Identification of oat (<Emphasis Type="Italic">Avena sativa</Emphasis>) and buckwheat (<Emphasis Type="Italic">Fagopyrum esculentum</Emphasis>) proteins and their prolamin fractions using two-dimensional polyacrylamide gel electrophoresis

Total (non-fractionated) kernel proteins and the prolamin fraction (soluble in 75% ethanol) were extracted from oat (Avena sativa) var. Flämingstern kernel and from buckwheat (Fagopyrum esculentum) var. Kora kernel. As for buckwheat, extraction was effective only after kernel dehulling which allowed the removal of tannins and phenolic compounds that form complexes with proteins during extraction. The extracted proteins were analyzed using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Gels of the prolamin fractions of oat and buckwheat were used as reference gels in order to detect prolamins on gels of total kernel proteins. The occurrence of 26 and 29 spots corresponding to prolamins was found on gels of total oat proteins and on gels of total buckwheat proteins, respectively. The electrophoretic images of oat and buckwheat prolamins revealed organized subregions containing spots with similar isoelectric points (pI) and various molecular weights (MW), mostly on oat prolamin gels and spots of similar molecular weights with various isoelectric points, mostly on buckwheat prolamin gels. Such organized subregions can be used as identifiers for the occurrence of prolamin fractions in total proteins (particularly as regards buckwheat proteins).     

A filtration-based real-time PCR method for the quantitative detection of viable Salmonella enterica and Listeria monocytogenes in food samples

We developed a novel filtration-based method that can eliminate dead or severally damaged Salmonella enterica and Listeria monocytogenes in food samples. This new method can recover all viable bacteria in less than 30 min, and can be coupled with a subsequent bacterial DNA extraction and real-time PCR. No statically significant differences (p < 0.01) were found between real-time PCR results obtained separately from S. enterica and L. monocytogenes when different ratios of living and dead cells were used. The analytical sensitivity in both cases was 1 genome equivalent (GE), and the quantification was linear (R² > 0.9969) over a 5-log dynamic range with PCR efficiencies >0.9754. When compared with the standard microbiological methods for the detection of these foodborne pathogens, the relative accuracy was excellent ranging from 95.72% to 104.48%. Finally, we applied the pre-treatment method to the direct detection of viable forms of these foodborne pathogens in food samples using yogurt as a model, the results being similar to those obtained using pure cultures.     

Effects of Fish Gelatin and Tea Polyphenol Coating on the Spoilage and Degradation of Myofibril in Fish Fillet During Cold Storage

Fish fillet is easily spoiled during storage. Antimicrobial edible coating of gelatin extracted from fish skins and bones and tea polyphenol (TP) was developed to inhibit the spoilage of fish fillet during cold storage. For coating containing 0.4 % TP and 1.2 % gelatin, the pH only slightly increased from 6.17 at day 0 to 6.32 at day 17 of cold storage, while the pH of control coating increased to 6.87 at day 17. Atomic force spectrometry was used to analyse the nanostructure of myofibril, which is the major component of fish muscle. The results showed that the length of myofibril from 0.4 % TP and 1.2 % gelatin group was greater than 15 μm, while the diameter and height were 3.38 and 0.59 μm, respectively, which exhibited the most intact nanostructure after 17 days of cold storage. Meanwhile, matrix-assisted laser desorption–ionisation–time-of-flight mass spectrometry result showed that TP delayed the degradation of myosin light chain 3 and troponin T in myofibril. Gas chromatography–mass spectrometry of volatile organic compounds (VOCs) also showed that 0.4 % TP and 1.2 % gelatin coating group had minimal production of spoilage markers, such as 1-octen-3-ol, 2-methyl propanoic acid and dimethyl sulfide. The microbial analysis showed that the aerobic mesophilic/psychrotrophic count, yeasts and moulds of 0.4 % TP and 1.2 % gelatin group were significantly lower than the control group. Therefore, 0.4 % TP and 1.2 % gelatin coating showed the best antimicrobial effect and can be used to maintain the nanostructure of fish fillet and prevent the spoilage during cold storage.     

Comparison of biota-sediment accumulation factors across ecosystems

Sets of biota-sediment accumulation factors (BSAFs) for fish were compared across ecosystems for nonionic organic chemicals. The sets of BSAFs, when plotted against each other, in log-log space, formed linear relationships and demonstrated that the relative scaling or ranking of the individual BSAFs within a set are consistent, if not the same, across ecosystems. This behavior holds for chemicals that either are, or are not, metabolized by fish. These results demonstrate that sets of BSAF values can differ but with parallel shifts in magnitude between ecosystems (for example, all of the BSAFs in the set are uniformly larger in one ecosystem, while in another they all are uniformly smaller) in response to underlying differences in ecosystem conditions and parameters such as trophic level, diet of the organisms, and distribution of the chemical between the sediment and water column.     

A one-pot ultrasound-assisted water extraction/cyclodextrin encapsulation of resveratrol from Polygonum cuspidatum

The ultrasound-assisted extraction (UAE) of resveratrol and other polyphenols from Polygonum cuspidatum has been carried out with the aim of developing an efficient and eco-friendly extraction process. The finely milled roots were sonicated (titanium horn, 19.5 kHz) in methanol, in different cyclodextrin water solutions [β-cyclodextrin (β-CD) or hydroxypropyl β-CD (HPβ-CD)] and also in pure water. UAE dramatically increased the yields and cut down extraction times compared to conventional extraction under stirring. Outstanding results have been achieved with the β-CD solution (1.5% w/w); in fact HPLC analysis showed that the selective inclusion properties of CDs toward phenolic stilbenes gave a much cleaner analytical extract profile. This green method gave 7.51 mg of total resveratrol (free + cleaved polydatin glucoside) per gram of dry plant. Thanks to polyphenol encapsulation within CDs, this extract showed excellent water dispersibility, higher stability and an antioxidant power which is comparable to that of the MeOH extract (DPPH, ORAC_FL). These important features should pave the road for its application in food supplements or phytochemical preparations.     

The effect of cocoa fermentation and weak organic acids on growth and ochratoxin A production by Aspergillus species

The acidic characteristics of cocoa beans have influence on flavor development in chocolate. Cocoa cotyledons are not naturally acidic, the acidity comes from organic acids produced by the fermentative microorganisms which grow during the processing of cocoa. Different concentrations of these metabolites can be produced according to the fermentation practices adopted in the farms, which could affect the growth and ochratoxin A production by fungi. This work presents two independent experiments carried out to investigate the effect of some fermentation practices on ochratoxin A production by Aspergillus carbonarius in cocoa, and the effect of weak organic acids such as acetic, lactic and citric at different pH values on growth and ochratoxin A production by A. carbonarius and Aspergillus niger in culture media. A statistical difference (ρ<0.05) in the ochratoxin A level in the cured cocoa beans was observed in some fermentation practices adopted. The laboratorial studies demonstrate the influence of organic acids on fungal growth and ochratoxin A production, with differences according to the media pH and the organic acid present. Acetic acid was the most inhibitory acid against A. carbonarius and A. niger. From the point of view of food safety, considering the amount of ochratoxin A produced, fermentation practices should be conducted towards the enhancement of acetic acid, although lactic and citric acids also have an important role in lowering the pH to improve the toxicity of acetic acid.     

Intra- and interspecific mineral composition variability of commercial instant coffees and coffee substitutes: Contribution to mineral intake

The present paper reports the amount and estimated daily mineral intake of nine elements (Ca, Mg, K, Na, P, Fe, Mn, Cr and Ni) in commercial instant coffees and coffee substitutes (n = 49). Elements were quantified by high-resolution continuum source flame (HR-CS-FAAS) and graphite furnace (HR-CS-GFAAS) atomic absorption spectrometry, while phosphorous was evaluated by a standard vanadomolybdophosphoric acid colorimetric method.Instant coffees and coffee substitutes are rich in K, Mg and P (>100 mg/100 g dw), contain Na, Ca and Fe in moderate amounts (>1 mg/100 g), and trace levels of Cr and Ni. Among the samples analysed, plain instant coffees are richer in minerals (p < 0.001), except for Na and Cr. Blends of coffee substitutes (barley, malt, chicory and rye) with coffee (20–66%) present intermediate amounts, while lower quantities are found in substitutes without coffee, particularly in barley.From a nutritional point of view the results indicate that the mean ingestion of two instant beverages per day (total of 4 g instant powder), either with or without coffee, cannot be regarded as important sources of minerals to the human diet, although providing a supplementation of some minerals, particularly Mg and Mn in instant coffees. Additionally, and for authentication purposes, the correlations observed between some elements and the coffee percentage in the blends, with particular significance for Mg amounts, provides a potential tool for the estimation of coffee in substitute blends.     

New insights on the metabolism of ricinoleic acid in ruminants

Dairy goats were fed a total mixed ration with or without the inclusion of castor oil [40 g/kg of dry matter (DM)] to study the metabolism of ricinoleic acid (12-OH,cis-9–18:1). Ten goats, at 39.7 ± 4.0 d in milk, were individually penned and allocated at random to the 2 experimental diets. Goats were manually milked twice a day. Milk fatty acids (FA) were analyzed as methyl esters and hydroxyl groups were derivatized in trimethylsilyl ethers. Apart from ricinoleic acid, 6 FA were only detected in the milk of the castor oil group. Ricinoleic acid composed 0.3% of total FA in milk of the castor oil group, whereas the hydroxy-FA (8-OH-14:0, 10-OH-16:0, and 12-OH-18:0) and oxo-FA (8-oxo-14:0, 10-oxo-16:0, and 12-oxo-18:0) reached 7.5% of total FA in milk. We anticipate that these FA were derived from the metabolism of ricinoleic acid, although it was not clear if they were produced in the rumen or in the tissues. To confirm that, we conducted in vitro batch incubations repeated for 3 consecutive weeks with castor oil (40 g/kg of DM) and strained rumen fluid from 2 fistulated sheep. To examine the products formed over time, incubation tubes were stopped at 0, 6, 12, 24, 48, and 72 h. The results of the in vitro experiment showed that ricinoleic acid was metabolized in the rumen at a slow rate and the main products formed were 12-OH-18:0 and 12-oxo-18:0, by hydrogenation of the cis-9 double bond, followed by oxidation of the hydroxyl group, respectively. Our results suggest that the 12-OH-18:0 and 12-oxo-18:0 escape rumen and are further metabolized through partial β-oxidation in ruminant tissues. We propose that the 10-OH-16:0 and 8-OH-14:0 found in goat milk of the castor oil group are successive products of the β-oxidation of 12-OH-18:0, and the 10-oxo-16:0 and 8-oxo-14:0 are successive products of the 12-oxo-18:0 in tissues. Overall, our results indicate that ricinoleic acid is extensively metabolized in the rumen and tissues, producing mainly oxo- and hydroxy-FA that are further excreted in milk.     

Development of Quality Index Method (QIM) scheme for Acoupa weakfish (Cynoscion acoupa)

A Quality Index Method (QIM) scheme for gutted and iced Acoupa weakfish (Cynoscion acoupa) was developed and its efficiency for freshness evaluation was compared with instrumental colour analysis, microbial and chemical methods. The QIM scheme comprises sensory evaluation of 14 parameters, reaching a total sum of 23 demerit points. The evolution of Quality Index (QI) was identical in two independent storage experiments and could be represented by the equation QI = 1.3593 × days − 0.3822 (R² = 0.9614, P < 0.0001). The QIM results indicated a shelf life of 8–9 days, when the sensory quality was considered unacceptable. The storage time affected a∗ and b∗ values for fish gill and caudal fin, respectively. Psychrotrophic bacteria counts (PBC) increased at different rates during the iced storage of fish in three independent storage experiments, reaching the maximum recommended limit for marginally acceptable quality products of 10⁷ CFU/g between 6 and 14 days. The shelf life of Acoupa weakfish estimated by QIM was slightly longer than that evaluated by PBC. The rate and pattern of total volatile base nitrogen and trimethylamine production was different in fish from three independent storage experiments. The use of QIM scheme was a reliable way of assessing freshness of Acoupa weakfish.     

Detection of the adulteration of olive oils by solid phase microextraction and multidimensional gas chromatography

The presence or absence of filbertone in 21 admixtures of olive oil with virgin and refined hazelnut oils obtained using various processing techniques from different varieties and geographical origins was evaluated by solid phase microextraction and multidimensional gas chromatography (SPME–MDGC). The obtained results showed that the sensitivity achievable with the proposed procedure was enough to detect filbertone and, hence, to establish the adulteration of olive oil of different varieties with virgin hazelnut oils in percentages of up to 7%. The very low concentrations in which filbertone occurs in some refined hazelnut oils made difficult its detection in specific admixtures. In any case, the minimum adulteration level to be detected depends on the oil varieties present in the adulterated samples. In the present study, the presence of R- and S-enantiomers of filbertone could be occasionally detected in olive oils adulterated with 10–20% of refined hazelnut oil.