poultry species [31].
<table><caption>**Table 2**

Human TDI and animal RP determined by EFSA for other *Fusarium* toxins.</caption><thead><tr><th>Group of mycotoxins</th><th>Species</th><th>TDI or RP</th><th>Endpoint</th></tr></thead><tbody><tr><td>Grouped ZEN and its modified forms<sup>a</sup></td><td>Human</td><td>0.25 µg/kg<br>b.w./day</td><td>Estrogenicity in pigs on uterus and vulva [23]</td></tr><tr><td rowspan="2">Grouped ZEN and its modified forms<sup>a</sup></td><td>Sheep</td><td>1.5 mg/kg feed<br>28 µg/kg<br>b.w./day</td><td>Alteration of ovulation rates and lambing percentages [25]</td></tr><tr><td>Pig</td><td>0.22 mg/kg feed<br>0.4 µg/kg<br>b.w./day</td><td>Estrogenic effects [25]</td></tr><tr><td></td><td>Broiler chicken</td><td>50 mg/kg feed<br>7500 µg/kg<br>b.w./day</td><td>Reduced lymphocyte counts [25]</td></tr><tr><td></td><td>Turkey</td><td>100 mg/kg feed<br>9100 µg/kg<br>b.w./day</td><td>Vent swelling [25]</td></tr><tr><td></td><td>Fish</td><td>0.3 mg/kg feed<br>9 µg/kg b.w./day</td><td>Alteration of blood formula; lipid peroxidation [25]</td></tr><tr><td></td><td>Dog</td><td>0.57 mg/kg feed<br>8.8 µg/kg<br>b.w./day</td><td>Degenerative lesions of myometrium and endometrium; enlargement and atrophy of uterine glands; changes in blood formula; alteration of biochemical parameters [25]</td></tr><tr><td>Grouped FB<sub>1</sub>, FB<sub>2</sub>, FB<sub>3</sub>, and FB<sub>4</sub></td><td>Human</td><td>1 µg/kg b.w./day</td><td>Increased incidence of megalocytic hepatocytes in mice [28]</td></tr><tr><td rowspan="10">Grouped FB<sub>1</sub>, FB<sub>2</sub>, and FB<sub>3</sub></td><td>Cattle</td><td>31 mg/kg feed<br>600 µg/kg<br>b.w./day</td><td>Increase in serum enzymes, cholesterol, and bilirubin; decrease in lymphocyte blastogenesis [27]</td></tr><tr><td>Pig</td><td>1 mg/kg feed<br>40 µg/kg<br>b.w./day</td><td>Effect on performances and lung lesions [45]</td></tr><tr><td>Chicken</td><td>1 mg/kg feed<br>2600 µg/kg<br>b.w./day</td><td>Decrease in crypt depth [45]</td></tr><tr><td>Duck</td><td>8 mg/kg feed<br>400 µg/kg<br>b.w./day</td><td>Alterations of liver enzymes [45]</td></tr><tr><td>Turkey</td><td>20 mg/kg feed<br>670 µg/kg<br>b.w./day</td><td>Feed consumption; increased Sa/So ratios in liver and kidney [27]</td></tr><tr><td>Fish</td><td>3 mg/kg feed<br>0.12 mg/kg<br>b.w./day</td><td>Reduced weight gain; neuronal apoptosis in brain [27]</td></tr><tr><td>Rabbit</td><td>40 µg/kg feed</td><td>Decreased performance; alteration of plasma markers and blood formula; congestion of liver and kidney; impaired spermatogenesis, delayed puberty; increased Sa/So ratio [27]</td></tr><tr><td>Horse, other solipeds</td><td>1 mg/kg feed</td><td>Histopathological brain lesions<br>Equine leucoencephalomalacia<br>Neurological and cardiovascular effects [45]</td></tr></tbody></table>

In italics RPs established from LOAELs.

## Aflatoxins

AFs (AFB<sub>1</sub>, AFB<sub>2</sub>, AFG<sub>1</sub>, AFG<sub>2</sub>), produced by *Aspergillus* fungi, mainly *A. flavus* and *A. parasiticus*, in hot and humid climates, contaminate corn, wheat, rice, nuts, oilseeds, and spices. Climate change is likely to have a negative impact on the contamination of cereal-based foodstuffs in Europe. AFB<sub>1</sub>, the most prevalent AF, is metabolized in the liver to a genotoxic epoxide that binds to DNA, causing mutations and cancer [29,32]. AFB<sub>1</sub> is also metabolized in mammals or poultry to AFM<sub>1</sub> and excreted in milk [33].

AFB<sub>1</sub> induces oxidative stress and apoptosis, modulates key nuclear receptor expression, and alters DNA methylation, affecting tumor suppressor genes. Exposure to high doses, which is not a concern in Europe, leads to acute hepatitis, causing vomiting, anorexia, abdominal pain, fever, diarrhea, jaundice, severe liver damage, hemorrhage, edema, and death. Exposure to subchronic doses of AFB<sub>1</sub> causes immunosuppression, nutritional dysfunctions, and growth impairment. AFB<sub>1</sub>, AFB<sub>2</sub>, AFG<sub>1</sub>, and AFG<sub>2</sub> induced hepatocarcinoma in humans and are classified as ‘carcinogenic to human’ (group 1) by

<footnote><sup>a</sup> ZEN modified forms (and their relative potency factors): ZENGics (1), ZEN Sulfs (1), α-ZEL (60), α-ZELGics (60), α-ZELSulfs (60), β-ZEL (0.2), β-ZELGics (0.2), β-ZELSulfs (0.2), ZAN (1.5), ZANGics (1.5), ZANSulfs (1.5), α-ZAL (4), α-ZALGics (4), α-ZALSulfs (4), β-ZAL (2), β-ZALGics (2), β-ZALSulfs (2), cis-ZEN (1), cis-ZENGics (1), cis-ZENSulfs (1), cis-a-ZEL (8), cis-α-ZELGics (8), cis-α-ZELSulfs (8), cis-β-ZEL (1), cis-β-ZELGics (1) and cis-β-ZELSulfs (1).</footnote>
<table><caption>**Table 3**</caption><caption>Human BMDL and animal RP determined by EFSA for OTA.</caption><thead><tr><th>Mycotoxin</th><th>Species</th><th>BMDL or RP</th><th>Endpoint</th></tr></thead><tbody><tr><td rowspan="2">OTA</td><td rowspan="2">Human</td><td>Neoplastic effects BMDL<sub>10</sub> = 4.73 µg/kg b.w./day (MOE of 10 000)</td><td>Microscopic kidney lesions in pigs [30]</td></tr><tr><td>Non-neoplastic effects BMDL<sub>10</sub> of 14.5 µg/kg b.w./day (MOE of 200)</td><td>Formation of kidney tumors in rats [30]</td></tr><tr><td rowspan="5">OTA</td><td>Pig</td><td>0.01 mg/kg feed</td><td>Alteration of the function and the structure of both kidney and liver [31]</td></tr><tr><td>Chicken for fattening</td><td>0.03 mg/kg feed</td><td>Increased liver and kidney weight associated with lesions, decreased thymus weight, immunosuppression [31]</td></tr><tr><td>Hen</td><td>0.03 mg/kg feed</td><td>Reduced egg mass production, deterioration feed to egg ratio [31]</td></tr><tr><td>Fish</td><td>0.5 mg/kg feed</td><td>Alteration of zootechnical performances and hepatopancreatic tissues [31]</td></tr><tr><td>Rabbit</td><td>0.01 mg/kg feed</td><td>Alteration of zootechnical performances, intestinal morphology, blood hematology, and chemistry [31]</td></tr></tbody></table>

In italics RPs established from LOAELs.

IARC. AFM1 is less potent and classified in group 2B (possible carcinogen) but still poses safety concerns in milk [34].

As AFB₁ is a genotoxic carcinogen, no TDI could be established for human. Based on hepatocellular carcinomas observed in male rat, a BMDL₁₀ of 0.4 µg/kg bw/day was set and used for risk characterization using an MOE (MOE ≥ 10 000) approach.

## Ergot alkaloids

EAs are mainly produced by *Claviceps*, *Aspergillus*, and *Penicillium* within compact mycelium mass called sclerotium. Over 50 different EAs are identified [35]. Those associated with wheat, rye, triticale, barley, millets, and oats are lysergic acid amides such as ergometrine, ergotamine, ergosine, ergocristine, α- and β-ergocryptine, and ergocornine. The C9 = C10 double bond easily epimerizes from '-ines' to '-inines' based on temperature and pH. Those 14 EAs are assumed to have the same toxicity [36,37].

EAs share a similar structure to neurotransmitters such as dopamine, noradrenaline, and serotonin. EAs are used or tested for prolactin inhibition, treatment of Parkinson's disease, cerebrovascular or venous insufficiency, thrombosis, migraine, and uterine stimulation [36]. They reduce feed intake, growth, and reproduction and cause convulsions, muscle spasms, circulatory disorders, and dry gangrene [38].

For humans, a grouped TDI for 14 EAs was set from a pivotal study dealing with rats receiving ergotamine and displaying muscle atrophy in tails, with no toxic effect at 20 mg EAs/kg diet (0.9 mg ergotamine/kg bw/day). RPs for the 14 EAs were also set for pig, broiler chickens, and cattle (Table 4).

## Discussion

This review presents the HBGVs and RPs determined at the European level by EFSA for the main mycotoxins of concern for humans and for farm and companion animals. It should be noted that in addition to the regulated mycotoxins, the so-called emerging mycotoxins are neither routinely tested nor regulated [3]. Among these toxins, some, such as versicolorin or NX toxins, are poorly documented, and further data are required to establish HBGV/RP [39,40]. Conversely, in light of the current research on other toxins, we can expect that HBGVs and RPs will be established for some emerging mycotoxins, especially beauvericin, enniatins, versicolorin, and *Alternaria* toxins.

Mycotoxins are natural contaminants and are thus not subject to regulatory testing. For substances with pre-market authorization, many tests are required to assess their safety in terms of absorption, distribution, metabolism, excretion, acute and short-term toxicity, genotoxicity, long-term toxicity, carcinogenicity, reproductive toxicity, neurotoxicity, and overall impact on human health and the environment [41]. On the contrary, as natural compounds, mycotoxins are not subject to voluntary use or mandatory testing for regulatory purposes. Health authorities rely on available published data from *in vivo* tests to establish toxicological reference doses. Unfortunately, these data are not always performed for this purpose and are not always of very good quality. They are rarely conducted according to Organisation for Economic Co-operation and Development guidelines. For the determination of human HBGVs, appropriate animal models should be selected based on the results of studies performed not only on laboratory species but also on relevant farm and companion animals. In addition, data obtained in one species cannot be generalized to another due to considerable species-related differences in TK and toxicodynamics (TD). So, poultry includes
<table><caption>**Table 4**

Human TDI and animal RP determined by EFSA for EAs.</caption><thead><tr><th>Group of mycotoxins</th><th>Species</th><th>TDI or RP</th><th>Endpoint</th></tr></thead><tbody><tr><td>EAs<sup>a</sup></td><td>Human</td><td>0.6 µg/kg bw</td><td>Tail muscular atrophy in rats [35]</td></tr><tr><td rowspan="5">EAs<sup>a</sup></td><td>Pig</td><td>0.6 mg/kg feed</td><td>Increased relative heart and spleen weight [36]</td></tr><tr><td>Chicken for fattening</td><td>2.1 mg/kg feed</td><td>Reduced weight gain [36]</td></tr><tr><td>Laying hen</td><td>3.7 mg/kg feed</td><td>Various effects on eggs [36]</td></tr><tr><td>Duck</td><td>0.2 mg/kg feed</td><td>Reduced weight gain [36]</td></tr><tr><td>Sheep</td><td>0.3 mg/kg feed</td><td>Reduced growth [36]</td></tr><tr><td></td><td>Cattle</td><td>0.1 mg/kg feed</td><td>Loss of tail switches and tips, early lameness, feet swelling, reduced growth, and feed intake [36]</td></tr></tbody></table>

In italics RPs established from LOAELs.

different genera (duck, chicken, turkey...) with different susceptibilities to mycotoxins. Similarly, generic word 'fish' groups different families, genera, and species (trout, salmon, carp...) whose exposure and sensitivity may differ. Furthermore, despite its importance, the possible gender-dependent response is often overlooked in toxicological studies on mycotoxins [42,43]. In most cases, it is a challenge to conduct comprehensive risk assessment for all animal species.

As shown in the various tables presented in this review, RPs for animal species are very often based on zoo-technical parameters (weight gain, feed consumption), which in most cases are not very specific and not very sensitive parameters. In addition, guidelines should provide a better definition of appropriate parameters/biomarkers (including at the molecular level), which could be unambiguously classified as adverse effects.

We can therefore imagine that in the future, more specific effects will be described for mycotoxins and that these effects will be observed at lower doses. This should lead to the establishment of lower RPs, as it was done in 2022 for DON in poultry. Indeed, for broiler chickens, the LOAEL decreased from 5 to 1.9 mg/kg feed when the effect of this mycotoxin on the intestine was taken into consideration [44].

In addition to ethical considerations, toxicity testing on large animals for regulatory purposes is costly. The use of published case reports is a recognized alternative procedure that should be implemented. We also believe that approaching the health impact of mycotoxins from a broader toxico-epidemiological perspective would enable us to refine the RPs for animals. Indeed, clinical cases of mycotoxin poisoning are only occasionally studied in detail. In cattle, for example, a species very frequently exposed [45], few studies established a correlation between the level of mycotoxin exposure and clinical signs in animals in relation to the onset or severity of livestock pathologies. Increased toxico-epidemiological data would enable a better correlation between animal exposure and

the onset of pathologies, thus improving mycotoxin risk assessment. Recently, the inclusion of clinical cases has made it possible to refine fumonisin RP for horses from 8.8 to 1 mg/kg feed [46].

With societal pressure to reduce the use of animals for scientific purposes, future toxicology testing is likely to rely more on *in vitro* and *in silico* testing using advanced analytical techniques, computational methods, high-throughput screening, and omics applications. Open access platforms such as TK plate 1.0 or EFSA TK plate 2.0, which allow the prediction of TK and TD of mycotoxins, including residues in animal products, are promising tools for improving the sensitivity of risk assessment for potentially harmful compounds, including mycotoxins [47].

Up to now, reference values have mainly been proposed for individual mycotoxins. However, human and animal are not exposed to a single toxin but to several simultaneously. Combined effects must therefore be taken into account to better reflect consumer exposure.

As we have pointed out in this review, when there is sufficient knowledge of their mechanism of toxicity, TK, or structural similarity [48], toxins with similar properties can be grouped together to obtain group HBGV/RP [49]. This has already been done for parent toxins and their modified forms as well as the different FBs and AFs. We believe that this approach represents a major evolution in the field and could be extended to make a grouped HBGV/RP for TCTs. It should be noted, however, that this approach assumes an additive effect, whereas some studies have shown synergistic effects even between TCTs [50].

In conclusion, the determination of HBGV/RP for mycotoxins is crucial for public and animal health, as it sets limits to guarantee food and feed safety. Data from animal testing, albeit limited due to the absence of regulatory constraints, serve as the basis. We believe that the evolution of evaluation criteria and technological advances will continually improve the assessment of

<footnote><sup>a</sup> EAs considered: ergocristine, ergotamine, ergocornine, α- and β-ergocryptine, ergometrine, ergosine, and their corresponding 'inine' epimers.</footnote>
mycotoxin toxicity and that understanding their mode of
action will enable toxins with similar properties to be
grouped together.

Data Availability

No data were used for the research described in the ar-
ticle.

**Declaration of Competing Interest**

The authors declare that they have no known competing
financial interests or personal relationships that could
have appeared to influence the work reported in this
paper.

Acknowledgements

This work was supported by grants from the French Agence Nationale de la Recherche project GenoMyc (ANR-22-CE34) and the INRAE-Inserm project "Exposome et Santé" - UriTox.

**References and recommended reading**

Papers of particular interest, published within the period of review, have been highlighted as:

• of special interest

•• of outstanding interest

1. Khan R: Mycotoxins in food: occurrence, health implications, and control strategies—a comprehensive review. *Toxicon* 2024, **248**:108038, https://doi.org/10.1016/j.toxicon.2024.108038

2. Payros D, Garofalo M, Pierron A, Soler-Vasco L, Al-Ayoubi C, Maruo VM, et al.: Mycotoxins in human food: a challenge for research. *Cah Nutr Diet* 2021, **56**:170-183, https://doi.org/10.1016/j.cnd.2021.02.001

3. Gruber-Dorninger C, Novak B, Nagl V, Berthiller F: Emerging mycotoxins: beyond traditionally determined food contaminants. *J Agric Food Chem* 2017, **65**:7052-7070, https://doi.org/10.1021/acs.jafc.6b03413

4. Casu A, Camardo Leggieri M, Toscano P, Battilani P: Changing climate, shifting mycotoxins: a comprehensive review of climate change impact on mycotoxin contamination. *Compr Rev Food Sci Food Saf* 2024, **23**:e13323, https://doi.org/10.1111/1541-4337.13323

5. Berthiller F, Crews C, Dall'Asta C, Saeger SD, Haesaert G, Karlovsky P, et al.: Masked mycotoxins: a review. *Mol Nutr Food Res* 2013, **57**:165-186, https://doi.org/10.1002/mnfr.201100764

6. Eskola M, Altieri A, Galobart J: Overview of the activities of the European Food Safety Authority on mycotoxins in food and feed. *World Mycotoxin J* 2018, **11**:277-289, https://doi.org/10.3920/WMJ2017.2270

7. More SJ, Bampidis V, Benford D, Bragard C, Halldorsson TI, Hernández-Jerez AF, et al.: Use of the benchmark dose approach in risk assessment. *EFSA J* 2022, **20**:7584, https://doi.org/10.2903/j.efsa.2022.7584

8. EFSA: Applicability of the Margin of Exposure approach for the safety assessment of impurities which are both genotoxic and carcinogenic in substances added to food/feed. EFSA J 2012, **10**:2578, https://doi.org/10.2903/j.efsa.2012.2578

9. Marin S, Ramos AJ, Cano-Sancho G, Sanchis V: **Mycotoxins: occurrence, toxicology, and exposure assessment. Food Chem Toxicol** 2013, **60**:218-237, https://doi.org/10.1016/j.fct.2013.07.047

10. Garreau de Loubresse N, Prokhorova I, Holtkamp W, Rodnina MV, Yusupova G, Yusupov M: **Structural basis for the inhibition of the eukaryotic ribosome. Nature** 2014, **513**:517-522, https://doi.org/10.1038/nature13737

11. Garofalo M, Payros D, Taieb F, Oswald E, Nougayrède JP, Oswald IP: **From ribosome to ribotoxins: understanding the toxicity of deoxynivalenol and Shiga toxin, two food borne toxins. Crit Rev Food Sci Nutr** 2025, **65**:193-205, https://doi.org/10.1080/10408398.2023.2271101

12. Mishra S, Dwivedi PD, Pandey HP, Das M: Role of oxidative stress in Deoxynivalenol induced toxicity. *Food Chem Toxicol* 2014, **72**:20-29, https://doi.org/10.1016/j.fct.2014.06.027

13. Terciolo C, Maresca M, Pinton P, Oswald IP: Role of satiety hormones in anorexia induction by Trichothecene mycotoxins. *Food Chem Toxicol* 2018, **121**:701-714, https://doi.org/10.1016/j.fct.2018.09.034 Epub 2018 Sep 20.

14. Knutsen HK, Alexander J, Barregard L, Bignami M, Bruschweiler B, Ceccatelli S, et al.: Risks to human and animal health related to the presence of deoxynivalenol and its acetylated and modified forms in food and feed. EFSA J 2017, 15:4718, https://doi.org/10.2903/j.efsa.2017.4718

15. Kumar P, Mahato DK, Gupta A, Pandey S, Paul V, Saurabh V, et al.: An overview on occurrence, impact on human and animal health and its detection and management strategies. *Toxins* 2022, **14**:527, https://doi.org/10.3390/toxins14080527

16. Knutsen HK, Barregard L, Bignami M, Bruschweiler B, Ceccatelli S, Cottrill B, et al.: Appropriateness to set a group health-based guidance value for nivalenol and its modified forms. EFSA J 2017, 15:4751, https://doi.org/10.2903/j.efsa.2017.4751

17. EFSA: Risks for animal and public health related to the presence of nivalenol in food and feed. EFSA J 2013, 11:3262, https://doi.org/10.2903/j.efsa.2013.3262

18. Knutsen HK, Barregård L, Bignami M, Brüschweiler B, Ceccatelli S, Cottrill B, et al.: Appropriateness to set a group health-based guidance value for T2 and HT2 toxin and its modified forms. EFSA J 2017, 15:4655, https://doi.org/10.2903/j.efsa.2017.4655

19. Wu Q, Qin Z, Kuca K, You L, Zhao Y, Liu A, et al.: An update on T-2 toxin and its modified forms: metabolism, immunotoxicity mechanism, and human exposure assessment. *Arch Toxicol* 2020, **94**:3645-3669, https://doi.org/10.1007/s00204-020-02899-9

20. Escrivá L, Font G, Manyes L: In vivo toxicity studies of fusarium mycotoxins in the last decade: a review. *Food Chem Toxicol* 2015, **78**:185-206, https://doi.org/10.1016/j.fct.2015.02.005

21. EFSA: Risks for animal and public health related to the presence of T-2 and HT-2 Toxin in food and feed. EFSA J 2011, 9:2481, https://doi.org/10.2903/j.efsa.2011.2481

22. Schrenk D, Bignami M, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, et al.: Assessment of information as regards the toxicity of T-2 and HT-2 toxin for ruminants. EFSA J 2022, 2022, **20**:7564, https://doi.org/10.2903/j.efsa.2022.7564

23. EFSA: Appropriateness to set a group health-based guidance value for zearalenone and its modified forms. EFSA J 2016, **14**:4425, https://doi.org/10.2903/j.efsa.2016.4425

24. Balló A, Busznyákné Székvári K, Czétány P, Márk L, Török A, Szántó Á, Máté G: Estrogenic and non-estrogenic disruptor effect of zearalenone on male reproduction: a review. *Int J Mol Sci* 2023, **24**:1578, https://doi.org/10.3390/ijms24021578

25. Knutsen HK, Alexander J, Barregard L, Bignami M, Bruschweiler B, Ceccatelli S, et al.: Risks for animal health related to the presence of zearalenone and its modified forms in feed. EFSA J 2017, 15:4851, https://doi.org/10.2903/j.efsa.2017.4851

26. Riley RT, Merrill AH Jr.: Ceramide synthase inhibition by fumonisins: a perfect storm of perturbed sphingolipid metabolism, signaling, and disease. *J Lipid Res* 2019, **60**:1183-1189, https://doi.org/10.1194/jlr.S093815

27. Knutsen HK, Alexander J, Barregard L, Bignami M, Bruschweiler B, Ceccatelli S: Risks for animal health related to the presence of fumonisins, their modified forms and hidden forms in feed. EFSA J 2018, **16**:5242, https://doi.org/10.2903/j.efsa.2018.5242

28. Knutsen H-K, Barregard L, Bignami M, Bruschweiler B, Ceccatelli S, Cottrill B, et al.: Appropriateness to set a group health-based guidance value for fumonisins and their modified forms. EFSA J 2018, **16**:5172, https://doi.org/10.2903/j.efsa.2018.5172
29. Nazareth TM, Soriano Pérez E, Luz C, Meca C, Quiles JM: **Comprehensive review of aflatoxin and ochratoxin a dynamics: emergence, toxicological impact, and advanced control strategies.** *Foods* 2024, 13:1920, https://doi.org/10.3390/foods13121920

30. Schrenk D, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, Hogstrand C, et al.: **Risk assessment of ochratoxin A in food.** *EFSA J* 2020, **18**:6113, https://doi.org/10.2903/j.efsa.2020.6113

31. Schrenk D, Bignami M, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, et al.: **Risks for animal health related to the presence of ochratoxin A (OTA) in feed.** *EFSA J* 2023, **21**:1-89, https://doi.org/10.2903/j.efsa.2023.8375.

As the previous assessment was almost 20 years old, the authors of this Scientific Opinion have reassessed the risks to animal health and transfer from feed to food of animal origin associated with the presence of OTA in feed. This work confirmed that OTA is most animal species absorbed in the gastrointestinal tract. The effects on performance, as well as on the liver and kidneys, led to the determination of RPs for pigs, chickens, rabbits, and laying hens.

32. Marchese S, Polo A, Ariano A, Velotto S, Costantini S, Severino L: **Aflatoxin B1 and M1: biological properties and their involvement in cancer development.** *Toxins* 2018, **10**:214, https://doi.org/10.3390/toxins10060214

33. Yuan Y, Li M, Qiu X: **Chicken CYP1A5 is able to hydroxylate aflatoxin B1 to aflatoxin M1** (https://doi.org/10.1016/j.toxicon.2024.107625, https://doi.org/10.1016/j.toxicon.2024.107625

34. Schrenk D, Bignami M, Bodin L, Chipman JK, Del Mazo J, Grasl-Kraupp B, et al.: **Risk assessment of aflatoxins in food.** *EFSA J* 2020, **18**:e06040, https://doi.org/10.2903/j.efsa.2020.6040.

Authors of this Scientific Opinion estimate European dietary exposure and evaluate the associated human health risks. They encourage the production of data to derive potency factors for different AFs and to quantify the relationship between individual exposure and biomarker levels. This demonstrates the vigilance of food safety agencies to the problem of the presence of AFs, in the light of potential increase due to climate change.

35. Orlando B, Maumene C: **Ergot and ergot alkaloids in French cereals: occurrence, pattern and agronomic practices for managing the risk.** *World Mycotoxin J* 2017, **10**:327, https://doi.org/10.3920/WMJ2017.2183

36. **EFSA: Ergot alkaloids in food and feed.** *EFSA J* 2012, **10**:2798, https://doi.org/10.2903/j.efsa.2012.2798

37. Schrenk D, Bignami M, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, et al.: **Risks for animal health related to the presence of ergot alkaloids in feed.** *EFSA J* 2024, **22**:e8496, https://doi.org/10.2903/j.efsa.2024.8496.

This Scientific Opinion focuses on the risks for animal health related to the presence of EAs in feed. Exposure to EAs leads to vasoconstriction and to a decrease in prolactin, resulting in a reduced milk production. Considering the sum of the EAs, the authors determined RPs for adverse animal health effects for pigs, chickens for fattening and hens, ducks, bovines and sheep.

38. Cherewyk JE, Blakley BR, Al-Dissi AN: **The C-8-S-isomers of ergot alkaloids — a review of biological and analytical aspects.** *Mycotoxin Res* 2024, **40**:1-17, https://doi.org/10.1007/s12550-023-00507-0

39. Pierron A, Neves M, Puel S, Lippi Y, Soler L, Miller JD, Oswald IP: **Intestinal toxicity of the new type A trichothecenes, NX and 3ANX.** *Chemosphere* 2022, **288**:132415, https://doi.org/10.1016/j.chemosphere.2021.132415

40. Al-Ayoubi C, Rocher O, Naylies C, Lippi Y, Vignard J, Puel S, et al.: **Moore tan a mutagenic Aflatoxin B1 precursor: the multiple cellular targets of Versicolorin A revealed by global gene expression analysis.** *Environ Pollut* 2024, **363**:125138, https://doi.org/10.1016/j.envpol.2024.125138

41. **European Union: Commission Regulation (EU) N°283/2013 of 1 March 2013 Setting Out the Data Requirements for Active Substances, in Accordance With Regulation (EC) N°1107/2009 of the European Parliament and of the Council Concerning the Placing of Plant Protection Products on the Market; OJEU 2013; L 93/1-84.**

42. Soler L, Oswald IP: **The importance of accounting for sex in the search of proteomic signatures of mycotoxin exposure.** *J Proteom* 2018, **178**:114-122, https://doi.org/10.1016/j.jprot.2017.12.017

43. Thenuwara G, Javed B, Singh B, Byrne HJ, Tian F: **Sex- and gender-specific considerations in mycotoxin screening: assessing differential exposure, health impacts, and mitigation strategies.** *Microbiol Res* 2024, **15**:2455-2492, https://doi.org/10.3390/microbiolres15040165.

Authors argues that, despite extensive literature on their general impacts, mycotoxin research lacks sex- and gender-specific data. They emphasize the need to consider sex- and gender-specific differences in exposure risk, susceptibility and health outcomes in mycotoxin screening and mitigation strategies.

44. Schrenk D, Bignami M, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, et al.: **Assessment of information as regards the toxicity of deoxynivalenol for horses and poultry.** *EFSA J* 2023, **21**:7806, https://doi.org/10.2903/j.efsa.2023.7806.

Authors re-evaluated existing studies and newly available literature to revise established benchmarks for DON in horses and poultry, considering not only its effects on reduced feed intake and weight gain but also those on the jejunum, including reduced villi height and histological lesions.

45. Chhaya RS, O'Brien J, Nag R, Cummins E: **Prevalence and concentration of mycotoxins in bovine feed and feed components: a global systematic review and meta-analysis.** *Sci Total Environ* 2024, **929**:172323, https://doi.org/10.1016/j.scitotenv.2024.172323

46. Schrenk D, Bignami M, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, et al.: **Assessment of information as regards the toxicity of fumonisins for pigs, poultry and horses.** *EFSA J* 2022, **20**:7534, https://doi.org/10.2903/j.efsa.2022.7534.

Authors review the information regarding the toxicity of FBs for horses with the aim, if necessary, of revising the established NOAEL of 8.8 mg/kg feed. They re-examine the data including case studies and recent case reports and conclude that the leukoencephalomalacia might be induced by fumonisin concentrations lower than the previously established NOAEL and derived a RP of 1 mg/kg feed.

47. Schmeisser S, Miccoli A, von Bergen M, Berggren E, Braeuning A, Busch W, et al.: **New approach methodologies in human regulatory toxicology - Not if, but how and when!** *Environ Int* 2023, **178**:108082, https://doi.org/10.1016/j.envint.2023.108082.

Authors review the evolution and limitations of new methodological approaches in the field of regulatory toxicology, in a context where the use of animals for scientific purposes is increasingly questioned.

48. More SJ, Bampidis V, Benford D, Bragard C, Hernandez-Jerez A, Bennekou SH, et al.: **Guidance Scientific criteria for grouping chemicals into assessment groups for human risk assessment of combined exposure to multiple chemicals.** *EFSA J* 2021, **19**:e07033, https://doi.org/10.2903/j.efsa.2021.7033

49. Steinkellner H, Binaglia M, Dall'Asta C, Gutleb AC, Metzler M, Oswald IP, Parent-Massin D, Alexander J: **Combined hazard assessment of mycotoxins and their modified forms applying relative potency factors: Zearalenone and T2/HT2 toxin.** *Food Chem Toxicol* 2019, **131**:110599, https://doi.org/10.1016/j.fct.2019.110599

50. Alassane-Kpembi I, Puel O, Pinton P, Cossalter AM, Chou TC, Oswald IP: **Co-exposure to low doses of the food contaminants deoxynivalenol and nivalenol has a synergistic inflammatory effect on intestinal explants.** *Arch Toxicol* 2017, **91**:2677-2687, https://doi.org/10.1007/s00204-016-1902-9