Exoglucanase‐encoding genes from three Wickerhamomyces anomalus killer strains isolated from olive brine

Wickerhamomyces anomalus killer strains are important for fighting pathogenic yeasts and for controlling harmful yeasts and bacteria in the food industry. Targeted disruption of key genes in β‐glucan synthesis of a sensitive Saccharomyces cerevisiae strain conferred resistance to the toxins of W. anomalus strains BS91, BCA15 and BCU24 isolated from olive brine. Competitive inhibition of the killing activities by laminarin and pustulan refer to β‐1,3‐ and β‐1,6‐glucans as the main primary toxin targets. The extracellular exoglucanase‐encoding genes WaEXG1 and WaEXG2 from the three strains were sequenced and were found to display noticeable similarities to those from known potent W. anomalus killer strains. Accession Nos for WaEXG1 genes for the strains in brackets are JQ734563 (BS91), JQ734564 (BCA15) and JQ734565 (BCU24); for WaEXG2 genes JQ734566 (BS91), JQ734567 (BCA15) and JQ734568 (BCU24), respectively. Copyright © 2012 John Wiley & Sons, Ltd.     

Effect of natural and synthetic antioxidants incorporation on the gas permeation properties of poly(lactic acid) films

Gas permeation properties (permeability, diffusivity and solubility coefficients) were determined for carbon dioxide and oxygen in poly(lactic acid) (PLA) films enriched with 0, 2, 4 and 10 wt.% of different antioxidants, at three temperatures, 284, 293 and 303 K, using a time-lag apparatus. Three antioxidants, a natural, α-tocopherol (AT), and two synthetic, butylated hydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ), were tested. DSC results show that the polymer glass transition temperature slightly decreases with the increase of the antioxidant content. The crystallinity degree of PLA films decreases with the addition of BHT and TBHQ, whereas the incorporation of AT increases the crystallinity of PLA films. The permeability towards water vapour, at 299 K and 45% of relative humidity, and surface energy show a decreased in the wettability of the prepared materials with the increase of the antioxidants content. The incapacity to measure gas permeation in PLA films with 10 wt.% of AT and BHT incorporated was due to phase separation, proved by SEM images. The CO₂ and O₂ permeation results show that PLA barrier properties can be improved by the incorporation of antioxidants but are strongly dependent on the amount and structure of the antioxidant added.     

Effect of cooking and germination on phenolic composition and biological properties of dark beans (Phaseolus vulgaris L.)

Legumes are the base´s diet in several countries. They hold a high nutritional value, but other properties related to human health are nowadays being studied. The aim of this work was to study the influence of processes (boiling or germination) on the phenolic composition of dark beans (Phaseolus vulgaris L. c.v. Tolosana) and their effect on their antioxidant, neuroprotective and anticancer ability. Phenolic composition of raw and processed dark beans was analysed by HPLC-PAD and HPLC–ESI/MS. The antioxidant activity was evaluated by ORAC. Astrocytes cultures (U-373) have been used to test their neuroprotective effect. Anticancer activities were evaluated on three different cell lines (renal adenocarcinoma (TK-10), breast adenocarcinoma (MCF-7) and melanoma (UACC-62)) by sulphorhodamine B method. Qualitative and quantitative differences in phenolic composition have been observed between raw and processed dark beans that influence the antioxidant activity, mainly for germinated samples which show a decrease of antioxidant capacity. Although every assayed extracts decreased reactive oxygen species release and exhibited cytotoxicity activities on cancer cell lines, raw beans proved to be the most active in neuroprotective and antitumoral effects; this sample is especially rich in phenolic compounds, mainly anthocyanins. This study further demonstrated that phenolic composition of dark beans is related with cooking process and so with their neuroprotective and anticancer activity; cooking of dark beans improves their digestion and absorption at intestinal level, while maintaining its protective ability on oxidative process at cellular level.     

Experimental approaches for the estimation of uncertainty in analysis of trace inorganic contaminants in foodstuffs by ICP-MS

Total Diet Studies to estimate dietary exposure to food contaminants need to evaluate laboratory measurements data variance. In this process it is critical that data from analytical methods are reliable to correctly scrutinize and compare values over time and between countries. In Europe it is widely recognized that the evaluation of measurement uncertainty is an important parameter when assessing the sources of analytical data variability. Two approaches are considered to estimate uncertainty in analytical measurement. Arsenic, Lead, Chromium and Cadmium, content in several food matrix determined by Inductively Coupled Mass Spectrometry (ICP-MS) microwave digestion assisted, are used as examples. The aim of the present research work is to compare both approaches accepted by Eurolab and GUM: Mathematical modeling to assess uncertainty components based on a classical model (bottom up) and an empirical method (top down), based on either experimental data obtained from a single laboratory validation data or inter-laboratory data from Proficiency Testing schemes. Relative expanded uncertainty calculated by both approaches agree when U (%) <20%. These values are concordant with RSD_R reported in collaborative studies of EN 15763 (2009), which were assumed as target uncertainty. The top down approach described is simple and easy to use when compared with the mathematical modeling approach providing considerable benefits to those who assess data produced by several laboratories.     

Changes on indigenous microbiota,colour, bioactive compounds and antioxidant activity of pasteurised pomegranate juice

Juices prepared from arils of ‘Mollar’ pomegranates were analysed for naturally occurring microorganisms, CIE Lab colour parameters, total phenols, anthocyanins and punicalagins, ellagic acid content and antioxidant capacity before and after low-, mild- and high-temperature pasteurisations (LTPs, MTPs and HTPs): 65, 80 and 90 °C for 30 or 60 s. Mean aerobic plate count (APC), yeast and mold count (YMC), and lactic acid bacteria (LAB) for fresh juices were 5.7, 5.36 and 4.0 log CFU/mL, respectively. MTPs and HTPs were sufficiently effective to decrease APCs to nil or negligible levels. An increase in CIE a values and decrease in CIE b values were the characteristic colour changes in heat-treated juices. The effect of pasteurisations showed that total phenols, punicalagins and ellagic acid were not much affected by thermal processing. Total anthocyanin content and antioxidant capacity were substantially and significantly influenced by the heat treatment applied. A linear relationship was observed between Trolox equivalent antioxidant capacity (TEAC) values and total anthocyanins, suggesting that they contributed strongly to the antioxidant capacity of pomegranate juice.     

Effect of different temperature–time combinations on physicochemical,microbiological, textural and structural features of sous-vide cooked lamb loins

Lamb loins were subjected to sous-vide cooking at different combinations of temperature (60, 70, and 80 °C) and time (6, 12, and 24 h). Different physicochemical, histological and structural parameters were studied. Increasing cooking temperatures led to higher weight losses and lower moisture contents, whereas the effect of cooking time on these variables was limited. Samples cooked at 60 °C showed the highest lightness and redness, while increasing cooking temperature and cooking time produced higher yellowness values. Most textural variables in a texture profile analysis showed a marked interaction between cooking temperature and time. Samples cooked for 24 h showed significantly lower values for most of the studied textural parameters for all the temperatures considered. Connective tissue granulation at 60 °C and gelation at 70 °C were observed in the SEM micrographs. The sous-vide cooking of lamb loins dramatically reduced microbial population even with the less intense heat treatment studied (60 °C–6 h).     

A SYBR Green real-time PCR assay to detect and quantify pork meat in processed poultry meat products

Species identification in meat products has grown in interest in recent years since these foodstuffs are susceptible targets for fraudulent labelling. In this work, a real-time PCR approach based on SYBR Green dye was proposed for the quantitative detection of pork meat in processed meat products. For the development of the method, binary meat mixtures containing known amounts of pork meat in poultry meat were used to obtain a normalised calibration model from 0.1 to 25% with high linear correlation and PCR efficiency. The method revealed high specificity by melting curve analysis, being successfully validated through its application to blind meat mixtures, which confirmed its adequacy for pork meat determination. The fully applicability of the method was further demonstrated in commercial meat products, allowing verification of labelling compliance and identification of meat species in processed foods.     

Effect of freezing/thawing conditions and long‐term frozen storage on the quality of mashed potatoes

The effects of freezing temperature (−80, −40 or −24 °C) and thawing mode (microwave or overnight at 4 °C) on quality parameters of mashed potatoes made from tubers (cv Kennebec) and from potato flakes were examined, as was the effect of long‐term frozen storage on the quality of mashed potatoes. Mashed potatoes were tested for texture profile analysis (TPA) and cone penetration, oscillatory and steady rheometry, colour, dry matter, Brix and sensory analyses. In natural mashed potatoes, TPA hardness and oscillatory parameters showed that processing resulted in a softer product than the fresh control. The parameters were lower in the samples thawed at 4 °C than in those thawed by microwave at all the freezing temperatures used, which may be ascribed to gelatinisation of the starch released from damaged cells. Differences from the freshly prepared product decreased when the samples were frozen at −80 °C and thawed by microwave. No difference was found in sensory acceptability between samples frozen at −80 and −40 °C, which probably reflects the panellists' mixed preferences for air‐thawed versus microwave‐thawed samples. Increasing the time in frozen storage led to a natural mash with a firmer texture, higher L*/b* value and Brix; nonetheless, panellists found the samples at 0, 3 and 12 months of frozen storage equally acceptable. In commercial mash, penetration and oscillatory parameters showed that processing made for a firmer product than the fresh control, probably owing to retrogradation of gelatinised starch. Thawing mode had a significant effect on parameters, which were lower in the samples thawed at 4 °C. The structure and quality of commercial mash was more detrimentally affected by freezing and, therefore, we would not recommend either freezing or frozen storage of this mashed potato in the used conditions. Natural mash made from Kennebec potatoes should be frozen quickly and thawed by microwave in the conditions described to obtain a product more similar to that freshly made. If the samples are frozen by air blasting at −40 °C, the product can withstand frozen storage for one year. Copyright © 2005 Society of Chemical Industry     

Microbiological hazards involved in fresh‐cut lettuce processing

BACKGROUND: The increasing consumption of produce all over the world has resulted in increasing concern by the regulatory agencies with respect to the level of safety performed by the processors. The objective of the present study was to investigate the hazards involved in the various steps of fresh‐cut lettuce processing (reception/selection of raw material, washing, rinsing, sanitisation and final product) by means of microbiological analyses of microbial groups used as indicators of hygienic conditions and of pathogens. RESULTS: High microbial loads of mesophilic and psychrotrophic bacteria and Pseudomonas spp. were found in the ram reception (～6 log colony‐forming units (CFU) g^?1), which were reduced by a single logarithmic cycle for the last two microbial groups after the sanitisation step (P < 0.05), the latter being ineffective against the first microbial group (P > 0.05). Lower counts of yeasts and moulds, total coliforms (35 °C) and faecal coliforms (44 °C) were observed in the initial step (3.49–4.53 log CFU g^?1, 0.65–1.55 log most probable number (MPN) g^?1 and 0.50–0.90 log MPN g^?1 respectively), these values increasing significantly after the sanitisation step for yeasts and moulds (～5 log CFU g^?1) but remaining unaltered for coliforms (P > 0.05). Salmonella spp. were not found in any of the experiments carried out, while the presence of Escherichia coli was observed in the final product. CONCLUSIONS : Practices compromising the hygienic quality of the final product during commercial storage were observed and corrective measures suggested. To the best of the authors' knowledge, these are the first data on microbiological safety in Brazilian fresh‐cut processing plants. Copyright © 2008 Society of Chemical Industry     

Validation of the performance of a GMO multiplex screening assay based on microarray detection

A new screening method for the detection and identification of GMO, based on the use of multiplex PCR followed by microarray, has been developed and is presented. The technology is based on the identification of quite ubiquitous GMO genetic target elements first amplified by PCR, followed by direct hybridisation of the amplicons on a predefined microarray (DualChip^® GMO, Eppendorf, Germany). The validation was performed within the framework of a European project (Co-Extra, contract no 007158) and in collaboration with 12 laboratories specialised in GMO detection. The present study reports the strategy and the results of an ISO complying validation of the method carried out through an inter-laboratory study. Sets of blind samples were provided consisting of DNA reference materials covering all the elements detectable by specific probes present on the array. The GMO concentrations varied from 1% down to 0.045%. In addition, a mixture of two GMO events (0.1% RRS diluted in 100% TOPAS19/2) was incorporated in the study to test the robustness of the assay in extreme conditions. Data were processed according to ISO 5725 standard. The method was evaluated with predefined performance criteria with respect to the EC CRL method acceptance criteria. The overall method performance met the acceptance criteria; in particular, the results showed that the method is suitable for the detection of the different target elements at 0.1% concentration of GMO with a 95% accuracy rate. This collaborative trial showed that the method can be considered as fit for the purpose of screening with respect to its intra- and inter-laboratory accuracy. The results demonstrated the validity of combining multiplex PCR with array detection as provided by the DualChip^® GMO (Eppendorf, Germany) for the screening of GMO. The results showed that the technology is robust, practical and suitable as a screening tool.