Maize kernels were analysed for FB1, FB2 and AFB1initial content and data are reported in Table 3.2.
Tab. 3.2 – Different mycotoxins content present in maize used for the experiment.
FB1(µg/Kg) FB2(µg/Kg) AFB1(µg/Kg)
Maize sample 13444 4807 2.1
Maize heap 5297 1424 0
Sub-samples (1-1.5 Kg) were analysed after 10, 20, 40, 60, 90,120,180 days.
Both mycological and mycotoxins analysis were carried out on samples as described in Annex 1 and Annex 2.
Statistically, kernels moisture content did not show significant differences during the 18 days. Relative humidity observed was between 19 and 21%. At 18 days post harvest, an important decrease in the incidence of kernels infected by F.
verticillioides and in the number of Fusarium CFU/g was observed, while until 96 hours the incidence of kernels infected by F. verticillioides and the number of Fusarium CFU/g did not differ significantly. Data recorded also showed a tendency toward decreased FB1 mean concentrations in the samples collected in both field monitored in this study (Table 3.3) with significant reduction after 18 days from harvest.
The incidence of A. flavus and AFB1content in maize were lower and did not change significantly during the pre-storage time (Table 3.3).
Tab. 3.3 – Analysis of variance of incidence of F. verticillioides, A .flavus, CFU/g of Fusarium, temperature (T) and relative humidity (RH) on maize kernels collected in Ferrara and Ravenna during pre-storage period of wet kernels before drying and storage (average data).
Time T
(°C)
RH (%)
F. verticillioides
(%) CFU_fus/ g A. flavus (%)
FB1
(ppb)
AFB1
(ppb)
After harvest 24.8 b 19.4 61.0 a 1.5E+07 a 3.0 21244 a 0.0
24 h 26.9 b 19.2 52.5 a 1.6E+06 a 0.5 13608 a 1.2
48 h 27.2 b 20.6 54.5 a 4.0E+05 a 0.5 9494 a 4.3
96 h 24.9 b 21.6 56.0 a 3.0E+05 a 6.0 11422 a 11.7
18 days 36.6 a 20.5 10.0 b 3.06 b 2.0 1213 b 1.3
Different letters indicate significant differences according to the Tuckey test (P≤0.01)
3.3.1.2 Storage trial - 2004
After drying and before storage, the kernels T was around 21 °C, but after one month of storage a decrease was detected. From one month to six months of storage T remained almost constant (Table 3.4).
Regarding RH, ANOVA highlighted differences from dried kernels to six months of storage: RH increased in time, from 12,27% to around 14%, during the storage.
F. verticillioides incidence decreased significantly over the six months of storage from about 42% of infected kernels after drying to 0% (Table 3.4). The production of FB1 by F. verticillioides increased by 30% after six months of storage.
From a statistical point of view, the number of Fusaria CFU did not show significant variations. The incidence of A. flavus and AFB1 content did not change during 6 months storage. Storage types did not influence any of the factors considered (Table 3.4).
Tab. 3.4 – Analysis of variance of incidence of F. verticillioides, A. flavus, CFU/g of Fusaria, temperature and relative humidity on maize kernels collected in Ferrara after 4 different times of storage and considering 3 different types of storage (average data).
Storage T
(°C) RH (%)
F.verticillioides
(%) CFU_fus/g A.flavus (%)
FB1
(ppb)
AFB1
(ppb)
After drying 21.3 12.3 b 42.0 a 86.4 1.3 2667 b 17.3
1 month 16.0 14.3 a 44.7 a 303.9 0.7 5691 ab 10.9
3 months 15.7 14.0 a 42.0 a 147.4 0.7 4634 ab 1.2
6 months 16.3 13.8 a 0.0 b 8.0 0.7 9018 a 3.6
Type of storage
silo 16.7 14.1 34.0 163.0 3.0 6798 16.9
horizontal store 16.1 13.8 − − 2.0 4927 0.0
Vertical cell 16.7 13.7 30.0 89.0 7.0 6484 2.5
3.3.1.3 Storage trial - 2005
Taking into account initial aflatoxins contamination, the factor considered influenced significantly AFB1 content and both Fusarium and Aspergillus CFU, except the FB1contamination (Table 3.5).
The population of Fusarium and Aspergillus and FB1 contamination did not change significantly based on the store type, horizontal or vertical cells. Only AFB1content was significantly lower in vertical than in horizontal stores (-21%).
Considering storage time, from time 0 to 6 months, both Fusarium and Aspergillus CFU did not show significant changes while an increase occurred in FB1content although not statistically significant.
Tab. 3.5 – Analysis of variance of CFU of Fusarium and Aspergillus and content of fumonisin B1 and aflatoxin B1 in maize kernels considering 2 different initial aflatoxins contamination, 2 different types of storage and 3 different storage time (average data).
Aflatoxins contamination CFU_fus/g CFU_asp/g FB1
(ppb)
AFB1
(ppb)
High 102.5 b 7.6E+02 a 5537 642.9 a
Low 2.4E+04 a 4.1 b 5883 4.4 b
Storage Type
Horizontal store 1.1E+03 4.5E+01 5457 364.8 a
Vertical Cell 1.3E+04 3.5E+02 7228 76.9 b
Time Storage
After drying 5.7E+02 1.2E+01 4392 181.1
1 month 1.4E+03 7.6E+01 2234 296.8
3 months 2.8E+03 6.2E+01 9711 437.6
6 months 6.5E+03 1.1E+03 8100 435.6
3.3.2 Maize in controlled conditions
3.3.2.1 Trial 2006 - Relative humidity of maize kernels
The relative humidity (RH) of maize kernels, during the 180 days of time incubation and under the different environmental conditions used, are given in table 3.6.
The RH of maize kernels, detected before harvesting, was 14%. This value remaind constant until the end of incubation. There were no important changes at all environmental conditions in RH of maize kernels kept at different and controlled temperature and humidity for 180 days.
Tab. 3.6 – Mean relative humidity of kernels detected at controlled environment with different levels of temperature and humidity after incubation from 0 to 180 days.
Humidity
Temperature 70% 85%
0 14.0-15.2 14.0-15.3
6 14.0-15.7
20 14.0-15.3
30 13.4-14.3
3.3.2.2 Maize samples at 0°C and 0.70-0.85 aw
The population of F. verticillioides (lnCFU) on stored maize grain at 0°C and 0.70 aw was small and remained below 10 with variable trend until the end of incubation (Figure 3.1).
At 0°C and 0.85 aw, the sample showed few CFU of F. verticillioides and this population remained stable until 60 days of incubation. From 60 to 120 days of
storage F. verticillioides increased, but at 180 days the trend showed a suddenly decrease (Figure 3.1).
Fig. 3.1 – Colony forming units (ln) of F. verticillioides in maize kernels during storage (0-180 days) at 0°C and 0.70 and 0.85 aw.
3.3.2.3 Maize samples at the other environmental conditions
Considering the environmental conditions usually present in the horizontal store and those selected in greenhouse, F. verticillioides populations were about ten times higher with respect to 0°C and 0.70-0.85 aw, mostly over 40 days of incubation.
A similar trend was observed at 6 °C and 20 °C where the F. verticillioidesCFU remained stable and low until 20 days and the populations increased after 40 days of incubation.
On the other hand the population presence at 30 °C showed a trend extremely changeable.
Fig. 3.2 – Colony forming units (ln) of F. verticillioides in maize kernels during storage (0-180 days) at different temperature and awconditions.
3.3.2.4 Maize mycotoxins analysis
Concentration of FUM throughout the incubation periods are shown in Figure 3.3 and are expressed as FB1 + FB2. In the samples at harvest, FB1+FB2
concentration was 18.3 µg/kg. In the sample incubated at 0°C and 0.70 awafter 10 days from harvest a significant decrease of fumonisins (-70%) was observed, followed by an increase at 20 days. From 40 days to the end of storage, a 50% constant reduction for FUM was observed.
The FUM concentration detected at 0°C and 0.85 aw showed a similar trend, except at 10 days where the FUM concentration showed an increase.
At all the other environmental conditions the FUM concentration trends were analogous at the trend observed at 0°C.
0 5 10 15 20 25
0 10 20 40 60 90
Time (days) FumonisinsB1+B2(µg/kg)
0°C 0.7aw 0°C 0.85aw 6 °C 0.85aw 20°C 0.7aw 30°C 0.8aw
Fig. 3.3 – Concentration of FB1+FB2(µg/kg) in maize stored at different incubation conditions and sampled at five subsequent times (10,20,40,60, 90 days).
Aspergillus flavus was also detected in the samples, but with limited CFU (data not showed). Considering the AFB1 concentration, it remained under 10 µg/kg and very variable data were found in different sampling time (Fig. 3.4).
0 5 10 15 20 25
0 10 20 40 60 90
Time (days) AflatoxinB1(µg/kg)
0°C 0.7aw 0°C 0.85aw 6°C 0.85aw 20°C 0.7aw 30°C 0.8aw
Fig. 3.4 – Concentration of AFB1(µg/kg) in A. flavus infected maize samples after harvesting from different samples incubated at five subsequent times (10,20,40,60, 90 days).