# Ochratoxin A and citrinin production and migration in Penicillium verrucosum molded strawberry jams

Monika Coton, Elsa Sielleur, Jean-Luc Jany, Franck Deniel, Elisabeth Poirier, Philippe Dantigny

► To cite this version:

Monika Coton, Elsa Sielleur, Jean-Luc Jany, Franck Deniel, Elisabeth Poirier, et al.. Ochratoxin A and citrinin production and migration in Penicillium verrucosum molded strawberry jams. Food Microbiology, 2026, 135, pp.104982. 10.1016/j.fm.2025.104982. hal-05469386

HAL Id: hal-05469386

https://hal.univ-brest.fr/hal-05469386v1

Submitted on 21 Jan 2026

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 CC BY 4.0 - Attribution - International License
<figure><img src="image_2.png" /></figure>

Check for
updates

# Ochratoxin A and citrinin production and migration in *Penicillium verrucosum* molded strawberry jams

Monika Coton* , Elsa Sielleur, Jean-Luc Jany, Franck Deniel, Elisabeth Poirier, Philippe Dantigny

Univ Brest, Laboratoire Universitaire de Biodiversité et Ecologie Microbienne, F-29280, Plouzané, France

ARTICLE INFO

Keywords:

Food safety

Moldy foods

Food waste

Secondary metabolites

Mycotoxins

Migration

LC-QTOF

A B S T R A C T

Food losses and waste have become a major worldwide challenge, partly due to mold spoilage at the consumer level. One possible way to reduce food waste due to moldy foods would be to avoid a too conservative approach where products are directly discarded if fungal growth is observed. However, a food safety risk exists as many fungal species produce potentially toxic mycotoxins that can migrate into foods, so this hazard needs to be considered to establish consumer recommendations. This study quantified citrinin and ochratoxin A accumulation and migration in strawberry jams after inoculation with *Penicillium verrucosum* UBOCC 109221 and incubation at 8 and 20 °C. The mold failed to grow after 28days of incubation at 20 °C on jam with 59% sugar content but exhibited a constant growth rate of about 1.15 mm/d on the other sugar concentrations. After 14 days of incubation, citrinin concentration (10000 ng/g) for the jam containing 34 % sugar was about twice the concentration observed for 39 and 44 % sugar. Mycotoxin migration experiments were then carried out for 39 % sugar content and showed that the maximum mycotoxin concentrations were obtained for the 5 cm lesion diameter. At 20 °C, citrinin concentration (30 000 ng/g) was about twice that obtained at 8 °C, while the maximum ochratoxin A concentration was about 100 ng/g. For 1 and 2 cm lesions with 8°C storage, mycotoxins were not detected at 3 cm depth, accordingly jam can be consumed after removing about 2 to 3 cm beyond the moldy area. For greater lesions, jam should be discarded because mycotoxins were detected at >4 cm depth.

1. Introduction

For the past decade, food losses and waste have become a major
global concern undermining the sustainability of food systems. This
occurs throughout the entire food chain as food can be lost or wasted at
different stages of the supply chain (pre-harvest, post-harvest, storage,
distribution etc.). Food can also be directly thrown away by consumers,
especially in developed countries. In the EU, roughly 10 % of food
available to consumers at retail, food services or households may be
wasted while this may reach 30 % at the global level in some industri-
alized countries (FAO, 2011; ADEME, 2022) and consumers are likely to
play a major role. Based on these values, there is a real need to reduce
food waste as this not only concerns the food products but also the
depletion of limited natural resources used to produce them thus having
a broader environmental impact.

On February 13, 2019, Theresa May, a former British Prime Minister, stated that she scrapes off the mold on jam instead of throwing it away

and considers that the rest is “perfectly edible” (BBC, 2019). However,
food experts have mixed views on how safe that is as some jams can host
mycotoxin-producing mold species that may produce one or multiple
mycotoxins simultaneously. Once these molds grow, the mycotoxins
they produce may migrate into the apparently unspoiled section of the
product. According to some microbiologists, moldy jams should be
discarded immediately to ensure consumer safety.

However, there is no unique solution. The decision to eat the un-
spoiled portion of the food product once the moldy zone has been
removed, or to directly throw away the entire product, will depend on
the type of food product, its intrinsic properties and the extent of mold
contamination. In fact, mycotoxin production is highly dependent on the
fungal species, environmental conditions, and food composition (Coton
and Dantigny, 2019). Although, some molds such as Mucorales are not
known to produce any mycotoxins, consumers cannot necessarily
identify the mold or distinguish molds between the very vast number of
known fungal species. Therefore, recommendations to consumers

<footnote>* Corresponding author.
E-mail address: monika.coton@univ-brest.fr (M. Coton).</footnote>
should not be based on identifying the mold species but rather rely on simple and understandable recommendations.

For example, *Penicillium expansum* is the causative agent of blue rot of pome fruits and between 70 % and 80 % of decay can be observed for stored fruits (Viñas et al., 1993). *P. expansum* is also a major producer of a regulated mycotoxin patulin (PAT) in fruits and related products (Morales et al., 2007) and almost all strains are PAT producers (Andersen et al., 2004); in fact, *P. expansum* strains simultaneously produce multiple mycotoxins (Tannous et al., 2018) which may all migrate into foods but PAT is the only one regulated for apple products in the food chain (Commission regulation (EU) 2023/915). Mold contamination of apples is most likely to be synonymous with PAT production and migration of one or more mycotoxins into the flesh. Recently, *P. expansum* molded apples were analyzed for mycotoxin migration using a 3D approach and for lesions less than or equal to 3 cm in diameter, consumers were recommended to cut off at least 1 cm beyond and below the molded spot to limit patulin exposure and ensure consumer safety (Coton et al., 2020).

Another recent mycotoxin migration study was dedicated to Comté cheese, a French semi-hard cheese, that was artificially contaminated by a *Penicillium verrucosum* isolate capable of producing both ochratoxin A (OTA) and citrinin (CIT) (Coton et al., 2019); both mycotoxins are regulated in the food chain for specific food categories or food supplements (Commission regulation (EU) 2023/915) due to their known toxicities inducing nephrotoxic, hepatotoxic, teratogenic, embryotoxic and/or immunotoxic effects (Hymery and Coton, 2024; Kitchen et al., 1977; Wichmann et al., 2002; Rasmussen et al., 2011; Bui-Klimke and Wu, 2015). These two mycotoxins have also been described to co-occur in diverse foods as cereals, cereal-based products as well as fruits and fruit-based products (Kumari and Nusrath, 1987; Vrabcheva et al., 2000; Ostry et al., 2013; Pleadin et al., 2018; Meerpeol et al. 2021) thus reinforcing their relevance to study them together in a food safety context. However, when it comes to regulations in foods, nothing is possible at the consumer level thus a worst-case approach should be applied for mycotoxin migration studies to determine recommendations and protect consumers as best as possible. Based on the results obtained in the case of *P. verrucosum* molded Comté cheese, as long as only a light white mycelium developed on the cheese surface, trimming the mold off the surface or removing the first mm of the cheese can be acceptable, but intense growth of blue mold (the blue color is due to fungal sporulation) that colonized the cheese surface was associated with the accumulation of significant amounts of mycotoxins into the cheese so the product should be discarded. It was assumed that the consumer did not know for how long the cheese started to turn blue. Therefore, the recommendation was again based on the worst-case scenario, the cheese had turned blue for one week rather than for a single day.

A few other migration studies have been published in breads (Franck, 1968; Reiss, 1981; Rychlik and Schieberle, 2001), fruits or fruit-based products and vegetables (Taniwaki et al., 1992; Paster et al., 1995; Marín et al., 2006; Rychlik and Schieberle, 2001; Bandoh et al., 2009; Montbaliu et al. 2010; Olsen et al., 2017; Olsen et al., 2019; Coton et al., 2020; Schamann et al., 2025) or animal based products (Escher et al., 1973; Olsen et al. 2017, 2019; Parussolo et al., 2019; Peromingo et al., 2019; Coton et al., 2019) but in most cases no link to fungal growth (for ex. diameter size, visual aspect) was carried out which makes it a challenge to provide recommendations. Mycotoxin migration in foods is also dependent on various factors linked not only to consumer habits (i. e. storage temperature) but also the intrinsic properties of foods (i.e. water activity, pH, biochemical composition ...) and the fungal agent contaminating the food commodity (Coton and Dantigny, 2019).

Foods in consumer households can also be contaminated by more than one mold that produces mycotoxins, and we showed that nearly 25 % of moldy foods from consumer households contained more than one mold (unpublished data). For this reason, both fungal and/or mycotoxin co-occurrences should be considered in the case of moldy foods and to determine to what extent mycotoxin migration occurs as not only can an

individual fungal isolate produce multiple mycotoxins, but fungal co-contaminations could exacerbate this situation which is clearly a concern for food safety. Mycotoxin co-exposure in humans may also induce additive, synergistic or antagonistic toxicological effects. Nowadays, mycotoxin co-occurrence is considered as the most common scenario for contaminated foods at different stages of the food chain (Smith et al., 2016; Coulet et al., 2024; Arce-Lopez et al., 2024) and likely happens in the case of moldy foods in consumer households. Therefore, due to the known mycotoxin toxicities of regulated mycotoxins (i.e. Commission regulation (EU) 2023/915), they should be privileged in migration studies in a food safety context.

In the present study, a multi-mycotoxin producing *P. verrucosum* strain was inoculated onto the surface of four different strawberry jams, containing different sugar contents, stored at either 8 and 20 °C. Thallus diameter was monitored in all cases and samples from the surface and in depth were taken from one strawberry jam when the mold colony reached 1, 2, 3 and 4 cm in diameter to quantify OTA and CIT mycotoxins based on a 3D migration strategy that we developed. The selected fungal isolate-food association for this study was thus based on our worst-case approach contributing to food safety evaluation.

## 2. Material and methods

### 2.1. *Penicillium verrucosum* conidial suspensions and growth tests

#### 2.1.1. Conidial suspensions

*P. verrucosum* strain UBOCC-A-109221 was provided by the Université de Bretagne Occidentale Culture Collection, UBOCC (https://nouveau.univ-brest.fr/ubocc/fr). This strain was isolated from sugar cane syrup. Fungal stock cultures (1 ml) were thawed at ambient temperature for about 1h. Malt extract agar (M2Lev) plates were inoculated with 0.1 ml of the stock culture and incubated at 25 °C for 7d. Fresh conidia were harvested using a sterile loop after adding 2 ml of distilled water containing 0.01 % Tween 80 (Sigma Aldrich, Saint Quentin Fal-lavier, France) onto the surface of plates and scraping conidia from the surface. The conidia suspension was recuperated in an Eppendorf tube, counted using a hemocytometer, then conidial suspensions were standardized to 2.10⁷ conidia.mL⁻¹. Aliquots of this suspension (250 µL) were then distributed into cryoconservation tubes and supplemented with 250 µL 40 % (v/v) sterile glycerol to obtain a final concentration of 1.10⁷ conidia.mL⁻¹. Before use, suspensions were diluted to obtain final inoculums of 1.10⁶ conidia.mL⁻¹ for the different experiments.

#### 2.1.2. Fungal growth follow-up on strawberry jams and mycotoxin production

For each jam, physicochemical analysis was first done to determine water activity (a<sub>w</sub>) values for each of the studied jams using an AQUA-LAB 4TEV (Meter, Germany).

Ten g portions of four different strawberry jams were placed in 55 mm sized Petri dishes. Then, 10 µL of a *P. verrucosum* conidial suspension, adjusted to 1.10⁶ conidia.mL⁻¹, were inoculated into the center, in triplicate. Radial growth was monitored daily by measuring colony diameter in two perpendicular directions and plates were incubated at 25 °C for 28 days.

After incubation, mycotoxin extractions and determinations were carried out on the entire portion of *P. verrucosum* molded jams. In parallel, Yeast Extract Sucrose (YES) agar medium (Frisvad and Filtenborg, 1983) buffered at pH 4.5 with phosphate-citrate buffer (Chang et al., 1998) was also used to evaluate and compare mycotoxin production in vitro as described by Gillot et al. (2017), as this medium is characterized by a high C/N ratio favoring mycotoxin production (Visagie et al., 2014). To do so, 1 µL of each spore suspension (1.10⁶ conidia.mL⁻¹) were inoculated in the center of YES plates, in triplicate, and incubated for 7 days at 25 °C (Coton et al., 2019; Gillot et al., 2017). After incubation, the entire contents of the agar plates were stored at -20 °C until mycotoxin extractions followed by analyses by Q-TOF LC-MS.
### 2.1.3. Jam inoculation, incubation and sampling

For migration experiments, strawberry jams (Carrefour brand strawberry jam; 39g sugar) were purchased from the local supermarket. The jam was first heated, then portioned into empty sterile yogurt containers (6.6 × 5.3 cm) and stored at a refrigerated temperature until use. In total, 30 jams were inoculated using five µl of prepared *P. verrucosum* conidial suspension (1.10⁶ conidia.mL⁻¹) onto the center of each jam before covering with a sterile Petri dish lid and 30 others by spreading 5 µL of the same conidial suspension over the entire surface. These experiments included triplicates. For each inoculation method, inoculated jams were incubated at either 8 or 20 °C in closed plastic containers to maintain constant water activities (c.a 0.99 *a*<sub>w</sub>), as previously described (Coton et al., 2019). *P. verrucosum* radial growth was monitored for point inoculations by measuring two perpendicular diameters every day, until 1, 2, 3, 4 and 5 cm diameter fungal colonies were obtained. For each time point, jams with *P. verrucosum* spread on the surface were also visually monitored for growth aspect. Once each colony size was reached, triplicates were taken for both conditions to determine mycotoxin migration. After slightly freezing jams, jam cylinders were removed from containers then cut into 1 cm sized sliced depths, from D1 (1 cm) to D5 (5 cm), using the device described in Coton et al., (2019). Each 1 cm width disk was stored at -20 °C until mycotoxin extraction and quantification.

### 2.1.4. Mycotoxin extraction from YES medium and jams

For YES fungal cultures, mycotoxins were extracted from a homogenized sample combining the medium and fungal mycelium using our previously developed method (Gillot et al., 2017). Briefly, mycotoxins were extracted from 2 g aliquots using 12.5 mL of 100 % acetonitrile (ACN) containing 0.1 % formic acid (v/v). After adding solvent, samples were vortexed for 30 s, followed by 15 min sonication, before vortexing again for 30 s. Samples were centrifuged at 5000 g for 10 min at 4 °C. The ACN phase was then collected and stored at -20 °C until Q-TOF LC-MS analyses. Before analysis, 1 mL was filtered with a 0.2 µm PTFE membrane syringe 4 mm filter (Phenomenex, Torrance, USA) into an amber vial.

For jam samples, mycotoxins were extracted from 2 g portions using 12.5 mL acetonitrile:water (84:16, v/v) supplemented with 2.5 % acetic acid/g of sample. Each sample was vortexed for 30 s before being placed on a Rotoflex apparatus for 1h, again vortexed 30 s before centrifuging at 5000 g for 10 min at 4 °C. One mL aliquots were filtered through a 0.45 µm PTFE membrane syringe 4 mm filter (Phenomenex, Torrance, USA) into an amber vial and stored at -20 °C until QTOF LC-MS analyses.

### 2.1.5. Mycotoxin detection and quantification by Q-TOF LC-MS

An Agilent 6530 Accurate-Mass Quadrupole Time-of-Flight (Q-TOF) LC-MS system (Agilent Technologies, Santa Clara, CA, USA) was used for detection and quantification of ochratoxin A (OTA) and citrinin (CIT). The HPLC system was equipped with a Binary pump 1260 and degasser, well plate autosampler set to 10 °C and a thermostated column compartment set to 35 °C. OTA and CIT standards were obtained from Sigma-Aldrich (St Louis, MO, USA). The stock solution (5 mg/mL) was prepared in dimethyl sulfoxide (DMSO) and stored at -20 °C in amber vials. The compound characteristics to detect and quantify both metabolites are provided in Supp Table 1 and were previously described in Coton et al., (2020).

Two µL filtered samples (0.45 µm PTFE membrane) were injected and separated using a ZORBAX Extend-C18 column (2.1 × 50 mm and 1.8 µm, 600 bar) maintained at 35 °C with the flow rate set to 0.2 mL/min. Mobile phases were as follows: solvent A was LC-MS grade water (Carlo Erba Reagents, France) and solvent B was 100 % LC-MS grade ACN. Solvent B was maintained at 10 % for the first 4 min, followed by a gradient of 10–100 % of B for 18 min and 5 min at 100 % solvent B. Finally, solvent B was maintained at 100 % for 5-min post-time. PAT was ionized in ESI- mode (ESI: Electrospray Ionization) and mass

spectrometer conditions were: capillary voltage, 4.5 kV; source temperature, 325 °C; nebulizer pressure, 50 psig; drying gas, 12 L/min and ion range, 100–1000 m/z. OTA and CIT detection and quantification were performed using the retention time value ± 1 min and the corresponding quantifier and qualifier ions listed in Supp Table 1. Standard (using ACN) and matrix matched (using blank jam extracts or YES blanks) calibration curves were obtained by plotting the peak area against mycotoxin concentrations using a 1/x2 weighted linear regression model. Specific mycotoxin production was expressed as ng per g jam samples and ng per g dry weight for YES medium.

Method performance and validation parameters including linearity (R²), limit of detection (LOD), limit of quantification (LOQ), recovery rates (from jams spiked with OTA + CIT) were also assessed according to the ICH guidelines (ICH Harmonized Tripartite Guideline, 2005) and are presented in Supp Table 1. LOD and LOQ values were determined by multiplying the standard deviation of y-intercepts of regression lines divided by the slope, by 3.3 and 10, respectively, for the matrix-matched calibration curve (Supp Table 1). Linear range calculations and OTA + CIT concentration determinations were carried out using the Agilent MassHunter Workstation Software (Agilent Technologies, Santa Clara, CA, USA). Finally, extraction yields were determined by spiking jam samples with either 1000 or 10000 ng/g final concentrations. The same extraction protocol as described above was used and OTA yields for 1000 and 10000 ng/g concentrations were 95 % and 102 % and for CIT 92 % and 102 %, respectively.

### 2.1.6. Statistical analysis

Statistical analyses were performed in R 4.4.3 (https://www.r-project.org/). For citrinin (CIT) and ochratoxin A (OTA), concentrations were analyzed separately for point inoculation and surface spread inoculation as a function of lesion diameter (1–5 cm), incubation temperature (8 °C vs 20 °C) and sampling depth within the jam (D1–D5; 0–5 cm). Three biological replicates per condition were available. Full-factorial linear models (lm) were fitted and their residuals assessed for normality (Shapiro–Wilk) and homogeneity of variance (Levene; car package version 3.1-3). Both assumptions were violated for CIT and OTA (*p* < 0.05), and data were therefore analyzed with a non-parametric aligned-rank transform ANOVA (ART; ARTool package version 0.11.2; Kay and Wobbrock, 2021; Elkin et al., 2021; Wobbrock et al., 2011). Type-III tests (anova) and pairwise contrasts (emmeans 1.10.0, Tukey adjustment) were reported at α = 0.05. Group statistics are given as mean ± SE and visualized with ggplot2 package 3.5.2. Global temperature effect (8°C vs 20°C) on CIT and OTA concentrations were also assessed using Wilcoxon test on pooled data (all depths and lesion diameters), separately for each inoculation method.

## 3. Results

### 3.1. *P. verrucosum* growth and mycotoxin production in different jams

*P. verrucosum* radial growth was first monitored after inoculation on 4 strawberry jams containing increasing concentrations of sugars ranging from 34g to 59g per 100g. After plotting mold diameter as a function of time for a 28d incubation period at 20 °C, fungal growth rates were determined. Results clearly indicated that growth rates were directly correlated to sugar contents (Fig. 1; Supp Table 2). Mold development was visualized after 14, 21 and 28 days to observe any relevant differences between samples such as thallus diameter or sporulation (Supp Fig. 1). Fastest growth rates were observed for jams with reduced sugar contents and higher water activities: growth rate was 1.12 mm/d for jam containing 34 % sugar and an *a*<sub>w</sub> = 0.927, 1.16 mm/d for 36 % sugar and *a*<sub>w</sub> = 0.927 followed by a slower growth rate of 0.96 mm/d for jam containing 44 % sugars and *a*<sub>w</sub> = 0.912 (Supp Table 2). On the other hand, no growth was observed on strawberry jam containing 59 % sugar content and for a water activity of 0.849, even after 28d incubation at 20 °C.
<figure><img src="image_3.png" /><figcaption>**Fig. 1.** *P. verrucosum* growth and mycotoxin content after 28d incubation at 20 °C. Lines represent radial growth follow-up while bars represent CIT content for each time. Light grey 34g sugar, dark grey 39g sugar, black 44g sugar.</figcaption></figure>

For the same jam samples, as well as yeast extract sucrose medium (YES) used as a control condition, mycotoxin content was determined. OTA was only detected in YES cultures (data not shown) as well as for one out of three replicates of jam containing 44g sugar (Supp Table 2). On the other hand, CIT was quantified at increasing concentrations between 14 and 28 days incubation reaching levels as high as ~17500 ng/g in the jam with the lowest sugar content (34g) and highest water activity (0.927), followed by 10835 ng/g (for 39g; 0.927) and 7867 ng/g (for 44g; 0.912) (Fig. 1; Supp Table 2); these results were thus well correlated to the highest to lowest calculated growth rates and according to jam water activities. These results also indicate the strong matrix effect for mycotoxin production as OTA was mainly produced under optimal growth conditions on synthetic medium while only sporadically produced in jams.

## 3.2. CIT and OTA production and migration in *P. verrucosum* molded strawberry jam

Strawberry jam with 39g sugar content per 100g was further used for mycotoxin migration experiments. Jams were inoculated using two methods in triplicate, either a central point inoculation of a calibrated spore suspension or by spreading the spore suspension over the entire surface, to mimic as closely as possible the way mold contamination of jams occurs in consumer households. After inoculation, jams were incubated at either 8 °C or 20 °C to mimic refrigerated or room temperature consumer storage conditions and *P. verrucosum* growth was monitored daily until the thallus size reached 1 cm, 2 cm, 3 cm, 4 cm or 5 cm in diameter for the point inoculation; at the same time, jams inoculated with the spread method were also tested to compare mycotoxin contents. Both CIT and OTA concentrations were determined in a depth wise manner by slicing jams (after slightly freezing them) using 1 cm thickness intervals from D1 (first 1 cm depth) to D5 (5 cm depth). For both metabolites and for both inoculation patterns, the ART-ANOVA revealed significant main effects of temperature, depth, lesion diameter as well as all possible two-way interactions (Supplementary Table 3).

For 8 °C incubation, CIT and OTA contents were quantified at increasing levels as the diameter size increased regardless of contamination method. For 1 cm diameter thallus development, only CIT was quantified at ~250 ng/g jam in the uppermost 1 cm slice portion containing the moldy zone for the spread method (Supp Fig. 2A) while no quantifiable levels of either mycotoxin were obtained for the point inoculation method in the same uppermost portions (Fig. 2A/B) which can be linked to limited fungal growth in these samples (Fig. 5). For 2 cm and 3 cm lesions and both inoculation methods, the highest quantifiable CIT levels accumulated in the uppermost 1 cm portion (~346 and

~1053 ng/g jam for point inoculations and ~793 and ~1788 ng/g for surface inoculations for 2 cm_D1 and 3 cm_D1 fractions, respectively). CIT content decreased from 2.5 to nearly 5-fold in the next depth below the moldy portion and no CIT was detected in the third depths (<LOD). For OTA, quantifiable levels were only detected for point inoculations at ~28 ng/g jam for the 2 cm_D2 portion and at ~4.3 and ~26 ng/g for the 3 cm_D2 and 3 cm_D3 portions thus suggesting limited production and diffusion; moreover, OTA was only quantified for the surface inoculation method in the 3 cm_D2 portion at ~150 ng/g. For 4 cm and 5 cm moldy lesions, CIT production and migration was intense with the highest levels systematically quantified in the uppermost portions containing the moldy zone (i.e. ~644 ng/g/~15457 ng/g for 4 cm_D1/5 cm_D1 fractions for point inoculations versus ~805 ng/g/~16870 ng/g for 4 cm_D1/5 cm_D1 fractions for surface inoculations), then successively decreasing contents were quantified in all depth wise portions including at 5 cm in depth, regardless of diameter size (4 cm versus 5 cm) or inoculation method (point versus spread) (Fig. 3A/B). For OTA, more limited production and diffusion was observed for lesion sizes up to 4 cm and mainly for point inoculation samples (Fig. 2B; Fig. 3C/D). OTA content was ~59 ng/g jam in the uppermost 4 cm_D1 portion (1 cm depth) and diffused to the 4 cm_D3 portion (3 cm depth) at a concentration ~16 ng/g jam.

At 20 °C, CIT and OTA production and migration were directly observed for the 1 cm thallus size and in both the 1 cm and 2 cm depths (1 cm_D1 and 1 cm_D2 for point inoculations) while only OTA was quantified in the 3 cm depth (1 cm_D3 for point inoculations) (Figs. 2 and 3; Supp Fig. 2). Highest mycotoxin contents were systematically quantified in the uppermost portions for CIT while highest OTA content was detected in the 3 cm depth (~38 ng/g versus ~9 ng/g in the uppermost 1 cm portion). For 2 cm lesions, a similar trend for CIT was observed with diffusion and decreasing concentrations in the 2 cm or 3 cm portions for point or spread inoculations, respectively. Surprisingly, OTA content was limited to the uppermost portion for this lesion size. For 3 cm–5 cm lesions and both inoculation methods, highest CIT content accumulated in the uppermost portions and ranged from ~8145 to 8795 ng/g for 3 cm lesions (3 cm_D1) up to 24785–29294 ng/g for 5 cm lesions (5 cm_D1). CIT content then diffused in a decreasing manner up to 4 cm in depth for 3 cm lesions (~198–220 ng/g for 3 cm_D4 samples) and up to 5 cm in depth for both 4 cm (~218–277 ng/g for 4 cm_D5 samples) and 5 cm (~198–302 ng/g for 5 cm_D5 samples) mold lesions. For OTA, quantifiable levels were only detected in a limited number of portions with most OTA content identified in the uppermost 2 cm, regardless of inoculation method (Fig. 2B, Supp Fig. 2B).

The overall citrinin (CIT) concentration showed a temperature-dependent trend: at 20 °C, CIT levels were higher than at 8 °C for both inoculation methods, with a statistical significance for point
A

<figure><img src="image_4.png" /></figure>

B

<figure><img src="image_5.png" /><figcaption>**Fig. 2.** Mycotoxin production and migration in *P. verrucosum* point inoculated molded strawberry jam. Results are presented for CIT (A), OTA (B) after incubation at either 8 °C (light grey bars ■) or 20 °C (dark grey bars ■).</figcaption></figure>

inoculation (Wilcoxon test, W = 1816, p = 0.010) and a near-significant
trend for surface spread (Wilcoxon test, W = 2054, p = 0.065) (Fig. 4).
Ochratoxin A (OTA) exhibited no significant temperature effect: p =
0.475 for surface spread and p = 0.061 for point inoculation (Fig. 4).
These univariate results, based on the temperature factor alone
(ignoring depth and thallus size) are in accordance with the main-effect
pattern revealed by the full ART-ANOVA.

Fungal lesions were also visually monitored over time for both
storage conditions (Fig. 5). At 8 °C, a light fluffy white mold was
observed for 1 cm and 2 cm lesions while a very faint blue color was
observed for the 3 cm lesion with the point inoculation and a more
intense blue color was observed for the spread inoculation due to fungal
sporulation and conidial production. At 20 °C, a fluffy white mold was
observed for 1 cm lesions for both inoculation methods and at 2 cm only
<figure><img src="image_6.png" /><figcaption>**Fig. 3.** Two-dimensional profiles of mycotoxin accumulation in jam. Heat-map tiles show the mean concentration of citrinin (CIT) (panels A–B) and ochratoxin A (OTA) (panels C–D) as a function of sampling depth with the jam (x-axis, D1–D5 = 0–5 cm) and lesion diameter (y-axis, 1–5 cm). Panels A and C correspond to the surface-spread inoculation, panels B and D to the central-point inoculation. Within each panel, the two vertical facets compare storage at 8 °C (left) versus 20 °C (right). Tile colour encodes mycotoxin level on a linear scale from light grey (low) to red (high); values exceeding the dataset-specific maximum are squished to the upper limit (see colour bar). Each cell represents the average of three biological replicates (n = 150 per toxin). Concentrations are expressed in ng.g⁻¹. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)</figcaption></figure>

for the point inoculation. In all other cases, fungal sporulation was observed and a blue color clearly visible. Fungal growth and sporulation were very intense, especially for spread inoculations incubated at 20 °C in comparison to 8 °C.

The total mycotoxin content, sum of CIT and OTA, that migrated into jam was also evaluated for all conditions and values were expressed as % relative abundances (Fig. 6). For all lesion sizes and both temperatures, total mycotoxin content was systematically at the highest levels within the first depth (D1 samples). Both mycotoxins were also detected simultaneously in D1 samples for all point inoculations but not for all surface inoculations. As lesion size increased, total mycotoxin contents also increased and highest contents were observed at 20 °C versus 8 °C. When evaluating diffusion patterns, a depth dependent decrease was clearly observed. Based on the calculated % relative abundances for samples stored at 20 °C, a decrease from 100 % in D1 samples to ~51 % in the "1 cm_D2" sample or even as low as ~21–27 % for the 2 cm–5cm depth "D2" samples were observed thus suggesting a rapid decrease in overall content in the 2 cm depth portions. For the same sampling points that were analyzed at 8 °C, the % relative abundances were generally like those observed at 20 °C except for 4 cm thallus sample set; in this case, total mycotoxin content remained at much higher levels. Indeed, around 70–100 % total mycotoxin content remained in the "D2" samples before progressively decreasing in the remaining depths to ~15 %–~48 %, even in the D5 portions. For all the other tested conditions (i.e. different thallus sizes 1, 2, 3, 4 and 5 cm or migration depths D3-D5), relative mycotoxin abundances dropped to levels below ~10 % in D3 samples and below ~4 % for D4 and D5 samples with some samples containing <1 % total mycotoxin content. Overall, although mycotoxins migrated in depth into the strawberry jam, a very large drop in total

content was observed, especially for 3–5 cm depths.

# 4. Discussion

## 4.1. *Mycotoxin migration in P. verrucosum molded jams*

Among molds identified on jams, some species may produce one or multiple mycotoxins which potentially migrate and accumulate in the food product. The most concentrated zones containing mycotoxins typically surround the fungal colony although diffusion will more or less intensely occur according to the chemical and structural properties of both the mycotoxins and the food itself (Coton and Dantigny, 2019; Olsen et al., 2019). To date, only a very limited number of studies have focused on exploring mycotoxin migration in fruit jams (Lindroth and Niskanen, 1978; Lindroth et al., 1978; Akerstrand and Andersson, 1979; Pensala et al., 1978; Ostry et al., 2004; Olsen et al. 2017, 2019; Schamann et al., 2025). In all these studies, similar fungal species known to contaminate this type of food product were used and mainly focused on four *Penicillium* species, *P. expansum*, *P. crustosum*, *P. roqueforti* and like in this study *P. verrucosum*. These species were all identified in moldy consumer jams that we recently collected (unpublished data) thus confirming their prevalence on this type of substrate. In general, *Penicillium* species are well-known mycotoxin producers, often simultaneously producing multiple mycotoxins. Thus, *Penicillium* mycotoxins can pose a food safety risk for human and animal health due to their known toxicological effects, thus some are currently regulated in certain food products, including fruit or fruit-based products, like patulin (produced by *P. expansum*) (EU, 2023/915) or ochratoxin A (produced by *P. verrucosum*) (EU, 2022/1370). On the other hand, other *Penicillium*
<figure><img src="image_7.png" /><figcaption>**Fig. 4.** Overall effect of incubation temperature on citrinin (CIT) and ochratoxin A (OTA) production in jam. Citrinin (CIT) (panels A and B) and ochratoxin A (OTA) (panels C and D) measured across all depths within the jam and lesion diameters for the two inoculation methods: central point inoculation (A, C) and surface spread inoculation (B, D). Blue violins correspond to samples stored at 8 °C, red violins to 20 °C (n = 75 observations per temperature). Filled circles represent individual measurements; the bold horizontal line marks the median, and the diamond denotes the mean. Wilcoxon rank-sum *p*-values (temperature as the sole factor) are reported in each panel. Concentrations are expressed in ng.g⁻¹ on a log10 scale. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)</figcaption></figure>

mycotoxins such as roquefortines, penitrem A, cyclopiazonic acid, iso-
fumigaclavine or PR toxin have not yet been regulated although
commonly produced by food related Penicillium species (Scott et al.,
1977; Lund et al., 1995; Fontaine et al., 2015; Crequer et al., 2024;
Ropars et al., 2020; Olsen et al., 2019). All these mycotoxins can thus be
found as contaminants in foods and may potentially migrate both
widthwise and depth wise as was already described for patulin in
P. expansum molded apples (Coton et al., 2020; Marín et al., 2006) or
ochratoxin A and citrinin for P. verrucosum molded cheese or cream
(Coton et al., 2019; Olsen et al., 2019).

In our study, we selected strawberry jam for mycotoxin migration
studies using a mycotoxin producing *P. verrucosum* isolate as strawberry
jam is among the three most consumed jams in France, along with
apricot and bitter orange jams (Femme Actuelle, 2022). We first wanted
to evaluate the impact of sugar content, which has a strong impact on
water activity, fungal growth and mycotoxin production. Four different
strawberry jams, with progressively increasing sugar contents but with
similar compositions (all four contained strawberries, water and pectin;
three also contained concentrated lemon juice; one contained apple),
were selected and *P. verrucosum* grow rates were monitored. As sugar
content progressively increased and water activity decreased, fungal
growth rates were reduced, and no growth was observed for the jam
containing the highest sugar content (59g per 100g) and lowest water
activity ~0.85. This reduced water activity most likely acted as a key
barrier to mold development over the 28 days and prevented subsequent
mycotoxin production. Minimal water activity values for *P. verrucosum*
germination and mycelial growth were previously determined to be 0.80
and 0.85, respectively, while optimal growth is between 0.95 and 0.99
(Pardo et al., 2006) thus confirming our results for no growth in reduced
water activity jam. This species also grows optimally in temperatures
ranging between 20 and 30 °C (Pardo et al., 2006) which is also well

correlated to our results. Lindroth et al. (1978) studied *P. expansum*
growth and patulin diffusion in blackberry, blueberry and strawberry
jams. Their results showed a strong but not complete inhibition of
patulin production in the presence of 44 g sugar per 100 g jam,
regardless of jam type, after incubated at 22 °C although these condi-
tions did not significantly affect *P. expansum* growth. Like the results of
our study, the highest patulin levels were detected in jams with less
added sugar (water activities >0.93). *Aspergillus flavus* molded apricot
jam was also more recently studied to determine aflatoxin diffusion
(Ostry et al., 2004). The highest contents were observed in the upper-
most portion (0.8 cm slice) after incubation at 19 °C while levels were
lower at 6 °C thus these results are well correlated to our data.

In *P. verrucosum* molded strawberry jam stored at 20 °C and for all
thallus diameter sizes that we monitored, CIT and OTA production and
migration was more intense than at 8 °C suggesting cold storage can act
as a key barrier to reduce mycotoxin production and diffusion while also
delaying fungal growth. Our data showed OTA production was signifi-
cantly lower in comparison to CIT and a more sporadic production
profile was observed at 8 °C in comparison to 20 °C. When comparing
the highest mycotoxin contents observed for the smallest to largest
thallus diameters (1 cm-5 cm) using the point inoculation method, CIT
production was produced at 25X to 300X the level of OTA. Different
environmental factors including temperature, water activity or pH have
already been shown to impact CIT biosynthesis in *P. verrucosum*
(Touhami et al., 2018) and higher levels of CIT versus OTA production
has also been observed by *P. verrucosum* at 6 °C on standard yeast extract
sucrose (YES) medium (Takahashi and Yazaki, 2007), at 20 °C on Comté
cheese (Coton et al., 2019) and has been suggested to maybe exceed OTA
levels in cereals (Vrabcheva et al., 2000; Meister, 2004; Zaied et al.,
2012; Kononenko and Burkin, 2013; Wawrzyniak and Waskiewicz,
2014). Although both OTA and CIT can be produced simultaneously by
A

<figure><img src="image_8.png" /></figure>

B

1cm

3cm

<figure><img src="image_9.png" /></figure>

1cm

2cm

3cm

4cm

5cm

<figure><img src="image_10.png" /><figcaption>**Fig. 6.** Percentage of remaining total mycotoxins (OTA + CIT) expressed as % for different thallus sizes (1 cm–5 cm) and different depths (D1 = 1 cm depth to D5 = 5 cm depth). Results are shown for incubation at 8 °C (●) or 20 °C (■).</figcaption></figure>

*P. verrucosum*, Schmidt-Heydt et al. (2011) observed that light conditions impacted production and inhibited OTA biosynthesis thus other regulation mechanisms are likely to play a role in the production of these two mycotoxins which may explain the different production profiles.

**5. Conclusions**

Regulations setting maximum limits for mycotoxins in foods can only be done at different steps of the food chain from production to retail. At
the consumer level, only simple recommendations can be provided to consumers to help them prevent or reduce food waste while ensuring food safety. Pinpointing at risk scenarios based on consumer behaviour towards moldy foods in households would also be useful. The main problem with moldy foods is the possible production of mycotoxins by some molds, and then the migration of one or more mycotoxins outside the moldy section. Many consumers, probably including Theresa May, are unaware of these realities. In this paper, it was shown that many experiments and analyses were necessary to provide a complete overview of the production and the migration of CIT and OTA in *P. verrucosum* molded strawberry jams.

It is very challenging to turn these scientific results into simple and understandable recommendations for consumers. We must emphasize that a typical consumer can only determine the size and the color of the moldy sections. They obviously will not be able to identify the actual contaminating fungal species and even less know whether mycotoxins can be produced. Therefore, recommendations should be based on the worst-case scenario, i.e., scientific data generated from the most mycotoxigenic mold. Consumer habits should also be considered prior to setting recommendations. In a recent survey in France (unpublished data), it was shown that only 3.2 % of spoiled jams found in consumer's homes were not opened. While about 50 % of French people (*Femme actuelle*, 2022) are producing their own homemade jams, we can thus recommend that unopened jams be stored at ambient temperature.

In contrast, jams should be systematically stored in the refrigerator after opening, because fungal growth and mycotoxin production are lower at 8 °C than at 20 °C. However, this recommendation is not relevant for jams containing more than 55 % sugar as no fungal growth occurs due to the low water activities. In the case of moldy jams, the "one, two, three, spoon" rule can be applied to consumers. For 1 cm diameter moldy jam, remove as much jam as you can, around the moldy section, with one heaping coffee spoon while for 2 cm moldy lesions, a tablespoon should be used. In the case of larger moldy spots, so 3 cm or bigger, no spoon would be large enough so the jam should be discarded as mycotoxins can diffuse far from the moldy lesion.

### CRediT authorship contribution statement

**Monika Coton:** Writing – review & editing, Writing – original draft, Validation, Supervision, Resources, Project administration, Methodology, Funding acquisition, Formal analysis, Conceptualization. **Elsa Sielleur:** Investigation, Formal analysis. **Jean-Luc Jany:** Writing – review & editing, Methodology, Investigation, Formal analysis. **Franck Deniel:** Writing – review & editing, Investigation, Formal analysis. **Elisabeth Poirier:** Investigation, Formal analysis. **Philippe Dantigny:** Writing – review & editing, Writing – original draft, Supervision, Methodology, Funding acquisition, Conceptualization.

### 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 public funding from the French Research Agency ANR for the MYNION project (ANR-22-CE21-0003-01). We thank the Université de Bretagne Occidentale Culture Collection, UBOCC (https://nouveau.univ-brest.fr/ubocc/fr), for providing fungal isolates.

### Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.fm.2025.104982.

## Data availability

Data will be made available on request.

## References

* ADEME (L'Agence de l'Environnement et de la Maîtrise de l'Energie), 2022. Éviter le gaspillage alimentaire dans les collectivités et les entreprises [WWW document]. https://economie-circulaire.ademe.fr/gaspillage-alimentaire. (Accessed 11 March 2025).

* Akerstrand, K., Andersson, A., 1979. Production and diffusion of patulin in jams, apple jam and apples. Var. foda 6, 357–374 (in Swedish).

* Andersen, B., Smedgaard, J., Frisvad, J.C., 2004. Penicillium expansum: consistent production of patulin, chaetoglobosins, and other secondary metabolites in culture and their natural occurrence in fruit products. J. Agri. Food Chem. 52, 2421–2428.

* Bandoh, S., Takeuchi, M., Ohsawa, K., Higashihara, K., Kawamoto, Y., Gotoa, T., 2009. Patulin distribution in decayed apple and its reduction. Int. Biodeterior. Biodegrad. 63, 379–382.

* BBC, 2019. Moldy jam: should you eat what's beneath? https://www.bbc.com/news/uk-47227477. (Accessed 17 April 2025).

* Bui-Klimke, T.R., Wu, F., 2015. Ochratoxin A and human health risk: a review of the evidence. Crit. Rev. Food Sci. Nutr. 55, 1860–1869.

* Chang, S.C., Lei, W.Y., Tsai, Y.C., Wei, Y.H., 1998. Isolation, purification, and characterization of the PR oxidase from Penicillium roqueforti. Appl. Environ. Microbiol. 64, 5012–5015.

* Coton, M., Dantigny, P., 2019. Mycotoxin migration in moldy foods. Curr. Opin. Food Sci. 29, 88–93.

* Coton, M., Auffret, A., Poirier, E., Debaets, S., Coton, E., Dantigny, P., 2019. Production and migration of ochratoxin-A and citrinin in conté cheese by an isolate of Penicillium verrucosum selected among penicillium spp. mycotoxin producers in YES medium. Food Microbiol. 82, 551–559.

* Coton, M., Bregier, T., Poirier, E., Debaets, S., Arnich, N., Coton, E., Dantigny, P., 2020. Production and migration of patulin in Penicillium expansum molded apples during cold and ambient storage. Int. J. Food Microbiol. 313, 108377.

* Crequer, E., Coton, E., Cueff, G., Cristiansen, J.V., Frisvad, J.C., Rodriguez De La Vega, R., Giraud, T., Jany, J.-L., Coton, M., 2024. Different metabolite profiles across Penicillium roqueforti populations associated with ecological niche specialisation and domestication. IMA Fungus 15, 38.

* Elkin, L.A., Kay, M., Higgins, J.J., Wobbrock, J.O., 2021. An aligned rank transform procedure for multifactor contrast tests. In: The 34th Annual ACM Symposium on User Interface Software and Technology, pp. 754–768.

* Escher, F.E., Koehler, P.E., Ayres, J.C., 1973. Production of ochratoxin A and B on country cured ham. Appl. Microbiol. 26, 27–30.

* FAO, 2011. Global Food Losses and Food Waste – Extent, Causes and Prevention. Rome.

* **Femme actuelle, 2022.** Quelle est la confiture préférée des français. https://www.femmeactuelle.fr/cuisine/recettes-de-cuisine/quelle-est-la-confiture-preférée-des-fra-ncais-la-recette-maison-pour-en-profiter-2138311. (Accessed 26 May 2025).

* Fontaine, K., Passero, E., Vallone, L., Hymery, N., Coton, M., Jany, J.-L., Mounier, J., Coton, E., 2015. Occurrence of roquefortine C, mycophenolic acid and aflatoxin M1 mycotoxins in blue-veined cheeses. Food Cont 47, 634–640.

* Franck, H.K., 1968. Diffusion of aflatoxins in foodstuffs. J. Food Sci. 33, 98–100.

* Frisvad, J.C., Filtenborg, O., 1983. Classification of terverticillate penicillia based on profiles of mycotoxins and other secondary metabolites. Appl. Environ. Microbiol. 46, 1301–1310.

* Gillot, G., Jany, J.-L., Poirier, E., Maillard, M., Debaets, S., Thierry, A., Coton, E., Coton, M., 2017. Functional diversity within the Penicillium roqueforti species. Int. J. Food Microbiol. 241, 141–150.

* Hymery, H., Coton, M., 2024. Autres toxines d'aspergillus spp. and penicillium spp. In: Oswald, I., Forget, F., Puel, O. (Eds.), Les Mycotoxines. Connaissances Actuelles Et Futurs Enjeux. Editions Quae, France, pp. 120–135.

* Kay, M., Wobbrock, J., 2021. ARTool: aligned rank transform for nonparametric factorial ANOVAs. R package version 0 11 (10.5281), 1.

* Kitchen, D.N., Carlton, W.W., Tuite, J., 1977. Ochratoxin A and citrinin induced nephrotoxicosis in beagle dogs. I. Clinical and clinicopathological features. Vet. Pathol. 14, 154–172.

* Kononenko, G.P., Burkin, A.A., 2013. Peculiarities of feed contamination with citrinin and ochratoxin A. Agri. Sci. 4, 34–38.

* Kumari, C.K., Nusrath, M., 1987. Natural occurrence of citrinin and ochratoxin A in coconut products. Natl. Acad. Sci. Lett. 10, 303–305.

* Lindroth, Niskanen, 1978. Comparison of potential patulin hazard in home-made and commercial apple products. J. Food Sci. 43 (2), 446–448.

* Lindroth, S., Niskanen, A., Pensala, O., 1978. Patulin production during storage of blackcurrant, blueberry and strawberry jams inoculated with Penicillium expansum mold. J. Food Sci. 43, 1427–1429.

* Lund, F., Filtenborg, O., Frisvad, J.C., 1995. Associated mycoflora of cheese. Food Microbiol. 12, 173–180.

* Marín, S., Morales, H., Hassan, A., Ramos, A.J., Sanchis, V., 2006. Patulin distribution in the tissue of penicillium expansum-contaminated fuji and golden apples. Food Add. Contam. 23, 1316–1322.

* Meerpoel, C., Vidal, A., Andjelkovic, M., De Boevre, M., Tangni, E.K., Huybrechts, B., Devreese, M., Croubels, S., De Saeger, S., 2021. Dietary exposure assessment and risk characterization of citrinin and ochratoxin A in Belgium. Food Chem. Toxicol. 147, 111914.
Monbaliu, S., Van Poucke, K., Heungens, K., Van Peteghem, C., de Saeger, S., 2010. Production and migration of mycotoxins in sweet pepper analyzed by multimycotoxin LC-MS/MS. J. Agri. Food Chem. 58, 10475-10479.

Morales, H., Sanchis, V., Rovira, A., Ramos, A.J., Marín, S., 2007. Patulin accumulation in apples during postharvest: effect of controlled atmospheres and fungicide treatments. Food Control 18, 1443-1448.

Olsen, M., Gidlund, A., Sulyok, M., 2017. Experimental mold growth and mycotoxin diffusion in different food items. World Mycotox. J. 10, 153-161.

Olsen, M., Lindqvist, R., Bakeeva, A., Leong, S.L., Sulyok, M., 2019. Distribution of mycotoxins produced by *penicillium* spp. inoculated in apple jam and crème fraîche during chilled storage. Int. J. Food Microbiol. 292, 13-20.

Ostry, V., Skarkova, J., Ruprich, J., 2004. The experimental contamination of foodstuffs with the spores of toxigenic micromycetes and the production of mycotoxins. Mycotox. Res. 20, 31-35.

Ostry, V., Malir, F., Ruprich, J., 2013. Producers and important dietary sources of ochratoxin A and citrinin. Toxins 5, 1574-1586.

Pardo, E., Malet, M., Marín, S., Sanchis, V., Ramos, A.J., 2006. Effects of water activity and temperature on germination and growth profiles of ochratoxigenic *Penicillium verrucosum* isolates on barley meal extract agar. Int. J. Food Microbiol. 106 (1), 25-31.

Parussolo, G., Oliveira, M.S., Garcia, M.V., Bernardi, A.O., Lemos, J.G., Stefanello, A., Mallmann, C.A., Copetti, M., 2019. Ochratoxin A production by *Aspergillus westerdijkiae* in italian-type salami. Food Microbiol. 83, 134-140.

Paster, N., Huppert, D., Steinmeier, U., Thrane, U., 1995. Production of patulin by different strains of *Penicillium expansum* in pear and apple cultivars stored at different temperatures and modified atmospheres. Food Add. Contam. 12, 52-58.

Pensala, O., Niskanen, A., Lindroth, S., 1978. Aflatoxin production in black currant, blue berry and strawberry jams. J. Food Prot. 41, 344-347.

Peromingo, B., Sulyok, M., Lemmens, M., Rodriguez, A., Rodriguez, M., 2019. Diffusion of mycotoxins and secondary metabolites in dry-cured meat products. Food Contam 101, 144-150.

Pleadin, J., Zadravec, M., Lesic, T., Vahcic, N., Frece, J., Mitak, M., Markov, K., 2018. Co-occurrence of ochratoxin A and citrinin in unprocessed cereals established during a three-year investigation period. Food Addit. Contam. Part B Surveill. 11, 20-25.

Rasmussen, R.R., Rasmussen, P.H., Larsen, T.O., Bladt, T.T., Binderup, M.L., 2011. *In vitro* cytotoxicity of fungi spoiling maize silage. Food Chem. Toxicol. 49, 31-44.

Reiss, J., 1981. Studies on the ability of mycotoxins to diffuse in bread. Appl. Microbiol. Biotechnol. 12, 239-241.

Ropars, J., Didiot, E., Rodríguez De La Vega, R.C., Bennetot, B., Coton, M., Poirier, E., Coton, E., Snirc, A., Le Prieur, S., Giraud, T., 2020. Domestication of the emblematic white cheese-making fungus *Penicillium camemberti* and its diversification into two varieties. Curr. Biol. 30, 1-13.

Rychlik, M., Schieberle, P., 2001. Model studies on the diffusion behavior of the mycotoxin patulin in apples, tomatoes, and wheat bread. Euro. Food Res. Technol. 212, 274-278.

Schamann, A., Caprio, A., Krpan, M., Malachová, A., Gössinger, M., Labuda, R., Sulyok, M., Krska, R., 2025. Mycotoxin migration in jam samples: a study on natural contamination and experimental inoculation. World Mycotoxin J. https://doi.org/ 10.1163/18750796-bja10028 (published online ahead of print 2025).

Schmidt-Heydt, M., Rüfer, C., Raupp, F., Bruchmann, A., Perrone, G., Geisen, R., 2011. Influence of light on food relevant fungi with emphasis on ochratoxin producing species. Int. J. Food Microbiol. 145, 229-237.

Scott, P.M., Kennedy, B.P.C., Harwig, J., Blanchfield, B.J., 1977. Study of conditions for production of roquefortine and other metabolites of Penicillium roqueforti. Appl. Environ. Microbiol. 33, 249-253.

Takahashi, H., Yazaki, H., 2007. Production and contamination of ochratoxin by *penicillium* species. Mycotoxins 57, 57-63.

Taniwaki, M., Hoenderboom, C., Vitali, A., Eiroa, M., 1992. Migration of patulin in apples. J. Food Prot. 55, 902-904.

Tannous, J., Keller, N., Atoui, A., El Khour, A., Lteif, R., Oswald, O., Pue, O., 2018. Secondary metabolism in *penicillium expansum*: Emphasis on recent advances in patulin research. Crit. Rev. Food Sci. Nutr. 58 (12), 2082-2098.

Touhami, N., Soukup, S.T., Schmidt-Heydt, M., Kulling, S.E., Geisen, R., 2018. Citrinin as an accessory establishment factor of *P. expansum* for the colonization of apples. Int. J. Food Microbiol. 266, 224-233.

Viñas, I., Vela, E., Sanchis, V., 1993. Capacidad productora de patulina de cepas Penicillium expansum procedentes de centrales hortofructicolas de Lleida. Rev. Iberoam. Micol. 10, 30-32.

Visagie, C.M., Houbraken, J., Frisvad, J.C., Hong, S.-B., Klaassen, C.H.W., Perrone, G., Seifert, K.A., Varga, J., Yaguchi, T., Samson, R.A., 2014. Identification and nomenclature of the genus *penicillium*. Stud. Mycol. 78, 343-371.

Vrabcheva, T., Usleber, E., Dietrich, R., Martlauber, E., 2000. Co-occurrence of ochratoxin A and citrinin in cereals from Bulgarian villages with a history of Balkan endemic nephropathy. J. Agri. Food Chem. 48, 2483-2488.

Wawrzyniak, J., Waskiewicz, A., 2014. Ochratoxin A and citrinin production by Penicillium verrucosum on cereal solid substrates. Food add. Contam. 31, 139-148.

Wichmann, G., Herbarth, O., Lehmann, I., 2002. The mycotoxins citrinin, gliotoxin, and patulin affect interferon-gamma rather than interleukin-4 production in human blood cells. Environ. Toxicol. 17, 211-218.

Wobbrock, J.O., Findlater, L., Gergle, D., Higgins, J.J., 2011. The aligned rank transform for nonparametric factorial analyses using only anova procedures. In: Proceedings of the SIGCHI Conference on Human Factors in Computing Systems, pp. 143-146.

Zaied, C., Zouaoui, N., Bacha, H., Abid, S., 2012. Natural occurrence of citrinin in Tunisian wheat grains. Food Cont 28, 106-109.