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دراسة التاثيرات السمية للـ Gliotoxin المنتج من العفن Aspergillus fumigatus باستخدام تخمرات الحالة الصلبة == Study Toxicity Effects of Gliotoxin Produced By Aspergillus Fumigatus Using Solid State Fermentation
Author name:
محمد عادل نوري
Supervisor name:
عبد الكريم جاسم هاشم
General topic:
Biology
Specific topic:
Microbiology - Fungi
Degree:
Master
University:
University of Baghdad - College Of Science - Department Of Biotechnology
Language:
English
University location:
Baghdad
First pages:
24T2688 - p.pdf
Abstract:
تم اختبار قابلية عشرة عزلات للعفن Aspergillus fumigatus على انتاج الغليوتوكسين بوساطة تخمرات الحالة الصلبة وباستخدام الرز كوسط زرعي (ركيزة). اشارت نتائج الغربلة ان العزلة AF - 5 كانت هي الافضل واعطت اعلى انتاجية.كانت الظروف المثلى لانتاج السم على وسط الر | The ability of Ten Aspergillus fumigatus isolates for gliotoxin production were screened by solid state fermentation (SSF) using rice medium as substrate. The results indicated that, the AF - 5 isolate was the highest gliotoxin producer.The optimum conditions for gliotoxin production by AF - 5 isolate on rice medium were : - inoculum size 6×106 spores, moisturizing ratio (5 : 1) (w : v) with distilled water and incubation at 37 ?C for 10 days. Gliotoxin was purified from crude extract of AF - 5 isolate after production under the optimum conditions using solid state fermentation SSF. Purification of gliotoxin was achieved by two steps including : filtration and solid phase extraction. These processes were accessed to remove debris from crude extract and gives pure gliotoxin. The gliotoxin concentration was (122.6 ppm), which was detected by Thin layer Chromatography and High Performance Liquid Chromatography.The result of cytotoxicity effect of both purified extract and standard gliotoxin on human lymphocytes with exposure time of 24 hours at four different concentrations 25, 50, 100 and 200 ppb showed growth inhibition percentage 21, 39.10, 61.99, 87.45% and 17.89, 34.92, 58.34, and 85.22% respectively. The cytotoxicity effects have been done using conversion of the MTT to MTT formazan. After DNA was extracted from lymphocyte treated with toxin and analyzed by electrophoresis on a 1% agarose gel, gliotoxin appeared to have ability to damage the DNA. Results showed that both growth inhibition and DNA damage were increased gradually with the increasing of gliotoxin concentration.
