Antifungal proteins (AFPs) offer a great potential as new biofungicides to control deleterious fungi. The phytopathogenic fungus Penicillium expansum encodes three phylogenetically distinct AFPs, PeAfpA, PeAfpB and PeAfpC. Here, PeAfpA, a potent in vitro self-inhibitory protein, was demonstrated to control the infection caused by P. expansum in Golden apple fruits. We determined the production of the three proteins in different growth media. PeAfpA and PeAfpC were simultaneously produced by P. expansum in three out of the eight media tested as detected by Western blot, whereas PeAfpB was not detected even in those described for class B AFP production. Regardless of the culture medium, the carbon source affected Peafp expression. Notably, the production of PeAfpA was strain-dependent, but analyses of PeafpA regulatory sequences in the three strains studied could not explain differences in protein production. None of the PeAFPs was produced during apple infection, suggesting no relevant role in pathogenesis. PeAfpA together with PeAfpB and also with Penicillium digitatum PdAfpB showed synergistic interaction. The highly active antifungal PeAfpA also showed moderate antibacterial activity. We conclude that there is not a general pattern for Peafp gene expression, protein production or antimicrobial activity and confirm PeAfpA as a promising compound for postharvest conservation.

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A number of foodborne outbreaks have occurred in the past decade, with higher incidences associated with romaine lettuce and strawberries. Contaminated agricultural water has been reported as the source of microbial contamination in most of these outbreaks. Maintaining the adequate and sanitary quality (0 E. coli/100 mL) of agricultural water can be challenging during post-harvest operations such as washing. The study focused on the attachment of generic E. coli (Rifampicin resistant) onto romaine lettuce and strawberries, mimicking the produce wash step. The produce was washed with contaminated water, air-dried, and stored in display units for 7 days. The produce was sampled randomly each day and analyzed for the surviving E. coli count. The results indicated that E. coli can survive in both lettuce and strawberries over extended periods. A survival population of 2.3 log CFU/cm2 (day 8) was observed on lettuce with an initial population of 2.8 log CFU/cm2 (day 0). On strawberries, the population reduced from 3.0 (day 0) to 1.7 log CFU/cm2 (day 7), with an initial E. coli concentration of approx. 6 log CFU/mL in the wash water. Strawberry leaves had a higher attachment of E. coli than the fruit (p < 0.05). In conclusion, romaine lettuce and strawberries washed with contaminated water can cause an outbreak affecting consumers and public health.

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