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The antimicrobial activity of zinc oxide nanoparticles (ZnONPs) and cinnamon oils (C.O.) was evaluated by conventional and molecular methods against Aspergillus flavus (A.flavus) and Escherichia coli 0157 (E.coli) that recovered from cattle mastitis. In agar well diffusion method (WD), Minimum inhibitory concentration (MIC) of ZnONPs and C.O. for A.flavus was (100 μg/ml; 0.25%) and for E.coli 0157 were (50 μg/ml; 0.25%), respectively. The synergistic effects of these materials caused higher significant inhibition of all microbial growth by low and high doses by agar method. But, the molecular detection of virulent genes of E. coli (stx1) and A. flavus (AflR) by polymerase chain reaction (PCR) and the real-time PCR (RT-PCR) yielded uncorrelated results with WD tests. It is concluded that no direct correlation between WD, PCR, and RT-PCR and the WD tests are still inexpensive, eco-friendly, and rapidly applicable for screening of antimicrobials activity than genetic methods.

In the past decades, the application of nanotechnology indicated significant improvements in animal health. In the present work, 60 samples of poultry feeds were examined, including 20 samples for each yellow corn, soya bean, and processed feed. The prevalence of total fungi was reported as 100%, 95%, and 100% in yellow corn, soya bean, and processed feed, respectively. Toxin-producing Aspergillus flavus represented 75% of isolates from yellow corn, 88% from soya bean meal, and 50% from processed feed. Aflatoxins were found in 88%, 60%, and 80% of yellow corn, soya bean, and processed feed with mean levels of 18.5 ± 3.216.0 ± 4.08.3 ± 1.7 ppm, respectively. The copper nanoparticles embedded with chitosan were green synthesized using an eco-friendly method, and their antifungal activity was evaluated against aflatoxigenic mold that recovered from poultry feeds. However, the molecular detection of virulent genes of Aspergillus flavus (aflR gene) after their exposure to high doses of copper-chitosan nanoparticles (CuCh-NPs) 150 µg/ml prevents aflR gene expression. The embedded chitosan with copper nanomaterial helps decrease their suspected toxicity to animals by reducing the used doses. Hence, the use of nanocomposites of nanomaterials with green benefits substances, such as chitosan, was the essential strategy of field application in veterinary.

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