**4. Contamination level in cereal and soybean-based food and feed products**

Food security strategy in the European Union (EU) includes the Rapid Alert System for Food and Feed. The RASFF was established by the European Parliament and Council Regu‐ lation No. 178/2002 laying down the general principles and requirements of food law, estab‐ lishing the European Food Safety Authority and specifying the procedures in matters concerning food safety [58].

In 2002 – 2011, the number of notifications to the RASFF system due to mycotoxin contami‐ nation of food was respectively: 302, 803, 880, 996, 878, 760, 933, 669, 688, 631 notifications identifying the presence of aflatoxin B1 (AFB1) and the amount of AFB1, B2, G1, G2, AFM1, ochratoxin A (OTA), fumonisins B1 and B2 (FB1, FB2), patulin, deoxynivalenol (DON) and zearalenone (ZEA) in such groups of foods, as nuts and milk, oilseeds, cereal, dried fruit, fruit, cocoa, coffee, herbs and spices, wine, milk, products for children. Approximately 95% of the notifications concerned foodstuffs contaminated with aflatoxins. During this period, the number of notifications regarding mycotoxin contamination of grains did not exceed 15% of the total number of notifications. The data in Figure 1 show that in 2002-2011 aflatox‐ ins, ochratoxin A and fumonisins were the main contaminants isolated from cereals [59].

In the research of [60], ninety-fife cereal samples from retail shops and local markets of dif‐ ferent locations in Pakistan were examined in terms of the presence of aflatoxins. The results showed the percentage of aflatoxin contamination samples in the commodities such as in:


#### **Table 3.** Mycotoxigenic fungi and mycotoxins

rice (25%), broken rice (15%), wheat (20%), maize (40%), barley (20%) and sorghum (30%), while in soybean (15%). The highest contamination levels of aflatoxins were found in one wheat sample (15.5 ppb), one maize sample (13.0 ppb) and one barley sample (12.6 μg/kg). In the research of [61], seventeen samples of wheat grain from Morocco were tested for OTA and DON contamination. The results show that only two samples (11.76%) out of 17 were contaminated with OTA, at the mean concentration of 29.4 ppb. However, seven samples (41.17%) were contaminated with DON at the mean concentration of 65.9 ppb.

ng/kg [63], whereas in the samples of barley, wheat, and oat grains from Sweden it was con‐

2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 **year**

**aflatoxins deoxynivalenol fumonisins ochratoxin A zearalenone**

Figure 1. The number of notifications received by RASFF on mycotoxins in cereals in 2002-2011

**Figure 1.** The number of notifications received by RASFF on mycotoxins in cereals in 2002-2011

barley wheat corn

Figure 2. Level of contamination with aflatoxins in grains coming from different regions of Poland

Central Poland Western Poland Eastern Poland Polish Noon

Mill 1 Mill 2 Mill 3 Mill 4 Mill 5 Mill 6

**Figure 2.** Level of contamination with aflatoxins in grains coming from different regions of Poland

barley wheat corn

Mill 1 Mill 2 Mill 3 Mill 4 Mill 5 Mill 6

Figure 2. Level of contamination with aflatoxins in grains coming from different regions of Poland

Central Poland Western Poland Eastern Poland Polish Noon

Figure 1. The number of notifications received by RASFF on mycotoxins in cereals in 2002-2011

2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 **year**

notifications regarding mycotoxin contamination of grains did not exceed 15% of the total number of notifications. The data in Figure 1 show that in 2002-2011 aflatoxins, ochratoxin A and fumonisins were the main contaminants isolated from cereals [59].

Mycotoxins in Cereal and Soybean-Based Food and Feed

**aflatoxins deoxynivalenol fumonisins ochratoxin A zearalenone**

In the research of [60], ninety-fife cereal samples from retail shops and local markets of different locations in Pakistan were examined in terms of the presence of aflatoxins. The results showed the percentage of aflatoxin contamination samples in the commodities such as in: rice (25%), broken rice (15%), wheat (20%), maize (40%), barley (20%) and sorghum (30%), while in soybean (15%). The highest contamination levels of aflatoxins were found in one wheat sample (15.5 ppb), one maize sample (13.0 ppb) and one barley sample (12.6 μg/kg). In the research of [61], seventeen samples of wheat grain from Morocco were tested for OTA and DON contamination. The results show that only two samples (11.76%) out of 17 were contaminated with OTA, at the mean concentration of 29.4 ppb. However, seven samples (41.17%) were contaminated with DON at the mean concentration of 65.9

The aim of our own research [15] was mycotoxic analysis of grains included in the standard mixtures used in feed formulations. Eighteen samples were tested containing seeds evenly divided into three types: barley, wheat and corn. The tested seeds were from randomly selected Polish mills: the central, western, eastern and south ones (Figure 2). The aflatoxins content in 51% of the screened barley samples and in 34% of the screened wheat and maize samples did not exceed the limit set in the European Union Regulation, i.e. 4 ppb [62]. In reference to the grain origin, it was established that grains from the central and western parts of Poland exhibited the highest extent of AFs contamination. To compare, the AFs level in wheat grains from various regions of Turkey was very low, ranging from 10.4 to 634.5 ng/kg [63], whereas in the samples of barley, wheat, and oat grains from Sweden it

notifications regarding mycotoxin contamination of grains did not exceed 15% of the total number of notifications. The data in Figure 1 show that in 2002-2011 aflatoxins, ochratoxin A and fumonisins were the main contaminants isolated from cereals [59].

In the research of [60], ninety-fife cereal samples from retail shops and local markets of different locations in Pakistan were examined in terms of the presence of aflatoxins. The results showed the percentage of aflatoxin contamination samples in the commodities such as in: rice (25%), broken rice (15%), wheat (20%), maize (40%), barley (20%) and sorghum (30%), while in soybean (15%). The highest contamination levels of aflatoxins were found in one wheat sample (15.5 ppb), one maize sample (13.0 ppb) and one barley sample (12.6 μg/kg). In the research of [61], seventeen samples of wheat grain from Morocco were tested for OTA and DON contamination. The results show that only two samples (11.76%) out of 17 were contaminated with OTA, at the mean concentration of 29.4 ppb. However, seven samples (41.17%) were contaminated with DON at the mean concentration of 65.9

http://dx.doi.org/10.5772/54470

197

The aim of our own research [15] was mycotoxic analysis of grains included in the standard mixtures used in feed formulations. Eighteen samples were tested containing seeds evenly divided into three types: barley, wheat and corn. The tested seeds were from randomly selected Polish mills: the central, western, eastern and south ones (Figure 2). The aflatoxins content in 51% of the screened barley samples and in 34% of the screened wheat and maize samples did not exceed the limit set in the European Union Regulation, i.e. 4 ppb [62]. In reference to the grain origin, it was established that grains from the central and western parts of Poland exhibited the highest extent of AFs contamination. To compare, the AFs level in wheat grains from various regions of Turkey was very low, ranging from 10.4 to 634.5 ng/kg [63], whereas in the samples of barley, wheat, and oat grains from Sweden it

tained between 50 and 400 ppb [64].

was contained between 50 and 400 ppb [64].

was contained between 50 and 400 ppb [64].

ppb.

**number of notifications**

0

**ppb**

4

8

12

16

**ppb**

20

24

28

ppb.

0

5

10

15

20

25

**number of notifications**

30

35

40

45

50

The aim of our own research [15] was mycotoxic analysis of grains included in the standard mixtures used in feed formulations. Eighteen samples were tested containing seeds evenly divided into three types: barley, wheat and corn. The tested seeds were from randomly se‐ lected Polish mills: the central, western, eastern and south ones (Figure 2). The aflatoxins content in 51% of the screened barley samples and in 34% of the screened wheat and maize samples did not exceed the limit set in the European Union Regulation, i.e. 4 ppb [62]. In reference to the grain origin, it was established that grains from the central and western parts of Poland exhibited the highest extent of AFs contamination. To compare, the AFs level in wheat grains from various regions of Turkey was very low, ranging from 10.4 to 634.5

**year**

ppb) and one barley sample (12.6 μg/kg). In the research of [61], seventeen samples of wheat grain from Morocco were tested for

randomly selected Polish mills: the central, western, eastern and south ones (Figure 2). The aflatoxins content in 51% of the screened barley samples and in 34% of the screened wheat and maize samples did not exceed the limit set in the European Union

notifications regarding mycotoxin contamination of grains did not exceed 15% of the total number of notifications. The data in Figure 1 show that in 2002-2011 aflatoxins, ochratoxin A and fumonisins were the main contaminants isolated from cereals [59].

Regulation, i.e. 4 ppb [62]. In reference to the grain origin, it was established that grains from the central and western parts of In the research of [60], ninety-fife cereal samples from retail shops and local markets of different locations in Pakistan were **Figure 1.** The number of notifications received by RASFF on mycotoxins in cereals in 2002-2011

0

5

10

15

20

25

**number of notifications**

30

35

40

45

50

rice (25%), broken rice (15%), wheat (20%), maize (40%), barley (20%) and sorghum (30%), while in soybean (15%). The highest contamination levels of aflatoxins were found in one wheat sample (15.5 ppb), one maize sample (13.0 ppb) and one barley sample (12.6 μg/kg). In the research of [61], seventeen samples of wheat grain from Morocco were tested for OTA and DON contamination. The results show that only two samples (11.76%) out of 17 were contaminated with OTA, at the mean concentration of 29.4 ppb. However, seven samples

**Mycotoxin Produced species Commodities**

Nuts, spices, Cereals, maize, soybean, rice

Cereals, fruits, spices, coffee, Food of animal origin

Oats, rice, corn, beans, fruits, fruit and vegetable juices, herbs and spices

Cereals, cheese

Maize, soybean, cereals

Vegetables, fruit, cereals, soybean

Vegetables, fruit, cereals, soybean

*A.ochraceoroseus, A.pseudotamari*

*P.nordicum, P.palitans, P.commune, P.variabile, Aspergillus ochraceus, A.melleus, A.niger, A.carbonarius, A.sclerotiorum, A.sulphureus*

*Monascus purpureus*

*A.aureolatus, A.quadrilineatus, Eurotium amstelodami*

*F.cerealis, F.equiseti, F.incarnatum*

*Alternaria alternata, A.brassicae, A.capsici-anui, A.citri, A.cucumerina, A.dauci, A.kikuchiana, A.solani, A.tenuissima, A.tomato, A.longipes, A.infectoria, A.oregonensis*

*A.kikuchiana, A.mali, A.solani, A.oryzae, A.porri, A.radicina, A.tenuissima, A.tomato, A.longipes*

Deoksynivalenol *Fusarium graminearum, F.culmorum, F.crokwellense* Maize, soybean, cereals Fumonisins *Fusarium proliferatum, F.verticillioides,* Maize, soybean, cereals

Aflatoxins *Aspergillus flavus, A.parasiticus, A.nomius, A.bombycis,*

Citrinin *Penicillium citrinum, P.verrucosum, P.viridicatum,*

Sterigmatocystin *Aspergillus versicolor, A.nidulans, A.chevalieri, A.ruber,*

Zearalenone *Fusarium graminearum, F.sporotrichoides, F.culmorum,*

Tenuazonic acid *Alternaria alternata, A.capsici-anui, A.citri, A.japonica,*

Alternariol, alternariol monomethyl ether

196 Soybean - Pest Resistance

**Table 3.** Mycotoxigenic fungi and mycotoxins

Ochratoxin A *Penicillium verrucosum, P.auriantiogriseum,*

The aim of our own research [15] was mycotoxic analysis of grains included in the standard mixtures used in feed formulations. Eighteen samples were tested containing seeds evenly divided into three types: barley, wheat and corn. The tested seeds were from randomly se‐ lected Polish mills: the central, western, eastern and south ones (Figure 2). The aflatoxins content in 51% of the screened barley samples and in 34% of the screened wheat and maize samples did not exceed the limit set in the European Union Regulation, i.e. 4 ppb [62]. In reference to the grain origin, it was established that grains from the central and western parts of Poland exhibited the highest extent of AFs contamination. To compare, the AFs level in wheat grains from various regions of Turkey was very low, ranging from 10.4 to 634.5

(41.17%) were contaminated with DON at the mean concentration of 65.9 ppb.

ng/kg [63], whereas in the samples of barley, wheat, and oat grains from Sweden it was con‐ tained between 50 and 400 ppb [64]. Poland exhibited the highest extent of AFs contamination. To compare, the AFs level in wheat grains from various regions of Turkey was very low, ranging from 10.4 to 634.5 ng/kg [63], whereas in the samples of barley, wheat, and oat grains from Sweden it was contained between 50 and 400 ppb [64]. examined in terms of the presence of aflatoxins. The results showed the percentage of aflatoxin contamination samples in the commodities such as in: rice (25%), broken rice (15%), wheat (20%), maize (40%), barley (20%) and sorghum (30%), while in soybean (15%). The highest contamination levels of aflatoxins were found in one wheat sample (15.5 ppb), one maize sample (13.0

Figure 1. The number of notifications received by RASFF on mycotoxins in cereals in 2002-2011

Figure 2. Level of contamination with aflatoxins in grains coming from different regions of Poland **Figure 2.** Level of contamination with aflatoxins in grains coming from different regions of Poland **ppb**

Mill 1 Mill 2 Mill 3 Mill 4 Mill 5 Mill 6

Figure 2. Level of contamination with aflatoxins in grains coming from different regions of Poland

Central Poland Western Poland Eastern Poland Polish Noon

ppb.

Figure 3. Level of contamination with ochratoxina A in grains coming from different regions of Poland **Figure 3.** Level of contamination with ochratoxina A in grains coming from different regions of Poland

The OTA level in the examined grains collected from mills in central, eastern and southern Poland was low and ranged from 0.5 to 2.5 ppb (Figure 3). Therefore, it did not exceed the permissible limit set by the European Union (Commission Regulation No. 105/2010), i.e. 5 ppb [65]. Only in barley coming from a mill located in western Poland, the OTA level exceeded the limits fivefold. The extent of OTA contamination of barley, wheat, and maize grain from various regions of Mexico was also low and recorded 0.17 ppb, 0.42 ppb, and 1.08 ppb, respectively. Only 1 out of 20 examined maize grains showed the OTA level of 7.22 [66]. To compare, the OTA concentration in barley and wheat grain from the UK equalled from 1 to 33 ppb [67]. In the research of [68], among others, the levels of AFs and OTA in 532 grain and feed samples from Poland from 2002 and 2003 were determined. The average mycotoxin concentration levels were similar and quite low, i.e. AFs - 0.3 ppb and OTA - 1.1 ppb in grains and feeds from 2002, and respectively, AFs 3.1 and 1.0 ppb and OTA 0.5 and 0.7 OTA in samples from 2003. The authors of the study stressed that in 2002 and 2003 the harvesting seasons were hot and dry, which might have resulted in the low extent of fungi contamination of the examined grain. Although the extent of mycotoxin contamination of grain in the quoted studies varies, their authors concur that it is a serious issue whose scale depends on the microclimate during arable farming and the subsequent phases, i.e. grain storage. It was reported that no mycotoxins were found in barley samples stored for 20 weeks at 15% seed humidity, whereas the samples of wheat stored for the same period of time at 19% humidity recorded relatively high concentration levels: OTA - 24 ppb, citrinin - 38 ppb, and sterigmatocystin even up to 411 ppb [69]. The aim of our research was the assessment of cereal products available in trade and meant for direct consumption as for contamination with selected mycotoxins. The research included corn flakes, corn flakes with nuts and honey, various kinds of The OTA level in the examined grains collected from mills in central, eastern and southern Poland was low and ranged from 0.5 to 2.5 ppb (Figure 3). Therefore, it did not exceed the permissible limit set by the European Union (Commission Regulation No. 105/2010), i.e. 5 ppb [65]. Only in barley coming from a mill located in western Poland, the OTA level ex‐ ceeded the limits fivefold. The extent of OTA contamination of barley, wheat, and maize grain from various regions of Mexico was also low and recorded 0.17 ppb, 0.42 ppb, and 1.08 ppb, respectively. Only 1 out of 20 examined maize grains showed the OTA level of 7.22 [66]. To compare, the OTA concentration in barley and wheat grain from the UK equalled from 1 to 33 ppb [67]. In the research of [68], among others, the levels of AFs and OTA in 532 grain and feed samples from Poland from 2002 and 2003 were determined. The average my‐ cotoxin concentration levels were similar and quite low, i.e. AFs - 0.3 ppb and OTA - 1.1 ppb in grains and feeds from 2002, and respectively, AFs 3.1 and 1.0 ppb and OTA 0.5 and 0.7 OTA in samples from 2003. The authors of the study stressed that in 2002 and 2003 the har‐ vesting seasons were hot and dry, which might have resulted in the low extent of fungi con‐ tamination of the examined grain. Although the extent of mycotoxin contamination of grain in the quoted studies varies, their authors concur that it is a serious issue whose scale de‐ pends on the microclimate during arable farming and the subsequent phases, i.e. grain stor‐ age. It was reported that no mycotoxins were found in barley samples stored for 20 weeks at 15% seed humidity, whereas the samples of wheat stored for the same period of time at 19% humidity recorded relatively high concentration levels: OTA - 24 ppb, citrinin - 38 ppb, and sterigmatocystin even up to 411 ppb [69].

breakfast cereal products and muesli containing dried fruit, nuts as well as cereal and coconut flakes (15 samples). None of the products was contaminated with AB1 on the level exceeding the acceptable limits (2 ppb). The presence of ochratoxin A exceeding the amount of 3 ppb was discovered in four samples (two kinds of corn flakes, exotic muesli and traditional muesli). The The aim of our research was the assessment of cereal products available in trade and meant for direct consumption as for contamination with selected mycotoxins. The research includ‐ ed corn flakes, corn flakes with nuts and honey, various kinds of breakfast cereal products

contamination with that toxin equalled 4.5 ppb on average. According to the current regulation, contamination of breakfast flakes with deoxynivalenol DON should not exceed 500 ppb. Four samples (containing corn) exceeded this limit by 50%. In case of one

and muesli containing dried fruit, nuts as well as cereal and coconut flakes (15 samples). None of the products was contaminated with AB1 on the level exceeding the acceptable lim‐ its (2 ppb). The presence of ochratoxin A exceeding the amount of 3 ppb was discovered in four samples (two kinds of corn flakes, exotic muesli and traditional muesli). The contami‐ nation with that toxin equalled 4.5 ppb on average. According to the current regulation, con‐ tamination of breakfast flakes with deoxynivalenol DON should not exceed 500 ppb. Four samples (containing corn) exceeded this limit by 50%. In case of one sample, DON contami‐

Mycotoxins in Cereal and Soybean-Based Food and Feed

http://dx.doi.org/10.5772/54470

199

Mycotoxin contamination of soybean is not considered a significant problem as compared to commodities such as corn, cottonseed, peanuts, barley and other grains. In the early surveys conducted by the U.S. Department of Agriculture (USDA), 1046 soybean samples collected from different regions of the United States were examined for aflatoxins contamination. Aflatoxin presence was confirmed at low levels (7-14 ppb) in only two of the tested samples [70]. In the research of [71], fifty-five samples of soybean meals were analysed for the con‐ tent of aflatoxins, deoxynivalenol (DON), zearalenone (ZEA) and ochratoxin A (OTA). Re‐ garding aflatoxins, only AFB1 was detected in 32 out of the 51 non-suspicious samples, but the maximal concentration found was only 0.41 ppb. ZEA was detected in 23 out of the 51 samples with a maximum concentration of 18 ppb. DON could be detected only in one sus‐ picious sample in a low concentration of 104 ppb. OTA was found in 5 samples, with the

The research of [72] tested 122 soybean samples that came from Asia and the Pacific region. Aflatoxin was found in only in 2% (maximum of 13 ppb, median 9 ppb), zearalenone in 17% (maximum 1078 ppb, median 57 ppb), ochratoxin in 13% (maximum 11 ppb, median 7 ppb), and DON and fumonisins each in 7% of the analyzed samples (DON: maximum 1347ppb, median 264 ppb; fumonisins: maximum 331 ppb, median 154 ppb). In maize and maize products, the levels of fumonisins varied from 0.07 to 38.5 ppm in Latin America, from 0.004 to 330 ppm in North America, from 0.02 to 8.85 ppm in Africa, and from 0.01 to 153 ppm in Asia. The data available for Europe varied from 0.007 to 250 ppm in maize, and from 0.008

Effects of mycotoxins on human and animal health are now increasingly recognised. Myco‐ toxins enter human and animal dietary systems mainly through ingestion, but increasing evidence also points to inhalation as another entry route. Mycotoxins exhibit a wide array of

**•** hematopoietic - aflatoxins and trichothecenes. Hemotopoiesis refers to the production of all types of blood cells from the primitive cells stem cells in the bone marrow. The dys‐

**5. Influence of mycotoxins on human and animal organisms**

**•** carcinogenic - aflatoxins, ochratoxins, fumonisins, and possibly patulin;

biological effects and individual mycotoxins can be [73]:

**•** mutagenic - aflatoxins and sterigmatocystin;

nation was very high, almost three times higher than the acceptable level [19].

greatest concentration being only 1 ppb.

to 16 ppm in maize products. [54].

Mycotoxin contamination of soybean is not considered a significant problem as compared to commodities such as corn, cottonseed, peanuts, barley and other grains. In the early surveys conducted by the U.S. Department of Agriculture (USDA), 1046 soybean samples collected from different regions of the United States were examined for aflatoxins contamination. Aflatoxin presence was confirmed at low levels (7-14 ppb) in only two of the tested samples [70]. In the research of [71], fifty-five samples of soybean meals were analysed for the content of aflatoxins, deoxynivalenol (DON), zearalenone (ZEA) and ochratoxin A (OTA). Regarding aflatoxins, only AFB1 was detected in 32 out of the 51 non-suspicious samples, but the maximal concentration found was only 0.41 ppb. ZEA was detected in 23 out of the 51 samples with a maximum concentration of 18 ppb. DON could be detected only in one suspicious sample in a low concentration of 104 ppb. OTA was found in 5 samples, with the greatest concentration being only 1

The research of [72] tested 122 soybean samples that came from Asia and the Pacific region. Aflatoxin was found in only in 2% (maximum of 13 ppb, median 9 ppb), zearalenone in 17% (maximum 1078 ppb, median 57 ppb), ochratoxin in 13% (maximum 11 ppb, median 7 ppb), and DON and fumonisins each in 7% of the analyzed samples (DON: maximum 1347ppb, median 264 ppb; fumonisins: maximum 331 ppb, median 154 ppb). In maize and maize products, the levels of fumonisins varied from 0.07 to 38.5 ppm in Latin America, from 0.004 to 330 ppm in North America, from 0.02 to 8.85 ppm in Africa, and from 0.01 to 153 ppm in Asia.

The data available for Europe varied from 0.007 to 250 ppm in maize, and from 0.008 to 16 ppm in maize products. [54].

**5. Influence of mycotoxins on human and animal organisms** 

sample, DON contamination was very high, almost three times higher than the acceptable level [19].

and muesli containing dried fruit, nuts as well as cereal and coconut flakes (15 samples). None of the products was contaminated with AB1 on the level exceeding the acceptable lim‐ its (2 ppb). The presence of ochratoxin A exceeding the amount of 3 ppb was discovered in four samples (two kinds of corn flakes, exotic muesli and traditional muesli). The contami‐ nation with that toxin equalled 4.5 ppb on average. According to the current regulation, con‐ tamination of breakfast flakes with deoxynivalenol DON should not exceed 500 ppb. Four samples (containing corn) exceeded this limit by 50%. In case of one sample, DON contami‐ nation was very high, almost three times higher than the acceptable level [19].

The OTA level in the examined grains collected from mills in central, eastern and southern Poland was low and ranged from 0.5 to Mycotoxin contamination of soybean is not considered a significant problem as compared to commodities such as corn, cottonseed, peanuts, barley and other grains. In the early surveys conducted by the U.S. Department of Agriculture (USDA), 1046 soybean samples collected from different regions of the United States were examined for aflatoxins contamination. Aflatoxin presence was confirmed at low levels (7-14 ppb) in only two of the tested samples [70]. In the research of [71], fifty-five samples of soybean meals were analysed for the con‐ tent of aflatoxins, deoxynivalenol (DON), zearalenone (ZEA) and ochratoxin A (OTA). Re‐ garding aflatoxins, only AFB1 was detected in 32 out of the 51 non-suspicious samples, but the maximal concentration found was only 0.41 ppb. ZEA was detected in 23 out of the 51 samples with a maximum concentration of 18 ppb. DON could be detected only in one sus‐ picious sample in a low concentration of 104 ppb. OTA was found in 5 samples, with the greatest concentration being only 1 ppb.

2.5 ppb (Figure 3). Therefore, it did not exceed the permissible limit set by the European Union (Commission Regulation No. 105/2010), i.e. 5 ppb [65]. Only in barley coming from a mill located in western Poland, the OTA level exceeded the limits fivefold. The extent of OTA contamination of barley, wheat, and maize grain from various regions of Mexico was also low and recorded 0.17 ppb, 0.42 ppb, and 1.08 ppb, respectively. Only 1 out of 20 examined maize grains showed the OTA level of 7.22 [66]. To compare, the OTA concentration in barley and wheat grain from the UK equalled from 1 to 33 ppb [67]. In the research of [68], among others, the levels of AFs and OTA in 532 grain and feed samples from Poland from 2002 and 2003 were determined. The average mycotoxin concentration levels were similar and quite low, i.e. AFs - 0.3 ppb and OTA - 1.1 ppb in grains and feeds from 2002, and respectively, AFs 3.1 and 1.0 ppb and OTA 0.5 and 0.7 OTA in samples from 2003. The authors of the study stressed that in 2002 The research of [72] tested 122 soybean samples that came from Asia and the Pacific region. Aflatoxin was found in only in 2% (maximum of 13 ppb, median 9 ppb), zearalenone in 17% (maximum 1078 ppb, median 57 ppb), ochratoxin in 13% (maximum 11 ppb, median 7 ppb), and DON and fumonisins each in 7% of the analyzed samples (DON: maximum 1347ppb, median 264 ppb; fumonisins: maximum 331 ppb, median 154 ppb). In maize and maize products, the levels of fumonisins varied from 0.07 to 38.5 ppm in Latin America, from 0.004 to 330 ppm in North America, from 0.02 to 8.85 ppm in Africa, and from 0.01 to 153 ppm in Asia. The data available for Europe varied from 0.007 to 250 ppm in maize, and from 0.008 to 16 ppm in maize products. [54].

#### examined grain. Although the extent of mycotoxin contamination of grain in the quoted studies varies, their authors concur that it is a serious issue whose scale depends on the microclimate during arable farming and the subsequent phases, i.e. grain storage. It **5. Influence of mycotoxins on human and animal organisms**

was reported that no mycotoxins were found in barley samples stored for 20 weeks at 15% seed humidity, whereas the samples of wheat stored for the same period of time at 19% humidity recorded relatively high concentration levels: OTA - 24 ppb, citrinin - 38 Effects of mycotoxins on human and animal health are now increasingly recognised. Myco‐ toxins enter human and animal dietary systems mainly through ingestion, but increasing evidence also points to inhalation as another entry route. Mycotoxins exhibit a wide array of biological effects and individual mycotoxins can be [73]:


Figure 3. Level of contamination with ochratoxina A in grains coming from different regions of Poland

Central Poland Western Poland Eastern Poland Polish Noon

Mill 1 Mill 2 Mill 3 Mill 4 Mill 5 Mill 6

The OTA level in the examined grains collected from mills in central, eastern and southern Poland was low and ranged from 0.5 to 2.5 ppb (Figure 3). Therefore, it did not exceed the permissible limit set by the European Union (Commission Regulation No. 105/2010), i.e. 5 ppb [65]. Only in barley coming from a mill located in western Poland, the OTA level ex‐ ceeded the limits fivefold. The extent of OTA contamination of barley, wheat, and maize grain from various regions of Mexico was also low and recorded 0.17 ppb, 0.42 ppb, and 1.08 ppb, respectively. Only 1 out of 20 examined maize grains showed the OTA level of 7.22 [66]. To compare, the OTA concentration in barley and wheat grain from the UK equalled from 1 to 33 ppb [67]. In the research of [68], among others, the levels of AFs and OTA in 532 grain and feed samples from Poland from 2002 and 2003 were determined. The average my‐ cotoxin concentration levels were similar and quite low, i.e. AFs - 0.3 ppb and OTA - 1.1 ppb in grains and feeds from 2002, and respectively, AFs 3.1 and 1.0 ppb and OTA 0.5 and 0.7 OTA in samples from 2003. The authors of the study stressed that in 2002 and 2003 the har‐ vesting seasons were hot and dry, which might have resulted in the low extent of fungi con‐ tamination of the examined grain. Although the extent of mycotoxin contamination of grain in the quoted studies varies, their authors concur that it is a serious issue whose scale de‐ pends on the microclimate during arable farming and the subsequent phases, i.e. grain stor‐ age. It was reported that no mycotoxins were found in barley samples stored for 20 weeks at 15% seed humidity, whereas the samples of wheat stored for the same period of time at 19% humidity recorded relatively high concentration levels: OTA - 24 ppb, citrinin - 38 ppb, and

The aim of our research was the assessment of cereal products available in trade and meant for direct consumption as for contamination with selected mycotoxins. The research includ‐ ed corn flakes, corn flakes with nuts and honey, various kinds of breakfast cereal products

barley wheat corn

**Figure 3.** Level of contamination with ochratoxina A in grains coming from different regions of Poland

ppb, and sterigmatocystin even up to 411 ppb [69].

sterigmatocystin even up to 411 ppb [69].

ppb.

0

4

8

12

16

**ppb**

20

24

28

198 Soybean - Pest Resistance

and 2003 the harvesting seasons were hot and dry, which might have resulted in the low extent of fungi contamination of the

contamination with that toxin equalled 4.5 ppb on average. According to the current regulation, contamination of breakfast flakes with deoxynivalenol DON should not exceed 500 ppb. Four samples (containing corn) exceeded this limit by 50%. In case of one

Mycotoxin contamination of soybean is not considered a significant problem as compared to commodities such as corn, cottonseed, peanuts, barley and other grains. In the early surveys conducted by the U.S. Department of Agriculture (USDA), 1046 soybean samples collected from different regions of the United States were examined for aflatoxins contamination. Aflatoxin presence was confirmed at low levels (7-14 ppb) in only two of the tested samples [70]. In the research of [71], fifty-five samples of soybean meals were analysed for the content of aflatoxins, deoxynivalenol (DON), zearalenone (ZEA) and ochratoxin A (OTA). Regarding aflatoxins, only AFB1 was detected in 32 out of the 51 non-suspicious samples, but the maximal concentration found was only 0.41 ppb. ZEA was detected in 23 out of the 51 samples with a maximum concentration of 18 ppb. DON could be detected only in one suspicious sample in a low concentration of 104 ppb. OTA was found in 5 samples, with the greatest concentration being only 1

The research of [72] tested 122 soybean samples that came from Asia and the Pacific region. Aflatoxin was found in only in 2% (maximum of 13 ppb, median 9 ppb), zearalenone in 17% (maximum 1078 ppb, median 57 ppb), ochratoxin in 13% (maximum 11 ppb, median 7 ppb), and DON and fumonisins each in 7% of the analyzed samples (DON: maximum 1347ppb, median 264 ppb; fumonisins: maximum 331 ppb, median 154 ppb). In maize and maize products, the levels of fumonisins varied from 0.07 to 38.5 ppm in Latin America, from 0.004 to 330 ppm in North America, from 0.02 to 8.85 ppm in Africa, and from 0.01 to 153 ppm in Asia.

The data available for Europe varied from 0.007 to 250 ppm in maize, and from 0.008 to 16 ppm in maize products. [54].

**5. Influence of mycotoxins on human and animal organisms** 

sample, DON contamination was very high, almost three times higher than the acceptable level [19].

breakfast cereal products and muesli containing dried fruit, nuts as well as cereal and coconut flakes (15 samples). None of the products was contaminated with AB1 on the level exceeding the acceptable limits (2 ppb). The presence of ochratoxin A exceeding the amount of 3 ppb was discovered in four samples (two kinds of corn flakes, exotic muesli and traditional muesli). The **•** hematopoietic - aflatoxins and trichothecenes. Hemotopoiesis refers to the production of all types of blood cells from the primitive cells stem cells in the bone marrow. The dys‐ function of hematopoiesis leads firstly to the decrease in the number of neutrophils, thus perturbing the animal's immune system and subsequently to the decrease in red blood cells, which leads to anemia;


Table 4 presents the groups of mycotoxins which are most harmful to human and animal organisms, together with the chosen disease symptoms they cause.

> found the level of aflatoxin B1 as high as 4400 ppb [76]. Research in Gambian children and adults reported a strong association between aflatoxin exposure and impaired immunocom‐ petence suggesting that the consumption of aflatoxin reduces resistance to infections in hu‐ man populations [77, 78]. In 1974, an epidemic of hepatitis in India affected 400 people resulting in 100 deaths. The death was due to consumption of corn that was contaminated

Aflatoxins I \* aflatoxicosis, primary liver cancer, lung neoplasm, lung

Ochratoxins II B \*\* renal diseases, nephropathy, anorexia, vomiting, intestinal

Fumonisins II B \*\* diseases of the nervous system, cerebral softening,

Zearalenone - reproduction disruptions, abortions, pathological changes in

Trichothecenes - nausea, vomiting, haemorrhages, anorexia, alimentary toxic

\*The agent (mixture) is carcinogenic to humans. The exposure circumstance entails exposures that are carcinogenic to

\*\*The agent (mixture) is possibly carcinogenic to humans. The exposure circumstance entails exposures that are possi‐

**Symptoms and diseases**

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cancer, failure of the immune system, vomiting, depression, hepatitis, anorexia, jaundice, vascular coagulation

haemorrhage, tonsillitis, dehydration

pulmonary oedema, liver cancers, kidney diseases, oesophagus cancers, anorexia, depression, ataxia, blindness, hysteria, vomiting, hypotension

the reproductive system

aleukia, failure of the immune system, infants' lung bleeding, increased thirst, skin rash

Ochratoxin A was the cause of epithelial tumours of the upper urinary tract in the Balkans [80, 81]. The condition is known as Balkan endemic nephropathy. Despite the seriousness of the problem, the study did not explain the mechanism of action and the size of OTA carcino‐ genicity in humans [82]. Ochratoxin has been detected in blood in 6-18% of the human pop‐ ulation in some areas where Balkan endemic nephropathy is prevalent. Ochratoxin A has also been found in human blood samples from outside the Balkan Peninsula. In some sur‐ vey, over 50% of the tested samples were contaminated. A highly significant correlation was observed between Balkan nephropathy and urinary tract cancers, particularly tumours of the renal pelvis and ureter. However, no data have been published that establishes a direct

with *A. flavus* containing up to 15000 ppb of aflatoxins [79].

**Table 4.** The list of adverse effects of the chosen mycotoxins.

**Mycotoxin Toxicity class according to**

humans

bly carcinogenic to humans.

**International Agency for Research on Cancer (IARC)**

causal role of ochratoxin A in the etiology of these tumours [81].

#### **5.1. Negative effects of mycotoxins on humans**

Mycotoxicoses can be divided into acute and chronic. Acute toxicity usually has a rapid on‐ set and obvious toxic response, chronic exposure is characterized by chronic doses over a long period of time and may lead to cancer and other effects that are generally irreversible. The symptoms of mycotoxicosis depend on the type, amount and duration of exposure, age, health and sex of the exposed individual, and many poorly understood synergistic effects involving genetics, dietary status, and interaction with other toxic contaminants. Thus, the severity of mycotoxin poisoning can be compounded by factors such as vitamin deficiency, caloric deprivation, alcohol abuse, and infectious disease status. Mycotoxicosis is difficult to diagnose because doctors do not have experience with this disease and its symptoms are so wide that it mimics many other conditions [74, 75].

Aflatoxicosis is toxic hepatitis leading to jaundice and, in severe cases, death. AFB1 has been extensively linked to human primary liver cancer and was classified by the International Agency for Research on Cancer (IARC) as a human carcinogen (Group 1A - carcinogens) [49]. Although acute aflatoxicosis in humans is rare, several outbreaks have been reported. In 2004, one of the largest aflatoxicosis outbreaks in Kenya, resulting in 317 cases and 125 deaths was observed. Contaminated corn was responsible for the outbreak, and officials


\*The agent (mixture) is carcinogenic to humans. The exposure circumstance entails exposures that are carcinogenic to humans

\*\*The agent (mixture) is possibly carcinogenic to humans. The exposure circumstance entails exposures that are possi‐ bly carcinogenic to humans.

**Table 4.** The list of adverse effects of the chosen mycotoxins.

function of hematopoiesis leads firstly to the decrease in the number of neutrophils, thus perturbing the animal's immune system and subsequently to the decrease in red blood

**•** hepatotoxic - aflatoxins, ochratoxins, fumonisins. All of them induce significant liver

**•** neurotoxic - ergot alkaloids, fumonisins, deoksynivalenol. The effects of mycotoxins are best evidenced by vomiting and taste aversion produced by DON, seizures, focal malata and liquefaction of the brain tissue, possibly mediated by sphingolipid synthesis under the influence of fumonisins, staggering and trembling produced by many tremorgenic penitrem mycotoxins seizures and other neural effects of ergot alkaloids and parasympa‐ thomimetic activity resulting from the effects of the metabolite slaframine for selected re‐

**•** immunosupresive - several mycotoxins. The predominant mycotoxins in this regard are aflatoxins, trichothecenes, and ochratoxin A. However, several other mycotoxins such as fumonisins, zearalenone, patulin, citrinin, and fescue and ergot alkaloids have been

Table 4 presents the groups of mycotoxins which are most harmful to human and animal

Mycotoxicoses can be divided into acute and chronic. Acute toxicity usually has a rapid on‐ set and obvious toxic response, chronic exposure is characterized by chronic doses over a long period of time and may lead to cancer and other effects that are generally irreversible. The symptoms of mycotoxicosis depend on the type, amount and duration of exposure, age, health and sex of the exposed individual, and many poorly understood synergistic effects involving genetics, dietary status, and interaction with other toxic contaminants. Thus, the severity of mycotoxin poisoning can be compounded by factors such as vitamin deficiency, caloric deprivation, alcohol abuse, and infectious disease status. Mycotoxicosis is difficult to diagnose because doctors do not have experience with this disease and its symptoms are so

Aflatoxicosis is toxic hepatitis leading to jaundice and, in severe cases, death. AFB1 has been extensively linked to human primary liver cancer and was classified by the International Agency for Research on Cancer (IARC) as a human carcinogen (Group 1A - carcinogens) [49]. Although acute aflatoxicosis in humans is rare, several outbreaks have been reported. In 2004, one of the largest aflatoxicosis outbreaks in Kenya, resulting in 317 cases and 125 deaths was observed. Contaminated corn was responsible for the outbreak, and officials

**•** teratogenic - aflatoxin B1, ochratoxin A, T-2 toxin, sterigmatocystin, and zearalenone;

**•** nephrotoxigenic - ochratoxins, citrinin, trichothecenes, and fumonisins;

cells, which leads to anemia;

200 Soybean - Pest Resistance

**•** oestrogenic - zearalenone;

ceptors in the nervous system

shown to produce some effects on the immune system.

**5.1. Negative effects of mycotoxins on humans**

wide that it mimics many other conditions [74, 75].

organisms, together with the chosen disease symptoms they cause.

damage when given to animals;

found the level of aflatoxin B1 as high as 4400 ppb [76]. Research in Gambian children and adults reported a strong association between aflatoxin exposure and impaired immunocom‐ petence suggesting that the consumption of aflatoxin reduces resistance to infections in hu‐ man populations [77, 78]. In 1974, an epidemic of hepatitis in India affected 400 people resulting in 100 deaths. The death was due to consumption of corn that was contaminated with *A. flavus* containing up to 15000 ppb of aflatoxins [79].

Ochratoxin A was the cause of epithelial tumours of the upper urinary tract in the Balkans [80, 81]. The condition is known as Balkan endemic nephropathy. Despite the seriousness of the problem, the study did not explain the mechanism of action and the size of OTA carcino‐ genicity in humans [82]. Ochratoxin has been detected in blood in 6-18% of the human pop‐ ulation in some areas where Balkan endemic nephropathy is prevalent. Ochratoxin A has also been found in human blood samples from outside the Balkan Peninsula. In some sur‐ vey, over 50% of the tested samples were contaminated. A highly significant correlation was observed between Balkan nephropathy and urinary tract cancers, particularly tumours of the renal pelvis and ureter. However, no data have been published that establishes a direct causal role of ochratoxin A in the etiology of these tumours [81].

Fumonisin B1 was classified by the IARC as a group 2B carcinogen (possibly carcinogenic for humans) [44]. Fumonisins, which inhibit the absorption of folic acid through the foliate re‐ ceptor, have also been implicated in the high incidence of neural tube defects in the rural population known to consume contaminated corn, such as the former Transkei region of South Africa and some areas of Northern China [75, 83].

or both were evaluated. Compared to the control group, the body weight gains were re‐ duced by 26, 24 and 52% for animals consuming diets containing AFs, OTA, or both, respec‐ tively. Additional symptoms in pig ochratoxicosis were anorexia, fainting, uncoordinated movements, and increased water consumption and urination. Pigs also are susceptible to other mycotoxins, such as fumonsins and ergot alkaloids. Fumonisin B1, for example, has been shown to cause pulmonary oedema and heart and respiratory dysfunction. The symp‐ toms of swine pulmonary oedema included dyspnoea, cyanosis, and death [89, 90]. Myco‐ toxic porcine nephropathy is a serious disease, often associated with pigs consuming feed contaminated with OTA, especially in Scandinavian region. In addition to the enlarged and pale kidneys (with vascular lesions and white spots), morphological changes include a prox‐ imal tubular injury, epithelial atrophy, fibrosis and hyalinization of renal glomerular [80, 81]. Negative effects of ZEA on pigs' reproductive function have also been demonstrated [91]. Oestrogenic effects of ZEA on gilts and sows include oedematous uterus and ovarian cysts, increased maturation of follicles, more numerous litters or decreased fertility [92].

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Aflatoxins affect the quality of the milk produced by dairy cows and result in a carry-over of AFM1 with AFB1-contaminated feed. Ten ruminally-canulated Holstein cows received AFB1 (13 mg per cow daily) through a hole in the rumen for 7 days. The AFM1 levels in the milk of the treated cows ranged from 1.05 to 10.58 ng/L. The carry-over rate was higher in early lac‐ tation (2-4 weeks) compared to late lactation (34 -36 weeks) [75, 93]. The T-2 toxin causes ne‐ crosis of the lymphoid tissues. Bovine infertility and natural abortion in the last trimester of pregnancy also result from consumption of feed contaminated with T-2 toxin. Calves con‐ suming T-2 toxin in the amount of 10-50 mg/kg of feed showed abomasal ulcers and slough‐

Since the discovery of aflatoxins in the 1960s, regulations have been established in many countries to protect consumers from harmful mycotoxins that can contaminate foods. Maxi‐ mum levels of mycotoxins have been established by the European Commission after consul‐ tations with the Scientific Committee for Food, based on the analysis of scientific data

**•** distribution of concentrations of mycotoxins in raw materials or a product batch

The first two factors provide the information necessary for risk assessment and exposure as‐ sessment, respectively. Risk assessment is the scientific evaluation of the likelihood of

ing of papillae in the rumen [75, 94, 95].

collected by EFSA and the Codex Alimentarius.

**•** toxicological properties of mycotoxins,

These data include [73, 96]:

**•** mycotoxin dietary exposure,

**•** availability of analytical methods,

**6. Current EU regulations concerning mycotoxins**

**•** regulations in other countries with which trade contacts exist.

Trichothecenes have been proposed as potential biological warfare agents. In the years 1975-1981, T-2 toxin was implicated as a chemical agent "yellow rain" used against the Lao Peoples Democratic Republic. A study conducted from 1978 to 1981 in Cambodia revealed the presence of T-2 toxin, DON, ZEA, and nivalenol in water and leaf samples taken from the affected areas [75, 84]. Clinical symptoms proceeding to death included vomiting, diar‐ rhoea, bleeding, and difficulty with breathing, pain, blisters, headache, fatigue and dizzi‐ ness. There also occurred necrosis of the mucosa of the stomach as well as the small intestine, lungs and liver [85]. One disease outbreak was recorded in China and was associ‐ ated with the consumption of scabby wheat containing 1000-40000 ppb of DON. The disease is characterized by gastrointestinal symptoms. Also, in India there took place a reported in‐ fection associated with the consumption of bread made from contaminated wheat (DON 350-8300 ppb, acetyldeoxynivalenol 640-2490 ppb, NIV 30-100 ppb and T-2 toxin 500-800 ppb). The disease is characterized by gastrointestinal symptoms and throat irritation, which developed within 15 minutes to one hour after ingestion of the contaminated bread [81].

#### **5.2. Negative effects of mycotoxins on animal**

Animals may show varied symptoms upon contact with mycotoxins, depending on the ge‐ netic factors (species, breed, and strain), physiological factors (age, nutrition) and environ‐ mental factors (climatic conditions, rearing and management). The natural contamination with mycotoxins in animal feed usually does not occur at the levels that may cause acute or overt mycotoxicosis, such as hepatitis, bleeding, nephritis and necrosis of the oral and enter‐ ic epithelium, and even death. It is often difficult to observe and diagnose the symptoms of the disease, but it certainly is the most common form of mycotoxicosis in farm animals, af‐ fecting such parameters as productivity, growth and reproductive performance, feed effi‐ ciency, milk and egg production.

The negative effects of mycotoxins on the performance of poultry have been shown in nu‐ merous studies. For example, feeding the broilers with feed containing an AFs mixture (79% AFB1, 16% AFG1, AFB2 4% and 1% AFG2) in the concentration of 3.5 ppm decreased their body weight and increased their liver and kidney weight [75, 86]. Feeding OTA (0.3-1 ppm) to broilers reduced glycogenolysis and dose-dependent accumulation of glycogen in the liv‐ er. These negative metabolic reactions were attributed to inhibition of cyclic adenosine 3',5' monophosphate-dependent protein kinase, and were reflected in reduced efficiency of feed utilization and teratogenic malformations [75].

*Fusarium* mycotoxins proved to be harmful to poultry. In addition to reduced feed intake and weight gain, sore mouth, cheeks and plaque formation was observed after 7-day-old chicks were exposed to T-2 toxin (4 or 16 ppm) [75, 87]. Pigs are among the most sensitive species to mycotoxins. In the study by [88], pigs in response to AFs (2 ppm), OTA (2 ppm), or both were evaluated. Compared to the control group, the body weight gains were re‐ duced by 26, 24 and 52% for animals consuming diets containing AFs, OTA, or both, respec‐ tively. Additional symptoms in pig ochratoxicosis were anorexia, fainting, uncoordinated movements, and increased water consumption and urination. Pigs also are susceptible to other mycotoxins, such as fumonsins and ergot alkaloids. Fumonisin B1, for example, has been shown to cause pulmonary oedema and heart and respiratory dysfunction. The symp‐ toms of swine pulmonary oedema included dyspnoea, cyanosis, and death [89, 90]. Myco‐ toxic porcine nephropathy is a serious disease, often associated with pigs consuming feed contaminated with OTA, especially in Scandinavian region. In addition to the enlarged and pale kidneys (with vascular lesions and white spots), morphological changes include a prox‐ imal tubular injury, epithelial atrophy, fibrosis and hyalinization of renal glomerular [80, 81]. Negative effects of ZEA on pigs' reproductive function have also been demonstrated [91]. Oestrogenic effects of ZEA on gilts and sows include oedematous uterus and ovarian cysts, increased maturation of follicles, more numerous litters or decreased fertility [92].

Aflatoxins affect the quality of the milk produced by dairy cows and result in a carry-over of AFM1 with AFB1-contaminated feed. Ten ruminally-canulated Holstein cows received AFB1 (13 mg per cow daily) through a hole in the rumen for 7 days. The AFM1 levels in the milk of the treated cows ranged from 1.05 to 10.58 ng/L. The carry-over rate was higher in early lac‐ tation (2-4 weeks) compared to late lactation (34 -36 weeks) [75, 93]. The T-2 toxin causes ne‐ crosis of the lymphoid tissues. Bovine infertility and natural abortion in the last trimester of pregnancy also result from consumption of feed contaminated with T-2 toxin. Calves con‐ suming T-2 toxin in the amount of 10-50 mg/kg of feed showed abomasal ulcers and slough‐ ing of papillae in the rumen [75, 94, 95].
