Isabel R. Amado

Influence of natural extracts on the shelf life of modified atmosphere-packaged pork patties

In this study four natural extracts from tea (TEA), grape (GRA), chestnut (CHE) and seaweed (SEA) with potential antioxidant activity were evaluated in pork patties. During 20 days of storage in modified atmosphere packs at 2°C, pH, colour, lipid oxidation and microbial spoilage parameters of raw minced porcine patties were examined and compared with a synthetic antioxidant (BHT) and control (CON) batch. Due to their higher polyphenol content, GRA and TEA extracts were the most effective antioxidants against lipid oxidation, also limiting colour deterioration. In addition, both natural extracts led to a decrease of total viable counts (TVC), lactic acid bacteria (LAB), Pseudomonas and psychotropic aerobic bacteria compared to the control. Among the four natural compounds tested, tea and grape extracts showed the most potential as alternatives to commercial antioxidants, for increasing the quality and extending the shelf-life of porcine patties.

Identification of the Major ACE-Inhibitory Peptides Produced by Enzymatic Hydrolysis of a Protein Concentrate from Cuttlefish Wastewater

The aim of this work was the purification and identification of the major angiotensin converting enzyme (ACE) inhibitory peptides produced by enzymatic hydrolysis of a protein concentrate recovered from a cuttlefish industrial manufacturing effluent. This process consisted on the ultrafiltration of cuttlefish softening wastewater, with a 10 kDa cut-off membrane, followed by the hydrolysis with alcalase of the retained fraction. Alcalase produced ACE inhibitors reaching the highest activity (IC50 = 76.8 ± 15.2 μg mL−1) after 8 h of proteolysis. Sequential ultrafiltration of the 8 h hydrolysate with molecular weight cut-off (MWCO) membranes of 10 and 1 kDa resulted in the increased activity of each permeate, with a final IC50 value of 58.4 ± 4.6 μg mL−1. Permeate containing peptides lower than 1 kDa was separated by reversed-phase high performance liquid chromatography (RP-HPLC). Four fractions (A–D) with potent ACE inhibitory activity were isolated and their main peptides identified using high performance liquid chromatography coupled to an electrospray ion trap Fourier transform ion cyclotron resonance-mass spectrometer (HPLC-ESI-IT-FTICR) followed by comparison with databases and de novo sequencing. The amino acid sequences of the identified peptides contained at least one hydrophobic and/or a proline together with positively charged residues in at least one of the three C-terminal positions. The IC50 values of the fractions ranged from 1.92 to 8.83 μg mL−1, however this study fails to identify which of these peptides are ultimately responsible for the potent antihypertensive activity of these fractions.

Cheese whey: A cost-effective alternative for hyaluronic acid production by Streptococcus zooepidemicus.

This study focuses on the optimisation of cheese whey formulated media for the production of hyaluronic acid (HA) by Streptococcus zooepidemicus. Culture media containing whey (W; 2.1g/L) or whey hydrolysate (WH; 2.4 g/L) gave the highest HA productions. Both W and WH produced high yields on protein consumed, suggesting cheese whey is a good nitrogen source for S. zooepidemicus production of HA. Polysaccharide concentrations of 4.0 g/L and 3.2g/L were produced in W and WH in a further scale-up to 5L bioreactors, confirming the suitability of the low-cost nitrogen source. Cheese whey culture media provided high molecular weight (>3000 kDa) HA products. This study revealed replacing the commercial peptone by the low-cost alternative could reduce HA production costs by up to a 70% compared to synthetic media.

Production of Hyaluronic Acid by Streptococcus zooepidemicus on Protein Substrates Obtained from Scyliorhinus canicula Discards

This work investigates the production of hyaluronic acid (H) by Streptococcus equi subsp. zooepidemicus in complex media formulated with peptones obtained from Scyliorhinus canicula viscera by-products. Initially, in batch cultures, the greatest productions were achieved using commercial media (3.03 g/L) followed by peptones from alcalase hydrolyzed viscera (2.32 g/L) and peptones from non-hydrolyzed viscera (2.26 g/L). An increase of between 12% and 15% was found in subsequent fed-batch cultures performed on waste peptones. Such organic nitrogen sources were shown to be an excellent low-cost substrate for microbial H, saving more than 50% of the nutrient costs.

Modelling the Biphasic Growth and Product Formation by Enterococcus faecium CECT 410 in Realkalized Fed-Batch Fermentations in Whey

The influence of initial pH on growth and nutrient (total sugars, nitrogen, and phosphorous) consumption by Enterococcus faecium CECT 410 was studied during batch cultures in whey. With these data, two realkalized fed-batch fermentations were developed using different feeding substrates. The shift from homolactic to mixed acid fermentation, the biphasic kinetics observed for cell growth and nitrogen consumption and the increase in the concentrations of biomass and products (lactic acid, acetic acid, ethanol, and butane-2,3-diol) were the most noteworthy observations of these cultures. Modelling the fed-batch growth of Ent. faecium with the Logistic and bi-Logistic models was not satisfactory. However, biomass production was best mathematically described with the use of a double Monod model, which was expressed in terms of biomass, product accumulation, and nitrogen utilization. Product formation was successfully modelled with a modified form of the Luedeking and Piret model developed in this study.

Production of Fish Protein Hydrolysates from Scyliorhinus canicula Discards with Antihypertensive and Antioxidant Activities by Enzymatic Hydrolysis and Mathematical Optimization Using Response Surface Methodology

Fish discards are of major concern in new EU policies. Alternatives for the management of the new biomass that has to be landed is compulsory. The production of bioactive compounds from fish protein hydrolysates (FPH) has been explored in recent years. However, the viability of Scyliorhinus canicula discards, which might account for up to 90–100% of captures in mixed trawler, gillnet, and longline industrial fisheries, to produce FPH from the muscle with bioactivities has still not been studied in terms of the optimization of the experimental conditions to enhance its production. The effect of pH and temperature on the hydrolysis of the S. canicula muscle was mediated by three commercial proteases using response surface methodology. Temperatures of 64.6 °C and 60.8 °C and pHs of 9.40 and 8.90 were established as the best hydrolysis conditions for Alcalase and Esperase, respectively. Optimization of the best conditions for the maximization of antihypertensive and antioxidant activities was performed. Higher Angiotensin-converting enzyme (ACE) activity was found with Esperase. The pH optimum and temperature optimum for antioxidants were 55 °C/pH8.0 for ABTS/DPPH-Esperase, 63.1 °C/pH9.0 for DPPH-Alcalase, and 55 °C/pH9.0 for ABTS-Alcalase. No hydrolysis was detected when using Protamex.

Optimization of Antimicrobial Combined Effect of Organic Acids and Temperature on Foodborne Salmonella and Escherichia coli in Cattle Feed by Response Surface Methodology

Despite the importance of feedstuffs and feed ingredients in the food chain, few studies have analyzed the effectiveness of usual methods for the microbial decontamination of feeds. This work aimed to study the combined effect of temperature and organic acids (formic or lactic) on the inactivation of 10 isolates of Salmonella enterica and Escherichia coli obtained from vegetable feed ingredients, including cereals and other processed materials, in cattle feed. A central composite design was used with acid concentrations ranging from 0 to 0.2% (vol/wt) and temperatures from 50°C to 70°C. Second-order models were obtained to predict the survival inhibitions. The results reported in the present study indicate that temperature affects the survivability more significantly than acid concentration, whose effect was isolate dependent. However, after 2 min of treatment, optimal microbial inhibitions were generally achieved at temperatures above 65°C, using 0.1% formic acid or 0.2% lactic acid for Salmonella isolates and around 0.1% lactic acid or 0.2% formic acid for E. coli isolates, respectively. This work provides a set of guidelines helpful to reduce microbial contamination of feeds and highlights the importance of feed-ingredient monitoring to reduce pathogen contamination risk during feed processing.

Pediocin SA-1: A selective bacteriocin for controlling Listeria monocytogenes in maize silages

In this study, we assessed the potential as silage additive of a bacteriocin produced by Pediococcus acidilactici Northern Regional Research Laboratory (NRRL) B-5627 (pediocin SA-1). Maize was inoculated either with a bacterial starter alone (I) or in combination with the bacteriocin (IP), and untreated silage served as control. We monitored the products of fermentation (ethanol, and lactic and acetic acids), the microbial population, and the presence of the indicator strain Listeria monocytogenes Colección Española de Cultivos Tipo (CECT) 4032 (1×10(5) cfu/g) after 1, 2, 5, 8, 16, and 30d of ensiling. Our results indicated antilisterial activity of the bacteriocin, anticipating the disappearance of L. monocytogenes in IP compared with I and control silages. The PCR-denaturing gradient gel electrophoresis analysis revealed the addition of the bacteriocin did not affect the bacterial communities of the spontaneous fermentation, and the inoculant-containing bacteria (Lactobacillus plantarum, Lactobacillus buchneri, and Enterococcus faecium) were found in addition to the bacterial communities of untreated maize silages in I and IP silages. Both treatments increased the concentration of antimicrobial compounds (acetic acid, ethanol, and 1,2-propanodiol) and led to lower residual sugar contents compared with the control, which would provide enhanced aerobic stability. The fact that the identified species L. plantarum, L. buchneri, and E. faecium produce some of these inhibitory compounds, together with their persistence throughout the 30d of fermentation, suggest these bacteria could actively participate in the ensiling process. According to these results, pediocin SA-1 could be used as an additive to control the presence of L. monocytogenes in maize silages selectively, while improving their fermentative quality and eventually their aerobic stability.

Chapter 28 – Smart Nanohydrogels for Controlled Release of Food Preservatives

The food industry faces great challenges. Consumers, for example, prefer fresh or minimally processed food, while market globalization lengthens storage and distribution times. Fortunately, innovative packaging strategies are emerging to ensure food safety and to minimize losses caused by spoilage microorganisms. This chapter reviews the research on antimicrobial active and smart packaging: its basis, categories of packaging devices and antimicrobial compounds used, mode of incorporation, and commercial applications. The chapter also addresses the role of nanotechnology in food packaging, closely examining the use of smart environmentally sensitive nanohydrogels in overcoming the limitations of the classical active packaging systems. Specifically, poly (N-isopropylacrylamide) nanohydrogels are key for their physicochemical properties, releasing mechanisms, and antimicrobial effectiveness. The chapter provides examples of their role in controlling food spoilage fungi and discusses future prospects.