Hasan Rafati

The online version contains supplementary material available at 10.1038/s41598-026-49846-9.

Tomato brown rugose fruit virus (ToBRFV) is a single-stranded positive-sense RNA virus that targets tomato and pepper plants and is causing significant damage to crops in some regions of the world. ToBRFV is a highly contagious virus that is stable and rapidly spreads by mechanical methods and seeds. As a result, it may spread both locally and over long distances, and it is now recognized as a pandemic in plants. This study investigates the effectiveness of the systems CRISPR-Cas12a and CRISPR-Cas9, in conjugation with recombinase polymerase amplification (RPA), to detect ToBRFV in tomato plant samples collected from the field. The trans-cleavage activity of both nucleases, Cas12a and Cas9, was exploited to process a probe labelled with fluorescein and biotin to be resolved on a lateral flow device, thereby enabling a visual readout. We were able to detect the RNA genome of the virus in about 1 h at a low constant temperature. These results could pave way to offer a rapid, sensitive, and specific method for on-site detection of ToBRFV.

Foodborne illnesses are often caused by microbial contamination during preparation or storage. In this work, stable nanoemulsions of clindamycin were prepared using Mentha piperita essential oil (MEO) as a nanocarrier delivery system. Response Surface Methodology was used to optimize the key variables for clindamycin nanoemulsion formulation, including 4.83, 2.83, and 0.14%w/w surfactant, essential oil, and clindamycin, respectively. The stability of MEO/clindamycin nanoemulsion (MEO/C NE) with a mean droplet size of 75.46 ± 3.2 nm was monitored over 3 months. The antibacterial activity of MEO/C NE and bulk compounds against E. coli bacterium was compared using a conventional method and a microfluidic chip. A significant difference in the antibacterial activity was observed by employing a microfluidic chip as compared to the conventional technique, probably due to a high contact surface area between the nanodroplets and bacterial membrane. In the microfluidic chip, the E. coli was completely inhibited in 30 min, whereas 3 h was needed for complete inhibition using the conventional method. The results of this study highlight the significance of nanoemulsion delivery systems to improve the antimicrobial activity of clindamycin and also microfluidic technology as a fast and reliable technique for examining antibiotics and nano delivery systems against microorganisms.