Abstract
Neem oil nanoemulsions (NO NEs) have gained attention as natural antibacterial agents due to toxicity concerns surrounding synthetic preservatives. This study aimed to prepare a response surface methodology (RSM)-optimized NO NE < 200 nm to achieve a stable dip solution to maintain the sensory quality of Oreochromis niloticus fillets. The NO NE achieved a stable formulation with a particle size of 160.2 ± 0.04 nm on average. The polydispersity index (PDI) was 0.1 ± 0.05, and the zeta potential was found to be 18.2 ± 0.09 mV. Gas chromatography confirmed the presence of nimbiol, nimbandiol, 6-deacetyl nimbinene, and azadirachtin in NO after ultrasonic homogenization for 10 min (alternating between 30 s rest and 30 s work time). The NE had a spherical shape with a smooth surface, as was evident from transmission electron microscopy (TEM). Furthermore, NO:PM (neem oil-potassium metabisulphite) had an MIC (minimum inhibitory concentration) value of 150 ppm, compared to 210 ppm for the NO NE alone, against Staphylococcus aureus. Time-kill dynamics revealed the more effective control of S. aureus until 72 h with NO:PM. Moreover, DNA and protein leakage also increased from 0.145 ± 0.001 to 0.769 ± 0.002 OD (optical density) and from 0.142 ± 0.002 to 0.740 ± 0.001 OD, respectively, with the co-formulation of NO:PM. Conclusively, NO:PM inhibited S. aureus at a lower dose compared to the NO NE alone. Time-kill dynamics revealed complete inhibition of S. aureus in vitro for a period of 72 h. On the other hand, a proximate analysis of O. niloticus fillets showed no alteration in pH, no protein loss, and juiciness/moisture retention during 30 days of storage (4 °C). Sensory panelists reported that O. niloticus fillets treated with NE NO had improved color, flavor, juiciness, aroma, and overall quality. These results show that NE NO is a suitable green preservative for fish and possibly other meat-based products.