تقييم فاعلية بعض الرواشح الفطرية العادية والنانوية ضد الفطرالأسود Rhizopus stolonifer خارج الجسم الحي

Evaluation of Some normal and nano filterates activity Against Black Molds In vitro Conditions

The study was conducted in the Graduate Studies Laboratory of the College of Education for Pure Sciences at the University of Karbala, during the period from 15/11/2021 to 15/8/2022. With the aim of controlling the fungus that causes black mold disease by using the phenomenon of fungal antagonism with fungi isolated from the sputum of some people who recovered from infection with Covid 19. With the possibility of converting silver and zinc from the metallic state to a nano-state using these fungal filtrates.
The study included the diagnosis of fourteen fungi isolated from the sputum of people who had recovered from infection with Covid 19 with a conventional diagnosis, then they were molecularly diagnosed using the polymerase chain reaction (PCR) technique, based on primers for molecular diagnosis.
Fungi have been registered in the global Genbank, and this study is considered the first study concerned with this work at the level of Iraq and the region. These are Rhizopus americanus has accession number ON981106, Rhizopus stolonifer has accession number ON981097, Aspergillus brasiliensis has accession number , Aspergillus costaricensis has accession number ON981102, Aspergillus flavus has accession number ON981297 and Aspergillus minisclerotigenehas accession number ON981101, Aspergillus niger 1 has accession number ON981098, Aspergillus niger2 has accession number ON981107 and fungus Aspergillus niger3 has accession number , Aspergillus oryzae, has accession number ON981295, Aspergillus piperis, has accession number ON981103, Aspergillus tubingensis1has accession number ON981101, Aspergillus tubingensis2, has accession number ON981296, Aspergillus welwitschiae has accession number ON981099 .
The results showed the affinity and similarity between the recorded fungi with the global isolates. The focus was on the phenomenon of fungal antagonism of fungi isolated from sputum, as well as some dermatophytes obtained from previous experiments.
The results of fungal antagonism showed that there was a inconsistency in the antagonistic ability of the fungi used in the study

with R. stolonifer , as it reached 3 according to the Bell scale, after growing them on Sabouraud dextrose agar)SDA) at a temperature of 28 0 C for five days. And then purified and converted into fungal filtrate.
The results showed that there were significant differences at the level of probability 0.05 for the type of fungi extract and its concentration, as it was observed that the effect of the fungal fluctuated compared to the control treatment, which gave a diameter growth rate of 8.0 cm after five days of incubation at a temperature of 28 0 C. The fungal filtrate at a concentration of 75% showed superiority in its inhibition effect over other concentrations of 100%, 50%, and 25% on the growth of R. stolonifer. And with significant differences at a probability level of 0.05, as it gave a fungus growth rate of 2.571 cm, with a rate of 68%, compared to the rest of the concentrations 100%, 50%, and 25%, which gave a growth rate for the fungus, respectively, 3.119, 6.143, 3.619 cm, with an inhibition ratio of 61%, 23%, 55 %.
All nanofilters were subjected after the use of fungal filtrate to convert metallic silver and zinc into nanoparticles, and the first point of transformation into a nanofilter was the chromatic function and a difference in the color of the nanofilter from the normal filter before treatment with zinc oxide or silver nitrate, and then other tests related to confirmation of obtaining were conducted. On the nanomaterial, including ultraviolet and infrared rays.
The results showed that there were significant differences at the level of 0.05 for the type of zinc nanofiltrate, its concentration, and the binary interactions in the average diameter of the R. stolonifer colony. The results showed a fluctuation in the effect of the filtrate, but in general all the fungal filtrate inhibited the growth of the R. stolonifer. As for the concentration of the fungal filtrate with zinc nanoparticles, the concentration of 25% showed superiority over the concentrations of 50%, 75%, and 100% in its inhibitory effect on the growth of the R. stolonifer, with significant differences at the level of 0.05, as it gave the fungus growth rate of 0.738 cm, with an inhibition rate of 91% compared to the rest. Concentrations of 50%, 75%, and 100% gave growth rates of 1.310, 1.857, and 1.833 cm, respectively, with inhibition rates of 84%, 77%, and 77%. As for the control treatment, it gave a diagonal growth rate of 8.0 cm.
The results also showed that there were significant differences at the level of probability 0.05 for the type of fungus nanofiltration with silver and its

concentration, as the results showed fluctuation in the effect of the filtrates, but in general all of them inhibited the growth of the R. stolonifer. In terms of the concentration of the fungal nanofiltrate, the concentration of 50% showed superiority over the concentrations of 25%, 75%, and 100% in its inhibitory effect on the growth of R. stolonifer, with significant differences at a probability level of 0.05, as it gave the fungus growth rate of 2.405 cm, with an inhibition rate of 70% compared to the rest of the concentrations. Which gave a growth rate of 2.714, 8, 5 cm, respectively, with zero inhibition rates, 66%, and 38%, respectively, compared to the control treatment, which gave a diagonal growth rate of 8.0 cm.
Several microscopic examinations were carried out on R. stolonifer treated with fungal filtrate and nano solutions and explained the effect of different treatments and the variation of this effect between decomposition or distortions in the mycelium and the fragmentation of the protoplasm and its agglomeration in other areas as well as the distortions in the sporangia and their different sizes and maturity.