Determination of melamine in milk and dairy products by high performance liquid chromatography

A simple, precise, accurate, and validated reverse-phase HPLC method was developed for the determination of melamine in milk (pasteurized and UHT milk) and dairy products (powdered infant formula, fruit yogurt, soft cheese, and milk powder). Following extraction with acetonitrile:water (50:50, vol/vol), samples were purified by filter (0.45 μm), separated on a Nucleosil C8 column (4.6 mm × 250 mm, 3 μm) with acetonitrile:10 mmol/L sodium L-octane sulfonate (pH 3.1; 15:85, vol/vol) as mobile phase at a flow rate of 1 mL/min, and determined by a photodiode array detector. A linear calibration curve was obtained in the concentration range from 0.05 to 5 mg/kg. Milk and dairy products were fortified with melamine at 4 levels producing average recovery yields of 95 to 109%. The limits of detection and quantification of melamine were 35 to 110 and 105 to 340 μg/kg, respectively. The method was then used to analyze 300 samples of milk and dairy products purchased from major retailers in Turkey. Melamine was not found in infant formulas and pasteurized UHT milk, whereas 2% of cheese, 8% of milk powder, and 44% of yogurt samples contained melamine at the 121, 694±146, and 294±98 μg/kg levels, respectively. These findings were below the limits set by the Codex Alimentarius Commission and European Union legislation. This is the first study to confirm the existence of melamine in milk and dairy products in Turkey. Consumption of foods containing these low levels of melamine does not constitute a health risk for consumers.     

Polyelectrolyte and silver nanoparticle modification of microfiltration membranes to mitigate organic and bacterial fouling

Membrane fouling remains one of the most problematic issues surrounding membrane use in water and wastewater treatment applications. Organic and biological fouling contribute to irreversible fouling and flux decline in these processes. The aim of this study was to reduce both organic and biological fouling by modifying the surface of commercially available poly(ether sulfone) (PES) membranes using the polyelectrolyte multilayer modification method with poly(styrenesulfonate) (PSS), poly(diallyldimethylammonium chloride) (PDADMAC), and silver nanoparticles (nanoAg) integrated onto the surface as stable, thin (15 nm) films. PSS increases the hydrophilicity of the membrane and increases the negative surface charge, while integration of nanoAg into the top PSS layer imparts biocidal characteristics to the modified surface. Fouling was simulated by filtering aqueous solutions of humic acid (5 and 20 mg L(-1)), a suspension of Escherichia coli (10(6) colony-forming units (CFU) mL(-1)), and a mixture of both foulants through unmodified and modified PES membranes under batch conditions. Filtration and cleaning studies confirmed that the modification significantly reduced organic and biological fouling.     

Yogurt-like beverages made of a mixture of cereals, soy and grape must: microbiology, texture, nutritional and sensory properties

Cereal (rice, barley, emmer and oat) and soy flours and concentrated red grape must were used for making vegetable yogurt-like beverages (VYLB). Two selected strains of Lactobacillus plantarum were used for lactic acid fermentation, according to a process which included the flour gelatinization. All VLYB had values of pH lower than 4.0 and both selected starters remained viable at ca. 8.4 log cfu/g throughout storage. All VLYB showed high values of apparent viscosity and water holding capacity. During fermentation, lactic acid bacteria consumed glucose, fructose, and malic acid, which was supplied with grape must. Compared to control vegetable yogurt-like beverages (CVYLB), without bacterial inoculum, an increase of total free amino acids (FAA) was found during fermentation and storage. Also the concentration of polyphenolic compounds and ascorbic acid (ASC) was higher in VLYB compared to CVYLB. This was reflected on the antioxidant activity. As determined by Solid Phase Micro-Extraction/Gas-Chromatography/Mass-Spectrometry analysis, several volatile compounds were identified. Beverages made with the mixture of rice and barley or emmer flours seemed to possess the best combination textural, nutritional and sensory properties.     

Milk production and somatic cell counts: a cow-level analysis

The objectives of this study were to quantify the relationship between 24-h milk loss and lactation milk loss due to mastitis at the cow level. For the year 2009, individual cow test-day production records from 2,835 Ontario dairy herds were examined. Each record consisted of 24-h milk and component yields, stage of lactation (days in milk, DIM), somatic cell count (SCC, ×10(3) cells/mL) and parity. The modeling was completed in 2 stages. In stage 1, for each animal in the study, the estimated slope from a linear regression of 24-h milk yield (kg), adjusted for DIM, the quadratic effect of DIM, and the 24-h fat yield (kg) on ln(SCC) was determined. In stage 2, the estimated slope were modeled using a mixed model with a random component due to herd. The fixed effects included season (warm: May to September, cool: October to April), milk quartile class [MQ, determined by the rank of the 24-h average milk yield (kg) over a lactation within the herd] and parity. The estimated slopes from the mixed model analysis were used to estimate 24-h milk loss (kg) by comparing to a referent healthy animal with an SCC value of 100 (×10(3) cells/mL) or less. Lactation milk loss (kg) was then estimated by using estimated 24-h milk loss within lactation by means of a test-day interval method. Lactation average milk loss (kg) and SCC were also estimated. Lastly, lactation milk loss (kg) was modeled on the log scale using a mixed model, which included the random effect of herd and fixed effects, parity, and the linear and quadratic effect of the number of 24-h test days within a lactation where SCC exceeded 100 (×10(3) cells/mL; S100). The effect of SCC was significant with respect to 24-h milk loss (kg), increasing across parity and MQ. In general, first-parity animals in the first MQ (lower milk yield animals) were estimated to have 45% less milk loss than later parity animals. Milk losses were estimated to be 33% less for animals in first parity and MQ 2 through 4 than later parity animals in comparable MQ. Therefore, the relative level of milk production was found to be a significant risk factor for milk loss due to mastitis. For animals with 24-h SCC, values of 200 (×10(3) cells/mL), 24-h milk loss ranged from 0.35 to 1.09 kg; with 24-h SCC values of 2,000 (×10(3) cells/mL), milk loss ranged from 1.49 to 4.70 kg. Lactation milk loss (kg) increased significantly as lactation average SCC increased, ranging from 165 to 919 kg. The linear and quadratic effect of S100 was a significant risk factor for lactation milk loss (kg), where greatest losses occurred in lactations with 5 or more 24-h test days where SCC exceeded 100 (×10(3) cells/mL).     

Development of a multilocus variable number of tandem repeat typing method for Oenococcus oeni

Oenococcus oeni is responsible for the malolactic fermentation of wine. Genomic diversity has already been established in this species. In addition, winemakers usually report varying starter culture efficiency. The monitoring of indigenous and selected strains is essential for understanding strain survival and implantation during the winemaking process. In this study, we report the development of the first typing scheme for O. oeni using multiple-locus variable number of tandem repeat analysis (VNTR). The discriminatory power of 14 out of 44 tandem repeat loci in the genome of the PSU-1 strain was initially evaluated with a test collection of 18 genotypically distinct starter strains. Then five VNTR loci, which can be easily scored with the technology used here, were identified and used to genotype a collection of 236 strains, previously classified by restriction endonuclease analysis-pulsed-field gel electrophoresis (REA-PFGE) and multilocus sequence typing (MLST) into 136 REA-PFGE types or 110 MLST types. The discriminatory power of VNTR (as determined by Simpson's index of discrimination) was higher than that of the other two methods, with 201 VNTR types. The targeted VNTR markers were found to be stable and did not change for the clones of the same strain deposited in a collection at intervals of several years. Strains isolated from the different wine producing areas or the products were assigned to phylogenetic groups and were statistically linked with the VNTR profiles. Another interesting observation was that the loci were found in sequences homologous to regions encoding for membrane-anchored proteins.     

Effect of physical properties on the stability of Lactobacillus bulgaricus in a freeze-dried galacto-oligosaccharides matrix

The ability of galacto-oligosaccharides (GOS) to protect Lactobacillus delbrueckii subsp. bulgaricus upon freeze drying was analyzed on the basis of their capacity to form glassy structures. Glass transition temperatures (T(g)) of a GOS matrix at various relative humidities (RH) were determined by DSC. Survival of L. bulgaricus in a glassy GOS matrix was investigated after freezing, freeze drying, equilibration at different RHs and storage at different temperatures. At 32 °C, a drastic viability loss was observed. At 20 °C, the survival was affected by the water content, having the samples stored at lower RHs, the highest survival percentages. At 4°C, no decay in the cells count was observed after 45 days of storage. The correlation between molecular mobility [as measured by Proton nuclear magnetic resonance (1H NMR)] and loss of viability explained the efficiency of GOS as cryoprotectants. The preservation of microorganisms was improved at low molecular mobility and this condition was obtained at low water contents and low storage temperatures. These results are important in the developing of new functional foods containing pre and probiotics.     

Inhibition of Listeria monocytogenes on bologna sausages by an antimicrobial film containing mustard extract or sinigrin

The ability of Listeria (L.) monocytogenes to convert glucosinolates into antimicrobial isothiocyanates was investigated. Mustard glucosinolates in pure (sinigrin) or extract forms (sinigrin, oriental; sinalbin, yellow mustard) were used in broth media and in a polyvinyl polyethylene glycol graft copolymer (PPG) packaging film with bologna to examine their value as antimicrobial precursors for the control of L. monocytogenes viability and extension of bologna shelf-life at 4 °C. During broth tests with deodorized (myrosinase-inactivated) mustard extracts (10 d at 20 °C) or with purified sinigrin (21 d at 20 °C) L. monocytogenes was only inhibited when exogenous myrosinase was added. None the less, the organism was able to hydrolyze almost half the pure sinigrin by 21 d in tests without added enzyme. Reductions in sinigrin levels were measured by reversed-phase liquid chromatography, and in the absence of L. monocytogenes or added myrosinase the glucosinolate was stable. When pure sinigrin, oriental or yellow mustard extracts were incorporated in PPG films containing 3, 5 and 6% (w/w) of the corresponding glucosinolate and used to package bologna inoculated with 4 log CFU/g L. monocytogenes, the pathogen became undetectable in bologna packed with the oriental mustard extract at 52 d storage and remained undetectable at 70 d. The yellow mustard extract was less inhibitory and the pure sinigrin was not antimicrobial. L. monocytogenes numbers reached >7 log CFU/g in the film and untreated controls at 17 d storage. At 35 d storage, samples packed with control film contained sufficient numbers of lactic acid bacteria (LAB) (>7 log CFU/g) to be considered spoiled, whereas treatments containing mustard or sinigrin remained <7 log CFU/g LAB for ≤ 70 d. L. monocytogenes played a key role in exerting control over its own viability in bologna by hydrolysis of the glucosinolate in the oriental mustard film, but other antimicrobials in treatments may have contributed.     

NMR-based metabonomics detection of differences in the metabolism of hydrolysed versus intact protein of similar amino acid profile

BACKGROUND: Proton nuclear magnetic resonance (NMR)‐based metabonomics has only recently been applied to nutritional research. The limitation of any analytical technique is its sensitivity in detecting the smallest variation. Alterations in nutrition often produce only subtle metabolic modulations. The objective of this study was to determine if NMR‐based metabonomics could detect variations in the metabolic profile of urine from pigs digesting either native casein (NC) or the same casein that had been enzymatically hydrolysed (EHC). NMR permits simultaneous detection of a large number of metabolites, thus allowing detection of unanticipated metabolic fluctuations that may otherwise have gone undetected with the use of only targeted analysis. RESULTS: Partial least squares discriminant analysis identified significantly (P < 0.05) higher urinary excretions of leucine, valine, taurine and glycine by pigs on the EHC‐based diet. CONCLUSION: NMR‐based metabonomics is a sensitive method that can uncover unanticipated metabolic changes brought about by physicochemical changes to the feedstock (i.e. hydrolysis). The data show a lower efficiency of retention by the kidney of some amino acids following ingestion of a hydrolysed protein. Copyright © 2012 Society of Chemical Industry