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Indigenous bacterial populations in fresh-cut produce processing facilities can have a profound effect on the survival and proliferation of inadvertently contaminating foodborne pathogens. In this study, environmental samples were collected from a variety of Zone 3 sites in a processing plant before and after daily routine sanitation. Viable mesophilic aerobic bacteria population was evaluated using both culturing method and quantitative real-time PCR (qPCR) after propidium monoazide treatment. Zone 3 surface microbiota were analyzed using 16S rRNA gene amplicon sequencing with the Qiime2 bioinformatic pipeline. Over 8000 bacterial species across 4 major phyla were identified in Zone 3 microbiomes in the processing facility. Overall, effective bacterial reduction was observed at the sampling sites on the production floor, while sanitation effect on peripheral surfaces was less evident. Effective sanitation resulted in both quantitative and qualitive shifts of Zone 3 microbiota. Several species were highly abundant at multiple sample sites for both winter and summer samplings. Based on the spatial and temporal distribution of the most abundant species, a Zone 3 core microbiome in the processing facility was tentatively described to included Cupriavidus sp., Pseudomonas sp., Ralstonia sp., Arthrobacter psychrolactophilus, Pseudomonas veronii, Stenotrophomonas sp., and an unknown species of the family Enterobacteriaceae.

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Irrigation water, particularly if applied overhead, could be an important source of bacterial contamination to fresh produce. The colonization, survival, and proliferation of exogenous bacterial pathogens can be strongly influenced by the produce microbiota. In this study, spinach grown in an organic field was irrigated with ground water and potential alternative irrigation water including reclaimed wastewater, and urban runoff water, over a period of 2 weeks. Water and spinach samples were collected before and after irrigation for bacterial plate count, qPCR, and community profiling using 16S rDNA high-throughput sequencing analyses. The average bacterial population densities on spinach (6.50 ± 0.04 log CFU/g, 7.40 ± 0.10 log 16S copies/g) were significantly higher than those in irrigation water (3.61 ± 0.12 log CFU/ml, 4.94 ± 0.13 log 16S copies/ml). The composition and relative abundance of spinach microbiomes varied with different types of irrigation waters; however, the most abundant microbial taxa on spinach were not significantly affected by the irrigation with different types of water. Shigella, Salmonella, Listeria, Campylobacter spp., and pathogenic Escherichia coli were not detected in this study. This study provides information on the microbial ecology of diverse bacterial communities on spinach surface after irrigation by different types of water, which can benefit future studies on the interaction of microbes on produce, and the prevention of foodborne pathogens and plant disease.