Polyphenolic profile of butterhead lettuce cultivar by ultrahigh performance liquid chromatography coupled online to UV-visible spectrophotometry and quadrupole time-of-flight mass spectrometry.

In the present study, the butterhead lettuce cultivar was analyzed by ultrahigh performance liquid chromatography (UHPLC) coupled online to diode array detection (DAD), electrospray ionization (ESI) and quadrupole time-of-flight mass spectrometry (QToF/MS) in the positive and negative ion mode in order to characterize its polyphenolic profile for the first time. The instrument acquisition mode MSE was used to collect automatic and simultaneous information of exact mass at high and low collision energies of precursor ions as well as other ions produced as a result of their fragmentation. One hundred eleven phenolic compounds were identified in the acidified hydromethanolic extract of freeze-dried leaves of butterhead lettuce cultivar: 40 hydroxycinnamic acid derivatives, 21 hydroxybenzoic acid derivatives, 2 hydroxyphenylacetic acid derivatives, 18 flavonols, 9 flavones, one flavanone, 7 coumarins, one hydrolysable tannin and 12 lignans. Forty-seven of these compounds have been tentatively identified for the first time in lettuce.

Characterization of phenolic compounds in green and red oak-leaf lettuce cultivars by UHPLC-DAD-ESI-QToF/MS using MSE scan mode.

Lettuce (Lactuca sativa) is one of the most popular leafy vegetables in the world and constitutes a major dietary source of phenolic compounds with health-promoting properties. In particular, the demand for green and red oak-leaf lettuces has considerably increased in the last years but few data on their polyphenol composition are available. Moreover, the usage of analytical edge technology can provide new structural information and allow the identification of unknown polyphenols. In the present study, the phenolic profiles of green and red oak-leaf lettuce cultivars were exhaustively characterized by ultrahigh-performance liquid chromatography (UHPLC) coupled online to diode array detection (DAD), electrospray ionization (ESI), and quadrupole time-of-flight mass spectrometry (QToF/MS), using the MSE instrument acquisition mode for recording simultaneously exact masses of precursor and fragment ions. One hundred fifteen phenolic compounds were identified in the acidified hydromethanolic extract of freeze-dried lettuce leaves. Forty-eight of these compounds were tentatively identified for the first time in lettuce, and only 20 of them have been previously reported in oak-leaf lettuce cultivars in literature. Both oak-leaf lettuce cultivars presented similar phenolic composition, except for apigenin-glucuronide and dihydroxybenzoic acid, only detected in the green cultivar; and for luteolin-hydroxymalonylhexoside, an apigenin conjugate with molecular formula C40 H54 O19 (monoisotopic MW = 838.3259 u), cyanidin-3-O-glucoside, cyanidin-3-O-(3″-O-malonyl)glucoside, cyanidin-3-O-(6″-O-malonyl)glucoside, and cyanidin-3-O-(6″-O-acetyl)glucoside, only found in the red cultivar. The UHPLC-DAD-ESI-QToF/MSE approach demonstrated to be a useful tool for the characterization of phenolic compounds in complex plant matrices.

Lactic acid as potential substitute of acetic acid for dissolution of chitosan: preharvest application to Butterhead lettuce.

Chitosan must be dissolved in acid solution to activate its antimicrobial properties. The objectives of present study were to determine whether acetic and lactic acids used to dissolve chitosan would influence its effectiveness to control the native microflora of Butterhead lettuce at harvest and during postharvest storage (7-8 °C, 5 days). Chitosan was applied as a SINGLE DOSE (14, 10, 7, 3 or 0 days previous to harvest) or in SUCCESSIVE DOSES (at 14 + 10 + 7+3 + 0 days prior to harvest). Although chitosan in acetic acid showed antimicrobial activity, treated plants showed dried brown stains which significantly reduced sensorial quality. Chitosan in lactic acid applied in a SINGLE DOSE at harvest or in SUCCESSIVE DOSES reduced microbial counts of all populations at harvest without affecting sensorial quality. After postharvest storage, lettuce treated with SUCCESSIVE APPLICATIONS of chitosan in lactic acid presented significant reductions in the microbial populations compared with untreated sample (-2.02 log in yeast and molds, -1.83 log in total coliforms, -1.4 log CFU g-1 in mesophilic bacteria and -1.1 log in psychrophilic bacteria). In conclusion, replacement of acetic by lactic acid did not affect the antimicrobial activity of chitosan, reducing microbial counts at harvest and after postharvest storage without affecting sensorial quality.

Freshly characterization and storability of mini head lettuces at optimal and abusive temperatures.

Selection of lettuce varieties less sensitive to quality deterioration and more tolerant to abusive temperatures during handling, transportation, and storage is essential to minimize economical and quality losses that affect both producers and consumers. This work was focused on the quality changes of four baby head lettuces (Lactuca sativa L.), two butter (red and green) and two oak-leaf (red and green) types, during storage at 0 ℃ and 10 ℃ for 10 days. Lettuce quality was determined by measuring bioactive content (ascorbic acid, total phenolics), physicochemical (total chlorophyll, browning potential), and microbiological indices. At harvest, red varieties presented lower browning potential and higher bioactive compounds but no differences were observed in microbial populations. During storage, ascorbic acid underwent first order degradation for all varieties, with a degradation rate at 10 ℃ twice faster than at 0 ℃. At 0 ℃, only the red oak-leaf lettuce exhibited chlorophyll degradation, while at 10 ℃ all varieties presented degradation. No changes were observed in total phenolics and browning potential of butter lettuces during storage at both temperatures. Microbial population counts were significant affected by the storage temperature. Red butter baby lettuce presented slightly better bioactive content and microbiological characteristics and then better storability.

Oregano essential oil-pectin edible films as anti-quorum sensing and food antimicrobial agents.

Edible films can be used as carriers for antimicrobial compounds to assure food safety and quality; in addition, pathogenesis of food bacteria is related to a cell to cell communication mechanism called quorum sensing (QS). Oregano essential oil (OEO) has proved to be useful as food antimicrobial; however, its food applications can be compromised by the volatile character of its active constituents. Therefore, formulation of edible films containing OEO can be an alternative to improve its food usages. QS inhibitory activity of OEO and pectin-OEO films was evaluated using Chromobacterium violaceum as bacterial model. Additionally, antibacterial activity was tested against Escherichia coli O157:H7, Salmonella Choleraesuis, Staphylococcus aureus, and Listeria monocytogenes. OEO was effective to inhibit bacterial growth at MIC of 0.24 mg/mL for all tested bacteria and MBC of 0.24, 0.24, 0.48, and 0.24 mg/mL against E. coli O157:H7, S. Choleraesuis, S. aureus, and L. monocytogenes, respectively. Pectin-films incorporated with 36.1 and 25.9 mg/mL of OEO showed inhibition diameters of 16.3 and 15.2 mm for E. coli O157:H7; 18.1 and 24.2 mm for S. Choleraesuis; 20.8 and 20.3 mm for S. aureus; 21.3 and 19.3 mm for L. monocytogenes, respectively. Pectin-OEO film (15.7 mg/mL) was effective against E. coli O157:H7 (9.3 mm), S. aureus (9.7 mm), and L. monocytogenes (9.2 mm), but not for S. Choleraesuis. All concentrations of OEO (0.0156, 0.0312, 0.0625 and 0.125 mg/mL) and pectin-OEO films (15.7, 25.9 and 36.1 mg/mL) showed a significant anti-QS activity expressed as inhibition of violacein production by C. violaceum. Additionally, the application of pectin-OEO films was effective reducing total coliforms, yeast, and molds of shrimp and cucumber slices stored at 4°C during 15 d. These results demonstrated the potential of pectin films enriched with OEO as food related microorganisms and QS inhibitors.

Brewer's spent grain: characterization and standardization procedure for the enzymatic hydrolysis by Bacillus cereus strain.

An important way to reuse agroindustrial by-products and to produce added-value products consists of the production of protein hydrolysates. In the current study, we used Brewer's spent grain (BSG), mainly because of its availability and cost, as a substrate for the enzymatic hydrolysis by Bacillus cereus. First, the physicochemical and microbiological characterization of BSG batches from three varieties was carried out. Furthermore, the optimal fermentation upstream processes for enzymatic hydrolysis by B. cereus were defined. Finally, the ability of B. cereus to hydrolyze different fractions of BSG was analyzed and possible synergistic effects of this bacterium along with other proteolytic bacteria were also investigated. Results showed that the naturally associated microflora was predominantly thermophilic aerobic bacteria and the drying process was the better alternative for BSG preservation. Water, lipids, and ash content differed significantly among the three varieties; however, no statistically significant differences were found in the protein content among them. After BSG characterization studies, the following protocol was set to obtain the fermentation substrate (FS): drying at 60°C for 24-48 H; sieving, grinding, and polyphenol extraction with an alcohol-water solution; and finally autoclaving. A synergistic effect was observed when B. cereus was inoculated with Pseudomonas strains in FS.

Effectiveness of edible coatings combined with mild heat shocks on microbial spoilage and sensory quality of fresh cut broccoli (Brassica oleracea L.).

UNLABELLED: The use of edible coatings and mild heat shocks is proposed as postharvest treatments to prevent microbial deterioration of refrigerated broccoli. Minimally processed broccoli was coated with either chitosan or carboxymethyl-cellulose (CMC) combined or not with a previous application of a mild heat shock. The evolution of microbial populations (mesophilic, psycrotrophic, Enterobacteriaceae, molds and yeast, and lactic acid bacteria) was studied during 20 d of storage and fitted to Gompertz and logistic models. Results revealed that, at the end of the storage, chitosan coating significantly reduced all microbiological population counts, except lactic acid bacteria; while higher reduction was observed with chitosan coating combined with a heat shock treatment. A significant delay at the beginning of the exponential phase was observed for all the bacterial populations analyzed. On the other hand, CMC coating, with and without a previous thermal treatment, did not exert any antibacterial effect. Excellent agreement was found between experimental microbial counts and predicted values obtained from Gompertz and logistic models. Kinetic modeling was found to be valuable for prediction of microbiological shelf life of broccoli during storage. Results showed that the application of chitosan coating effectively maintained microbiological quality and extended shelf life of minimally processed broccoli. According to these results, the use of the edible chitosan coating alone or in combination with a heat mild shock appear to be a viable alternative for controlling microbiological growth and sensory attributes in minimally processed broccoli. PRACTICAL APPLICATION: The continuous consumer interest in high quality and food safety, combined with environmental concern has induced to the development and study of edible coatings that avoid the use of synthetic materials. The edible coatings, formed from generally recognized as safe materials, have the potential to reduce weight loss, respiration rate, and improve food appearance and integrity. It is one of the most effective methods to maintain food quality. On the other hand, heat treatments have been demonstrated to be effective as a nonchemical means of improving postharvest quality for a variety of horticultural products. The applications of mild heat shocks combined with edible coatings constitute an alternative for the natural preservation of crops for which the use of synthetic chemicals is objectionable.

Essential oils as biopreservatives: different methods for the technological application in lettuce leaves.

Investigations were carried out to assess the efficiency of 3 essential oils, clove, tea tree, and rosemary, as natural preservatives during the postharvest of lettuce leaves. The effect of different concentration (1 and 0.5 MIC) of plant essential oils applied in 3 forms (spray, immersion, and capsules) was studied on lettuce leaves. The evolution of different microbial populations was evaluated during refrigerated storage. The application forms of the biopreservatives were shown to be an important factor in determining the effectiveness of the essential oils. Clove and tea tree essential oils at 1 MIC and applied embedded in lactose capsules presented a significant inhibition on mesophilic, psicrotrophic, and coliforms populations, while rosemary in none of the 3 technological applications forms exerted inhibitory effect on all microbial populations evaluated. Essential oils (at 0.5 MIC) applied by spray, immersion, and embedded in lactose capsules exerted lower inhibitory effects, with respect to 1 MIC, on the different microbial populations present on lettuce leaves. At the end of the storage (7 d), lettuce samples treated with tea tree, clove, and rosemary (at 1 and 0.5 MIC) by spray were the only organoleptically acceptable. It is concluded that clove and tea tree essential oils can control different microbial population present in lettuce. Practical Application: The exploration of naturally occurring antimicrobials in food preservation receives increasing attention due to consumer awareness of natural food products. Biopreservatives are useful in extending the shelf life of foods, reducing or eliminating pathogenic bacteria and increasing overall quality of food products. The effectiveness of essential oil application in foods is the result of factor associations such as applications forms, concentration applied, the way of action, storage temperatures. The application methods (spray, immersion, and embedded in lactose capsules) and the concentration of essential oils have been shown to be important factors in determining the effectiveness of these biopreservatives. The oil concentrations required to produce a certain level of inhibition in actual foods could be questionable due to the organoleptic impact. However, these novel natural preservatives in combination with other factors in obstacle technologies are an alternative to control the pathogen growth minimizing undesirable changes in organoleptic characteristics.

Antimicrobial effectiveness of bioactive packaging materials from edible chitosan and casein polymers: assessment on carrot, cheese, and salami.

Antimicrobial packaging is one of the most promising active packaging systems for controlling spoilage and pathogenic microorganisms. In this work, the intrinsic antimicrobial properties of chitosan (CH) were combined with the excellent thermoplastic and film-forming properties of sodium caseinate (SC) to prepare SC/CH film-forming solutions and films. The antimicrobial effectiveness of SC, CH, and SC/CH coatings on the native microfloras of cheese, salami, and carrots was evaluated. In vitro assays through the test tube assay indicated that the most significant antimicrobial effect was achieved by CH and SC/CH solutions on carrot and cheese native microfloras. SC film-forming solutions did not exert antimicrobial activity on any of the native microflora studied. SC, CH, and SC/CH films stored in controlled environments showed that the retention of the antimicrobial action was observed until 5-d storage, at 65% relative humidity in both temperatures (10 °C and 20 °C). In vivo assays were also performed with SC, CH, and SC/CH applied as coatings or wrappers on the 3 food substrates. CH and SC/CH applied at both immersion and wrapper exerted a significant bactericidal action on mesophilic, psychrotrophic, and yeasts and molds counts, showing the 3 microbial populations analyzed a significant reduction (2.0 to 4.5 log CFU/g). An improvement of the bactericidal properties of the CH/SC blend respect to those of the neat CH film is reported. The ionic interaction between both macromolecules enhances its antimicrobial properties. Practical Application: The continuous consumer interest in high quality and food safety, combined with environmental concerns has stimulated the development and study of biodegradable coatings that avoid the use of synthetic materials. Among them, edible coatings, obtained from generally recognized as safe (GRAS) materials, have the potential to reduce weight loss, respiration rate, and improve food appearance and integrity. They can be used in combination with other food preservation techniques in order to extend the effectiveness of the food preservation chain. Moreover, antimicrobial films and coatings have innovated the concept of active packaging and have been developed to reduce, inhibit, or delay the growth of microorganisms on the surface of food in contact with the package. The use of antimicrobials packaging films to control the growth of microorganisms in food can have a significant impact on shelf-life extension and food safety. In addition, antimicrobial films can be prepared by the combination of inherent antimicrobial materials (that is, CH), with good film-forming protein-based ones (that is, SC). Therefore, the objective of this work is to study the performance of 2 biodegradable and edible biopolymers and their combination as natural packages for selected food products.

Quality characterization of celery (Apium graveolens L.) by plant zones and two harvest dates.

The aim of this study was to characterize the quality of celery petioles (Apium graveolens L. cv Golden Clause) from the external, middle, and internal zones of each plant. Harvest dates were 93 (HD1) and 124 (HD2) days after transplantation. Total weight (TW), total length (TL), total leaves number (LN), and petiole length of leaves (PL) for the 3 zones were measured. Physicochemical (color, b* and h; texture; total soluble solids, TSS; titratable acidity, TA; water content, WC), nutraceutical (ascorbic acid, AA; total quinones, TQ; browning potential, BP), and sensorial analysis (flavor, fibrosity, crunchiness) were done at harvest on petioles of each zone. No differences between harvesting dates were found in TW, TL, LN, and PL. Middle leaves had the highest PL. Harvest dates were not different in b*, h, TA, AA, and WC. Texture, TSS, BP, and TQ resulted higher in petioles of HD2 than in HD1. Similar TSS and TA were found in leaves of different plant zones. The yellow color of both external and internal zones was significantly higher than in the middle zone. The texture and BP was similar between the external and middle zones but the WC was lower in the external zone. Similarly, the AA content as well as the TQ was also lower than in the middle zone. Harvest delay improved the nutraceutical value in terms of BP and TQ, even though it also resulted in pithiness and fibrosity of the leaves. This study therefore suggests that the petioles of the middle zone when harvested at HD1 are the most suitable for consumption. Practical Application: Celery is a vegetable reduced in calories, has a high nutritional value and its fresh petioles are mainly consumed in salads. The texture and flavor are the most important attributes that define consumers' acceptability. As nutritional value, texture, and flavor may change with plant age and different zones of the plant, harvest date plays an important role on quality. Results indicate that harvest delay improved the nutraceutical value even though it also resulted in pithiness and fibrosity of the stalks. Petioles of the middle zone, when harvested at 93 d after transplanting, are the most suitable for consumers' consumption.

Isolation and identification of Bacillus spp. and related genera from different starchy foods.

UNLABELLED: Samples of butternut squash, potatoes, rice, and wheat flour were analyzed. Bacillus spp. and related species belonging to Paenibacillus and Brevibacillus genera were found in 96% of the samples. In butternut squash, predominant species were Bacillus pumilus and Paenibacillus polymyxa together with other Bacillus spp. species (B. cereus, B. licheniformis, B. sphaericus, and B. subtilis). In all the potato samples, Bacillus species were detected (B. cereus, B. mycoides, and B. licheniformis). Also, Bacillus spp. were detected in 100% of the unhusked rice samples, while incidence in white rice samples was 83%. In total rice samples, B. pumilus, Brevibacillus brevis, and Paenibacillus macerans were the main species and B. cereus, P. polymyxa, B. subtilis, and Brevibacillus laterosporus had the lower percentage. The most important species found in wheat flour was P. polymyxa with colony forming units per gram of about 10(2). As the identified species were potentially causatives of foodborne diseases, attention should be given to sanitary and temperature conditions as critical factors that influence the safety and shelf life of these products. PRACTICAL APPLICATION: Foodborne illness produce by B. cereus have been associated with a wide variety of food. In addition, some other Bacillus species have been related to foodborne disease in humans. Information about the virulence mechanisms of other Bacillus spp. is scanty and their risk is underestimated. Identifying the group of food and the food processes in which Bacillus cereus or other Bacillus spp. would be hazardous for human health is vital for the prevention of foodborne outbreak. In this study, we determined the incidence of Bacillus spp. and related genera in some food items of agriculture origin from Argentina. This research is relevant to identify the presence of potentially pathogen Bacillus species and related genera in this type of food.