Effect of ethanolic leaf extract of Pinus eldarica on inhibiting the growth of Aspergillus flavus and Aspergillus niger in corn biomass under in vitro conditions

This experiment was conducted as completely randomized design (CRD) with factorial arrangement (4×3) including three levels of extract and four incubation times (10, 20, 30 and 40 days) to evaluate the effect of alcoholic extract of pine tree leaves under in vitro conditions to inhibit the growth of Aspergillus flavus and Aspergillus Niger on the biomass of wet corn. Pine tree leaves were collected in autumn and their extract prepared using ethanol and used at concentrations of 0, 1 and 2 percent. Mass commercial corn was purchased from the factory. Ten grams of corn were mixed with 90 ml of saline solution and stirred several times, then serially diluted and the volume of 0.1 ml was placed on the culture medium. The samples were incubated at 25oC for one week and the growth of fungi were checked every 24 hours from the second day of incubation. The fungus colonies were counted visually. The results indicated that 1 and 2% of pine leaf extract effectively reduced the number of fungi colonies at days 10 and 20 of incubation and the growth of fungi were completely inhibited on days 30 and 40 of incubation. The results were similar for both species of fungi.According to the results, pine leaf extract at the level of 1% is recommended for inhibition of fungi growth.