Widely distributed lysogeny in probiotic lactobacilli represents a potentially high risk for the fermentative dairy industry

Prophages account for most of the genetic diversity among strains of a given bacterial species, and represent a latent source for the generation of virulent phages. In this work, a set of 30 commercial, collection and dairy-isolated Lactobacillus casei group strains were used. A species-specific PCR assay allowed a reclassification, mainly of strains previously considered Lactobacillus casei, into either Lactobacillus paracasei or Lactobacillus rhamnosus. All the strains were induced with mitomycin C, allowing direct recovering of phage DNA in 25 cases, which corroborates the widely occurrence of lysogeny on Lactobacillus genomes, including probiotic strains of Lactobacillus casei group. Ten out of 11 commercial strains studied contained prophages, evidencing the potential risks of their use at industrial scale. Strains were also induced by treatment with different concentrations of hydrogen peroxide but, however, this agent was not able to evidence a prophage release for any of the strains tested. According to a RAPD-PCR fingerprinting with M13, 1254 and G1 primers, most of the commercial strains presented a high degree of homology and, regarding BglII- and BamHI-restriction profiles of phage DNA, six of them harboured the same prophage. Surprisingly, both Lactobacillus paracasei ATCC 27092 and Lactobacillus paracasei ATCC 27139 shared a second prophage with both an INLAIN collection and a commercial Lactobacillus paracasei strains, whereas two collection strains shared a third one. On the other hand, mitomycin C-inducible prophages were detected only on about a half of the strains isolated from dairy products, which had (with only one exception) from moderate to high correlation coefficients according to RAPD-PCR fingerprinting. After induction, supernatants were filtered and tested against nine Lactobacillus strains of the set sensitive to previously assayed virulent phages, allowing isolation of two new virulent phages: ф iLp1308 and ф iLp84. Both phages were able to lyse all but one strains sensitive to previously assayed phage MLC-A.     

Headspace solid‐phase microextraction analysis of volatile compounds from spice essential oils in dry flavourings

The composition of volatile compounds in nine spices was analysed by means of headspace solid‐phase microextraction (HS‐SPME) from dry flavourings of the spice essential oils, to devise an effective method to determine the content of the essential oils. The HS‐SPME was combined with GC‐MS and GC‐FID for identification and quantification of the volatile compounds. The optimisation of the extraction conditions was carried out by response surface methodology. The evaluation of the analytical parameters indicated that the method had a high precision (coefficient of variations lower than 3%), it was linear (response factors were smaller than 5%) and recoveries were higher than 89%. In the ruggedness test, sample quantity and pre‐extraction time were the factors that stood out as the causing of the greater effects on the analytical results.     

Polyphenol oxidase activity,phenolic acid composition and browning in cashew apple (Anacardium occidentale, L.) after processing

This study describes the extraction and characterisation of cashew apple polyphenol oxidase (PPO) and the effect of wounding on cashew apple phenolic acid composition, PPO activity and fruit browning. Purification factor was 59 at 95% (NH₄)₂SO₄ saturation. For PPO activity, the optimal substrate was catechol and the optimum pH was 6.5. PPO K_m and V_max values were 18.8 mM and 13.6 U min⁻¹ ml⁻¹, respectively. Ascorbic acid, citric acid, sodium sulphite and sodium metabisulphite decreased PPO activity, while sodium chloride increased PPO activity. Wounding at 2 °C and 27 °C for 24 h increased PPO activity but storage at 40 °C reduced PPO activity. Gallic acid, protocatechuic acid and cinnamic acid (free and conjugate) were identified in cashew apple juice. Cutting and subsequent storage at 40 °C hydrolysed cinnamic acid. 5-Hydroxymethylfurfural content in cashew apple juice increased after injury and storage at higher temperatures, indicating non-enzymatic browning.     

Caffeine (1, 3, 7-trimethylxanthine) in Foods: A Comprehensive Review on Consumption,Functionality, Safety,and Regulatory Matters

ABSTRACT: Caffeine ranks as one of the top most commonly consumed dietary ingredients throughout the world. It is naturally found in coffee beans, cacao beans, kola nuts, guarana berries, and tea leaves including yerba mate. The total daily intake, as well as the major source of caffeine varies globally; however, coffee and tea are the 2 most prominent sources. Soft drinks are also a common source of caffeine as well as energy drinks, a category of functional beverages. Moderate caffeine consumption is considered safe and its use as a food ingredient has been approved, within certain limits, by numerous regulatory agencies around the world. Performance benefits attributed to caffeine include physical endurance, reduction of fatigue, and enhancing mental alertness and concentration. Caffeine has also been recently linked to weight loss and consequent reduction of the overall risks for developing the metabolic syndrome. However, the caloric contribution of caffeine-sweetened beverages needs to be considered in the overall energy balance. Despite all these benefits the potential negative effects of excessive caffeine intake should also be considered, particularly in children and pregnant women.     

The role of K+ and H+ transport systems during glucose‐ and H2O2‐induced cell death in Saccharomyces cerevisiae

Glucose, in the absence of additional nutrients, induces programmed cell death in yeast. This phenomenon is independent of yeast metacaspase (Mca1/Yca1) and of calcineurin, requires ROS production and it is concomitant with loss of cellular K⁺ and vacuolar collapse. K⁺ is a key nutrient protecting the cells and this effect depends on the Trk1 uptake system and is associated with reduced ROS production. Mutants with decreased activity of plasma membrane H⁺‐ATPase are more tolerant to glucose‐induced cell death and exhibit less ROS production. A triple mutant ena1‐4 tok1 nha1, devoid of K⁺ efflux systems, is more tolerant to both glucose‐ and H₂O₂‐induced cell death. We hypothesize that ROS production, activated by glucose and H⁺‐ATPase and inhibited by K⁺ uptake, triggers leakage of K⁺, a process favoured by K⁺ efflux systems. Loss of cytosolic K⁺ probably causes osmotic lysis of vacuoles. The nature of the ROS‐producing system sensitive to K⁺ and H⁺ transport is unknown. Copyright © 2010 John Wiley & Sons, Ltd.     

Simulation and analysis of a sugarcane juice evaporation system

The production of sugar and alcohol is the main objective of the sugarcane processing industry. The evaporation of sugarcane juice has a high energetic cost and is usually performed in multiple-effect evaporators. The loss of performance during operation due to fouling makes the process more complex. In this study, modeling, simulation, validation, and analysis were performed for a sugarcane juice industrial evaporation system (IES) composed of a falling film evaporator followed by three short vertical-tube evaporators arranged in parallel. The IES model was developed using a commercial process simulator and validated with data from the plant. The IES had marked performance losses in the first 14 days of operation, mainly due to fouling in the first effect, with a 30% decrease in the evaporation rate.     

Volatile compounds on the surface and within Iberian dry-cured loin

The profile of volatile compounds from external and internal areas of Iberian dry-cured loin was studied by headspace solid-phase microextraction. Higher levels of 16 volatile compounds (hexane, decane, 3-methylbutanal, several sulphur compounds and some aromatic hydrocarbons) were detected in the outer part of the loins. These differences could be owing to several factors, such as greater exposure to oxygen and dehydration conditions in the surface of the product, which favour oxidation reactions and Strecker degradation of amino acids. Moreover, the addition of spices on the surface and the proximity of a mould layer growing on the surface of the product could also contribute to the higher levels of these compounds on the external layer. However, other sulphur compounds coming from spices showed similar levels on the surface and within the loins. This different behaviour could be a consequence of different diffusion rates depending on the features of the compound and the matrix.     

Effect of slurry ice on chemical changes related to quality loss during European Hake (Merluccius merluccius) chilled storage

Slurry ice is a biphasic system consisting of small spherical ice crystals surrounded by seawater at subzero temperature. Its employment was evaluated in the present work as a new chilled storage method for whole European Hake (Merluccius merluccius) and compared to traditional flake icing. Different chemical analyses (nucleotide degradation, formaldehyde formation, lipid hydrolysis and oxidation, interaction compound formation, and electrophoretic protein profiles) related to quality changes were analysed and compared to sensory evaluation. An inhibitory effect on quality loss mechanisms was observed for the slurry ice treatment, according to nucleotide degradation (K value), free formaldehyde content, browning development, and sarcoplasmic protein profiles. No differences in lipid hydrolysis and oxidation could be distinguished by a comparison between both icing conditions. The sensory analysis showed a higher shelf-life time for slurry icing than flake icing (12 days and 5 days, respectively). Results confirm the practical advantages of using slurry ice as a chilled storage method.     

Phage types, virulence genes and PFGE profiles of Shiga toxin-producing Escherichia coli O157:H7 isolated from raw beef, soft cheese and vegetables in Lima (Peru)

The present study was conducted in Lima Metropolitana to evaluate the prevalence of Shiga toxin-producing Escherichia coli (STEC) O157:H7 in raw beef, raw ground beef, soft cheese and fresh vegetables, sampled at different markets in the city. Between October 2000 and February 2001, 407 food samples were collected from different markets in the 42 districts of Lima Metropolitana. Samples were assayed for E. coli O157 by selective enrichment in modified Tryptic Soy Broth containing novobiocin, followed by immunomagnetic separation (IMS) and plating onto sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Fifty (12.3%) of 407 food samples resulted positive for E. coli O157 isolation (23 of 102 ground beef; 15 of 102 beef meat; eight of 102 soft cheese and four of 101 fresh vegetables). Thirty-five E. coli O157 isolates were further analysed for the presence of virulence genes. All 35 were positive by PCR for O157 rfbE, fliCh7, eae-gamma1 and ehxA genes. In addition, genes encoding Shiga toxins were detected in 33 of 35 isolates, five isolates (14%) encoded stx(1), stx(2), and 28 (80%) stx2 only. The isolates were of seven different phage types (PT4, PT8, PT14, PT21, PT34, PT54, and PT87) with three phage types accounting for 80% of isolates: PT4 (15 isolates), PT14 (8 isolates), and PT21 (5 isolates). Interestingly, the majority (31 of 35; 89%) of E. coli O157:H7 isolates characterized in this study belonged mainly to the phage types previously found in STEC O157:H7 strains associated with severe human disease in Europe and Canada. Pulsed-field gel electrophoresis (PFGE) of 32 isolates revealed 14 XbaI-PFGE groups (I to XIV) of similarity >85%, with 23 (72%) isolates grouped in five clusters. Some isolates from different districts presented a high clonal relatedness. Thus, PFGE group VIII clustered eleven strains from nine different districts. The broad range of PFGE subtypes found in this study demonstrates the natural occurrence of many genetic variants among STEC O157:H7 spread in Lima.