Investigations on advance effects of boron-containing compounds have gained attention the last decade. This study was carried out to investigate the effect of hexagonal boron nitride nanoparticles (BNNPs) on Bovine Coronavirus (BCoV) and Staphylococcus aureus (S. aureus) by different methods. First, biological effects of different BNNPs concentrations lower than 0.5 mg/mL examined on HRT-18 (Human Rectal Tumor) for 5 days. Different concentrations of hBN mixed with BCoV in liquid, on membrane or directly on cells and examined for differences of titers or replications. And also Bacterial Filtration Efficiency (BFE) test of hBN powders coated on polypropylene fabric by spray method was applied against S. aureus. The compound was found slightly toxic on the HRT-18 cell line by live cell counting, while no remarkable morphological difference was observed. BNNPs treatment with 0.025 or 0.3mg/mL concentrations did not reduce the infective titer and create no inhibitory effect on in vitro replication. Stability of virus titer after treatment of BNNPs coated fabric also indicated no antiviral efficiency. But hBN applied fabric formed a barrier of ?90.3%, while non hBN applied fabric formed ?64.6% barrier. The present study demonstrate that, BNNPs alone is not a good candidate for disinfectant or drug for BCoVs, while it could be valuable to use as coated fabric in the areas needing easy sanitation especially for S. aureus.