2.9 Parallel Streak Test
The assay was performed following American Association of Textile Chemists and Colorists (AATCC TM147) standard (Rejendra et al., 2011). Nutrient agar (Merck) was prepared in 90 mm Petri dishes for bio-assay. Parallel lines were drawn at the back of the petri dish with measurements of 6.5 cm in length with 1 cm space between each line. Before proceeding with the test, both the negative control and nanocomposite hydrogel with phomopsidione were excised to square hydrogels of 20 mm × 20 mm and sterilized under UV light for 30 min. Using sterile cotton swabs, the microbial inoculum was streaked on the nutrient agar (Merck) based on the parallel lines drawn. Next, the square nanocomposite hydrogels were placed symmetrically on the parallel lines, then gently pressed to allow proper contact between the hydrogel and the inoculated agar medium. The experiment was done in triplicate in separate occasions. All plates were incubated at 37˚C for 24 hours. After the incubation period, the diameters of inhibition zone surrounding the hydrogel were observed and measured.