Development of a PCR assay for the detection of animal tissues in ruminant feeds.

The European Community ban on use of meat and bone meal in ruminant feed, as a consequence of the spread of bovine spongiform encephalopathy in Europe, has prompted a number of investigations about the possibility of detecting animal tissues in feedstuff. In this paper, a study on vertebrate primers, designed in the 16S rRNA gene of mitochondrial DNA, is described. These primers were able to amplify fragments that contained between 234 and 265 bp. The fragments were specific for bovine, porcine, goat, sheep, horse, rabbit, chicken, trout, and European pilchard and were confirmed by sequence analysis amplicons. The primers were used in a PCR assay applied to five samples of meat and blood meals of different species and subjected to severe rendering treatments (134.4 to 141.9 degrees C and 3.03 to 4.03 bar for 24 min). The presence of vertebrate tissues was detected in all samples. The assay proved to be rapid and sensitive (detection limit 0.0625%). It can be used as a routine method to detect animal-derived ingredients in animal feedstuff

A portable setup for the voltammetric determination of total mercury in fish with solid and nanostructured gold electrodes

A simple procedure for field fish sample pretreatment was developed. This treatment in combination with square wave anodic stripping voltammetry (SW-ASV) with solid gold electrodes (SGE) and gold nanoparticle-modified glassy carbon electrodes (AuNPs-GCE) was applied for the determination of total mercury content. A certified reference material (CRM, Tuna Fish BCR 463), ten freeze-dried samples of canned tuna and two fresh fish samples were analysed both with a bench-top voltammetric analyser after microwave digestion and with a portable potentiostat after mild eating using a small commercial food warmer. The results obtained by the two SW-ASV approaches and by a Direct Mercury Analyser (DMA), the official method for mercury determination, were in very good agreement. In particular, (i) the results obtained with in field procedure are consistent with those obtained with the conventional microwave digestion; (ii) the presence of gold nanoparticles on the active electrode surface permits an improvement of the analytical performance in comparison to the SGE: the Limit of Quantification (LOQ) for mercury in fish-matrix was 0.1 μg L−1 (Hg cell concentration), corresponding to 0.06 mg kg−1 wet fish, which is a performance comparable to that of DMA. The pretreatment proposed in this study is very easy and applicable to fresh fish; in combination with a portable potentiostat, it proved to be an interesting procedure for on-site mercury determination

Occurrence of rare earth elements in water, sediment, and freshwater fish of diverse trophic levels and feeding ecology: Insights from the Po river (northwest Italy)

To date, the occurrence of rare earth elements (REEs) in freshwater ecosystems has garnered limited attention in the scientific literature. Furthermore, a dearth of data exists regarding their potential bioaccumulation in freshwater fish. To fill this knowledge gap, we studied REEs concentrations in water, sediment, and fish specimens collected along the Po River (northwest Italy) and calculated biota-sediment accumulation (BSAF) and bioconcentration (BCF) factors, while taking into account fish feeding behavior and trophic level effects on the overall content of total REEs (ƩREEs). The fish communities were composed of native and non-native species. Remarkably low concentrations of REEs (<0.0003 mg/L) were detected in the water samples, indicating REEs insolubility. In contrast, sediment samples were found to be a good sink for REEs, with a higher mean ƩREEs recorded for the samples from the Moncalieri station (70.93 mg/kg). Notably, no significant differences in ƩREEs concentration were observed in the muscle tissue of fish samples from the three stations. The highest mean ƩREEs was recorded in the samples from the Murazzi station (0.027 mg/kg). The BSAF was very low, consistently below the unit, indicating an absence of bioaccumulation in fish muscle from sediment. In contrast, the BCF was high for several REEs, mainly for Sc and Y. While feeding ecology did not appear to affect REEs accumulation in muscle, there was a significant negative relationship between the trophic level and ΣREEs, indicating a trophic dilution of REEs from predator (Silurus glanis) to planktivorous (Alburnus arborella) fish. This study provides baseline concentrations, trophic transfers, and patterns of REEs in a river system. Further studies are needed to understand the transfer of REEs to other biotic components of lotic ecosystems.The authors would like to thank Paolo Lo Conte (Città Metropolitana di Torino) for the fish sampling and for his technical support. This study was funded by Fondazione CRT “Richieste Ordinarie”, project: “Come stanno i pesci in città? Valutazione della comunità ittica nel tratto metropolitano del Fiume Po” (RF = 2018.2374).Peer reviewe