María del Rosario Moreira

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.

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.

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.

Isolation and characterization of bacterial strains with a hydrolytic profile with potential use in bioconversion of agroindustial by-products and waste

There is a trend towards the use of novel technologies nowadays, mainly focused on biological processes, for recycling and the efficient utilization of organic residues that can be metabolized by different microorganisms as a source of energy. In the present study the isolation of bacterial strains from six different agro-industrial by-products and waste was performed with the objective of evaluating their hydrolytic capacities and suitability for use in bioconversion of specific substrates. The 34 isolated strains were screened in specific culture media for the production of various hydrolytic enzymes (lipase, protease, cellulase, and amylase). It was found that 28 strains exhibited proteolytic activity, 18 had lipolytic activity, 13 had caseinolytic activity, 15 had amylolytic activity, and 11 strains exhibited cellulolytic activity. The strains that showed the highest hydrolytic capacities with biotechnological potential were selected, characterized genotipically, and identified as Bacillus, Serratia, Enterococcus, Klebsiella, Stenotrophomonas, Lactococcus, and Escherichia genera. It was concluded that the strain isolates have a high potential for use in the bioconversion of agro-industrial waste, both as a pure culture and as a microbial consortium.

Effects of pulsed light treatments and pectin edible coatings on the quality of fresh-cut apples: a hurdle technology approach

BACKGROUND Pulsed light (PL) treatments stand as an alternative for the shelf-life extension of fresh-cut products. The antimicrobial effects of PL are well known; however, its influence on quality attributes needs to be assessed. This study was aimed at evaluating the application of PL treatments in combination with pectin-based edible coatings enriched with dietary fiber for the preservation of fresh-cut apples. RESULTS Dipping of fresh-cut apples in ascorbic acid/calcium chloride solution prior to pectin coating and PL treatments was effective to minimize browning and softening of apple surfaces. Incorporation of fiber in the pectin coating did not cause any change in microbial loads and sensory acceptability of apple cubes. Pectin-coated PL-treated apple pieces exhibited significantly higher antioxidant activity values than fresh and PL control samples. At the end of storage, the combination of both treatments resulted in an almost 2 log CFU g-1 reduction of microbial counts. Sensory attribute scores did not fall below the rejection limit throughout 14 days, although the presence of off-odors limited the acceptability of the pectin-coated samples. CONCLUSION The results demonstrate that PL treatments applied to pectin-coated fresh-cut apples may be used to maintain quality attributes, thus conferring prebiotic potential and extending the shelf-life of the product.

Assessment of lettuce quality during storage at low relative humidity using Global Stability Index methodology

Abstract During postharvest, lettuce is usually exposed to adverse conditions (e.g. low relative humidity) that reduce the vegetable quality. In order to evaluate its shelf life, a great number of quality attributes must be analyzed, which requires careful experimental design, and it is time consuming. In this study, the modified Global Stability Index method was applied to estimate the quality of butter lettuce at low relative humidity during storage discriminating three lettuce zones (internal, middle, and external). The results indicated that the most relevant attributes were: the external zone - relative water content, water content , ascorbic acid, and total mesophilic counts; middle zone - relative water content , water content, total chlorophyll , and ascorbic acid; internal zone - relative water content, bound water, water content, and total mesophilic counts. A mathematical model that takes into account the Global Stability Index and overall visual quality for each lettuce zone was proposed. Moreover, the Weibull distribution was applied to estimate the maximum vegetable storage time which was 5, 4, and 3 days for the internal, middle, and external zone, respectively. When analyzing the effect of storage time for each lettuce zone, all the indices evaluated in the external zone of lettuce presented significant differences (p < 0.05). For both, internal and middle zones, the attributes presented significant differences (p < 0.05), except for water content and total chlorophyll.

Development of A Microbial Consortium For Dairy Wastewater Treatment

The wastewater from the dairy industries usually contains high concentrations of contaminants and, since the volume generated is also high, the total contaminant load is very significant. Among the available options for treatment, biological degradation looks like the most promising one. Furthermore, the supplementation of the native microbial populations with external microorganisms with high specific degradation rates (bio-augmentation) has demonstrated to improve the performance of treatment. The main objective of this research was to select a combination of bacteria to improve the aerobic treatment of dairy processing wastewater. For this purpose, eleven fat/protein-degrading microorganisms belonging to the genera Bacillus, Serratia, Lactococcus, Enterococcus, Stenotrophomonas, Klebsiella and Escherichia, were evaluated as potential degrading bacteria using a Plackett-Burman design. Assays were carried out to select the strains that most significantly influenced the degradation of wastewater and biomass yield, in terms of COD removal. A simulated dairy industry effluent was used as culture medium. Four strains were selected as potential members of the microbial consortium: Lactococcus garvieae, Bacillus thuringiensis, Escherichia coli and Stenotrophomonas sp. The optimal operation temperature and pH range of the selected consortium were 32°C and 6 ∼ 8, respectively. The degradation percentages reached with the selected consortium were 80.67 and 83.44% at 24 and 48 h, respectively. The selected consortium significantly improved the degradation of the dairy wastewater, and the degradation degree achieved by this consortium was higher than by using the strains individually.

Clove (Syzygium aromaticum) Oils

Abstract Clove essential oil (Syzygium aromaticum) has been used as a topical anesthetic and flavoring for years. It is known to have antimicrobial, antiinflammatory, and antioxidant activity, mostly related to its content of eugenol and other polyphenolic compounds. Other uses of clove have also arisen, like insect repellent or growth promoter agent. Clove essential oil has a controlling effect over native microflora and over pathogenic microorganisms such as Escherichia coli O157:H7. Clove has been successfully employed as a surface sanitizer agent in lettuce seeds and as a biopreservative in leafy green vegetables (applied either preharvest or postharvest). The application of clove on leafy vegetables also reduced peroxidase activity. Clove essential oil should be considered as viable alternative to chlorine as a sanitizer agent due to its low toxicity and low environmental impact.

Casein and Chitosan Polymers: Use in Antimicrobial Packaging

Abstract Minimal processing operations can alter the integrity of fruits and vegetables and bring about negative effects on quality. Also, the presence of microorganisms on the surface of produce may compromise food safety. Traditionally, edible coatings have been used as a strategy to reduce the deleterious effects that minimal processing imposes on intact vegetable tissues. Biologically active molecules, such as those in chitosan and edible coating from milk protein, have significant potential to curb contaminations associated with food, as well as increasing concerns regarding the negative environmental impact of conventional packaging materials. The protein-polysaccharide complexes could exhibit better functional properties than proteins and polysaccharides alone. 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. This chapter aims to summarize the most important information on chitosan and casein films from a bioactivity point of view, and to highlight their potential for food preservation technology.

Peroxidase activity and sensory quality of ready to cook mixed vegetables for soup: combined effect of biopreservatives and refrigerated storage

Enzymatic senescence processes and browning of fresh cut vegetables negatively affect their sensory properties and nutritional value and finally result in the rejection of affected products by consumers. In order to prevent quality decay, the combined effects of natural antioxidants and storage temperature on peroxidase activity and sensory attributes (overall visual quality, browning and odor) of individual and mixed vegetables for soup (butternut squash, leek and celery) were evaluated. Fresh cut vegetables were treated with antioxidant solutions as tea tree essential oil (15 μl/mL), propolis extract (15 μl/mL) and gallic acid (2 mg/mL) and stored at optimal (5 °C) and abusive (15 °C) temperature for a maximum of 14 days. The application of natural preservatives, plus optimal storage conditions, exerted significant inhibitory effects in peroxidase activity of squash, celery and mixed vegetables throughout the storage. Furthermore, propolis treatment applied on mixed vegetables retarded browning appearance and preserved the visual quality for a longer period when compared to untreated product.