The continued agricultural exposure of bacteria to metals such as copper and zinc may result in an increased copper tolerance through the food chain. The aim of this study was to determine the Cu and Zn tolerance of bacteria from fresh produce (cucumber, zucchini, green pepper, tomato, lettuce, vegetable salad, broccoli, cabbage, carrot, green onion, onion, and mango). Isolates (506 aerobic mesophiles) from 12 different food produce products were tested for growth in a range of Cu and Zn concentrations. Selected isolates were identified using 16S rDNA sequencing, and the presence of metal resistance genes was studied using PCR amplification. More than 50% of the isolates had MICs for copper sulfate greater than 16 mM, and more than 40% had MICs greater than 4 mM for zinc chloride. Isolates with high levels of tolerance to Cu and Zn were detected in all the produce products investigated. A selection of 51 isolates with high MICs for both Cu and Zn were identified as belonging to the genera Pseudomonas (28), Enterobacter (7), Serratia (4), Leclercia (1), Bacillus (10), and Paenibacillus (1). A study of the genetic determinants of resistance in the selected gram-negative isolates revealed a high incidence of genes from the pco multicopper oxidase cluster, from the sil cluster involved in Cu and silver resistance, and from the chromate resistance gene chrB. A high percentage carried both pco and sil. The results suggest that Cu and Zn tolerance, as well as metal resistance genes, is widespread in bacteria from fresh produce.
Keywords: Copper; Enterobacter; Fresh produce; Metal tolerance; Pseudomonas; Serratia.