María I. Gil

Fresh-cut product sanitation and wash water disinfection: Problems and solutions

It is well known that fresh-cut processors usually rely on wash water sanitizers to reduce microbial counts in order to maintain quality and extend shelf-life of the end product. Water is a useful tool for reducing potential contamination but it can also transfer pathogenic microorganisms. Washing with sanitizers is important in fresh-cut produce hygiene, particularly removing soil and debris, but especially in water disinfection to avoid cross-contamination between clean and contaminated product. Most of the sanitizing solutions induce higher microbial reduction after washing when compared to water washing, but after storage, epiphytic microorganisms grow rapidly, reaching similar levels. In fact, despite the general idea that sanitizers are used to reduce the microbial population on the produce, their main effect is maintaining the microbial quality of the water. The use of potable water instead of water containing chemical disinfection agents for washing fresh-cut vegetables is being advocated in some European countries. However, the problems of using an inadequate sanitizer or even none are considered in this manuscript. The need for a standardized approach to evaluate and compare the efficiency of sanitizing agents is also presented. Most new alternative techniques accentuate the problems with chlorine suggesting that the industry should move away from this traditional disinfection agent. However, the use of chlorine based sanitizers are presented as belonging to the most effective and efficient sanitizers when adequate doses are used. In this review improvements in water disinfection and sanitation strategies, including a shower pre-washing step and a final rinse of the produce, are suggested.

Characterisation of polyphenols and antioxidant properties of five lettuce varieties and escarole.

Salad vegetables could be relevant as dietary sources of natural antioxidants. A better knowledge of their composition can be useful for understanding their potential bioavailability and biological activities. The antioxidant compounds, polyphenols and vitamin C, have been determined in five varieties of lettuce (iceberg, romaine, continental, red oak leaf, lollo rosso) and one variety of escarole (frissé). The polyphenol study by HPLC-DAD-MS/MS ESI allowed the identification of two compounds previously not reported in lettuce; quercetin and luteolin rhamnosyl-hexosides. Qualitative and quantitative differences were observed between the polyphenol profiles. Caffeic acid derivatives were the main phenolics in green varieties, while flavonols were detected in higher quantities in red varieties and escarole, and anthocyanins were only present in red-leafed varieties. The highest total phenolic content was observed in red-leafed varieties while the highest level of vitamin C was detected in the continental variety. The red varieties showed the highest antioxidant activity by all the methods assayed.

HPLC-MS Analysis of Proanthocyanidin Oligomers and Other Phenolics in 15 Strawberry Cultivars†

The phenolic compounds of 15 strawberry cultivars grown in Spain were analyzed and quantified: anthocyanins (20.2-47.4 mg/100 g of fw) (cyanidin 3-glucoside and pelargonidin 3-glucoside, 3-rutinoside, and 3-malonyl glucoside), flavonols (1.5-3.4 mg/100 g of fw) (quercetin 3-glucuronide and kaempferol 3-glucoside and 3-p-coumaroyl-glucoside), proanthocyanidins (53.9-163.2 mg/100 g), p-coumaroyl-glucose (0.84-6.70 mg/100 g), ellagitannins (9.67-22.86 mg/100 g) (sanguiin H-6, lambertianin C, and galloyl bis-HHDP-glucose), and ellagic acid glycosides (0.88-2.06 mg/100 g of fw) (two ellagic acid deoxyhexosides). Proanthocyanidins, the main phenolic compounds, were characterized by phloroglucinol degradation. Their mean degree of polymerization ranged from 3.4 for cv. Chiflon to 5.8 for cv. Ventana, the average value being 4.3. The terminal unit of proanthocyanidin oligomers was always (epi)catechin (17.36-29.93%) and (epi)catechin (61.66-75.39%) or (epi)afzelechin (4.50-10.54%) as extension units. Different combinations of (epi)catechin and (epi)afzelechin were detected, and their sequence of linkage was characterized by HPLC-MS-MS. Relative percentages of dimers, trimers, tetramers, and pentamers were evaluated by the extracted ion chromatogram (EIC) analysis.

Keeping quality of fresh-cut tomato

The effects of calcium chloride washings and passive or active modified atmosphere packaging (MAP) on the keeping quality of fresh-cut tomato (Lycopersicon esculentum Mill., cv. Durinta) were investigated. Fresh-cut tomatoes were washed in chlorinated water (0.7 mM) with and without CaCl2 (0.09 M). Slices were placed in plastic trays, sealed with perforated or non-polymeric films and stored at 2 and 10°C. Firmness, colour, visual quality, aroma, texture, defects and overall quality were determined at harvest and after 7 and 10 days of storage. Soluble solids content (SSC), pH and titratable acidity (TA) were analyzed and a maturity index was designed. The initial CO2 production at 10°C was 120 nmol kg 1 s 1 while at 2°C it was 70 nmol kg 1 s 1 for both whole and tomato slices. During storage, at 10°C, CO2 production for whole tomato declined to 80 nmol kg 1 s 1 whereas for tomato slices after 2 days a continuous increase up to 490 nmol kg 1 s 1 was observed. Ethylene production was maintained at between 6 and 24 pmol kg 1 s 1 for whole and tomato slices at 2°C. Ethylene production by tomato slices reached 120 pmol kg 1 s 1 after 2‐4 days of storage at 10°C followed by a decrease to 12 pmol kg 1 s 1 after 7 days. In the passive MAP packages O2 decreased to 14 and 2.5% and CO2 increased to 6 and 20% on day 10 at 2 and 10°C, respectively. However, when active MAP was used (7.5% O2 0% CO2), O2 decreased to 6 and 1.5% and CO2 increased to 6 and 14% on day 10 at 2 and 10°C, respectively. After 10 days of storage, the quality attributes of tomato slices were maintained better at 2 than at 10°C. When slices were stored at 10°C, both passive and active MAP reduced the rate of ripening. The best results were found during storage at 2°C under active or passive MAP. Calcium chloride dips were useful only at 2°C in maintaining quality of tomato slices held in the perforated film. Active MAP should be used for maintaining fresh-cut tomato when stored at 10°C.

Prevention of Escherichia coli cross-contamination by different commercial sanitizers during washing of fresh-cut lettuce.

The efficacy of fresh-cut produce sanitizers has mainly been evaluated by measuring microbial reductions on produce. However, its suitability to ensure that pathogens are rapidly killed avoiding cross-contamination of subsequent product also needs to be considered. The efficacy of chlorine, Tsunami, Citrox and Purac on non pathogenic Escherichia coli reductions in processing water and on fresh-cut lettuce were studied. Selection of minimum effective doses was carried out in processing water, which contained a chemical oxygen demand (COD) within the range of 700-1000 mg/l and a total mesophilic load of about 7 log CFU/ml. The processing water was inoculated with two inoculum levels (3 and 5 log CFU/ml). It was observed that 40 mg/l of chlorine and 500 mg/l of Tsunami were effective in reducing the inoculum levels in the processing water to the detection limit (5 and 4 log units). However, Citrox and Purac were not effective in reducing E. coli population even at the highest manufacturers recommended doses. Evaluation of cross-contamination in fresh-cut lettuce was carried out by measuring E. coli transfer from inoculated (~5 log CFU/g) to uninoculated lettuce after washing the contaminated product in the water containing different sanitizing agents. Chlorine and Tsunami were able to inactivate E. coli in wash water, avoiding cross-contamination between contaminated and non-contaminated product. However, Citrox and Purac at the recommended doses did not prevent transfer of E. coli cells between inoculated and uninoculated fresh-cut lettuce and therefore indicating cross-contamination. The results obtained show that chlorine and Tsunami are recommended as water disinfection agents preventing E. coli cross-contamination of produce during processing.

Disinfection potential of ozone, ultraviolet-C and their combination in wash water for the fresh-cut vegetable industry

The purpose of this research was to investigate the disinfection efficacy of ozone (O(3)) and UV-C illumination (UV), and their combination (O(3)-UV) for reducing microbial flora of fresh-cut onion, escarole, carrot, and spinach wash waters collected from the industry. Furthermore, the influence of water physicochemical parameters on the decontamination efficacy and the effect of these technologies on physicochemical quality of wash water were analyzed. O(3), UV, and O(3)-UV were effective disinfection treatments on vegetable wash water, with a maximum microbial reduction of 6.6 log CFU mL(-1) after 60 min treatment with O(3)-UV. However, maximum total microbial reductions achieved by UV and O(3) treatments after 60 min were 4.0 and 5.9 log CFU mL(-1), lower than by O(3)-UV treatment. Furthermore, turbidity of wash water was reduced significantly by O(3) and O(3)-UV treatments, while UV treatment did not affect the physicochemical quality of the water. Conclusions derived from this study illustrate that O(3) and O(3)-UV are alternatives to other sanitizers used in the fresh-cut washing processes. The use of these technologies would allow less frequent changing of spent water and the use of much lower sanitizer doses. Nevertheless, in specific applications such as carrot wash water, where levels of undesirable microbial and chemical constituents are lower than other vegetable wash water, UV treatment could be an appropriate treatment considering cost-effectiveness criteria.

Cross-contamination of fresh-cut lettuce after a short-term exposure during pre-washing cannot be controlled after subsequent washing with chlorine dioxide or sodium hypochlorite

Chlorine dioxide (ClO(2)) has been postulated as an alternative to sodium hypochlorite (NaClO) for fresh-cut produce sanitization to avoid risks associated with chlorination by-products. Experiments were performed to determine the prevention of cross-contamination of fresh-cut lettuce by Escherichia coli using chlorine dioxide (3 mg/L) or sodium hypochlorite (100 mg/L) as sanitation agents. The efficacy of these sanitation solutions was evaluated simulating as much as possible the conditions of a fresh-cut processing line. Thus, to evaluate the potential risk of cross-contamination during pre-washing, inoculated fresh-cut lettuce was pre-washed and after that non-inoculated lettuce was then pre-washed in the same water. After this pre-washing, non-inoculated lettuce was cross-contaminated, changing from 0 to 3.4 log units of E. coli cells. During washing with sanitizers, none of the tested sanitation agents significantly reduced E. coli counts in both inoculated and cross-contaminated lettuce. These results suggest that when cross-contamination occurs, even if the event is recent, subsequent sanitation steps are inefficient for inactivating E. coli cells on the vegetable tissue. However, chlorine dioxide and sodium hypochlorite solutions were able to inactivate most E. coli cells that passed from inoculated product to wash water. Therefore, they might be able to avoid cross-contamination between clean and contaminated product during the washing step. Scanning electron microscopy micrographs indicated that bacterial cells were mainly located in clusters or tissue stomata where they might be protected, which explains the low efficacy of sodium hypochlorite and chlorine dioxide solutions observed in this study.

Pre- and Postharvest Preventive Measures and Intervention Strategies to Control Microbial Food Safety Hazards of Fresh Leafy Vegetables

This review includes an overview of the most important preventive measures along the farm to fork chain to prevent microbial contamination of leafy greens. It also includes the technological and managerial interventions related to primary production, postharvest handling, processing practices, distribution, and consumer handling to eliminate pathogens in leafy greens. When the microbiological risk is already present, preventive measures to limit actual contamination events or pathogen survival are considered intervention strategies. In codes of practice the focus is mainly put on explaining preventive measures. However, it is also important to establish more focused intervention strategies. This review is centered mainly on leafy vegetables as the commodity identified as the highest priority in terms of fresh produce microbial safety from a global perspective. There is no unique preventive measure or intervention strategy that could be applied at one point of the food chain. We should encourage growers of leafy greens to establish procedures based on the HACCP principles at the level of primary production. The traceability of leafy vegetables along the chain is an essential element in ensuring food safety. Thus, in dealing with the food safety issues associated with fresh produce it is clear that a multidisciplinary farm to fork strategy is required.

Floral nectar phenolics as biochemical markers for the botanical origin of heather honey

In order to find out biochemical markers for the botanical origin of heather (Erica) honey, the phenolic metabolites present in heather floral nectar, collected from the honey-stomach of bees gathering nectar from these flowers, were analysed. The flavonoid fraction of nectar contained four main flavonoids. Two of them were quercetin and kaempferol 3-rhamnosides, and the other two were tentatively identified as myricetin 3′-methyl ether and isorhamnetin 3-rhamnosides. Since the natural glycosides are hydrolysed by bee enzymes to render the corresponding aglycones, which are the metabolites detected in honey, acid hydrolysis of the nectar glycosides was achieved. The aglycones quercetin, myricetin 3′-methyl ether, kaempferol and isorhamnetin were identified, as well as the gallic acid derivative ellagic acid. The analysis of Portuguese heather honey samples showed that ellagic acid was present in all the samples in significant amounts ranging between 100 μg and 600 μg per 100 g honey. The other nectar-derived flavonoids were also present, although some of them in very variable amounts. Ellagic acid and myricetin 3′-methyl ether, which have not been detected in any of the monofloral honey samples investigated so far, with the only exception being a French honey sample of the botanically relatedCalluna (Ericaceae) which also contained ellagic acid, seem to be the most useful potential markers for the floral origin of heather honey. However, more detailed and extensive investigations are needed to prove the utility of these markers.

A chemotaxonomic study of flavonoids from european teucrium species

Abstract A survey of aerial tissues of 42 European taxa of the genus Teucrium has revealed the widespread presence of five surface flavonoids: cirsiliol, cirsimaritin, cirsilineol, salvigenin and 5-hydroxy-6,7,3′,4′-tetramethoxyflavone. The latter two compounds are useful taxonomic markers in that salvigenin is characteristic of species of section Polium, while 5-hydroxy-6,7,3′,4′-tetramethoxyflavone is completely confined to species of the other five sections surveyed. Eleven flavone glycosides, four flavonol glycosides and the glycoflavone vicenin-2 were found to occur as vacuolar constituents. One of the flavone glycosides, cirsimaritin 4′-glucoside, only occurs in the species T. arduini, while two others, hypolaetin and isoscutellarein 7-acetyl-allosylglucosides, are characteristic of the closely related T. chamaedrys and T. webbianum. 6-Hydroxyluteolin is widely present as the 7-glucoside and 7-rhamnoside, the latter compound being a new glycoside. In general, the chemical results are correlated with sectional classification and usefully indicate that at least one taxon, T. compactum, is misplaced within the genus. Phyletically, the restriction of flavonol glycosides mainly to section Teucrium suggests that this may be the basic group within the genus.