Effects of an Eco-Friendly Sanitizing Wash on Spinach Leaf Bacterial Community Structure and Diversity

Ready-to-eat (RTE) spinach is considered a high-risk food, susceptible to colonization by foodborne pathogens; however, other microbial populations present on the vegetable surface may interact with foodborne pathogens by inhibiting/inactivating their growth. In addition, sanitizers applied to minimally processed salad leaves should not disrupt this autochthonous barrier and should be maintained throughout the shelf life of the product. This investigation aimed at comparing the effects of a pH neutral electrochemically activated solution (ECAS), a peroxyacetic acid (PAA)-based commercial sanitizer (Ecolab Tsunami® 100), and tap water wash on the minimally processed spinach leaf microbiome profile for 10 days after washing. The bacterial microbiota composition on spinach samples was assessed by 16S rRNA pyrosequencing and downstream analyses. Predominant phyla observed in decreasing order of abundance were Proteobacteria, Bacteroidetes, Actinobacteria and Firmicutes corresponding with the dominant families Micrococcaceae, Clostridiales Family XII, Flavobacteriaceae, Pseudomonadaceae, and Burkholderiaceae. Bacterial species richness and evenness (alpha diversity) and bacterial community composition among all wash types were not significantly different. However, a significant difference was apparent between sampling days, corresponding to a loss of overall heterogeneity over time. Analysis of composition of microbiome (ANCOM) did not identify any amplicon sequence variants (ASVs) or families having significantly different abundance in wash types; however, differences (17 ASVs and five families) were found depending on sampling day. This was the first bacterial microbiome composition study focused on ECAS and PAA-based wash solutions. These wash alternatives do not significantly alter microbial community composition of RTE spinach leaves; however, storage at refrigerated temperature reduces bacterial species heterogeneity.