Content of mycotoxins in the wheat grain of evaluated wheat species (average data of both cultivars).
Abstract
The effect of Fusarium spp. contamination on baking quality of winter common wheat and spelt wheat from different growing systems (organic and conventional) was evaluated by the standard technological quality characteristics and by the rheological system Mixolab. The content of Fusarium spp. mycotoxins [deoxynivalenol (DON), deoxynivalenol-3-β-d-glucoside (D3G), 3-acetyldeoxynivalenol (3-ADON), and Zearalenones (ZON)] was determined too. Significantly worse standard technological quality parameters and rheological parameters were determined for artificially inoculated variants of both evaluated wheat species. Statistically significant negative correlation coefficients were discovered between content of mycotoxins and many of technological characteristics, for example, DON content and Zeleny sedimentation for common wheat and spelt (−0.60*; −0.66*) and also between DON content and volume weight (−0.63*; −0.95**) for both wheat species. Resulted Mixolab parameters confirmed that Fusarium spp. infection worsens both protein and starch characteristics for both wheat species. However, effect of Fusarium spp. contamination in spelt wheat was generally less pronounced in comparison with common wheat. Despite of visible shifts of Mixolab curves of samples from organic and conventional growing systems, resulted Mixolab characteristics were statistically comparable.
Keywords
- common wheat
- spelt wheat
- Fusarium spp. contamination
- mycotoxins
- baking quality
- organic and conventional growing systems
1. Introduction
The largest group of
Regarding huge consumption of cereal product understanding of
By this time, there are not many studies about a capability of Mixolab to predict rheological parameters of common wheat with various grade of fungi infestation [6]; studies about a capability of Mixolab to predict rheological parameters of other wheat species with various grade of fungi infestation are almost not available.
The study was focused on the detection of baking quality changes in common wheat and spelt wheat with a different grade of
2. Methodology
2.1. Field experiments
Two winter common wheat cultivars (Bohemia and Darwin, both quality group A) and two winter spelt wheat cultivars (Ceralio and Rubiota) from the exact field plot trials, conducted in the years 2010/2011 and 2011–2012 at the experimental station of the Czech University of Life Sciences in Prague (295 m above sea level, average annual temperature 8.4°C, average sum of precipitation 575 mm), were used for the evaluation of the effect of FHB infestation on the
2.2. Artificial inoculation
The isolates of
2.3. Fusarium mycotoxins determination
A modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure was used for the isolation of analytes from the wheat grain. It is the multiresidue determination of wide range of mycotoxins applicable within different matrices (cereals, feed, and others), based on the extraction of analytes with acetonitrile; water and further purification of the extracts consists of the division between the two phases by means of inorganic salts (NaCl, MgSO4). Analytes were transported into the upper acetonitrile layer, while the polar co-extract matrix (e.g. sugars or amino acids) remained in the aqueous phase [15]. The ultra-high performance liquid chromatograph Acquity UPLC System (Waters, USA), coupled with the tandem mass spectrometer LCT Premier XE (Waters, USA) with analyzer time-of-flight MS (TOFMS) was used for the identification and detection of analytes. Wider scale of
2.4. Standard technological quality parameters
Crude protein content (CP) in grain dry matter according to the Kjeldahl method (EN ISO 20483; ICC-Standard No. 105/2), wet gluten content (WG) in grain dry matter and gluten index (GI) using the apparatus Glutomatic Perten (ISO 5531), falling number (FN)—ISO 3093, sedimentation index—Zeleny test (ZS)—ISO 5529, volume weigh (VW)—ISO7971-2, and TKW (thousand kernels weight) were determined with the frame of the baking quality.
2.5. Rheological characteristics
Protein and starch characteristics of the wheat flour (dough development, protein weakening, starch gelatinization, diastatics activity, and anti-stalling effect) were determined by the apparatus Mixolab (Chopin, Tripette et Renaud, Paris, France) according to the Mixolab protocol Chopin+ [16]. Evaluated flour was obtained by milling the cereal grain samples on a Bühler mill automat MLU 202.
A typical Mixolab curve, which is shown in Figure 1, is separated to the five stages represented by five (C1–C5) points [16].
The two first stages of the Mixolab curve correspond to the rheological characteristics of proteins—stability, elasticity, and water absorption, whereas the other stages relate mainly to the starch and amylolytic activity. Evaluated characteristics from measured Mixolab curve are C1 (Nm) marks maximum torque during mixing, used to determine water absorption; C1 (min) time required to achieve the maximum torque; C2 (Nm) measures the weakening of the protein based on the mechanical work and the increasing temperature; C3 (Nm) indicates the rate of starch gelatinization; C4 (Nm) represents the stability of the hot-formed gel; C5 (Nm) expresses starch retrogradation during the cooling period; difference C1–C2 represents the protein network strength under the increasing heating; difference C3–C4 shows diastatic activity and relates with falling number; difference C5–C4 correlates with the anti-stalling effects, represents the shelf life of end products; and DS indicates the stability of the dough before weakening [16].
Results were statistically evaluated by the analysis of variance (ANOVA) and correlation analysis with the statistical significance expression on the level α = 0.05 and α = 0.01 in the “Statistica 9.0 CZ”.
3. Results
3.1. Content of mycotoxins and standard technological quality parameters
An average content of evaluated
Treatment | Growing system | DON (μg kg−1) | D3G (μg kg−1) | 3-ADON (μg kg−1) | ZON (μg kg−1) |
---|---|---|---|---|---|
Artificial inoculation | Organic | 19411.1b | 2704.6b | 472.3ab | 2965.3b |
Conventional | 26729.4b | 4141.3b | 1051.9b | 3191.4b | |
Natural contamination | Organic | 304.7a | 153.8a | 7.5a | 19.0a |
Conventional | 257.5a | 66.5a | 7.5a | 22.3a | |
Artificial inoculation | Organic | 12648.7b | 2154.5b | 149.8b | 117.1b |
Conventional | 14433.7b | 2797.8b | 169.8b | 237.0b | |
Natural contamination | Organic | 25.6a | 22.6a | 7.5a | 4.0a |
Conventional | 260.1a | 142.4a | 7.5a | 7.1a |
Significant decrease of volume weight and thousand kernels weight but increase of protein and wet gluten content was observed in artificially inoculated variants for both wheat species—higher protein content in smaller grain is expectable. At the same time, general reductions of Zeleny sedimentation and gluten index in artificially inoculated variants were observed (Table 2). Similar situation—decreased falling number values were determined in artificially inoculated variants too.
Treatment | Growing system | CP (%) | WG (%) | GI | ZS (ml) | FN (s) | VW (kg hl−1) | TKW (g) |
---|---|---|---|---|---|---|---|---|
Artificial inoculation | Organic | 13.7ab | 30.0a | 42.0a | 34.0a | 250.5a | 50.4a | 23.9a |
Conventional | 14.1b | 32.5a | 58.5ab | 33.1a | 259.3ab | 47.2a | 22.4a | |
Natural contamination | Organic | 12.0a | 26.1a | 88.3b | 49.3b | 283.8b | 72.8b | 47.3b |
Conventional | 12.5ab | 27.3a | 83.8b | 54.8b | 279.8b | 74.1b | 49.5b | |
Artificial inoculation | Organic | 19.6b | 52.5a | 24.5a | 25.7a | 307.0a | 54.1a | 22.1a |
Conventional | 20.6b | 53.2a | 21.0a | 27.1a | 321.0a | 50.1a | 20.9a | |
Natural contamination | Organic | 16.7a | 49.1a | 38.0a | 39.1b | 343.0a | 73.0b | 40.9b |
Conventional | 18.8ab | 52.8a | 33.0a | 34.7b | 313.5a | 74.0b | 41.2b |
Above mentioned findings were also confirmed by determined correlations between mycotoxins content and evaluated technological parameters (Table 3). Negative correlation coefficients were found between content of mycotoxins and most of the technological parameters for both wheat species. The most evident negative effect of mycotoxins content was seen on VW and TKW for the spelt wheat (correlation: −0.95**; −0.97**). On the other hand, positive correlation coefficients were found between content of mycotoxins, crude protein content, and wet gluten content in grain.
DON (μg kg−1) | D3G (μg kg−1) | 3-ADON (μg kg−1) | ZON (μg kg−1) | |
---|---|---|---|---|
CP (%) | 0.48 | 0.37 | 0.44 | 0.22 |
WG (%) | 0.30 | 0.21 | 0.27 | 0.06 |
GI | −0.26 | −0.36 | −0.15 | −0.14 |
ZS (ml) | −0.60* | −0.75** | −0.58* | −0.64* |
FN (s) | −0.53* | −0.56* | −0.56* | −0.64* |
VW (kg hl−1) | −0.63* | −0.71* | −0.56* | −0.47 |
TKW (g) | −0.61* | −0.70* | −0.55* | −0.45 |
CP (%) | 0.69* | 0.64* | 0.64* | 0.35 |
WG (%) | 0.18 | 0.18 | 0.22 | 0.17 |
GI | −0.66* | −0.66* | −0.68* | −0.51 |
ZS (ml) | −0.68* | −0.78* | −0.77** | −0.72* |
FN (s) | −0.11 | −0.27 | −0.27 | −0.50 |
VW (kg hl−1) | −0.95** | −0.88** | −0.88** | −0.48 |
TKW (g) | −0.97** | −0.92** | −0.93** | −0.54 |
3.2. Mixolab
3.2.1. Triticum aestivum L.
Resulted common wheat Mixolab parameters confirmed that
Treatment | Growing system | C1 (min) | C2 (Nm) | C3 (Nm) | C4 (Nm) | C5 (Nm) |
---|---|---|---|---|---|---|
Artificial inoculation | Organic | 2.40a | 0.18a | 2.04a | 1.66a | 2.12a |
Conventional | 1.77a | 0.15a | 1.99a | 1.70a | 2.28a | |
Natural contamination | Organic | 3.21a | 0.44b | 2.66b | 1.96a | 2.50a |
Conventional | 3.00a | 0.40b | 2.63b | 1.97a | 2.59a | |
Treatment | Growing system | C1C2 (Nm) | C3C4 (Nm) | C5C4 (Nm) | DS (min) | |
Artificial inoculation | Organic | 0.90b | 0.38a | 0.46a | 6.3a | |
Conventional | 0.93b | 0.29a | 0.57a | 5.0a | ||
Natural contamination | Organic | 0.69a | 0.66b | 0.54a | 9.9b | |
Conventional | 0.74a | 0.70b | 0.62a | 9.5b |
It is evident from Figure 2 that the inferior effect of infection on protein part of curve is especially obvious for variety Bohemia in 2011, where after dough development, there is rapid fall of the curve which implies low quality of gluten.
Dough heating and thus swelling of starch granules and increasing viscosity cause the increase of the curve—point C3. It was evident from our results that values of artificially inoculated variants were markedly lower than values of variants with natural
Deteriorated rheological quality of inoculated variants was also confirmed by rated strong negative correlation coefficients between mycotoxins content and Mixolab parameters (Table 5).
DON (μg kg−1) | D3G (μg kg−1) | 3-ADON (μg kg−1) | ZON (μg kg−1) | |
---|---|---|---|---|
C1 (min) | −0.38 | −0.40 | −0.36 | −0.36 |
C2 (Nm) | −0.82** | −0.85** | −0.77** | −0.67* |
C3 (Nm) | −0.74** | −0.66* | −0.72* | −0.54* |
C4 (Nm) | −0.50* | −0.40 | −0.50* | −0.35 |
C5 (Nm) | −0.43 | −0.35 | −0.44 | −0.31 |
C1–C2 (Nm) | 0.73** | 0.74** | 0.69* | 0.56* |
C3–C4 (Nm) | −0.49 | −0.52* | −0.45 | −0.38 |
C5–C4 (Nm) | −0.22 | −0.20 | −0.26 | −0.18 |
DS (min) | −0.89** | −0.93** | −0.90** | −0.89** |
3.2.2. Triticum spelta L
Final Mixolab characteristics of
Treatment | Growing system | C1 (min) | C2 (Nm) | C3 (Nm) | C4 (Nm) | C5 (Nm) |
---|---|---|---|---|---|---|
Artificial inoculation | Organic | 5.12a | 0.14a | 1.64a | 1.25a | 1.83a |
Conventional | 4.20a | 0.14a | 1.65a | 1.33a | 1.89a | |
Natural contamination | Organic | 4.82a | 0.30b | 2.33b | 1.10a | 1.70a |
Conventional | 3.72a | 0.28b | 1.78a | 0.85a | 1.30a | |
Treatment | Growing system | C1C2 (Nm) | C3C4 (Nm) | C5C4 (Nm) | DS (min) | |
Artificial inoculation | Organic | 0.97a | 0.39a | 0.58a | 6.3c | |
Conventional | 0.95a | 0.32a | 0.56a | 4.3a | ||
Natural contamination | Organic | 0.83b | 1.23b | 0.60a | 6.0bc | |
Conventional | 0.83b | 0.93b | 0.45a | 4.9ab |
Average value of C2, which represents the weakening of the protein, was lower in artificially inoculated variants for both wheat species, but in spelt, the difference between naturally contamined and artificially inoculated variants was not so high. At the same time, higher rate of protein thermal weakening (C1C2) and thus shorter time of dough stability were found in inoculated variants for both wheat species, but in spelt, the difference between naturally contamined and artificially inoculated variants was slightly lower compared to common wheat.
Correlation between Mixolab parameters and mycotoxins content for
DON (μg kg−1) | D3G (μg kg−1) | 3-ADON (μg kg−1) | ZON (μg kg−1) | |
---|---|---|---|---|
C1 (min) | 0.18 | 0.12 | 0,20 | 0,02 |
C2 (Nm) | −0.94** | −0.92** | −0.90** | −0.56* |
C3 (Nm) | −0.69* | −0.68* | −0.66* | −0.40 |
C4 (Nm) | 0.28 | 0.25 | 0.30 | 0.19 |
C5 (Nm) | 0.19 | 0.17 | 0.22 | 0.13 |
C1–C2 (Nm) | 0.94** | 0.90** | 0.91** | 0.54* |
C3–C4 (Nm) | −0.51 | −0.49 | −0.52 | −0.32 |
C5–C4 (Nm) | 0.05 | 0.03 | 0.08 | 0.03 |
DS (min) | −0.13 | −0.21 | −0.11 | −0.27 |
It is evident from Figure 3 that despite the shifts of individual curves for variety Ceralio, majority of resulting Mixolab parameters for various type of treatment were statistically insignificant. Just characteristics C2 and dough stability for the control from organic treatment were preferable to the conventional variant.
4. Discussion
4.1. Content of mycotoxins and standard technological quality parameters
There are contradictory reports on the relationship between the
Several authors mentioned negative effects of
Zeleny sedimentation index and gluten index measure swelling potential of kernel protein. At the same time, general reductions of Zeleny sedimentation and gluten index in artificially inoculated variants were observed. This indicates that
Decrease of falling number value in artificially inoculated variants was observed too. According to Refs. [27] and [14], fungal infection is expected to increase the degradation of starch due to the activity of enzymes as α—amylase in kernels, which is measurable by means of falling number.
Our results showed negative correlation between content of the most mycotoxins and technological quality parameters for both wheat species. The most evident negative effect of mycotoxins content was seen on volume weight and thousand kernels weight. These results are in accordance with [28]—these authors mentioned that FHB can lead to the production of small-sized grains.
4.2. Mixolab
Mixolab parameters show, based on the results published up to now, high compatibility with standard rheological analysis (for example, farinograph, extensograph, or amylograph). Consequently, it is possible to anticipate a potential prediction of bread making quality of wheat from these parameters [10]. But there are not many studies on the efficiency of this Mixolab system to predict rheological parameters of wheat with changed characteristics caused by fungi species, which have become a serious problem in the wheat cultivation during recent years.
4.2.1. Triticum aestivum L
Some authors [14, 17–20] mentioned that
4.2.2. Triticum spelta L
Despite the fact that common wheat is the most widespread of all cultivated wheat species, at the present time, spelt wheat has growing popularity thanks to the nutritive and pro-health properties. The consummation of spelt products helps to reduce the cholesterol level in blood and fosters the circulatory system [32]. This wheat species has been also known for the high resistance to unsupportive environmental factors. Due to a higher stalk and hard adherent husks, spelt has poor fungal infestation in comparison with common wheat (
Final Mixolab characteristics of
5. Conclusions
The negative effect of
Our results confirmed that some of the problems related to the rheological properties of flour during processing, which regularly occur in some years and in some areas may be caused by
Acknowledgments
Supported by the Ministry of Agriculture of the Czech Republic, Project NAZV QI111B154.
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