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عزل وتشخيص مركب ايضي ثانوي جديد لكل من الفطرين Fusarium Trichoderma harzianum وsolani المعزولين من السواحل الرسوبية في محافظة البصرة وتقييم فعاليتهما الحياتية == Isolation and Identification of Novel Metabolic Compound From Both Fusarium solani and Trichoderma harzianum Isolated From Sedimentary Coast of Al - Basrah City and Estimate their Activity

Author name: فردوس نوري جعفر
Supervisor name: محمد لطيف علي | نيران جاسم الصالحي
General topic: Biology
Specific topic: Microbiology - Fungi
Degree: Doctorate
University: University Of Basrah - College Of Education For Pure Sciences
Language: Arabic
University location: Basrah
First pages:
Abstract: with the interest of medical and pharmaceutical uses of the natural products from marine fungi , thirty species of fungi belong to ninteen genus were isolated and identified from sea coast sediment of Al - Faw and Al - Seeba .The fungi Acremonium strictium , Geotricum candidum, Fusarium solani ,F.oxysporum , Penicilium sp. ,Trichoderma harzianum exhibited high inhibition activity toward growth of two reference strains of bacteria Escherichia coli (ATCC25922) and Staphylococcus aureus (NCtC 6571). The six selected fungi were grown in liquid culture media PDB at 25°C for two weeks ethyl acetate was used to extract the active compounds from thefermentation liquid media while ethanol was used to extract the active compounds from fungal mycelia .The crud extracts (fermentation and mycelium ) showed different inhibition activity against the pathogenic bacteria , Proteus mirabilis , S. aureus, isolates E. coli, Pseudomonas aeruginosa Klebsella pneumoniae. and pathoginic yeastes Candida albicans, C.tropicals,Cryptococcus neoformans and dermatophytic fungi Trichophyton verrucosum and Microsporium canis . The results revealed that the fermental fungi extracts has more inhibition activity than mycelial fungal extracts.The minimal inhibitory concentration (MIC) of fungal crude extracts were determined against gram positive and gram negative bacteria and pathogenic yeasts and fungi . antagonisum between studied fungi were evaluated , and the fungus T.harzianum has been the more antagonistic effect against other fungi. The toxicity of the fungal crude extracts were tested by using of human RBC at different concentration the result of this experiment showed that all extracts are non toxic except F.oxysporum extract.From all the above results the both F.solni and T.harzianum extracts showed higher activities ,thus these two extracts chosen for further studies. The anticancer activity of both extracts were studied against Hela cell line and the anticancer activity appeared at 250 Mg/ml and 62.5Mg/ml respectivily. The antioxidant activity of extracts were examined by three different methods ,B - carotene ,hydrogen peroxide scavenger and ferrus ion reduction , the results of three methods showed that both fungal extracts have antioxidant behaviours and the T.harzianum extract had more antioxident activity than F.solani extract and the antioxidant activity of both fungal extracts were increased as the concentration of the extracts increase.An optimization of the fungal growth and bioactivity of both fungi were conducted, by using different Nitrogen sources ( Asparagine , Yeast extract , peptone , Ammonium chloride , Sodium nitrate , Ammonium sulphate ) and carbon sources ( Maltose , Galactose , Glucose , Dextrose , Sucrose ,xylose and Starch ) the results elucidated that yeast extract was the best for growth of both fungi on solid and liquid media and high inhibition activity appear with T.harzianum against bacteria isolates , the carbon source starchenriched the growth of F.solani with more activity in inhibition the bacterial growth while the sucrose give high enrichment to the growth of T.harzianum in solid and liquid media with high activity toward bacterial growth. The study also indicates that the beast temperature and pH which gives the higher activity and growth 25°C and pH6 . The effect of different concentration of salinity were studied as well (1%,2%,3%,4%,5%,6% NaCl)and 2% showed the best concentration .The activity of two studied fungal extracts were superior than commercial antibiotic toward bacterial isolates .Preliminary qualitative chemical tests were carried out on the extracts and their chemical nature were identified .TLC technique were used and the chemical analysis showed that F.solani extract consist of two components with constant Rf values, the components marked as A1 and A2 . A1 shows higher activity against bacterial strains while T.harzianum extract consist of one component.The chemical structures of the isolated compound were

دراسة تصنيفية وحياتية للفطريات الناقصة الملونة في البصرة == Taxonomical and Biological study of Dematiaceous imperfect - fungi in Basrah

Author name: زينب خلف عبد الله
Supervisor name: توفيق محمد محسن
General topic: Biology
Specific topic: Microbiology - Fungi
Degree: Doctorate
University: University Of Basrah - College Of Education For Pure Sciences
Language: Arabic
University location: Basrah
First pages:
Abstract: During this study 23 species belong to 10 genera of imperfect fungi were isolated from different natural sources ( plants, soil , and ari) from Basrah govermanete using different isolation methods.Among the isolated fungi the genus Alternaria was selected to identify its species by using the morphological characteristis by light microscopy and Scaning Electron in Microscope (SEM) and verified by molecular and chemical methods. Eight species of Alternaria were identified namely; A.alternata (isolate1 and 2) A.chlamydospora A.citri, A.raphani , A.longipes , A.infectoria A.tenuissima and A.chrysanthemi based on light microscope examination.Taxonomic relegation of these species were confirmed by SEM image. SEM revealed that there was differences in the conidial wall ornamentation .The molecular analysis using PCR depending on the loci ITS1, and ITS4 700bp which confirmed the morphological identification of most of species namely A.alternata (isolate 1 and 2) , A.chlamydospora , A.longipes , A.infectroia and A.tenuissima. However , A.citri was related to A.alternata, A.raphani was belonged to A.infectoria and A.chrysanthemi is belonging to A.lolii. Nevertheless ,by using the loci OPA1 - 3 there was no variation among the species of Alternaria ,this is may be due to the inadequate method applied.The genetic variation of 36 isolates of A.alternata was studied using 9 - primers by RAPD - PCR technique. All primes gave an amplification of DNA for all the fungal isolates. Among the primers OPC08 exhibited a highest efficacy reaching 235 total bands .In the present study the antibacterial activity of fungal crude extract was tested. It appeared that the extract exhibit an antibacterial bioactivity against E.coli , S.aureus , S.epidermidis and S.saprophyticus.However, the extract of A.alternata (isolate 1 and 2) showed an inhibition growth against all the tested bacterial strains. This study was also revealed that the fungal filtrates of Alternaria species possessed various chemical compounds. However, there were differences in the chemical composition among the fungal filtrates.The results showed that Alternaria species ( A.alternata isolate 1 and 2, A.chlamydospora A.citri, A.raphani , A.longipes) were capable to biosynthesize silver nanoparticles, when the fungal filtrate were treated with AgNO3 solution after72 hrs incubation .Silver nanoparticles synthesis was checked by the change of colour from yellow to brown and confirmed by Uv - vis spectrophotometry at wave length (420, 440 nm). SEM image showed that the silver nanoparticles with spherical shape and size ranged between (20 - 72nm). Aconclusion can be derived that application of SEM,chemical analysis of fungi extracts ,molecular analysis and the silver nanoparticles characteristics are useful in the taxonomy of fungi to solve the overlapping of the morphological identification .Taxonomic key were prposed for the species of the genes Alternaria based on the morphology , chemical compound with the fungal extracts and size of silver nanoparticles synthesized by fungi .Melanin pigment identify by TEM microscope and it appeare like dark drop inside the cell .

استخدام عوامل استحثاث لانتاج مؤيضات ثانوية من بعض عزلات انواع الفطر واختبار فعاليتها الضد بكتيرية والخلايا السرطانية Penicillium والكولسترول == Use of inducing agents in secondary metabolites production from some Penicillium isolates and assay their activity against bacteria , cancer cells and cholesterol

Author name: امل صالح عبد الرضا
Supervisor name: توفيق محمد محسن
General topic: Biology
Specific topic: Microbiology - Fungi
Degree: Doctorate
University: University Of Basrah - College Of Education For Pure Sciences
Language: Arabic
University location: Basrah
First pages:
Abstract: The current study was designed to investigate active compounds from different Penicillium isolates ,that isolated from different sources , using different media . Eleven isolates related to five species : P. chrysogenum, P.paxilli , P. citrinum, P. cosmopolitanum, P. digitatum were isolated. After primary screening was performed , the best isolates that have high antibacterial activity against gram positive and gram negative bacterial was selected. The selected isolates were cultured on fermentation media according to the suitable media for each isolate, and the active compounds were extracted using ethyl acetate as a solvent. P. cosmopolitanum and P. chrysogenum were cultured on penicillin production medium (PPM) , P. paxilli was cultured on Aspergillus complete medium (ACM) and P. digitatum and P. citrinum were cultured on solid state fermentation media consist of wheat bran . The efficiency of fungal crude extracts from each isolate against the reference strains S. aureus NCTC 6571 and E. coli ATCC 25922 and against Candida albicans (pathogenic and reference strain)was examined, and the Minimum inhibitory concentration (MIC) was determinated for each extracts.The inhibition zone of fungal extracts against E. coli and S. aureus ranged between (20 - 50) mm , and they had high activity against C. albicans ( reference and pathogenic isolate) ranged between 25 - 45 mm.Induction of secondary metabolite production by the selected fungi was carried out using biological factor (Streptomyces sp.) , chemical agent (colchicine) and physical agent by UV radiation. Apparently that the biological induction by Streptomyces isolate was the best compared with the other treatments.The results revealed that the two fungal extracts ALO1 and Apax exhibit a high antioxidant activity at low concentrations and volumes of the extract compared to ascorbic acid as control. ALO1 had radical scavenging activity (RSA%) reach 99.9% at volume 3μl , and Apax had RSA 91.6% at concentration 500μg/ml. Also, the fungal extract ,ALO1, showed a significant activity as anticancer agent against RD - cell line. The IC50 of ALO1 was 8.5μl after 24h and 23.76μl after 72h. The hypercholesterolemia ability of ALO1 in vivo and in vitro was assayed . In vivo experiment designed using female albino mice and divided into three groups each one contain 8 mice : group 1 treated with 10μl , group 2 treated with 20μl of ALO1 , and group 3 treated with normal saline as control group. The results showed that total cholesterol , triglycerides, LDL and VLDL were significantly reduced when compared with control group while the HDL level was increased in comparing with control group. The chemical analysis of the crude extracts showed that Apen2 contained 259 μg/g of penicillin G when analyzed with HPLC apparatus . Also the GC mass analysis of ALO1 showed that Docasadionic acid initialize area76.18% and Palmitic acid initialize area 15.09% from the total identified compounds.Whereas the GC mass analysis of Apax showed that Hexandioic acid initialize area40% and Stearolic acid initialize area 35.57% from the total identified compounds.In addition , it was detected the presence of penicillin biosynthetic gene cluster : pcbAB, pcbC and penDE in P. cosmopolitanum and P. chrysogenum.The result showed that gene pcbC and penDE were found in the genomic DNA but the gene pcbAB was not amplified by PCR.