Total of 28 fungal isolates from wheat seeds of Al-Mahmoudia variety obtained from grain stores within Nineveh Province during the 2024–2025 season were collected and diagnosed. Most common fungus in wheat seeds was Aspergillus niger 46.42%, followed by Aspergillus terrus 14.28%, Penicillium spp. 14.28%, Rhizoctonia solani 10.71%, followed by pests like, Lewia scrophulariae 7.14%, and A. flavus 7.14% Fungus isolation Biological strategies for control are environmentally friendly methods that seek to support sustainable development goals by reducing the use of chemical pesticides that may have long-term negative impacts on human health. To detect the antifungal effects of Trichoderma harzianum against Lewia scrophulariae, we evaluated in vitro antagonistic properties using a dual culture method and measured antibiotic efficacy using non-volatile metabolites. Results showed that the dual culture method showed statistically significant inhibition up to scale 1, with antibiotic efficacy greater than 70%. The morphological classification of selected pathogenic fungi Lewia scrophulariae was confirmed. This procedure was effectuate using a customized removal kit. Samples were then sent to Psomagene, Inc. in the U.S.A for sequencing, and the sequences were warranty in GenBank. The fungi isolated from the identified locations as a result of the alignment showed a nucleotide sequence characteristic of Lewia scrophulariae with accession number PV114987.