Search

In recent decades, fresh and minimally processed produce items have been associated with an increasing proportion of food-borne illnesses. Most pathogens associated with fresh produce are enteric (fecal) in origin, and contamination can occur anywhere along the farm-to-fork chain. Microbial source tracking (MST) is a tool developed in the environmental microbiology field to identify and quantify the dominant source(s) of fecal contamination. This study investigated the utility of an MST method based on Bacteroidales 16S rRNA gene sequences as a means of identifying potential fecal contamination, and its source, in the fresh produce production environment. The method was applied to rinses of fresh produce, source and irrigation waters, and harvester hand rinses collected over the course of 1 year from nine farms (growing tomatoes, jalapeño peppers, and cantaloupe) in Northern Mexico. Of 174 samples, 39% were positive for a universal Bacteroidales marker (AllBac), including 66% of samples from cantaloupe farms (3.6 log10 genome equivalence copies [GEC]/100 ml), 31% of samples from tomato farms (1.7 log10 GEC/100 ml), and 18% of samples from jalapeño farms (1.5 log10 GEC/100 ml). Of 68 AllBac-positive samples, 46% were positive for one of three human-specific markers, and none were positive for a bovine-specific marker. There was no statistically significant correlation between Bacteroidales and generic Escherichia coli across all samples. This study provides evidence that Bacteroidales markers may serve as alternative indicators for fecal contamination in fresh produce production, allowing for determination of both general contamination and that derived from the human host.

Although RT-qPCR is a powerful tool for human norovirus (HuNoV) detection, low virus concentrations in potentially large sample volumes necessitate the use of inefficient sample processing step(s) prior to detection. Process control viruses (PCVs) are used to monitor the efficiency of these virus concentration steps. This study compared five PCVs [Mengovirus (Mengo), murine norovirus (MNV-1), MS2 coliphage, Tulane virus, and turnip crinkle virus (TCV)] to two HuNoV strains for recovery during the steps of elution, polyethylene glycol precipitation (PEG), and RNA extraction from select foods (lettuce and sliced deli ham). Results demonstrate high recovery efficiencies of HuNoV GI.6 and GII.4 using the methods described in this study: combined (sequential) losses during processing from sliced deli ham and lettuce were < 1 log 10 genome equivalent copies (GEC). When considering the processing steps separately, HuNoV loss was negligible after elution, and low after PEG precipitation (mean 0.5 log 10 GEC) and RNA extraction (mean 0.1 log 10 GEC). The virus that least mimicked the behavior of HuNoV during sample processing was MNV-1. Of the viruses tested, a commercial mengovirus strain gave recovery efficiencies closest to HuNoV, showing combined losses from sliced deli ham and lettuce of < 1 log 10 GEC and ~ 1 log 10 GEC, respectively. All PCVs do not behave equivalently and validation of their performance is recommended before their routine use on an application-by-application basis. [ABSTRACT FROM AUTHOR]

Hand hygiene interventions are critical for reducing farmworker hand contamination and preventing the spread of produce-associated illness. Hand hygiene effectiveness may be produce-commodity specific, which could influence implementation strategies. This study’s goal was to determine if produce commodity influences the ability of handwashing with soap and water or two-step alcoholbased hand sanitizer (ABHS) interventions to reduce soil and bacteria on farmworker hands. Farmworkers (n)326) harvested produce (cantaloupe, jalapeño, and tomato) for 30 to 90 minutes before engaging in handwashing, two-step ABHS (jalapeño and cantaloupe), or no hand hygiene. Hands were rinsed to measure amounts of soil (absorbance at 600 nm) and indicator bacteria (coliforms, Enterococcus sp., generic Escherichia coli, and Bacteroidales universal [AllBac] and human-specific [BFD] 16S rRNA gene markers). Without hand hygiene, bacterial concentrations (0.88 to 5.1 log10 CFU/hand) on hands significantly differed by the produce commodity harvested. Moderate significant correlations (ρ = -0.41 to 0.56) between soil load and bacterial concentrations were observed. There were significant produce-commodity-specific differences in the ability of handwashing and two-step ABHS interventions to reduce soil (P < 0.0001), coliforms (P=0.002), and Enterococcus sp. (P=0.003), but not the Bacteroidales markers AllBac (P=0.4) or BFD (P=0.3). Contamination on hands of farmworkers who harvested cantaloupe was more difficult to remove. Overall, we found that a two-step ABHS intervention was similar to handwashing with soap and water at reducing bacteria on farmworker hands. In summary, produce commodity type should be considered when developing hand hygiene interventions on farms. [ABSTRACT FROM AUTHOR]