Aspergillus Carbonarius Growth Research Articles

Effect of gallic acid on Aspergillus carbonarius growth and ochratoxin A production

The aim of the present study was to investigate the gallic acid influence on Aspergillus carbonarius growth and ochratoxin A (OTA) biosynthesis. A mixed inoculum was used in Czapek Yeast Extract Agar and growth and OTA production was observed at 100 to 500 mg/l of the phenolic compound. Slower colony growth rates and longer lag phases were observed with increasing concentration and 500 mg/l was totally inhibitory for A. carbonarius growth. OTA production decrease was observed even at the lower concentration assayed. Gallic acid could be used as a natural antimicrobial for the control of ochratoxigenic A. carbonarius.

Inhibition of growth and ochratoxin A biosynthesis in Aspergillus carbonarius by flavonoid and nonflavonoid compounds

The effect of naturally occurring phenolic compounds on Aspergillus carbonarius growth and ochratoxin A (OTA) production was studied. Caffeic acid and the flavonoids, rutin and quercetin, were added to Czapek Yeast Extract agar at concentrations ranging between 50 and 500mg/l. All phenolic compounds had a significant influence on growth rate and lag phase of A. carbonarius at 250mg/l. The growth was completely inhibited with 500mg/l. In comparison with the control, a significant decrease in OTA production was observed with all phenolic compounds. In general, effect on growth was less evident than effect on toxin production. An inhibitory effect on growth and OTA production, as concentration was increased was observed in all cases. The response of A. carbonarius to the flavonoids, rutin and quercetin, was similar. The inhibitory effect of these natural phenolic compounds on fungal growth and OTA production could be an alternative to the use of chemical fungicides.

Mycelial growth and ochratoxin A production byAspergillussectionNigrion simulated grape medium in modified atmospheres

To evaluate the impact of modified atmosphere packaging on in vitro growth of Aspergillus carbonarius and Aspergillus niger, and possible effects on ochratoxin A (OTA) biosynthesis. Ochratoxigenic isolates belonging to the species A. carbonarius and A. niger were grown on a synthetic grapejuice medium (SNM) and packaged in combinations of controlled O(2) (1% and 5%) and CO(2) levels (0% and 15%), and in air as a control. Colony diameters were recorded every 3 days up to 21 days, and OTA was analysed after 7, 14 and 21 days. The greatest reductions in mycelial growth rate were observed at 1% O(2) followed by 1% O(2)/15% CO(2), whereas 5% O(2) stimulated the growth of all isolates. OTA production by A. carbonarius and A. niger isolates was minimized at 1% O(2)/15% CO(2) and 1% O(2), respectively, after 7 days of incubation. Maximal OTA accumulation after 7 days was observed for all isolates in the control pack and at 5% O(2). Of the atmospheres tested, only 1% O(2) combined with 15% CO(2) consistently reduced fungal growth and OTA synthesis by A. carbonarius and A. niger. Storage under modified atmospheres is unlikely to be suitable as the sole method for OTA minimization and grape preservation; other inhibitory factors are necessary.

Effect of carbendazim and physicochemical factors on the growth and ochratoxin A production of Aspergillus carbonarius isolated from grapes

Carbendazim is a systemic fungicide that is commonly used on several crops (tobacco, fruit, vegetables, cereals, etc.). This fungicide is used to control fungal infections in vineyards. It is indicated against Botrytis cinerea, Uncinula necator, Plasmopara viticola and other fungi and can be used either alone or coupled with other fungicides. However, there is a lack of in-depth studies to evaluate its effectiveness against growth of Aspergillus carbonarius isolated from grapes and OTA production. A medium based on red grape juice was used in this study. Preliminary studies were performed at 0.98 a w and 25 °C using carbendazim concentrations over a wide range (1–2000 ng/ml medium) to control both growth of a strain of A. carbonarius isolated from grape and its ability to produce OTA. As the lag phase increased considerably at levels > 1000 ng/ml of medium, detailed studies were carried out in the range 50–450 ng/ml of medium at 0.98–0.94 a w and 20–28 °C. Statistical analysis (multifactor ANOVA) of the data revealed that the three factors assayed and the interactions a w–carbendazim concentration and a w–temperature had significant effects on lag phase duration. The highest lag-times were observed at 0.94 a w, 20 °C, and with 450 ng carbendazim/ml. The three factors also had significant effects of the growth rate and there was an interaction between a w and temperature. The growth rate of A. carbonarius in these cultures is favoured by high water availability and relatively high temperatures. However, addition of carbendazim at the assayed levels did not significantly influenced fungal growth rate. Accumulation of OTA was studied as a function of four factors (the three previously considered, and time). All factors had significant effects on the accumulation of OTA. There were also two significant interactions ( a w–temperature and temperature–time). On the basis of the results obtained, carbendazim does not increase the lag phase of A. carbonarius except at relatively low a w and temperatures. It does not substantially decrease fungal growth rate once growth is apparent but it appears to cause an increase in OTA accumulation in the medium at the doses assayed. Carbendazim, which is widely used against fungal infections in grape, can positively influence OTA production by A. carbonarius in field, which can increase OTA content in grape juices and wines.

Efficacy of natamycin for control of growth and ochratoxin A production by Aspergillus carbonarius strains under different environmental conditions

To examine the efficacy of natamycin produced by Streptomyces natalensis against strains of Aspergillus carbonarius growth and ochratoxin A (OTA) production under different environmental factors on a grape juice-based medium. Detailed studies in the range 0-20 ng ml(-1) for control of growth and ochratoxin production by strains of A. carbonarius at 0.98, 0.96 and 0.94 water availabilities (a(w)) and 15-25 degrees C on a fresh red grape extract medium were examined. Inhibition of growth was depending on temperature and a(w) level. At 15 degrees C, 5-10 ng ml(-1) natamycin was effective in reducing growth almost completely. However, at 20-25 degrees C and all the three a(w) levels, growth was only slightly inhibited by 5-10 ng ml(-1) natamycin. There were strain differences with regard to inhibition of OTA production. At 15 degrees C and 0.98 a(w), 10 ng ml(-1) was required to inhibit production by >90%. However, at 0.96 and 0.94 a(w), almost complete inhibition occurred. At 20 degrees C, OTA production was only significantly inhibited by 10 ng ml(-1) natamycin at 0.94 a(w). At 0.96 and 0.98 a(w), some inhibition occurred with 5-10 ng ml(-1), but greater concentrations would be required for effective inhibition. At 25 degrees C, 5 ng ml(-1) was effective at all a(w) levels. However, at 15 degrees C and 25 degrees C and a wide range of a(w) levels, natamycin effectively controlled OTA production. Natamycin appears to be a very effective for controlling growth and OTA production by strains of A. carbonarius over a range of a(w) and temperature conditions on grape-based media. This is the first detailed study to demonstrate the impact of natamycin against A. carbonarius. This study suggests that use of natamycin at 50-100 ng ml(-1) can give complete inhibition of growth of A. carbonarius and OTA production over a range of environmental conditions. Natamycin could be an important component of a system to prevent OTA contamination of wine as well during the drying and production of vine fruits.

Effect of preharvest fungicides and interacting fungi on Aspergillus carbonarius growth and ochratoxin A synthesis in dehydrating grapes

To evaluate the effect of preharvest grape pesticides in Aspergillus section Nigri infection in dehydrating grapes and the final ochratoxin A (OTA) content. Additionally, the effect of coinoculation of moulds frequently isolated from grapes and raisins on Aspergillus section Nigri infection was studied. Fungicide-treated grapes were inoculated with Aspergillus carbonarius, Aspergillus niger aggregate, Eurotium amstelodami and Penicillium janthinellum in different combinations, then dehydrated by reducing a(w) for 20 days. The percentages of colonized grapes treated with fungicides were, in general, lower, but no differences were observed among fungicides. The untreated grapes always showed higher concentrations of OTA, regardless of the inoculum applied. In general, Chorus was the most effective antifungal treatment in reducing OTA accumulation in grapes during dehydration. Penicillium janthinellum reduced Aspergillus section Nigri colonization and OTA accumulation in grapes during dehydration. The four preharvest fungicides studied reduced the Aspergillus section Nigri growth and OTA production by A. carbonarius during dehydration of grapes. The success of these chemical treatments might depend on the mycobiota composition of grapes.

Impact of fungicides on Aspergillus carbonarius growth and ochratoxin A production on synthetic grape-like medium and on grapes

A study was undertaken to evaluate the impact of the application of several fungicide treatments used in Spanish vines on Aspergillus carbonarius growth and ochratoxin A production. Three trials were designed in order: (1) to screen 26 fungicides at the doses recommended by manufacturers on grape-like synthetic medium at 20 and 30°C; (2) to find out the minimum inhibitory concentration of each fungicide for A. carbonarius growth on synthetic medium; and (3) to investigate the effect of several fungicides on A. carbonarius-inoculated grapes. In synthetic medium nine fungicides significantly reduced A. carbonarius growth rate. Meanwhile, 13 fungicides completely inhibited its growth. In general, growth was faster at 30°C than at 20°C, contrary to ochratoxin A production. Fungicides that stopped fungal growth also inhibited ochratoxin A production, but not all the fungicides that reduced growth reduced the ochratoxin A synthesis. In general, fungicides that contained copper or strobilurins reduced both growth and ochratoxin A production, contrary to sulphur fungicides. At the optimum temperature for A. carbonarius growth of 30°C, higher amounts of fungicide were needed to prevent fungal growth than at 20°C. Among the fungicides that inhibited A. carbonarius growth on synthetic medium at the initial doses, cyprodinil seemed to be the active ingredient more effective at stopping fungal growth when testing reduced doses. The fungicide effect on grapes was similar to that on synthetic medium. Both infection and ochratoxin A production were reduced when using cyprodinil (37.5%) plus fludioxonil (25%) and azoxystrobin (25%). Penconazole (10%) also showed a clear reduction in ochratoxin A production at both temperatures, although infection was only reduced at 20°C. Ochratoxin A reduction was strain and temperature-dependent. In general, fenhexamid (50%), mancozeb (80%) and copper hydroxide (80%) plus copper (50%) enhanced infection and ochratoxin A production.

Survival and growth of Aspergillus carbonarius on wine grapes before harvest

Aspergillus carbonarius, the primary OTA-producing species in Australia, was inoculated onto the surface of Chardonnay and Shiraz bunches at pre-bunch closure, veraison and pre-harvest during the 2002–03 and 2003–04 seasons. Mean A. carbonarius counts decreased between pre-bunch closure and veraison, and increased between veraison and pre-harvest. Increases in A. carbonarius counts from veraison onwards were most marked in Chardonnay bunches during 2003–04; such bunches comprised more berries and were heavier than in 2002–03. Bunches with no berry damage yielded low A. carbonarius counts at pre-harvest and harvest. Exposure to direct sunlight over several days reduced viability of A. carbonarius spores supported on filter membranes by 10 5, despite the spores having thick, heavily melanised walls. The estimated cumulative UV exposure for that period was 10 mWh. Thus, UV radiation may be a contributory factor to the decline of A. carbonarius spores on berry surfaces, particularly in the early stages of berry development.

Effect of photoperiod and day–night temperatures simulating field conditions on growth and ochratoxin A production of Aspergillus carbonarius strains isolated from grapes

The effect of light and temperature regimes simulating day and night in the field in the months preceding grape harvest on Aspergillus carbonarius growth and ochratoxin A (OTA) production were investigated. Twelve-hour photoperiod affected positively A. carbonarius growth with no differences between incubating the mould at day temperature (28 °C) or alternating day/night temperatures (28 °C/20 °C). Slower growth, however, was observed with constant incubation at 20 °C. Under 12h-alternation periods of day and night temperatures, growth was faster at continuous darkness than under continuous light conditions. Light did not cause any morphological changes in the aspect of the colonies. No significant differences on OTA production were detected due to either fluctuating temperature or photoperiod. However, as photoperiod enhanced the growth of colonies, it also enhanced OTA accumulation. The ability of A. carbonarius to produce OTA reported in previous laboratory studies has been demonstrated to be stimulated in field conditions.

Aspergillus carbonarius growth and ochratoxin A production on a synthetic grape medium in relation to environmental factors

The effects of water activity (0.90-0.99 a(w)), temperature (15-37 degrees C), and their interaction on growth and ochratoxin A (OTA) production by eight isolates of Aspergillus carbonarius were investigated on synthetic nutrient medium (SNM) with composition similar to grapes. Growth data were modelled by an multiple linear regression and response surface models were obtained. Aspergillus carbonarius grew much faster at 30 degrees C than at the other temperature levels tested and its growth rate increased with increasing a(w), maximum growth rate being between 0.95 and 0.99 a(w). In general, isolates grew faster at 35-37 degrees C than at 20 degrees C, although no significant differences were found between these temperatures. OTA accumulation was also favoured by high a(w) levels, and although it was observed in the whole range of temperatures, maximum amounts were detected at 20 degrees C. No OTA was found at the most unfavourable growth conditions. Optimum a(w) level for growth seems to correspond with optimum for OTA production, meanwhile the most propitious temperature for the toxin production was below the best one for growth. Prediction of A. carbonarius growth would allow estimating their presence and therefore, the OTA production, as it was found that conditions for the toxin production were more limited than those permitting growth.