Summary In Fall 2006, four separate outbreaks of foodborne illness associated with the consumption of fresh produce occurred in the United States. In follow‐up investigations, spinach, lettuce, and tomatoes were identified as the vehicles of illness. Epidemiologic investigations subsequently focused on finding the specific growing regions using traceback records. While the areas most likely involved in the outbreaks have been identified, the specific mode of contamination remains unconfirmed. Suspected risk factors in these cases include: proximity of irrigation wells and surface waterways exposed to faeces from cattle and wildlife; exposure in fields to wild animals and their waste materials; and improperly composted animal manure used as fertilizer. Difficulty in deciphering these and other on‐farm routes of contamination is due to the sporadic nature of these events. Hence, evidence to support these contamination modes is based largely on experimental studies in the laboratory and field. Still at issue is the relevance of internalization of pathogens, whether this occurs through the roots and plant vascular tissues of vegetables and fruits or through plant surfaces into cracks and crevices. Potential for these events, conditions under which the events occur, and pathogen survival following these events, are questions that still need to be answered. Answers to these questions will ultimately affect the type of interventions needed for application postharvest. Currently, many chemical and biological interventions can reduce surface pathogens and minimize cross‐contamination, however, they are largely ineffective on internalized pathogens. In the event internalization is a significant route of contamination in the field, physical interventions (irradiation and high pressure) may be needed to minimize risk. Ultimately, risk assessment studies will be useful tools in developing risk management strategies for the produce industry.

Green fluorescent protein-labeled Escherichia coli O157:H7 and Salmonella enterica serovar Enteritidis were inoculated at 10(7) CFU/g into cow, hog, or chicken manure. Ten- or 11-day-old soldier fly larvae (Hermetia illucens L.) (7 to 10 g) were added to the manure and held at 23, 27, or 32 degrees C for 3 to 6 days. Soldier fly larvae accelerated inactivation of E. coli O157:H7 in chicken manure but had no effect in cow manure and enhanced survival in hog manure. The initial pH values of the hog and chicken manure were 6.0 to 6.2 and 7.4 to 8.2, respectively, and it is surmised that these conditions affected the stability of the larval antimicrobial system. Reductions of E. coli O157:H7 populations in chicken manure by larvae were affected by storage temperature, with greater reductions in samples held for 3 days at 27 or 32 degrees C than at 23 degrees C. Pathogen inactivation in chicken manure by larvae was not affected by the indigenous microflora of chicken manure, because Salmonella Enteritidis populations in larvae-treated samples were approximately 2.5 log lower than control samples without larvae when either autoclaved or nonautoclaved chicken manure was used as the contaminated medium during 3 days of storage. Extending the storage time to 6 days, larvae again accelerated the reduction in Salmonella Enteritidis populations in chicken manure during the first 4 days of storage; however, larvae became contaminated with the pathogen. After 2 days of feeding on contaminated manure, Salmonella Enteritidis populations in larvae averaged 3.3 log CFU/g. Populations decreased to 1.9 log CFU/g after 6 days of exposure to contaminated chicken manure; however, the absence of feeding activity by the maggots in later stages of storage may be responsible for the continued presence of Salmonella Enteritidis in larvae. Transfer of contaminated larvae to fresh chicken manure restored feeding activity but led to cross-contamination of the fresh manure.

Numerous field studies have revealed that irrigation water can contaminate the surface of plants; however, the occurrence of pathogen internalization is unclear. This study was conducted to determine the sites of Escherichia coli O157:H7 contamination and its survival when the bacteria were applied through spray irrigation water to either field-grown spinach or lettuce. To differentiate internalized and surface populations, leaves were treated with a surface disinfectant wash before the tissue was ground for analysis of E. coli O157:H7 by direct plate count or enrichment culture. Irrigation water containing E. coli O157:H7 at 10(2), 10(4), or 10(6) CFU/ml was applied to spinach 48 and 69 days after transplantation of seedlings into fields. E. coli O157:H7 was initially detected after application on the surface of plants dosed at 10(4) CFU/ml (4 of 20 samples) and both on the surface (17 of 20 samples) and internally (5 of 20 samples) of plants dosed at 10(6) CFU/ml. Seven days postspraying, all spinach leaves tested negative for surface or internal contamination. In a subsequent study, irrigation water containing E. coli O157:H7 at 10(8) CFU/ml was sprayed onto either the abaxial (lower) or adaxial (upper) side of leaves of field-grown lettuce under sunny or shaded conditions. E. coli O157:H7 was detectable on the leaf surface 27 days postspraying, but survival was higher on leaves sprayed on the abaxial side than on leaves sprayed on the adaxial side. Internalization of E. coli O157:H7 into lettuce leaves also occurred with greater persistence in leaves sprayed on the abaxial side (up to 14 days) than in leaves sprayed on the adaxial side (2 days).

Several foodborne pathogens, including Salmonella species and campylobacters, are common contaminants in poultry and livestock. Typically, these pathogens are carried in the animal's intestinal tract asymptomatically; however, they can be shed in feces in large populations and be transmitted by other vectors from feces to animals, produce, or humans. A wide array of interventions has been developed to reduce the carriage of foodborne pathogens in poultry and livestock, including genetic selection of animals resistant to colonization, treatments to prevent vertical transmission of enteric pathogens, sanitation practices to prevent contamination on the farm and during transportation, elimination of pathogens from feed and water, feed and water additives that create an adverse environment for colonization by the pathogen, and biological treatments that directly or indirectly inactivate the pathogen within the host. To successfully reduce the carriage of foodborne pathogens, it is likely that a combination of intervention strategies will be required.

Recent studies addressing the internalization of fresh produce by foodborne pathogens arose in response to the growing number of recent and high profile outbreaks involving fresh produce. Because chemical sanitizing agents used during harvest and minimal processing are unlikely to reach enteric pathogens residing within plant tissue, it is imperative that paths for pathogen entry be recognized and minimized. Using both microscopy and microbial enumeration tools, enteric pathogens have been shown to enter plant tissues through both natural apertures (stomata, lateral junctions of roots, flowers) and damaged (wounds, cut surfaces) tissue. In studies revealing preharvest internalization via plant roots or leaf stomata, experimental conditions have primarily involved exposure of plants to high pathogen concentrations (≥ 6 log g⁻¹ soil or 6 log ml⁻¹ water), but those pathogens internalized appear to have short-term persistence. Postharvest internalization of pathogens via cut surfaces may be minimized by maintaining effective levels of sanitizing agents in waters during harvesting and minimal processing.