Quantitative analysis of N-phenylpropenoyl-L-amino acids in roasted coffee and cocoa powder by means of a stable isotope dilution assay

Since recent reports on the role of N-phenylpropenoyl-L-amino acids as powerful antioxidants and key contributors to the astringent taste of cocoa nibs, there is an increasing interest in the concentrations of these phytochemicals in plant-derived foods. A versatile analytical method for the accurate quantitative analysis of N-phenylpropenoyl-L-amino acids in plant-derived foods by means of HPLC-MS/MS and synthetic stable isotope labeled N-phenylpropenoyl-L-amino acids as internal standards was developed. By means of the developed stable isotope dilution assay (SIDA), showing recovery rates of 95-102%, 14 N-phenylpropenoyl-L-amino acids were quantified for the first time in cocoa and coffee samples. On the basis of the results of LC-MS/MS experiments as well as cochromatography with the synthetic reference compounds N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-tryptophan, N-[4'-hydroxy-(E)-cinnamoyl]-L-tryptophan, and N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-tyrosine, respectively, were detected for the first time in cocoa powder, and (-)-N-[4'-hydroxy-(E)-cinnamoyl]-L-tyrosine, (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-tyrosine, N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-tyrosine, (+)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-aspartic acid, (+)-N-[4'-hydroxy-(E)-cinnamoyl]-L-aspartic acid, N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-tryptophan, N-[4'-hydroxy-(E)-cinnamoyl]-L-tryptophan, and N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-tryptophan, respectively, were detected for the first time in coffee beverages.     

Impact of hardness of model fresh cheese on aroma release: in vivo and in vitro study

Using atmospheric pressure chemical ionization-mass spectrometry, aroma release was investigated in vivo and in vitro from three cheese-like gels with different hardnesses. In vivo, nosespace experiments were performed with 14 subjects. Results showed that the harder gel induced a greater and a faster release of all aroma compounds. In vitro, aroma release was followed in a mouth simulator where breakdown was mechanically produced. The same rate of stirring was applied to the three gels. In these conditions, we found that the amount of aroma released from the three gels was not discriminant. Thus, modification of gel structure had a strong impact on in vivo aroma release, but structural variations alone were not sufficient to induce a greater release. Natural breakdown provided by panelists during food consumption and adapted to the texture of the food was proposed to be the key parameter affecting in vivo aroma release.     

Contribution of Fimbrial and Afimbrial Adhesins of Xylella fastidiosa to Attachment to Surfaces and Virulence to Grape

ABSTRACT The role of fimbrial and afimbrial adhesins of Xylella fastidiosa in biofilm formation was assessed by visualization of cell aggregates of mutant strains after incubation on glass surfaces. FimA- or FimF- fimbrial mutants adhered as solitary cells at a slightly lesser frequency to glass surfaces than the parental strain; however, cell aggregates were not formed, unlike the wild-type strain. Conversely, whereas the XadA- and HxfB- nonfimbrial mutants also exhibited a much lower frequency of adherence to glass surfaces than the wild-type strain, most of the cells retained on the surfaces were in cell aggregates of different sizes, much like that of the parental strain. Neither fimbrial or afimbrial mutants formed a mature biofilm on the sides of flasks of broth cultures, unlike the dense biofilm formed by the wild-type strain. Although FimA- and FimF- mutants did not form cell aggregates on glass surfaces when incubated as individual strains, aggregates of a FimA- or FimF- mutant were observed when co-incubated with either a XadA- mutant or HxfB- mutant, respectively. These results are consistent with a model in which the fimbrial adhesins FimA and FimF are involved preferentially in cell-to-cell aggregate formation whereas the afimbrial adhesions XadA and HxfB preferentially contribute to initial cell binding to surfaces, whereupon further cell aggregation can occur. In each of five separate experiments, FimA, FimF, XadA, and HxfB mutants of X. fastidiosa all were less virulent to grape than the corresponding wild-type strain. Fimbrial and afimbrial mutants might produce a reduced biofilm within vessels of grape and, hence, be deficient in various cell-density-dependent traits required for movement through the plant and, thus, virulence.     

Collembolan dietary specialisation on soil grown fungi

Preferences of six collembolans for eight species of microfungi grown in soil have been investigated. Collembolans (Folsomia fimetaria, Isotoma anglicana, Isotama notabilis, Heteromurus nitidus, Protaphorura armata, Pseudosinella alba) and microfungi where chosen for the experiment on the basis of their abundance and vertical distribution in an organically grown field where all organisms and soil were collected. Collembolans were tested with pairs of different species of microfungi and a control (sterile soil) in petri dishes and their position determined after 90 min. Based on the first two axes in a Principal Component Analysis we could identify two main feeding patterns in the collembolans: (1) their general acceptance of the fungi (the mean value of the preference indices for all eight fungi), and (2) their alternating preference for Cladosporium herbarum and Fusarium culmorum (high preference index for C. herbarum and low for F. culmorum or vice versa). The six collembolan species in our study combined these two feeding patterns with an intraspecific preference for the eight fungal species and seemed to minimise food competition among collembolans co-occurring in the same soil horizons. Our data suggest that differences in preference between collembolan species may help to explain the coexistence of many species in the same microhabitats of the soil.     

Combining conservative and variable markers to infer the evolutionary history of Prunus subgen. Amygdalus s.l. under domestication

The genus Prunus L. is large and economically important. However, phylogenetic relationships within Prunus at low taxonomic level, particularly in the subgenus Amygdalus L. s.l., remain poorly investigated. This paper attempts to document the evolutionary history of Amygdalus s.l. and establishes a temporal framework, by assembling molecular data from conservative and variable molecular markers. The nuclear s6pdh gene in combination with the plastid trnSG spacer are analyzed with bayesian and maximum likelihood methods. Since previous phylogenetic analysis with these markers lacked resolution, we additionally analyzed 13 nuclear SSR loci with the δµ² distance, followed by an unweighted pair group method using arithmetic averages algorithm. Our phylogenetic analysis with both sequence and SSR loci confirms the split between sections Amygdalus and Persica, comprising almonds and peaches, respectively. This result is in agreement with biogeographic data showing that each of the two sections is naturally distributed on each side of the Central Asian Massif chain. Using coalescent based estimations, divergence times between the two sections strongly varied when considering sequence data only or combined with SSR. The sequence-only based estimate (5 million years ago) was congruent with the Central Asian Massif orogeny and subsequent climate change. Given the low level of differentiation within the two sections using both marker types, the utility of combining microsatellites and data sequences to address phylogenetic relationships at low taxonomic level within Amygdalus is discussed. The recent evolutionary histories of almond and peach are discussed in view of the domestication processes that arose in these two phenotypically-diverging gene pools: almonds and peaches were domesticated from the Amygdalus s.s. and Persica sections, respectively. Such economically important crops may serve as good model to study divergent domestication process in close genetic pool.     

Population sizes, site fidelity and residence patterns of Australian snubfin and Indo-Pacific humpback dolphins: Implications for conservation

Very little is known about the ecology of snubfin Orcaella heinsohni and Indo-Pacific humpback dolphins Sousa chinensis in Australian waters. We used photo-identification data collected between 1999 and 2002 in Cleveland Bay, northeast Queensland, to estimate abundance, site fidelity and residence patterns of these species in order to make recommendations for their effective conservation and management. Our abundance estimates indicate that less than a hundred individuals of each species inhabit this coastal area. Even with relatively unbiased and precise abundance estimates population trends will be extremely difficult to detect in less than three years unless changes in population size are very high (>20% p.a.). Though both species are not permanent residents in Cleveland Bay, they both used the area regularly from year to year following a model of emigration and reimmigration. Individuals of both species spend periods of days to a month or more in coastal waters of Cleveland Bay before leaving, and periods of over a month outside the study area before entering the bay again. Because of their small population sizes and movement patterns, snubfin and humpback dolphins are particularly vulnerable to local extinction. Our results illustrate that: (1) detection of population trends should not be a necessary criterion for enacting conservation measures of both species in this region, and (2) efforts to maintain viable populations of both species in Cleveland Bay must include management strategies that integrate anthropogenic activities in surrounding areas.     

Principal component analysis applied to Fourier transform infrared spectroscopy for the design of calibration sets for glycerol prediction models in wine and for the detection and classification of outlier samples

Principal component analysis (PCA) was used to identify the main sources of variation in the Fourier transform infrared (FT-IR) spectra of 329 wines of various styles. The FT-IR spectra were gathered using a specialized WineScan instrument. The main sources of variation included the reducing sugar and alcohol content of the samples, as well as the stage of fermentation and the maturation period of the wines. The implications of the variation between the different wine styles for the design of calibration models with accurate predictive abilities were investigated using glycerol calibration in wine as a model system. PCA enabled the identification and interpretation of samples that were poorly predicted by the calibration models, as well as the detection of individual samples in the sample set that had atypical spectra (i.e., outlier samples). The Soft Independent Modeling of Class Analogy (SIMCA) approach was used to establish a model for the classification of the outlier samples. A glycerol calibration for wine was developed (reducing sugar content < 30 g/L, alcohol > 8% v/v) with satisfactory predictive ability (SEP = 0.40 g/L). The RPD value (ratio of the standard deviation of the data to the standard error of prediction) was 5.6, indicating that the calibration is suitable for quantification purposes. A calibration for glycerol in special late harvest and noble late harvest wines (RS 31-147 g/L, alcohol > 11.6% v/v) with a prediction error SECV = 0.65 g/L, was also established. This study yielded an analytical strategy that combined the careful design of calibration sets with measures that facilitated the early detection and interpretation of poorly predicted samples and outlier samples in a sample set. The strategy provided a powerful means of quality control, which is necessary for the generation of accurate prediction data and therefore for the successful implementation of FT-IR in the routine analytical laboratory.     

Geographical distribution of salmonella infected pig,cattle and sheep herds in Sweden 1993-2010

Background

The Swedish salmonella control programme covers the entire production chain, from feed to food. All salmonella serotypes are notifiable. On average, less than 20 cases of salmonella in food-producing animals are reported every year. In some situations, the cases would be expected to cluster geographically. The aim of this study was to illustrate the geographic distribution of the salmonella cases detected in pigs, cattle and sheep.

Methods

Data on all herds with pigs, cattle and sheep found to be infected with salmonella during the time period from 1993 to 2010 were obtained from the Swedish Board of Agriculture. Using the ArcGIS software, various maps were produced of infected herds, stratified on animal species as well as salmonella serotype. Based on ocular inspection of all maps, some were collapsed and some used separately. Data were also examined for temporal trends.

Results

No geographical clustering was observed for ovine or porcine cases. Cattle herds infected with Salmonella Dublin were mainly located in the southeast region and cattle herds infected with Salmonella Typhimurium in the most southern part of the country. Some seasonal variation was seen in cattle, but available data was not sufficient for further analyses.

Conclusions

Analyses of data on salmonella infected herds revealed some spatial and temporal patterns for salmonella in cattle. However, despite using 18 years'' of data, the number of infected herds was too low for any useful statistical analyses.