Development and validation of rapid multiresidue and multi-class analysis for antibiotics and anthelmintics in feed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry

A new multi-residue method for the analysis of veterinary drugs, namely amoxicillin, chlortetracycline, colistins A and B, doxycycline, fenbendazole, flubendazole, ivermectin, lincomycin, oxytetracycline, sulfadiazine, tiamulin, tilmicosin and trimethoprim, was developed and validated for feed. After acidic extraction, the samples were centrifuged, purified by SPE and analysed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry. Quantitative validation was done in accordance with the guidelines laid down in European Commission Decision 2002/657/CE. Matrix-matched calibration with internal standards was used to reduce matrix effects. The target level was set at the authorised carryover level (1%) and validation levels were set at 0.5%, 1% and 1.5%. Method performances were evaluated by the following parameters: linearity (0.986 < R² < 0.999), precision (repeatability < 12.4% and reproducibility < 14.0%), accuracy (89% < recovery < 107%), sensitivity, decision limit (CCα), detection capability (CCβ), selectivity and expanded measurement uncertainty (k = 2).This method has been used successfully for three years for routine monitoring of antibiotic residues in feeds during which period 20% of samples were found to exceed the 1% authorised carryover limit and were deemed non-compliant.     

Peptides in rainbow trout (Oncorhynchus mykiss) muscle subjected to ice storage and cooking

Although proteolysis during post mortem storage is an important factor affecting fish texture, little is known about degradation end-products. This study was performed to investigate the occurrence of low molecular weight peptides (<5 kDa) in post mortem rainbow trout muscle, during ice storage, and to evaluate their stability during cooking. It combined quantitative (amino acid analysis) and qualitative approaches (mass spectrometry). The results showed that muscle of trout was poor in peptides. These were mainly anserine and glutathione. Their concentration was almost unaffected by the seven days of ice storage and vacuum cooking for 5 min at 70 °C. MS analysis revealed a limited but highly reproducible appearance of small peptides in trout muscle during the ice storage and after cooking. The compounds detected by MS analysis remain to be characterised.     

Influence of whip speed ratios on the inclusion of air into a bakery foam produced with a planetary mixer device

In the present study, dimensional analysis has been developed in order to identify the ratios governing the foam development network, with mixers combining dual whip revolution speeds. It is first suggested that, for a given planetary whip mixer and foaming solution, the set of dimensionless ratios characterizing the course of gas hold-up can be reduced to modified Froude and revolution numbers, as in the case of a classical mixing system. This requires the introduction of a characteristic velocity, instead of the two individual rotational speeds of the whip, in the corresponding list of governing parameters. An analytical expression for this characteristic velocity is then proposed, and gas hold-up measurements are made at various whip speed ratios, using a given food model-recipe, containing whole eggs and sugar. Our experimental results were evaluated, in order to validate the proposed dimensional analysis. It is shown that the aeration process requires a very low, modified revolution number to reach steady state values. It is also shown that the asymptotic values of gas hold-up are influenced by the modified Froude number. Depending on the values of the modified Froude number, it is noted that coalescence can occur, which then contributes to a reduction in gas hold-up. It has also been established that the modified Froude number can account for the eccentricity of planetary mixers, and consequently, that this number is a robust indicator for the determination of the optimum gas hold-up value, when air incorporation is produced by various planetary whipping devices. The results described in this paper enable foaming processes using planetary whipping mixers to be optimized, by proposing a framework in which the impeller speed ratio and aeration time can be set in such a way as to control the simultaneous entrainment and disentrainment of gas. Finally, this work can be further extended to other gas/liquid applications in which planetary mixers are used, such as surface aeration in wastewater treatment plants.     

Variations of DOM quality in inflows of a drinking water reservoir: linking of van Krevelen diagrams with EEMF spectra by rank correlation

Elevated concentrations of dissolved organic matter (DOM) such as humic substances in raw water pose significant challenges during the processing of the commercial drinking water supplies. This is a relevant issue in Saxony, Central East Germany, and many other regions worldwide, where drinking water is produced from raw waters with noticeable presence of chromophoric DOM (CDOM), which is assumed to originate from forested watersheds in spring regions of the catchment area. For improved comprehension of DOM molecular composition, the seasonal and spatial variations of humic-like fluorescence and elemental formulas in the catchment area of the Muldenberg reservoir were recorded by excitation emission matrix fluorescence (EEMF) and ultrahigh-resolution mass spectrometry (FT-ICR-MS). The Spearman rank correlation was applied to link the EEMF intensities with exact molecular formulas and their corresponding relative mass peak abundances. Thereby, humic-like fluorescence could be allocated to the pool of oxygen-rich and relatively unsaturated components with stoichiometries similar to those of tannic acids, which are suspected to have a comparatively high disinfection byproduct formation potential associated with the chlorination of raw water. Analogous relationships were established for UV absorption at 254 nm (UV(254)) and dissolved organic carbon (DOC) and compared to the EEMF correlation.     

Development of a rapid, specific and efficient procedure for the determination of proteolytic activity in dry-cured ham: definition of a new proteolysis index

A method was adapted to determine proteolytic activity in dry-cured ham using fluorescamine-specific labelling of N-terminal α-amino groups of peptides and amino acids. Fluorescence of the complex was measured using a microplate procedure and optimum excitation and emission wavelengths of 375nm and 475nm, respectively. A new proteolysis index (PI) was defined as the percentage ratio of the N-terminal α-amino group content to the total protein content of the ham extract. The robustness of the method was evaluated by measuring PI in pork meat samples subjected to standardized processing conditions and in samples extracted from industrial hams taken at different stages of processing. For the industrial samples, a comparison with the classic nitrogen procedure of PI determination was performed and a formula relating the two PIs was established. The rapidity, sensitivity and specificity of the procedure make it a good candidate for a screening test to evaluate ham quality in industry.     

Milk β-lactoglobulin complexes with tea polyphenols

The effect of milk on the antioxidant capacity of tea polyphenols is not fully understood. The complexation of tea polyphenols with milk proteins can alter the antioxidant activity of tea compounds and the protein secondary structure. This study was designed to examine the interaction of β-lactogolobulin (β-LG) with tea polyphenols (+)-catechin (C), (−)-epicatechin (EC), (−)-epicatechin gallate (ECG) and (−)-epigallocatechin gallate (EGCG) at molecular level, using FTIR, CD and fluorescence spectroscopic methods as well as molecular modelling. The polyphenol binding mode, the binding constant and the effects of polyphenol complexation on β-LG stability and secondary structure were determined. Structural analysis showed that polyphenols bind β-LG via both hydrophilic and hydrophobic interactions with overall binding constants of K_C–β-LG = 2.2 (±0.8) × 10³ M⁻¹, K_EC–β-LG = 3.2 (±1) × 10³ M⁻¹, K_ECG–β-LG = 1.1 (±0.6) × 10⁴ M⁻¹ and K_EGCG–β-LG = 1.3 (±0.8) × 10⁴ M⁻¹. The number of polyphenols bound per protein molecule (n) was 1.1 (C), 0.9 (EC), 0.9 (ECG) and 1.3 (EGCG). Molecular modelling showed the participation of several amino acid residues in polyphenol–protein complexation with extended H-bonding network. The β-LG conformation was altered in the presence of polyphenols with an increase in β-sheet and α-helix suggesting protein structural stabilisation. These data can be used to explain the mechanism by which the antioxidant activity of tea compounds is affected by the addition of milk.     

Antifungal activity of 2 lactic acid bacteria of the Weissella genus isolated from food

Abstract: In the present study, a total of 116 lactic acid bacteria (LAB) strains isolated from Mill flour and fermented cassava were screened for their antifungal activity. Three strains among 116 were selected for their strongest inhibitory activity against food molds. These 3 strains were Lactobacillus plantarum VE56, Weissella cibaria FMF4B16, and W. paramesenteroides LC11. The compounds responsible for the antifungal activity were investigated. The strains displayed an inhibitory activity against targeted molds at acidic pH. However, the influence of organic acids was rejected according to the calculated minimal inhibitory concentration (MIC). Antifungal compounds were investigated in the cell‐free supernatants and phenyllactic acid (PLA) was detected in different amounts with a maximal concentration for Lb. plantarum VE56 (0.56 mM). Hydroxy fatty acid, such as 2‐hydroxy‐4‐methylpentanoic acid, was also produced and involved in the inhibitory activity of Lb. plantarum VE56 and W. paramesenteroides LC11. Antifungal LAB are known to produce PLA and 3‐hydroxy fatty acids and other organic acids with antifungal activity. This short communication focuses on antifungal activity from Weissella genus. The antifungal activity was attributed to antifungal compounds identified such as PLA, 2‐hydroxy‐4‐methylpentanoic acid, and other organic acids. Nevertheless, the concentration produced in the cell‐free supernatant was too low to compare to their MIC, suggesting that the inhibitory activity was caused by a synergy of these different compounds. Practical Application: Antifungal LAB are interesting to prevent food spoilage in fermented food and prolong their shelf life. In this way, chemical preservatives could be avoided and replaced by natural preservatives.     

Effect of diesel oxidation catalysts on the diesel particulate filter regeneration process

A Diesel Particulate Filter (DPF) regeneration process was investigated during aftertreatment exhaust of a simulated diesel engine under the influence of a Diesel Oxidation Catalyst (DOC). Aerosol mass spectrometry analysis showed that the presence of the DOC decreases the Organic Carbon (OC) fraction adsorbed to soot particles. The activation energy values determined for soot nanoparticles oxidation were 97 ± 5 and 101 ± 8 kJ mol(-1) with and without the DOC, respectively; suggesting that the DOC does not facilitate elementary carbon oxidation. The minimum temperature necessary for DPF regeneration was strongly affected by the presence of the DOC in the aftertreatment. The conversion of NO to NO(2) inside the DOC induced the DPF regeneration process at a lower temperature than O(2) (ΔT = 30 K). Also, it was verified that the OC fraction, which decreases in the presence of the DOC, plays an important role to ignite soot combustion.     

Evaluation of Ba2(In0.8Ti0.2)2O5.2−n(OH)2n as a potential electrolyte material for proton-conducting solid oxide fuel cell

Electrochemical measurements of fuel cells based on proton conductor electrolyte Ba₂(In_0.8Ti_0.2)₂O_5.2−n(OH)_2n and prepared through a tape casting process and a co-pressing of anode-composite powder and electrolyte tape were performed at 500 °C under wet H₂. The varying parameter between the prepared cells was the thickness of the electrolyte that can be controlled during the tape casting process. The maximum power density was obtained for the cell with the thinnest electrolyte (35 μm) and was about 22 mW cm⁻² with an ohmic resistance about 2 Ω cm² at 500 °C.     

Comparative analysis of CRISPR loci in lactic acid bacteria genomes

Clustered regularly interspaced short palindromic repeats (CRISPR) are hypervariable loci widely distributed in bacteria and archaea, that provide acquired immunity against foreign genetic elements. Here, we investigate the occurrence of CRISPR loci in the genomes of lactic acid bacteria (LAB), including members of the Firmicutes and Actinobacteria phyla. A total of 102 complete and draft genomes across 11 genera were studied and 66 CRISPR loci were identified in 26 species. We provide a comparative analysis of the CRISPR/cas content and diversity across LAB genera and species for 37 sets of CRISPR loci. We analyzed CRISPR repeats, CRISPR spacers, leader sequences, and cas gene content, sequences and architecture. Interestingly, multiple CRISPR families were identified within Bifidobacterium, Lactobacillus and Streptococcus, and similar CRISPR loci were found in distant organisms. Overall, eight distinct CRISPR families were identified consistently across CRISPR repeats, cas gene content and architecture, and sequences of the universal cas1 gene. Since the clustering of the CRISPR families does not correlate with the classical phylogenetic tree, we hypothesize that CRISPR loci have been subjected to horizontal gene transfer and further evolved independently in select lineages, in part due to selective pressure resulting from phage predation. Globally, we provide additional insights into the origin and evolution of CRISPR loci and discuss their contribution to microbial adaptation.