Transepithelial transport across Caco-2 cell monolayers of antihypertensive egg-derived peptides. PepT1-mediated flux of Tyr-Pro-Ile

This paper examines the in vitro transepithelial transport of antihypertensive peptides derived from egg proteins using Caco-2 cell monolayers. Ovokinin (FRADHPFL) was absorbed intact through the Caco-2 cell epithelium, although it was also susceptible to the action of brush-border aminopeptidases that yielded shorter fragments prior to their transport. The tripeptide YPI was resistant to cellular peptidases and transported through the monolayer, what suggests that the reduction in systemic blood pressure caused by this peptide may be mediated by effects at tissue level. Its pathway for transepithelial absorption was examined using inhibitors of the different mechanisms for oligopeptide transport in the intestinal tract. The main route involved in the transepithelial flux of YPI is probably the peptide H(+)-coupled transporter PepT1. These results highlight the potential of antihypertensive peptides to be used in the formulation of functional foods.     

Effect of the packaging method and the storage time on lipid oxidation and colour stability on dry fermented sausage salchichón manufactured with raw material with a high level of mono and polyunsaturated fatty acids

The effects of fatty acid composition, two packaging methods (vacuum and 20% CO₂/80% N₂) and storage under refrigeration for 210 days were evaluated on a dry fermented sausage (salchichón), manufactured with raw material enriched in monounsaturated or polyunsaturated fatty acids. Fatty acid composition was determined on sausage mixtures and on ripened sausages and lipid oxidation and colour stability was determined on ripened sausage at different times during storage. The modification of fatty acid composition of the sausages raised the nutritional quality, slightly affecting the colour properties. Dry fermented sausages enriched in polyunsaturated and monounsaturated fatty acids presented higher lipid oxidation values than the control ones. Both packaging methods (vacuum and 20% CO₂/80% N₂) during 210 days of chilled storage had minor effects on the colour and the lipid oxidation stability.     

Effect of ractopamine level, gender, and duration of ractopamine on belly and bacon quality traits

The objective was to determine belly and bacon quality traits in pigs fed ractopamine (RAC) for various durations during finishing. A 2 × 3 × 2 factorial arrangement was used with barrows and gilts, fed RAC levels of 0.0, 5.0, or 7.4 ppm, for 21 or 28 d prior to harvest. Bellies were fabricated and measured for length, thickness, firmness, and processing yields. Once processed, 1.27 cm slices were removed at 25%, 50%, and 75% the distance from the blade end, packaged and digitally imaged using a Chem1 Genius² Bio Imaging System. Total slice area (TA), total slice length (TL), secondary lean length (SL), secondary lean area (SA), and percent lean area (TA – all lean components = LA) were determined by tracing images in Adobe Photoshop Elements. A composite sample from the three slices was used for proximate analysis to determine moisture and fat composition for each belly. Feeding RAC increased belly yield, TA, TL, SA, and LA (P < 0.05), but did not alter moisture or fat composition (P > 0.05). Gilts had decreased firmness and higher pump uptakes compared to barrows (P < 0.05). Additionally gilts had increased TL, SL, and LA with lower fat and higher moisture content (P < 0.05). RAC feeding duration had no significant effect on belly or bacon quality traits (P > 0.05), furthermore, no interactions were found to be significant (P > 0.05). RAC administration during finishing resulted in improved belly and bacon yields with no negative effects on the quality traits evaluated.     

Microbial, physical-chemical and sensory spoilage during the refrigerated storage of cooked pork loin processed by the sous vide method

The aim was to study spoilage during the refrigerated storage of cooked pork loin processed by the sous vide method. Samples were packaged under vacuum into polyamide-polypropylene pouches, cooked at an oven temperature/time of 70 °C/12 h, chilled at 3 °C and stored at 2 °C for 0, 5 or 10 weeks. Microbial (psychrotrophs, lactic acid bacteria, Enterobacteriaceae, moulds and yeasts), physical–chemical (pH, water activity, TBARS, acidity, L^*a^*b^* colour, texture profile analysis and shear force) and sensory (appearance, odour, flavour, texture and acceptance) parameters were determined. The results showed that sensory spoilage preceded microbiological spoilage of sous vide pork loin. Counts bellow 1 log cfu/g of psychrotrophs, anaerobic psychrotrophs, Enterobacteriaceae and lactic acid bacteria were detected in any control week, while moderate counts (2–3 log cfu/g) of moulds and yeasts were found. Minor changes in water activity, lipid oxidation, CIELab colour, hardness, cohesiveness or gumminess were associated with spoilage of pork loin, only decreases of lactic acid, springiness and shear force were observed. The pork loin was unacceptable after 10 weeks. This loss of acceptance was mainly due to the deterioration of meaty flavour and odour, although the loss of appearance, juiciness and firmness also contributed. Moderate warmed-over and rancidity were detected. The sensory analysis was the most effective method for determining the shelf life of the sous vide pork-based dishes.     

European interlaboratory trial regarding the official microscopic method for the detection of the presence of animal constituents in feedstuffs

The bovine spongiform encephalopathy epidemic is thought to have occurred as a consequence of feeding prion-infected material to cattle. To avoid the risk of bovine spongiform encephalopathy diffusion, the European Commission (Directive 2003/126/EC) established an official method to detect the presence of animal-derived constituents in feedstuffs, using microscopic examination. This method allows easy identification of bone fragments among other animal constituents. The analysis is based on morphological conformation of the fragments and their characterization (mainly of the shape of lacunae) to discriminate among mammalian, poultry, and fish tissues. The aim of this study was to assess the performances of nine European laboratories through a ring trial of the official microscopic method, and to calculate accuracy and reproducibility of the method. In general the reproducibility of the microscopic method was very good (kappa overall = 0.83), with a high sensitivity for all laboratories. Concerning the analysis on the different animal-derived constituents, the results show values of sensitivity with large variability between fish and poultry or mammal. It was generally more difficult to discriminate between mammalian and poultry tissues than fish tissue.     

Antimicrobial activity of sub- and supercritical CO2 extracts of the green alga Dunaliella salina

The objective of this research was to evaluate the antimicrobial activity of carbon dioxide extracts of the unicellular biflagellated green alga Dunaliella salina against Escherichia coli, Staphylococcus aureus, Candida albicans, and Aspergillus niger. The effects of different extraction pressures ranging from 185 to 442 bar and extraction temperatures ranging from 9.8 to 45.2 degrees C on the extracts' composition and consequently on their antimicrobial activities were investigated. The extracts were analyzed by gas chromatography-mass spectrometry in order to identify the compounds responsible for the antimicrobial activity detected. Fourteen different volatile compounds and several fatty acids were identified. The highest antimicrobial activity was obtained using 314 bar and 9.8 degrees C. Under these conditions, the presence of an indolic derivative that had never been reported in D. salina was detected in the extract, together with polyunsaturated fatty acids and compounds related to carotene metabolism, such as beta-ionone and neophytadiene, with known antimicrobial activity.     

Activity against Listeria monocytogenes of human milk during lactation. A preliminary study

Human milk samples from three healthy donors were investigated in order to evaluate the antibacterial activity during lactation against Escherichia coli ATCC 25922 and Listeria monocytogenes. The concentration of the main human-milk antimicrobial proteins (lactoferrin (LF), lysozyme (LZ) and secretory immunoglobulin A (sIgA)) was determined by ELISA. Results showed that human milk exhibited antibacterial activity against List. monocytogenes, although it was weakly active against Esch. coli ATCC 25922. The observed antilisterial activity was positively correlated with LZ concentration. In addition, the effect of gastrointestinal proteases, at different pH conditions, that prevail in the stomach of infants (pH 2.0-6.5), on antilisterial activity and protein degradation was evaluated. Hydrolysis with pepsin at pH 4.0-6.5, followed by treatment with pancreatic enzymes, resulted in a decreased hydrolysis of LZ, LF and sIgA and an enhanced antibacterial activity against List. monocytogenes. It is suggested that partial degradation of certain milk proteins at the gastrointestinal level may produce peptides that could act synergistically with the remnant intact proteins.     

Characterization of homofermentative lactobacilli isolated from kefir grains: potential use as probiotic

Considering that several health promoting properties are associated with kefir consumption and a reliable probiotic product requires a complete identification of the bacterial species, the present work evaluates several proved markers of probiotic potential of eleven isolates of homofermentative lactobacilli isolated from kefir grains and molecular identification and genotypic diversity. Using restriction analysis of amplified ribosomal DNA (ARDRA) and analysis of the 16S-23S rRNA internal spacer region we confirmed that all homofermentative lactobacilli belong to the species Lactobacillus plantarum. RAPD-PCR analysis allowed the discrimination of lactobacilli in five clusters. All isolates exhibited high resistance to bile salt. High survival after one hour of exposure to pH 2.5 was observed in Lb. plantarum CIDCA 8313, 83210, 8327 and 8338. All isolates were hydrophilic and non autoaggregative. Isolate CIDCA 8337 showed the highest percentage of adhesion among strains. All tested lactobacilli had strong inhibitory power against Salmonella typhimurium and Escherichia coli. Seven out of eleven isolates showed inhibition against Sal. enterica and five isolates were effective against Sal. gallinarum. Only CIDCA 8323 and CIDCA 8327 were able to inhibit Sal. sonnei. We did not find any correlation between the five clusters based on RAPD-PCR and the probiotic properties, suggesting that these isolates have unique characteristics.     

Antimicrobial activity of nisin, reuterin, and the lactoperoxidase system on Listeria monocytogenes and Staphylococcus aureus in cuajada, a semisolid dairy product manufactured in Spain

The inhibitory activity of nisin (N), reuterin (R), and the lactoperoxidase system (LPS), added individually or in combination, against Listeria monocytogenes and Staphylococcus aureus was investigated in “cuajada” (curdled milk), a semisolid dairy product manufactured in Spain. Cuajada was manufactured from UHT skim milk separately inoculated with L. monocytogenes and Staph. aureus, each at approximately 4 log cfu/mL, and held under conditions of temperature abuse (10°C). On d 3, a synergistic bactericidal activity was observed for the combinations of biopreservatives assayed, with L. monocytogenes counts of only 0.30 log cfu/mL in cuajada made with N + R + LPS vs. 8.31 log cfu/mL in control cuajada. After 12 d, L. monocytogenes could not be detected in cuajada made with added N + LPS or N + R + LPS. Staphylococcus aureus was more resistant than L. monocytogenes to biopreservatives added individually. On d 3, the synergistic effect of the 3 biopreservatives against Staph. aureus resulted in counts of 3.03 log cfu/mL in cuajada made with N + R + LPS vs. 6.40 in control cuajada. After 12 d, Staph. aureus counts were 2.61 log cfu/mL in cuajada made with N + R + LPS, whereas they ranged from 6.11 to 7.70 log cfu/mL in control cuajada and in cuajada made with other combinations of biopreservatives. The most pronounced decrease in pathogen counts was achieved by the triple combination N + R + LPS, which acted synergistically on the inactivation of L. monocytogenes and Staph. aureus in cuajada over 12 d at 10°C. The treatment combining these 3 natural biopreservatives at low concentrations, within the hurdle concept of food preservation, might be a useful tool to control the growth of pathogenic microorganisms in nonacidified dairy products.     

Evaluation of exopolysaccharide production by Leuconostoc mesenteroides strains isolated from wine

ABSTRACT:  Exopolysaccharide (EPS)-producing lactic acid bacteria are responsible for the alteration of wine and other fermented beverages. The potential to produce EPS was investigated for Leuconostoc mesenteroides strains isolated from Spanish grape must and wine. Most strains were able to produce EPS from sucrose containing media. Based on their EPS-producing phenotype and on their EPS monosaccharide composition, the L. mesenteroides strains analyzed could be arranged in 2 groups. One group comprises mucoid strains producing a glucan polymer, and the other group includes strains producing a fructan polymer. The presence of a glucosyltransferase encoding gene in the glucan producing L. mesenteroides strains was assayed by PCR. Two primer sets, PF1-PF8 and GTFF-GTFR, were used to amplify internal fragment of known glucosyltransferase genes. None of the glucan-producing strains gave a positive amplicon by the primer sets used. Therefore, new tools need to be developed to broaden the range of potentially spoiling agents detected by PCR in fermented beverages.