Triacylglycerol analysis for the quantification of cocoa butter equivalents (CBE) in chocolate: feasibility study and validation

A new European legislation (2000/36/CE) has allowed the use of vegetable fats other than cocoa butter (CB) in chocolate up to a maximum value of 5% in the product. The vegetable fats used in chocolate are designated as cocoa butter replacements and are called cocoa butter equivalents (CBE). The feasibility of CBE quantification in chocolate using triacylglycerol (TAG) profiles was conducted by analyzing 55 samples of CBs and 31 samples of CBEs using a liquid chromatograph equipped with an evaporative light scattering detector (HPLC-ELSD). Statistical evaluation of the data obtained has been performed, and a simulation study has been carried out to assess the viability to use this method for quantifying the amount of CBE in real mixtures and in chocolates. The TAGs POP, POS, PLS, and the ratios POP/PLS, POS/PLP (P, palmityl; O, oleyl; S, stearyl; L, linoleyl) are particularly significant to discriminate between CB and CBE. Analysis of 50 mixtures between 5 different CBEs and 10 different CBs at 2 different concentration levels is presented. The data are visualized and interpreted. A mathematical model has been developed to assess the amount of CBE in real mixtures. This predictive model has been successfully applied and validated on dark chocolates including authorized CBE. The results are affected by +/-2.1% absolute average error. In particular, estimations between 10 and 20% of CBE show a very good match. On the other hand, values equal to or smaller than 5% show a larger prediction error (detection limit of the method). For the main purpose of this method (i.e., quantification of CBE at 5% max in chocolate, which represents about 15% of the total fat) this model shows very good results. For milk chocolate, the mathematical model can also be used if TAG are integrated from partition number (PN) 46 to 54. Consequently, the model proposed provides sufficient information to verify the real application of the European legislation.     

Copper Accumulation in a Reservoir Ecosystem Following Copper Sulfate Treatment (St. Germain Les Belles, France)

Copper sulfate (CuSO₄) addition to freshwater for phytoplankton control has been practiced for decades, and remains the most effective algicidal treatment for numerous managed water bodies. A reservoir in the centre of France was the site for an investigation of copper distribution in aquatic systems after a copper sulfate treatment. Results of monitoring showed a rapid conversion of dissolved Cu to particulate forms, with significant accumulation in the sediments. Total sediment Cu content increased from approximately 37.7 to 45.4 μg g^-1 dry weight after the first treatment. Sequential extraction suggested that a significant portion of the sediment-borne Cu was associated with the organic fraction which may release Cu to the water column, although significant release would occur only under extreme changes in water chemistry. Based upon measured Cu concentrations, flows at the down-stream water, and known mass applied during treatment, mass balance calculations indicated that approximately 17% of the Cu was exported from the reservoir over a 70 day period following a 196 μg L^-1 Cu²⁺ (as CuSO₄·5H₂O) treatment.Copper bioaccumulation by a moss, Fontinalis antipyretica in the down-stream water showed that it was possible to distinguish between a treated and an untreated area. The impact of copper treatmentin the down-stream reservoir could be followed using mosses. There is a distance effect which could be exploited to determine potential copper impact for example. The largest amount of copper was probably adsorbed on downstream sediment or lost in running water. Thirty days after copper sulfate addition, Fontinalis still indicatedcopper exposure.     

Harmonisation, standardisation for soil and sediment fractionation studies, and usefulness in assessment and risk management

Single and sequential extraction procedures are often used to study the environmental fate of trace elements from soils and sediments. Although these schemes are quite popular, they are prone to various sources of errors, which require them to be thoroughly controlled. Owing to their operationally-defined character, the only way to compare analytical data is to stick closely to procedures that have been collaboratively studied and written in the form of an operating protocol. Groups of scientists have hence developed ‘harmonised’ procedures that are used as a reference for fractionation studies. A higher level is the formal standardisation of the procedures by official standardisation bodies, which also fulfils the need for achieving the comparability of data obtained by different laboratories from different countries. This paper discusses aspects of harmonisation (consensus obtained by groups of scientists) and standardisation (adoption of standards as mandated by an international organisation) as applied to single and extraction schemes for trace element fractionation studies, including quality control aspects.     

Evaluation of Compost Maturity,Hydrophysical and Physicochemical Properties: Indicators for Use as a Component of Growing Media

The cocomposting of agricultural waste is a new management priority in Tunisia. In this study, four composts were evaluated by comparing the changes in measured hydrophysical and physicochemical properties and phytotoxicity. The organic wastes used were almond shell (AS), sesame bark (SB), olive husk (OH), and green and wood wastes (GW and WW, respectively). Composts CI and CII were composed of AS/SB and OH/SB, respectively, at a ratio of 75/25 (wet weight basis). CIII consisted of OH, SB and CAS (coarsely-ground almond shell used as a bulking agent) at a ratio of 55/25/20. Finally, CIV was composed of 25%SB+9%CAS+18%GW+48%WW. The composts studied were characterized by basic pH and an electric conductivity (EC) value ranging from 1.6 to 2.4 mS/cm. The organic matter contents (OM) and C/N ratios of composts ranged from 20 to 46% and from 10 to 21%, respectively. Based on hydrophysical analyses, composts CI, CIII and CIV, containing AS, were shown to have a porosity and a water content of 10-26% and 10-20%, respectively. The phytotoxicity of composts was studied on the basis of cress seed germination. Results revealed that differences in properties are mainly related to the nature of composted waste and that some of these composts are compatible for use as constituents in growing media for horticultural soilless cultures.     

Surrogate Correlations and Near-Infrared Diffuse Reflectance Sensing of Trace Metal Content in Soils

Near-infrared diffuse reflectance sensing (NIRS) of soils has been the object of considerable interest and research in the last few years. This has been motivated by the prospect that this method seems to provide a cheap, convenient alternative to conventional, time-consuming methods for the measurement of a wide range of soil parameters. In particular, various authors have advocated that NIRS could be used to measure rapidly and non-destructively the concentration of trace metals in surface soils. Correlation analyses between NIRS spectra and trace metal concentration have yielded inconclusive results to date, suggesting that trace metal concentration may belong to a class of “tertiary” soil parameters, linked to NIRS spectra through “surrogate”, or indirect, correlations, involving some other primary or secondary parameter like clay or organic matter content, to which NIRS spectra are very sensitive. To assess the validity of this surrogate correlation hypothesis in the case of trace metals, experiments were carried out with soil samples varying only in the amount of trace metals they contain. Field-aged Hudson and Arkport soil pots spiked with Cu and Zn, freshly spiked samples of the same soils, and samples of a metalliferous peat soil from Western New York naturally rich in Cd and Zn were subjected to NIRS under laboratory conditions. Detailed analysis indicates that the NIR spectrum is sensitive to sample handling, including the orientation of the samples in the NIRS instrument, but that, at the same time, there is no discernable effect of the presence of trace metals on any part of the NIR spectrum. These results provide strong experimental support to the hypothesis of “surrogate” correlation for trace metals, and indicate that trace metals, even in severely contaminated soils, should not interfere with the NIR sensing of primary or secondary parameters, like organic matter content. Further work is needed to determine if this feature of NIR spectra extends to other soil chemical parameters.     

Molecular characterization and PCR detection of a nitrogen-fixing Pseudomonas strain promoting rice growth

Nitrogen-fixing plant growth-promoting rhizobacteria (PGPR) from the genus Pseudomonas have received little attention so far. In the present study, a nitrogen-fixing phytohormone-producing bacterial isolate from kallar grass (strain K1) was identified as Pseudomonas sp. by rrs (16S ribosomal RNA gene) sequence analysis. rrs identity level was high with an uncharacterized marine bacterium (99%), Pseudomonas sp. PCP2 (98%), uncultured bacteria (98%), and Pseudomonas alcaligenes (97%). Partial nifH gene amplified from strain K1 showed 93% and 91% sequence similarities to those of Azotobacter chroococcum and Pseudomonas stutzeri, respectively. The effect of Pseudomonas strain K1 on rice varieties Super Basmati and Basmati 385 was compared with those of three non-Pseudomonas nitrogen-fixing PGPR (Azospirillum brasilense strain Wb3, Azospirillum lipoferum strain N4 and Zoogloea strain Ky1) used as single-strain inoculants. Pseudomonas sp. K1 was detected in the rhizosphere of inoculated plants by enrichment culture in nitrogen-free growth medium, which was followed by observation under the microscope as well as by PCR using a rrs-specific primer. For both rice varieties, an increase in shoot biomass and/or grain yield over that of noninoculated control plants was recorded in each inoculated treatment. The effect of Pseudomonas strain K1 on grain yield was comparable to those of A. brasilense Wb3 and Zoogloea sp. Ky1 for both rice varieties. These results show that nitrogen-fixing pseudomonads deserve attention as potential PGPR inoculants for rice.     

No-till only increases N2O emissions in poorly-aerated soils

Denitrification rates are often greater in no-till than in tilled soils and net soil-surface greenhouse gas emissions could be increased by enhanced soil N₂O emissions following adoption of no-till. The objective of this study was to summarize published experimental results to assess whether the response of soil N₂O fluxes to the adoption of no-till is influenced by soil aeration. A total of 25 field studies presenting direct comparisons between conventional tillage and no-till (approximately 45 site-years of data) were reviewed and grouped according to soil aeration status estimated using drainage class and precipitation during the growing season. The summary showed that no-till generally increased N₂O emissions in poorly-aerated soils but was neutral in soils with good and medium aeration. On average, soil N₂O emissions under no-till were 0.06 kg N ha⁻¹ lower, 0.12 kg N ha⁻¹ higher and 2.00 kg N ha⁻¹ higher than under tilled soils with good, medium and poor aeration, respectively. Our results therefore suggest that the impact of no-till on N₂O emissions is small in well-aerated soils but most often positive in soils where aeration is reduced by conditions or properties restricting drainage. Considering typical soil C gains following adoption of no-till, we conclude that increased N₂O losses may result in a negative greenhouse gas balance for many poorly-drained fine-textured agricultural soils under no-till located in regions with a humid climate.     

Effect of the Pietrain breed used as terminal boar on Post-weaning Multisystemic Wasting Syndrome (PMWS) in the offspring in four PMWS-affected farms

The aim of this study was to determine the potential risk factors for PMWS at the individual pig level and assess the effect of the Pietrain paternal genetic background of the animals in a cohort study. The survey was set up in four PMWS-affected farms with 2 repetitions (batches) per farm. A representative sample of 60 pigs per batch, stratified according to the paternal genetic background (Pietrain: yes vs. no), was randomly selected after farrowing. The representative cohort was divided into 8 batches and the pigs were individually monitored from birth to slaughter. Survival analysis was used to determine the factors related to the time to PMWS. The litter-cluster effect was taken into account using the marginal Cox model (robust estimation of the covariance matrix) and the gamma shared frailty model which were compared.No protective effect of the Pietrain breed on the time to PMWS and the proportion of affected pigs in the offspring was found in this study. Piglets showing low circovirus type 2 (PCV2) titres at 7 weeks-old with no subsequent seroconversion and piglets from PCV2 negative sows were most likely to be affected by PMWS (HR=7.0 and 2.8, respectively). Active infection of the pregnant dams with parvovirus was related to an increased risk of PMWS in the offspring (HR=2.3). Neck injuries due to poorly performed injections in the dams were associated with an increased risk of PMWS with the marginal model (HR=2.1). Oxytocin injection (dams) during farrowing was protective against PMWS in the offspring (HR=0.6).