The role of Paecilomyces Lilacinus (Thom) Samson and other fungal species in biodegradation of ochratoxin A

Nine isolates of fungi of genera Aspergillus, Fusarium, Paecilomyces and Penicillium were cultured on the modified Vogel’s medium with the addition of crude ochratoxin A (OTA) extract. This crude OTA extract was derived from a natural solid substrate on which Aspergillus ochraceus strain CBS 108.08 was cultivated. OTA was isolated, partially purified, dried by evaporating and dissolved in ethanol (1 mg/ml), and added to the test medium up to the final concentration of 10 microg/ml. The presence of OTA residues was determined after 7 and 14 day cultivation of fungi in the test medium at 27+/-1 deg C. The Paecilomyces lilacinus isolate (Inf. 2/A), which completely degraded OTA (150 microg) after only seven days, was selected for further studies. Wet sterile rice grains (50 g + 25 ml distilled water) were inoculated with individual isolates of fungi A. ochraceus (strain CBS 108.08) and P. lilacinus (isolate Inf. 2/A), and with their combination. In the case of P. lilacinus monoculture, 0.9 mg of crude OTA was also added into cultivation substrate. Each test was done in three replications. After the four week cultivation of individual and combined fungi at 27+/-1 deg C, inoculated rice grains were dried to the constant weight and pulverized. OTA was determined in these samples by the application of standard TLC method for fodder analysis. OTA in the amount of 61.310 microg/kg dry matter (DM) was determined only in the samples inoculated with a producer of ochratoxin A (A. ochraceus, strain CBS 108.08). On the other hand, a much smaller amount of OTA (80 microg/kg DM) was detected in samples inoculated with combined cultures of A. ochraceus and P. lilacinus isolates. Gained results indicate that P. lilacinus degraded, on average, 99.8% of OTA. After four week cultivation, the same fungal isolate in the samples of wet sterile rice kernels with the addition of 0.9 mg of crude OTA, completely degraded added crude OTA (less than 8 microg/kg).