Toxicon 218 (2022) 13–18

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Toxicon
journal homepage: www.elsevier.com/locate/toxicon

Total aflatoxin, aflatoxin B1, ochratoxin A and fumonisin in dry dog food: A
risk assessment for dog health
Hüsamettin Ekici a, Mustafa Yipel b, *
a
Kırıkkale University, Faculty of Veterinary Medicine, Department of Pharmacology and Toxicology, 71450, Kırıkkale, Turkey
b
Hatay Mustafa Kemal University, Faculty of Veterinary Medicine, Department of Pharmacology and Toxicology, 31040, Hatay, Turkey

A R T I C L E I N F O A B S T R A C T

Handling Editor: Dr. Ray Norton The aim of this study was to measure total aflatoxin (AFT), aflatoxin B1 (AFB1), ochratoxin A (OCA) and
fumonisin (FUM) concentrations in dry dog feed and to evaluate the risk to animal health posed by their
Keywords: increased levels. A total of 90 dry food samples, which were commercially available to the owner, were collected
Mycotoxins from different shops in Turkey. Some of the food samples were collected from open packages, from which the dry
Dry dog food
food was sold in smaller amounts. Using commercial Enzyme-Linked Immunosorbent Assay test kits, all samples
Health risk assessment
were examined for AFT, AFB1, OCA, and FUM concentrations. High-performance liquid chromatography was
ELISA
HPLC used for confirmation of measured parameters in 30 samples. The ELISA tests found AFT, AFB1, OCA, and FM
concentrations (ng g − 1) as 1.66, 0.64, 2.14, and 87.06, respectively. In terms of risk assessment, consumption of
the dry foods, which are contaminated by AFT, AFB1 and OCA due possibly to the fact that the dry foods are
produced from inappropriate raw material or sold in open packages in smaller amounts, poses a significant
health risk for dogs. As a result, it is necessary to monitor the mycotoxin load in dry dog food as the use of raw
materials of poor quality and selling the feed in smaller amounts from open packages over an uncertain time
period predispose the dry feed to the growth of mycotoxin, especially when the storage conditions are favorable.

1. Introduction
Dog breeds vary considerably in body weight. Adult body weights of
dogs vary between 1 kg (exp. Chihuahua) and 100 kg (exp. St. Bernard)
according to breed. This wide scale of body weight changes both the
digestive physiology and the amount of food consumption of dogs. These
differences between breeds are highly effective on exposure, which is
also an important parameter in terms of toxicology (Altınok-Yipel et al.,
2022; Weber et al., 2017; Zhao et al., 2021). Dog breeds are classified as
small breeds up to 15 kg, medium breeds between 15 and 25 kg, and
large breeds over 25 kg according to body weight (Mila et al., 2015;
Weber et al., 2017). All around the world, the first preference of dog
owners in nutrition is commercial dry foods. This is why, on average,
more than half of dog diets consist of dry food (Davies et al., 2017; Di
Donfrancesco et al., 2014; Laflamme et al., 2008).
Food and feed contaminants that cause health problems to both
animals and humans are either mainly natural (mycotoxins, metals etc.)
or anthropogenic (pesticides, persistent organic pollutants, endocrine
disruptors, metals, human/veterinary pharmaceuticals, personal care
products, nano-pollutants, and food/feed additives) (Altınok-Yipel et al.,
2022; Dorne and Fink-Gremmels, 2013; Yarsan and Yipel, 2013). Among
them, due to climate change, mycotoxin contamination in food and feed
has gained more importance and interest in studies has increased
(Martínez-Martínez et al., 2021; Twarużek et al., 2021; Zhao et al.,
2021). Mycotoxins, which cause toxic effects in humans and animals
consuming contaminated food/feeds, are secondary metabolites pro­
duced by fungi (Martínez-Martínez et al., 2021). Aflatoxin, ochratoxin,
zearalenone, fumonisin, patulin, trichothecene, and T-2 toxin are toxi­
cologically well-defined mycotoxins (Janik et al., 2020; Zhao et al.,
2021). Temperature, humidity, and damage to the crop it grows on
significantly change the production of these secondary metabolites by
fungi (EFSA, 2006; Janik et al., 2020; Magnoli et al., 2019). Aflatoxins
(Total, B1), ochratoxin (A), trichothecenes, zearalenone, and fumonisins
are the most investigated mycotoxins that are related to animal feed/­
food contamination like that in dry pet foods (Böhm et al., 2010; EFSA,
2006; PFIAA, 2020; Yang et al., 2020).
Among the aflatoxins which are known to be toxic to animals, AFB1
stands out with its potential carcinogenic and genotoxic effects. The
target organ is the liver and while hepatocellular damage occurs in acute
toxic exposures, hepatic fibrosis occurs in chronic exposures. In

* Corresponding author.
E-mail addresses: mustafa.yipel@mku.edu.tr, mustafa.yipel@mku.edu.tr, musyip@gmail.com, musyip@hotmail.com (M. Yipel).

https://doi.org/10.1016/j.toxicon.2022.08.013
Received 20 July 2022; Received in revised form 10 August 2022; Accepted 14 August 2022
Available online 20 August 2022
0041-0101/© 2022 Elsevier Ltd. All rights reserved.
H. Ekici and M. Yipel
addition, organisms can become vulnerable to pathogens due to the
immunosuppressive effect of mycotoxin (Deng et al., 2018; EFSA, 2004;
Zhao et al., 2021). Although its toxic effect levels have not been deter­
mined exactly, OCA is considered to be a potent renal toxin in many
animal species, especially for dogs, pigs, and poultry. It has carcino­
genic, teratogenic, and immunotoxic effects (EFSA, 2004). Although the
toxic mechanism is not fully elucidated, fumonisins inhibit ceramide
synthase, leading to disruption of sphingolipid metabolism and patho­
logical changes. FUM is known to cause tissue, organ (liver, kidney,
brain, etc.), and system damage (immune, respiratory, cardiovascular,
and reproduction) in cattle, pigs, and poultry, but the information about
its effects in dogs and cats is insufficient (EFSA, 2018).
The concentrations of mycotoxins, which are undesirable contami­
nants in animal feed/food have been limited by official authorities to
legal limits due to their toxic effects. In animal feeds, the MRL value has
been specified by FAO as 10.0 μg kg− 1 for AFB1, 0.05 mg kg− 1 for AFB1,
0.25 mg kg− 1 for OCA, and 5 mg kg− 1 for FUM by EC (EC., 2002; EC.,
2006; FAO, 2003). The European Petfood Federation (FEDIAF, 2013)
has set the maximum levels (mg kg− 1) in dog food to be 0.01 for AFB1
and OCA, and 5 for FUM. In 2021, the FDA issued a warning and recalled
the products as a result of the poisoning of more than 300 dogs, of which
more than 110 died due to the consumption of aflatoxin-contaminated
dry food (FDA, 2021).
Risk assessment is becoming a common approach in studies to
determining the possible risks to human health due to the consumption
of contaminated food (EFSA, 2012). On the other hand, studies are still
focused on the number of contaminants and legal limits for contami­
nated animal feeds with similar risk potential (Shao et al., 2018).
However, in current studies, it is reported that risk assessment, which is
a scientific tool, is a suitable approach, especially in pet animals such as
dogs, due to their high contamination risk diet and their long lifespan
(EFSA, 2012). Margin of Exposure (MoE) and provisional tolerable
weekly intake (PTWI) approaches were adopted for risk assessment of
contaminated food by mycotoxins (for AFB1 and OTA, respectively)
(EFSA, 2007; JECFA, 2007).
In this respect, parameters such as 1) dry foods make up the majority
of the dog’s diet, which increases the frequency of exposure, 2) the in­
clusion of cereals in dry dog food formulas, which are the primary source
of mycotoxin growth, 3) inadequate storage conditions during the
storage or sale of dry dog foods, which promote mycotoxin growth 4)
dog’s long life spans, which increase exposure time and potentially in­
crease health risks (Altınok-Yipel et al., 2022; Davies et al., 2017; Di
Donfrancesco et al., 2014; Laflamme et al., 2008; Weber et al., 2017).
Therefore, revealing the risk of mycotoxin related to the consumption of
dry food in dogs will directly contribute to the prevention of many
health risks. On the other hand, because dog breed differences affect life
span and food consumption, we determined which breeds are at risk for
which mycotoxins. Thus, in the study, outputs were revealed in the
determination of the risk posed by the storage conditions during the sale
and consumption of the food, the diet formation at the breed group level;
and the adjustment of the feeding frequency and amount. The aim of this
study was to evaluate contamination levels of total aflatoxin (AFT),
aflatoxin B1, (AFB1), ochratoxin A (OCA) and fumonisin (FUM) in dry
dog foods (lowest priced and packaged of ≥10 kg) from Turkey and to
assess the potential animal health risks.
2. Material and methods
2.1. Samples and analysis
A total of 90 commercial (opened original package, repackaged in
smaller weights or for sale in special stands as open) dry dog foods were
purchased from markets and pet shops. A microwave (Cem, USA) pro­
cessed 0.5 g of each milled food sample at 200 ◦ C for 15 min ramp time
and hold time, under 800 psi and 900 W with HNO3 and HCI. The
particle free digestion was then diluted up to 50 mL.
14
Toxicon 218 (2022) 13–18
AFT, AFB1, OCA, and FUM in dry food samples were analysed by
Enzyme-Linked Immunosorbent Assay (ELISA) test kits according to the
manufacturer’s guide (HELICA Biosystems, Inc., USA) and expressed as
ng g− 1. The method quality was validated in terms of recovery,
repeatability, and reproducibility. All chemicals and standards used in
the study were analytical grade (Merck, Germany).
High-performance liquid chromatography (HPLC) was used to
confirm the results of the samples obtained from the ELISA method
under the conditions given in Table 1. The results of 30 samples obtained
from ELISA were analysed by HPLC. The HPLC apparatus (Shimadzu LC-
20 A, HPLC, Shimadzu, Tokyo, Japan) had a fluorescence detector. The
AFT and AFB1 concentrations were determined with the method of Ghali
et al. (2009), OTA concentration was determined with the method of
Teixeira et al. (2010), and the FUM was determined with the method of
Pestka et al. (1994).
The ELISA and HPLC methods were validated in terms of LOD, LOQ,
recovery, and r2 parameters. The results of ELISA and HPLC were sta­
tistically analysed by a student-t test in package program (IBM, SPSS,
20.0, USA). The data was expressed as arithmetic mean ± standard
error, and the minimum and maximum values were recorded. All P
values less than 0.05 were considered significant for all statistical
calculations.
2.2. Animal health risk assessment
The potential health risk of mycotoxin contamination was assessed
according to consumption rates of commercial dry dog foods. In terms of
food consumption and lifespan, which are important parameters for risk
assessment, consumers (dogs) were classified as small (<15 kg), medium
(15–25 kg), and large (>25 kg) breeds. Estimated daily intake (EDI),
estimated weekly intake (EWI), Margin of Exposure (MOE) (EFSA ,
2020) (for AFT and AFB1) and provisional tolerable weekly intake
(PTWI) (EFSA, 2020) (for OCA and FUM) approaches were used to assess
the health risks of mycotoxin contamination in samples. The average
body weight (kg) of a dog was considered as 7.5, 20, and 35 for small,
medium, and large breeds, respectively. The daily dry food consumption
of dogs was calculated by the average of the recommended consumption
amounts according to the body weight of the purchased commercial
foods (200, 450, and 650 g day-1 for small, medium, and large breed
dogs, respectively).
3. Results and discussion
The validation results are given in Table 2. The AFT, AFB1, OCA, and
FUM contamination levels determined by ELISA are given in Table 3 and
confirmed results by HPLC are given in Table 4. All the samples (n = 90)
were positive for all the analysed mycotoxins in both the ELISA and
HPLC methods. The AFT contamination levels (ng g− 1) in the samples
analysed with the ELISA method were between 0.54 and 3.66 with a
mean of 1.66 and in samples analysed with the HPLC method were be­
tween 0.61 and 3.08 with a mean of 1.42. The AFB1 contamination
levels (ng g− 1) in the samples analysed with the ELISA method were
between 0.27 and 1.26 with a mean of 0.64 and in samples analysed
with the HPLC method were between 0.28 and 0.75 with a mean of 0.55.
The OCA contamination levels (ng g− 1) in the samples analysed with the
ELISA method were between 0.84 and 9.93 with a mean of 2.14 and in
samples analysed with the HPLC method were between 0.71 and 3.25
with a mean of 1.60. The FUM (ng g− 1) contamination levels in the
samples analysed with the ELISA method were between 23.75 and
365.82 with a mean of 87.06 and in samples analysed with the HPLC
method were between 21.38 and 93.96 with a mean of 48.84. The
positive AFB1 concentrations exceeded legal/recommended (FAO: 0.01,
EC: 0.05, FEDIAF: 0.01) limits in all samples. The positive OCA con­
centrations exceeded legal/recommended (EC: 0.25, FEDIAF: 0.01)
limits in all samples. The positive FUM concentrations exceeded legal/
recommended (EC: 5, FEDIAF: 5) limits in all samples. The average
H. Ekici and M. Yipel Toxicon 218 (2022) 13–18

Table 1
High-performance liquid chromatography conditions for Aflatoxins analyses.
Mycotoxins Mobile phase Fow-rate Dedector Wave-lengths Column
excitation-emission

AFT and water, acetonitrile and methanol mixture (60:20:20, v/v/v) 1 mL/ Fluorescence 365–435 nm C18 5 μm, column (4,6 mm ×
AFB1 min 250 mm length)
OTA water, acetonitrile and methanol mixture (60:20:20, v/v/v) 1,0 mL/ Fluorescence 333–460 nm C18 5 μm, column (4,6 mm ×
min 250 mm length)
FUM methanol:0.1 M sodium dihydrogen phosphate (75 : 25) adjusted to 0.1 mL/ Fluorescence 335–440 nm C18 5 μm, 3.2 × 250 mm
pH 3.35 with orthophosphoric acid min

AFT: Total aflatoxin; AFB1: Aflatoxin B1; OCA: Ochratoxin A; FUM: Fumonisin.

Table 2
Results of validation parameters.
Mycotoxin ELISA HPLC

LOD (ng g− 1) LOQ (ng g− 1) Recovery (%) r2 LOD (ng g− 1) LOQ (ng g− 1) Recovery (%) r2

AFT 0.1 0.332 95.3 0.970 0.025 0.083 98.44 0.996

AFB1 0.1 0.331 95.0 0.960 0.05 0.156 97.73 0.987
OCA 0.1 0.333 91,67 0.930 0.01 0.032 96.36 0.956
FUM 10 33.25 94.0 0.950 1.25 3.96 92.79 0.912

AFT: Total aflatoxin; AFB1: Aflatoxin B1; OCA: Ochratoxin A; FUM: Fumonisin; LOD: Limit of detection; LOQ: Limit of quantitation; r2: Linearity; ELISA: Enzyme-
linked immunosorbent assay; HPLC: High-performance liquid chromatography.

Table 3
Mycotoxin concentration of dry dog food samples and statistical compare of Enzyme-Linked Immunosorbent Assay (n = 90) (ng g− 1).
Arithmetic mean Standard error Median Minimum Maximum Limits (ppm), positive samples (%, positive samples numbers/total)

FAO EC FEDIAF

AFT 1.66 0.07 1.63 0.54 3.66 – – –

AFB1 0.64 0.02 0.62 0.27 1.26 0.01 100% (90/90) 0.05 100% (90/90) 0.01 100% (90/90)
OCA 2.14 0.15 1.80 0.84 9.93 – 0.25 100% (90/90) 0.01 100% (90/90)
FUM 87.06 7.07 69.84 23.75 365.82 – 5 100% (90/90) 5 100% (90/90)

AFT: Total aflatoxin; AFB1: Aflatoxin B1; OCA: Ochratoxin A; FUM: Fumonisin; FAO: Food and Agriculture Organization; EC: European Commission; FEDIAF: Eu­
ropean Petfood Federation.
Each sample was analysed in triplicate (n = 3).

Table 4
Statistical compare of Enzyme-Linked Immunosorbent Assay (n = 30) and High-performance liquid chromatography (n = 30) methods (ng g-1).
Mycotoxin Metot Arithmetic mean Standard error Median Minimum Maximum P value

AFT ELISA 1.68 0.12 1.41 0.72 3.66 0.11

HPLC 1.42 0.10 1.19 0.61 3.08
AFB1 ELISA 0.62 0.14 0.62 0.31 0.85 0.06
HPLC 0.55 0.13 0.55 0.28 0.75
OCA ELISA 1.84 0.14 1.68 0.84 3.86 0.10
HPLC 1.60 0.12 1.41 0.71 3.25
FUM ELISA 59.43 4.31 56.03 26.02 114.35 0.06
HPLC 48.84 3.55 46.04 21.38 93.96

AFT: Total aflatoxin; AFB1: Aflatoxin B1; OCA: Ochratoxin A; FUM: Fumonisin.
Each sample was analysed in triplicate (n = 3).

concentrations were ranked as FUM > OCA > AFT > AFB1.
The animal health risk assessment results of the MOE and PTWI
methods are given in Table 5. In terms of animal health risk assessment,
a risk was determined for small, medium, and large breed dogs by the
consumption of dry foods due to AFT, AFB1, and OCA contamination
according to the considered average body weight (kg) of breeds (7.5, 20,
and 35 respectively) and daily food consumption (g day− 1) (200, 450,
and 650 respectively). Only the level of FUM contamination did not pose
a potential health risk to the consumer dogs.
Mycotoxin contamination of feed is a major health concern for dogs
and also a threat to the food industry. Since commercial foods are the
major source of acute mycotoxicosis cases, monitoring and controlling
the contamination levels in dry pet foods is necessary (Böhm et al., 2010;
Wouters et al., 2013). For determination of the mycotoxin
contamination levels in dog foods and other animal feeds, the ELISA
method that was confirmed by HPLC analyses is used due to the detec­
tion sensitivity, result accuracy, analysis precision, reproducibility, and
high recovery rate, as well as selective, sensitive, fast, and easy-to-use
advantages (Böhm et al., 2010; Ekici et al., 2016).
AF’s, which have nephrotoxic, immunotoxic, and carcinogenic ef­
fects, are known as hepatotoxic mainly (Wouters et al., 2013). In the
current study, AFT (ELISA: 0.54–3.66, HPLC: 0.61–3.08) and AFB1
(ELISA: 0.27–1.26, HPLC: 0.28–0.75) concentrations (ng g− 1) deter­
mined by both ELISA and HPLC were above the legal/recommended
levels (0.01–0.05). In HPLC analysis, concentrations (ng g− 1) in 62 su­
permarkets and premium quality dog foods (≤4 kg) were 0.23 and 0.16
for AFB1, 0.38 and 0.25 for AFT, respectively (Macías-Montes et al.,
2020).

15
H. Ekici and M. Yipel Toxicon 218 (2022) 13–18

Table 5 in dog food (Razzazi et al., 2001). In a study conducted in 32 dry foods in
Risk assessment results. China, the OCA concentration was above the LOD in 2 samples at the
Breeds EDI ng kg day− 1
MOE PTWI ug kg− 1
Risk Limits levels of 15.1 and 17.3 μg kg− 1 (Shao et al., 2018).
FUMs are hepatotoxic, immunotoxic, and have toxic effects on the
AFT Small 44.27 3.84 – <10,000
Medium 37.35 4.55 lungs (Macías-Montes et al., 2020). In the current study, FUM concen­
Large 30.83 5.51 trations (ELISA: 87.06, HPLC: 48.84) determined by both ELISA and
AFB1 Small 17.07 9.96 HPLC were above the legal/recommended levels (0.01–0.25) (Böhm
Medium 14.40 11.81 et al., 2010). FUM concentrations (ng g− 1) were reported as 570.55
Large 11.89 14.30
OCA Small 21.40 – 0.15 >0.1 μg kg− 1 (supermarket brands) and 1450.32 (premium brands), as a result of
Medium 27.51 0.19 HPLC analysis in 62 dog foods (Macías-Montes et al., 2020). In a study
Large 39.74 0.28 conducted on 32 dry foods in China, 30 samples were positive and the
mean FUM contamination level was reported as 87.2 μg kg− 1 (Shao
1
FUM Small 2.32 – 0.02 >2 μg kg−
Medium 1.96 0.01
et al., 2018). While no nephrotoxic findings were observed in dogs given
Large 1.62 0.01
0.1–0.2 mg kg OCA for 14 days, tissue and organ damage were reported
AFT: Total aflatoxin; AFB1: Aflatoxin B1; OCA: Ochratoxin A; FUM: Fumonisin: when 5–10 mg kg was given with feed (EFSA, 2004).
EDI: Estimated daily intake; MOE: Margin of exposure; PTWI: Provisional In the present study, the values less than 10,000 of MOE indicate that
tolerable weakly intakes.
the consumption of the purchased commercial (the lowest priced) dry
dog foods poses a risk to the specified individuals (small, medium, and
In a study conducted in Austria, AFT and AFB1 contamination were large breed dogs) in terms of AFT and AFB1 contamination according to
below the LOD in dog foods in all (76) samples (Böhm et al., 2010). In the consumption scenarios recommended on the labels. It is thought that
the case of aflatoxin B1 poisoning, which caused the death of 60 dogs out this risk may be due to conditions that create a favorable environment
of 65 Concentrations (ng g) in 62 supermarkets and premium quality for rapid mycotoxin growth, such as the sale of food in packages
dog foods (≤4 kg) were reported as 0.23 and 0.16 for AFB1, 0.38 and weighing over 10 kg in open packaging or improper storage during
0.25 for AFT respectively in HPLC analysis. Contaminated corn prod­ consumption. In the risk assessment of dry foods sold in Spain, the MOE
ucts, the contamination level was determined as 1.64–1,77 ppb in the values of ng kg day− 1 (105.9 for AFT, 143.5 for AFB1) were found below
HPLC analysis of the products (Wouters et al., 2013). These levels are 10,000, similar to the current study.
approximately 3 times higher than the current study results. This result Amounts higher than 0.1 μg kg− 1 indicate that the consumption of
shows that consuming 3 times or more dry food that is sold open or not the purchased commercial (the lowest priced) dry dog foods poses a risk
stored well may pose a risk in terms of chronic AF poisoning in the risk to the specified individuals (small, medium, and large breed dogs) in
assessment and can cause acute poisoning that can result in death. In the terms of OCA according to the consumption scenarios. In a risk assess­
case where nine dogs with liver failure were poisoned by commercial ment of dry foods sold in Spain, the PTWI value was found to be 15 for
dog food contaminated with AF, AFB1 levels were determined to be OCA, which was above 0.1 μg kg− 1, similar to the current study.
between 222.8 and 579 μg kg− 1 in HPLC analysis (Newman et al., 2007). Amounts higher than 0.2 μg kg− 1 indicate that the consumption of
In the analysis made by HPLC, in 15 dog foods, AFB1 was determined at the purchased commercial (the lowest priced) dry dog foods poses a risk
a maximum level of 39.7 and an average of 5.00 ng g (Sharma and to the specified individuals (small, medium, and large breed dogs) in
Márquez, 2001). AFB1 level was determined as 6.69 μg kg− 1 in a study terms of FUM according to the consumption scenarios. In the risk
conducted with ELISA in 21 pellet dog foods in Turkey (Basalan et al., assessment of dry foods sold in Spain, the PTWI value was found to be
2004). In a study conducted on commercial dog foods in Nigeria, it was 2000 for FUM and above 0.1 μg kg− 1, similar to the current study.
reported that average AFB1 concentrations were 5.12–7.14 μg kg-1 and
average AFT concentrations were 9.61 μg kg− 1, in all samples that were 4. Conclusions
detected positive (Akinrinmade and Akinrinde, 2012). In a study con­
ducted on 180 commercial dog foods in Brazil, it was reported that AFB1 The topics about the undesirable substances in feed is generally
was determined at an average concentration of 6.00–7.60 ng g− 1 focused on farm animals because of their potential toxic effects by way
(Campos et al., 2009). In a study conducted on 32 dry foods in China, 28 of the food chain. However, dogs are the most susceptible animal species
samples were positive and the mean FUM contamination level was re­ to these substances due to their longer lifespan and frequency of expo­
ported as 47.7 μg kg− 1 (Shao et al., 2018). sure. In particular, dogs fed with dry food are very susceptible to the
The main reason for this is thought to be excessive mycotoxin growth negative effects that this type of nutrition can cause for most of their
due to unsuitable storage conditions and the time passed by until the lives. While these negativities can be manifested by nutritional de­
whole food is sold or consumed, especially dry foods sold by open ficiencies, the main risk is life-threatening diseases such as cancer,
packages or large packages to make the kg unit of the product cheaper which may occur due to the toxic consequences of the contaminants in
(>10 kg). It has been reported that the amount of aflatoxin in foods the food. Among the toxins that can be found in dog foods, mycotoxins
increases (several times in 2 months) rapidly depending on the storage are among the most important due to their high probability of occur­
condition (FAO, 1992; Villers, 2014). rence and toxicity. Some sellers and dog owners can prefer to sell or buy
OCA is known as primarily nephrotoxic (Shao et al., 2018). In the large packaged foods that are sold in open packages because they are
current study, OCA concentrations (ELISA: 2.14, HPLC: 1.60) deter­ more economical. On the other hand, dog owners, shelter staff, and
mined by both ELISA and HPLC were above the legal/recommended those who feed stray animals prefer large packaged foods but can not
levels (0.01–0.25). Mean OCA concentrations (ng g1) in low packages of store them under suitable conditions during a long consumption time. In
weighted dog foods were reported as 0.44 (supermarket brands) and dogs, choosing dry food in the diet can increase the frequency and
0.39 (premium brands) as a result of HPLC analysis in low packages of duration of exposure to possible contaminants. While the daily, weekly,
weighted dog foods (Macías-Montes et al., 2020). In a study conducted and monthly diets of humans vary considerably in terms of the food
in Austria, 2007); OCA contamination (ELISA: 3.5 μg kg− 1) was higher in groups, pet animals such as cats and dogs consume dry food at a major
76 samples (Böhm et al., 2010). In the study conducted in Austria with level once or several times a day throughout life, in line with the com­
the ELISA method in 29 samples, OCA contamination was determined to mercial packaging recommendation. However, it should be taken into
be between 7 and 40 μg kg− 1 in 3 samples (Songsermsakul et al., 2007). account that feeds sold in open packaging will pose a long-term risk to
In the study performed by ELISA method on 17 cat and dog dry food animal health. For this reason, it is necessary to monitor the contami­
samples, the highest OCA contamination (13.1 μg kg− 1) was determined nation levels of mycotoxins in dog dry food and to evaluate the potential

16
H. Ekici and M. Yipel
health risks they pose.
Authorship contribution statement
Hüsamettin Ekici: Concept, Design, Data Collection or Processing,
Analysis or Interpretation, Literature Search, Writing. Mustafa Yipel:
Concept, Design, Data Collection or Processing, Analysis or Interpreta­
tion, Literature Search, Writing.
Funding
This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
Ethical statement
The authors declare that the article does not contain any studies with
humans or animals performed by the authors.
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.
Data availability
Data will be made available on request.
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