Lactic acid bacteria as structural building blocks in non-fat whipping cream analogues

Lactic acid bacteria as food ingredients, show the potential of being exploited as structural building blocks in the formulation of colloidal foods such as emulsion and foam. The present work provides approaches to using lactic acid bacteria combined with two components, hydroxypropyl methylcellulose (HPMC) and casein sodium (CS) salt, to fully replace the saturated fat content in whipping cream analogues. By involving both hydrophobic and hydrophilic strains, the whipped cream exhibited comparable overrun (107%) and drainage stability (drainage area 1.4 mm2) to the commercial dairy whipping cream (30% and 2.7 mm2, respectively), where the foam stability was greatly affected by the Pickering capability and aggregating properties of the used strains. All the whipped cream displayed solid-like behaviors (G’>G″) and standing properties to different degrees (G’ ≈ 30–491 Pa), depending on the strength of bacterial aggregation jointly determined by both the intrinsic surface properties and the influence of added HPMC and CS components. No negative impacts on bacterial viability was found for the added components and the whipping process. The idea of involving edible lactic acid bacteria as fat replacers can thus provide possible alternatives to using nature-derived components as active structural building blocks for colloidal food systems such as whipping cream

The effect of surface properties of polycrystalline, single phase metal coatings on bacterial retention

In the food industry microbial contamination of surfaces can result in product spoilage which may lead to potential health problems of the consumer. Surface properties can have a substantial effect on microbial retention. The surface characteristics of chemically different coatings (Cu, Ti, Mo, Ag, Fe) were defined using white light profilometry (micro-topography and surface features), atomic force microscopy (nano-topography) and physicochemical measurements. The Ag coating had the greatest topography measurements and Fe and Mo the least. Mo was the most hydrophobic coating (lowest γAB,γ+, γ−) whilst Ag was the most hydrophilic (greatest γAB,γ+, γ−). The physicochemical results for the Fe, Ti and Cu coatings were found to lie between those of the Ag and Mo coatings. Microbiological retention assays were carried out using Listeria monocytogenes, Escherichia coli and Staphylococcus aureus in order to determine how surface properties influenced microbial retention. It was found that surface chemistry had an effect on microbial retention, whereas the shape of the surface features and nano-topography did not. L. monocytogenes and S. aureus retention to the surfaces were mostly affected by surface micro-topography, whereas retention of E. coli to the coatings was mostly affected by the coating physicochemistry. There was no trend observed between the bacterial cell surface physicochemistry and the coating physicochemistry. This work highlights that different surface properties may be linked to factors affecting microbial retention hence, the use of surface chemistry, topography or physicochemical factors alone to describe microbial retention to a surface is no longer adequate. Moreover, the effects of surface parameters on microbial retention should be considered individually for each bacterial genus

Enzymatic modification and adsorption of hydrophobic zein proteins on lactic acid bacteria stabilize Pickering emulsions

The effect of enzymatic and physical modifications of the surface of two different strains from lactic acid bacteria, Lactobacillus rhamnosus (LGG) and Lactobacillus delbruekii subs. lactis ATCC 4797 (LBD), to stabilize medium-chain triglyceride (MCT) oil based Pickering emulsions were investigated. A section of cell wall degrading enzymes, lysozyme from chicken egg white and human, lysostaphin, mutanolysin from Streptomyces globisporus and proteinase k and the hydrophobic protein zein were used for enzymatic and physical surface modifications. Cell surface modifications were characterized by optical microscopy, scanning electron cryo-microscopy (Cryo-SEM), transmission electron microscopy (TEM), microbial adhesion to hexadecane (MATH) test and zeta potential measurements. The modified cell hydrophobicity in terms of MATH values were increased (around four times) by the enzymatic and physical modifications for LBD and LGG compared to the control. Emulsions stabilized by modified bacterial cells showed higher stability in comparison with unmodified samples, especially for the samples modified with chicken egg lysozyme. Confocal microscopy revealed that the modified bacterial cells were absorbed at the interface between oil and water and preventing the oil particles from coalescence. Thus, modified bacterial cells can be used to formulate food-grade stable Pickering emulsions. Such Pickering emulsions can potentially be clean label alternatives to replace the conventional emulsion preparations

Multi-species colloidosomes by surface-modified lactic acid bacteria with enhanced aggregation properties

Hypothesis: Surface modification of lactic acid bacteria enhances their adsorption and aggregation at air–water interface and enables stabilization of microbubbles that spontaneously transform into water-filled colloidosomes, which can be further modified using LBL formulations. Experiments: The bacterial physicochemical properties were characterized using water contact angle (WCA) measurement, bacterial aggregation assay and zeta potential measurement. Cell viability was enumerated using plate-counting method. The LBL reinforcement of colloidosomes was examined by zeta potential measurement and the formed microstructure was investigated using bright-field microscopy, confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Shell permeability of colloidosomes was evaluated using a dye release study. Findings: Bacteria surface-modified using octenyl succinic anhydride (OSA) expressed strong adsorption and aggregation at air–water interface when producing microbubbles. Bacteria with enhanced aggregation ability formed stable shells, enabling complete removal of air and air–water interface without shell disintegration. The formed colloidosomes were studied as they were, or were further reinforced by LBL deposition using polymer or hybrid formulations. Hybrid coating involved assembly of two bacterial species producing colloidosomes with low shell porosity. The findings can be exploited to organize different living bacteria into structured materials and to encapsulate and release substances of diverse sizes and surface properties

Efficient chemical hydrophobization of lactic acid bacteria – one-step formation of double emulsion

A novel concept of stabilizing multiple-phase food structure such as emulsion using solely the constitutional bacteria enables an all-natural food grade formulation and thus a clean label declaration. In this paper, we propose an efficient approach to hydrophobically modifying the surface of lactic acid bacteria Lactobacillus rhamnosus (LGG) using lauroyl ahloride (LC) in non-aqueous media. Compared to the unmodified bacteria, cell hydrophobicity was dramatically altered upon modification, according to the higher percentages of microbial adhesion to hexadecane (MATH) and water contact angles (WCA) of LC-modified bacteria. No evident changes were found in bacterial surface charge before and after LC modification. By using one-step homogenization, all the modified bacteria were able to generate stabile water-in-oil-in-water (W/O/W) double emulsions where bacteria were observed on oil–water interfaces of the primary and secondary droplets. Modification using high LC concentrations (10 and 20 w/w%) led to rapid autoaggregation of bacteria in aqueous solution. A long-term lethal effect of modification primarily came from lyophilization and no apparent impact was detected on the instantaneous culturability of modified bacteria

Methodological approaches to determining the marine radiocarbon reservoir effect

The marine radiocarbon reservoir effect is an offset in 14C age between contemporaneous organisms from the terrestrial environment and organisms that derive their carbon from the marine environment. Quantification of this effect is of crucial importance for correct calibration of the <sup>14</sup>C ages of marine-influenced samples to the calendrical timescale. This is fundamental to the construction of archaeological and palaeoenvironmental chronologies when such samples are employed in <sup>14</sup>C analysis. Quantitative measurements of temporal variations in regional marine reservoir ages also have the potential to be used as a measure of process changes within Earth surface systems, due to their link with climatic and oceanic changes. The various approaches to quantification of the marine radiocarbon reservoir effect are assessed, focusing particularly on the North Atlantic Ocean. Currently, the global average marine reservoir age of surface waters, R(t), is c. 400 radiocarbon years; however, regional values deviate from this as a function of climate and oceanic circulation systems. These local deviations from R(t) are expressed as +R values. Hence, polar waters exhibit greater reservoir ages (δR = c. +400 to +800 <sup>14</sup>C y) than equatorial waters (δR = c. 0 <sup>14</sup>C y). Observed temporal variations in δR appear to reflect climatic and oceanographic changes. We assess three approaches to quantification of marine reservoir effects using known age samples (from museum collections), tephra isochrones (present onshore/offshore) and paired marine/terrestrial samples (from the same context in, for example, archaeological sites). The strengths and limitations of these approaches are evaluated using examples from the North Atlantic region. It is proposed that, with a suitable protocol, accelerator mass spectrometry (AMS) measurements on paired, short-lived, single entity marine and terrestrial samples from archaeological deposits is the most promising approach to constraining changes over at least the last 5 ky BP

Direct evidence of milk consumption from ancient human dental calculus.

Milk is a major food of global economic importance, and its consumption is regarded as a classic example of gene-culture evolution. Humans have exploited animal milk as a food resource for at least 8500 years, but the origins, spread, and scale of dairying remain poorly understood. Indirect lines of evidence, such as lipid isotopic ratios of pottery residues, faunal mortality profiles, and lactase persistence allele frequencies, provide a partial picture of this process; however, in order to understand how, where, and when humans consumed milk products, it is necessary to link evidence of consumption directly to individuals and their dairy livestock. Here we report the first direct evidence of milk consumption, the whey protein β-lactoglobulin (BLG), preserved in human dental calculus from the Bronze Age (ca. 3000 BCE) to the present day. Using protein tandem mass spectrometry, we demonstrate that BLG is a species-specific biomarker of dairy consumption, and we identify individuals consuming cattle, sheep, and goat milk products in the archaeological record. We then apply this method to human dental calculus from Greenland's medieval Norse colonies, and report a decline of this biomarker leading up to the abandonment of the Norse Greenland colonies in the 15(th) century CE

Disequilibrium, adaptation and the Norse settlement of Greenland

This research was supported by the University of Edinburgh ExEDE Doctoral Training Studentship and NSF grant numbers 1202692 and 1140106.There is increasing evidence to suggest that arctic cultures and ecosystems have followed non-linear responses to climate change. Norse Scandinavian farmers introduced agriculture to sub-arctic Greenland in the late tenth century, creating synanthropic landscapes and utilising seasonally abundant marine and terrestrial resources. Using a niche-construction framework and data from recent survey work, studies of diet, and regional-scale climate proxies we examine the potential mismatch between this imported agricultural niche and the constraints of the environment from the tenth to the fifteenth centuries. We argue that landscape modification conformed the Norse to a Scandinavian style of agriculture throughout settlement, structuring and limiting the efficacy of seasonal hunting strategies. Recent climate data provide evidence of sustained cooling from the mid thirteenth century and climate variation from the early fifteenth century. Archaeological evidence suggests that the Norse made incremental adjustments to the changing sub-arctic environment, but were limited by cultural adaptations made in past environments.Publisher PDFPeer reviewe

Reduction of volatile acidity of wines by selected yeast strains

Herein we isolate and characterize wine yeasts with ability to reduce volatile acidity of wines using a refermentation process, which consists in mixing the acidic wine with freshly crushed grapes or musts or, alternatively, in the incubation with the residual marc. From a set of 135 yeast isolates, four strains revealed ability to use glucose and acetic acid simultaneously. Three of them were identified as Saccharomyces cerevisiae and one as Lachancea thermotolerans. Among nine commercial S. cerevisiae strains, strains S26, S29 and S30 display similar glucose and acetic acid initial simultaneous consumption pattern and were assessed in refermentation assays. In a medium containing an acidic wine with high glucose/low ethanol concentrations, under low oxygen availability, strain S29 is the most efficient one, whereas L. thermotolerans 44C is able to decrease significantly acetic acid similar to the control strain Zygosaccharomyces bailii ISA 1307, but only under aerobic conditions. Conversely, for low glucose/high ethanol concentrations, under aerobic conditions, S26 is the most efficient acid degrading strain, while under limited-aerobic conditions, all the S. cerevisiae strains studied display acetic acid degradation efficiencies identical to Z. bailii. Moreover, S26 strain also reveals capacity to decrease volatile acidity of wines. Together, the S. cerevisiae strains characterized herein appear promising for the oenological removal of volatile acidity of acidic wines.Fundação para a Ciência e a Tecnologia (FCT) - Programa POCI 2010 (FEDER/FCT, POCI/AGR/56102/2004, PTDC/AGRALI/71460/2006

The fate of acetic acid during glucose co-metabolism by the spoilage yeast Zygosaccharomyces bailii

Zygosaccharomyces bailii is one of the most widely represented spoilage yeast species, being able to metabolise acetic acid in the presence of glucose. To clarify whether simultaneous utilisation of the two substrates affects growth efficiency, we examined growth in single- and mixed-substrate cultures with glucose and acetic acid. Our findings indicate that the biomass yield in the first phase of growth is the result of the weighted sum of the respective biomass yields on single-substrate medium, supporting the conclusion that biomass yield on each substrate is not affected by the presence of the other at pH 3.0 and 5.0, at least for the substrate concentrations examined. In vivo(13)C-NMR spectroscopy studies showed that the gluconeogenic pathway is not operational and that [2-(13)C]acetate is metabolised via the Krebs cycle leading to the production of glutamate labelled on C(2), C(3) and C(4). The incorporation of [U-(14)C]acetate in the cellular constituents resulted mainly in the labelling of the protein and lipid pools 51.5% and 31.5%, respectively. Overall, our data establish that glucose is metabolised primarily through the glycolytic pathway, and acetic acid is used as an additional source of acetyl-CoA both for lipid synthesis and the Krebs cycle. This study provides useful clues for the design of new strategies aimed at overcoming yeast spoilage in acidic, sugar-containing food environments. Moreover, the elucidation of the molecular basis underlying the resistance phenotype of Z. bailii to acetic acid will have a potential impact on the improvement of the performance of S. cerevisiae industrial strains often exposed to acetic acid stress conditions, such as in wine and bioethanol production.This work was supported by Fundacao para a Ciencia e Tecnologia (FCT), Portugal Grant PTDC/AGR-ALI/102608/2008 and by project FCOMP-01-0124-FEDER- 007047 and by FEDER through POFC - COMPETE and national funds from FCT - project PEst-C/BIA/UI4050/2011. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript