Photobleaching with phloxine B sensitizer to reduce food matrix interference for detection of Escherichia coli serotype O157:H7 in fresh spinach by flow cytometry

A flow cytometric method (RAPID-B™) with detection sensitivity of one viable cell of Escherichia coli serotype O157:H7 in fresh spinach (Spinacia oleracea) was developed and evaluated. The major impediment to achieving this performance was mistaking autofluorescing spinach particles for tagged target cells. Following a 5 h non-selective enrichment, artificially inoculated samples were photobleached, using phloxine B as a photosensitizer. Samples were centrifuged at high speed to concentrate target cells, then gradient centrifuged to separate them from matrix debris. In external laboratory experiments, RAPID-B and the reference method both correctly detected E. coli O157:H7 at inoculations of ca. 15 cells. In a follow-up study, after 4 cell inoculations of positives and 6 h enrichment, RAPID-B correctly identified 92% of 25 samples. The RAPID-B method limit of detection (LOD) was one cell in 25 g. It proved superior to the reference method (which incorporated real time-PCR, selective enrichment, and culture plating elements) in accuracy and speed.     

Common wheat (Triticum aestivum L.) and its use as a brewing cereal – a review

Wheat (Triticum aestivum L.) has a long tradition as a raw material for the production of malt and beer. Nevertheless, it has been studied to a much lesser extent than barley, which is the number one brewing cereal. The protein content of wheat ranges from about 6 to 20%, depending on the variety and baking characteristics, as well as on environmental conditions during growth. Since wheat is the most used cereal in the baking industry, the focus of wheat breeding and research has been about optimization for baking purposes (i.e. high protein content, stable falling numbers, constant baking qualities). It is well known that wheat varieties with a high protein content lead to problems in the brewing process. Therefore, varieties with a low protein content and with low viscosity values are favoured for malting and brewing. Since wheat beer yield has nearly doubled from 1990 to 2009, and is still increasing, more focus has been placed on conducting research on wheat for the malting and brewing industry. Currently, every tenth beer sold in Germany is a wheat beer. Therefore, it is of major interest to screen wheat varieties for brewing processability and to give more focus to wheat as a brewing cereal. In this review, a detailed characterization of wheat is given, particularly in regard to carbohydrates, pentosans, protein fractions and enzymes. The impact of wheat and its quality on the malting and brewing process is reviewed. Copyright © 2014 The Institute of Brewing & Distilling     

The application of dextran compared to other hydrocolloids as a novel food ingredient to compensate for low protein in biscuit and wholemeal wheat flour

Wheat is primarily used for bread-making. However, fungal diseases, grain moisture at harvest and low-protein contents strongly influence the quality of the wheat flour, thus creating challenges for traders, millers and commercial bakers who struggle to produce consistently high-quality products. This paper address the replacement of low-protein/wholemeal flour functionality for bread-making purposes. Three hydrocolloids, xanthan gum, dextran and hydroxypropyl methylcellulose, were incorporated into bread recipes based on high-protein flours, low-protein flours and coarse wholemeal flour. Hydrocolloid levels of 0–5 % (flour basis) were used in bread recipes to test the water absorption. The quality parameters of dough (farinograph, extensograph, rheofermentometre) and bread (specific volume, crumb structure and staling profile) were determined. Results showed that xanthan had negative impact on the dough and bread quality characteristics. HPMC and dextran generally improved dough and bread quality and showed dosage dependence. Volume of low-protein flour breads were significantly improved by incorporation of 0.5 % of the latter two hydrocolloids. However, dextran outperformed HPMC regarding initial bread hardness and staling shelf life regardless the flour applied in the formulation.     

Development and application of Multiple-Locus Variable number of tandem repeat Analysis (MLVA) to subtype a collection of Listeria monocytogenes

In this study we report the development and application of a Multiple-Locus Variable number of tandem repeat Analysis (MLVA) strategy for subtyping Listeria monocytogenes. Genome profiles of a collection of forty-five food-borne L. monocytogenes isolates were compared using MLVA. These isolates were obtained as part of an active surveillance programme of foods in the south-east region of Ireland. MLVA successfully discriminated amongst the isolates. The method was easy to perform, relatively fast and could be deployed in any molecular laboratory with basic laboratory equipment. This approach is a valuable tool, which has the capability to provide comparable results when compared with other more established typing methods, including pulsed-field gel electrophoresis (PFGE).     

Moisture migration in idealized bilayer systems: relationships among water-associated properties, structure, and texture

The effects of composition and thermal treatment on the water sorption and diffusional properties of idealized protein gels arranged in bilayer configurations were determined; these water binding/migration properties were related to the mechanical characteristics of the gels. Samples were prepared from whey protein concentrate (WPC), they consisted of water:WPC ratios of 1.5 to 5.67, and were thermally set for 20–60 min. Moisture migration rates from samples interfaced with filters were determined, as were moisture sorption capacities of samples immersed in water. The physical properties of the gels were assessed by uniaxial compression and microscopy. Results showed that gel strength and consequent extent of protein interaction—as affected by thermal treatment—controlled the ability of the gel structure to absorb water. Sorption was exponentially correlated with gel modulus and linearly correlated with a function of protein content, heating time, and immersion time. Rates of diffusion from interfaced gels were dependent solely on water content. It was concluded that the degree of protein interaction, whether influenced by concentration or thermal treatment, affected network extensibility and thus the capacity of the gels to act as receptors of moisture. Results have implications for the functionality of shelf-stable sandwiches and other multicomponent foods.     

Effect of UV irradiation on organic matter extracted from treated Ohio River water studied through the use of electrospray mass spectrometry

Ohio River water was treated by settling, sand filtration, and granular activated carbon filtration. It was then irradiated by low-pressure (monochromatic) and medium-pressure (polychromatic) UV lamps to investigate the effects of UV irradiation on the extracted organic matter (EOM). When the EOM, collected by solid phase extraction cartridges, was analyzed by conventional UV spectroscopy and size exclusion chromatography (SEC), no significant changes in the EOM were revealed for various UV doses. Positive and negative electrospray ionization mass spectrometry (ESI-MS) of the EOM produced mass spectra that vary significantly with UV dose. The UV dosage conditions also appear to affect the reactivity of the EOM to subsequent chlorination. The magnitude of the spectral changes is generally greater for medium-pressure lamps than for low pressure and increases with UV exposure. Based on the observed MS peaks, the changes may be due to the presence of lignin, resulting perhaps from photooxidation and/or photo rearrangement of macromolecules in the sample. When chlorination is used for secondary disinfection, these results suggest that it may be important to consider the effects of UV irradiation on the organic matter in the water before applying UV disinfection technology to a particular source water.     

Future developments in assisted reproduction in humans

The advent of human in vitro fertilization (IVF) over 30 years ago has made the oocyte and preimplantation embryo uniquely accessible. This accessibility has given rise to new micromanipulation techniques, such as intracytoplasmic sperm injection for treatment of male infertility, as well as embryo biopsy for preimplantation diagnosis of both genetic disease and aneuploidy, a major cause of early embryo demise and miscarriage. In the UK, average pregnancy rates after IVF and embryo transfer are < 25%, even after transfer of several embryos. Unfortunately, a third of these pregnancies involve multiple gestations. Research is currently focusing on methods to improve IVF success rates while reducing twin and triplet pregnancies and their associated increased morbidity and mortality. One approach is to develop screening methods to identify the most viable embryos, so that transfer of fewer healthy embryos will result in a higher proportion of singleton pregnancies. Screening methods include optimizing culture conditions for prolonged culture and selection of viable blastocysts for transfer, or embryo biopsy and aneuploidy screening. Assisted reproduction is also increasingly important in other branches of medicine: survival rates for cancer sufferers are improving continually and there is now a significant need for approaches to preserve fertility after sterilizing chemo-and radiotherapy treatment. Techniques for cryopreserving male and female gametes or gonadal tissue are being developed, although systems to grow and mature these gametes are in their infancy. Finally, there are also concerns regarding the safety of these new assisted reproductive technologies.     

Alternate and new brominated flame retardants detected in U.S. house dust

Due to the voluntary withdrawals and/or bans on the use of two polybrominated diphenyl ether (PBDE) commercial mixtures, an increasing number of alternate flame retardant chemicals are being introduced in commercial applications. To determine if these alternate BFRs are present in indoor environments, we analyzed dust samples collected from 19 homes in the greater Boston, MA area during 2006. Using pure and commercial standards we quantified the following brominated flame retardant chemicals using GC/ECNI-MS methods: hexabromocyclododecane (sigma HBCD), bis(2,4,6,-tribromphenoxy)ethane (BTBPE), decabromodiphenyl ethane (DBDPE), and the brominated components found in Firemaster 550 (FM 550): 2-ethylhexyl 2,3,4,5-tetrabromobenzoate (TBB) and (2-ethylhexyl)tetrabromophthalate (TBPH), the latter compound being a brominated analogue of di(2-ethylhexyl)phthalate (DEHP). The concentrations of all compounds were log-normally distributed and the largest range in concentrations was observed for HBCD (sum of all isomers), with concentrations ranging from <4.5 ng/g to a maximum of 130,200 ng/g with a median value of 230 ng/g. BTBPE ranged from 1.6 to 789 ng/g with a median value of 30 ng/g and DBDPE ranged from <10.0 to 11,070 ng/g with a median value of 201 ng/g. Of the FM 550 components, TBB ranged from <6.6 to 15,030 ng/g with a median value of 133 ng/g; whereas TBPH ranged from 1.5 to 10,630 ng/g with a median value of 142 ng/g. Furthermore, the ratio of TBB/TBPH present in the dust samples ranged from 0.05 to 50 (average 4.4), varying considerably from the ratio observed in the FM 550 commercial mixture (4:1 by mass), suggesting different sources with different chemical compositions, and/or differential fate and transport within the home. Analysis of paired dust samples collected from different rooms in the same home suggests HBCD, TBB, and TBPH are higher in dust from the main living area compared to dust collected in bedrooms; however, BTBPE and DBDPE levels were comparable between rooms. This study highlights the fact that numerous types of brominated flame retardants are present in indoor environments, raising questions about exposure to mixtures of these contaminants.     

Partitioning and bioaccumulation of PBDEs and PCBs in Lake Michigan

Water from Lake Michigan and fish from all five Great Lakes have been sampled and analyzed for a suite of six polybrominated diphenyl ether (PBDE) congeners and 110 polychlorinated biphenyl congeners (PCBs). The Lake Michigan dissolved phase PBDE congener concentrations (0.2 to 10 pg/L) are similar to dissolved phase PCB congener concentrations (nondetected to 13 pg/L). Partitioning of PBDEs between the particulate and dissolved phases exhibits behavior similar to that of PCBs. Organic-carbon-normalized water-particle partition coefficients (log K(OC)s) ranged from 6.2 to 6.5. Lake trout are depleted in BDE-99 relative to dissolved phase concentrations, and in contrast to what is expected from the PCB congener patterns. This reflects suspected debromination of BDE-99 in the food web of Lake Michigan. A regression of the log of the bioaccumulation factor (BAF) and the log of the octanol-water partition coefficent (K(OW)) indicated a positive relationship for both PCB congeners and PBDE congeners. BDE-99 does not appear to followthe same trend, a further indication that it is subject to biotransformation. Using the PBDE BAFs for Lake Michigan and the PBDE fish concentrations from the other Great Lakes it is expected that the dissolved phase concentrations of congeners in the other lakes would range from 0.04 to approximately 3 pg/L.