Enhanced disinfestation in grain spawn production through cold plasma and sodium hypochlorite synergy

Heat-resistant fungal conidia are a common source of contamination and can cause significant difficulties in producing spawns. Through the use of PCR method, Aspergillus tubingensis and Aspergillus flavus as common microbial contaminants found in wheat grain spawn were identified that had been sterilized at 120 ºc for 2 h. Since these conidia are highly resistant to standard sterilization techniques, alternative methods were used to treat them with NaOCl and cold plasma and evaluate their effectiveness in reducing contamination. Optical emission spectroscopy (OES) analysis of the plasma showed dominant emissions from the N₂ second positive system and N₂⁺ first negative system, while reactive oxygen species (ROS) spectral lines were undetected due to collision-induced quenching effects. Field Emission Scanning Electron Microscopy (FESEM) and Energy Dispersive X-ray Spectroscopy (EDXS) analyses revealed notable alterations in the elemental makeup of conidia surfaces, as evidenced by a marked rise in levels of Na, O, Cl (in the case of NaOCl treatment) and N (in the case of plasma treatment). The conidia size was reduced at lower levels of NaOCl, but with increased concentrations and plasma treatment, the conidia underwent rupture and, in some cases, pulverization. The research suggests that utilizing a combined approach can be highly effective in eliminating heat-resistant fungal conidia and drastically cutting down the sterilization time for producing wheat spawn to only 30 s.