Abstract:
This study examined black aspergilli, especially A. carbonarius and A. niger and
ochratoxin A (OTA) contamination of grapes, during drying and industrial processing
of dried vine fruits. This was complemented by studies on potential control using
preservatives and physical factors such as modified atmospheres. Fungal population
kinetics were determined in relation to grapes at harvest, and during drying at three
different altitudes (sea level: 0-200 m; medium level: 250-500 m; high level: >500 m)
in two seasons. At pre-harvest, A. niger aggregate species were the predominant
fungal species while A. carbonarius was occasionally isolated, in both years studied.
Both altitude and bunch position affected black aspergilli population dynamics.
Overall, they were increased during drying. However, both black aspergilli groups
were mostly isolated, at low and medium altitudes (<500 m). OTA contamination
was influenced by bunch position, although altitude did not significantly influenced
amounts. The fungal biodiversity was decreased during sun-drying of sultanas. The
widest diversity of species occurred at the sea level. However, A. niger aggregate,
were dominant during drying. Time of drying and altitude significantly influenced
fungal loads of black aspergilli. In contrast, OTA production (ca 0.001 - 0.0025 μg g
-
1
) was not significantly influenced by altitude and drying time. Shannon Index of
Biodiversity (H), for pre-harvest and pos-harvest studies, was determined for the first
time.
A. niger aggregate (ca 5.0 Log10 CFUs g
-1
) was predominant during industrial
processing, while A. carbonarius was only isolated at low levels (1.5-2.0 Log10 CFUs
g
-1
). Heat treatment (up to 90
o
C) appeared to be the key-procedure for the
elimination of fungal populations. In the contrary, SO2 treatment did not statistically
alter fungal population dynamics. OTA contamination was not significantly affected
by industrial processing.
In vitro studies conducted on both White Grape Juice Medium (WGJM) and in
sultanas with strains of A. carbonarius originated from Cretan sultanas and compared with a strain isolated from Italian wine grapes. They examined the impact of sodium
metabisulphite (NaMBS), elevated CO2 (up to 50%) concentrations and aw levels, on
black aspergilli spore germination, growth and OTA production. Moreover, fungal
interactions in vitro and in situ were also investigated.
In general, spore germination occurred over a wide range of sodium metabisulphite
concentrations, although germ tube extension was significantly controlled. At ≥ 750
mg L
-1
NaMBS, no spore germination was observed while both mycelial growth and
OTA production were completely inhibited. Medium concentrations of NaMBS (≤
250 mg L
-1
) enabled optimum spore germination, growth and OTA production (x
0.965 aw). The efficacy of controlled atmospheres x aw showed that there was very
little inhibitory effect on spore germination. However, both germ tube extension and
fungal growth were inhibited by 50% CO2. After 10 days, growth was not as
effectively controlled. Aw had a bigger effect on OTA production than modified
atmospheres. In situ experiments on sultanas confirmed these results. Competition
and dominance of A. carbonarius over other fungal species showed that aw and
temperature influenced Indices of Dominance and OTA production. In vitro and in
situ, OTA production by A. carbonarius was significantly influenced by the fungal
competitor used.