Molecular approaches are increasingly used to help in species diagnostics. These approaches have been recently and successfully applied to assess some taxonomic questions within the mite family Phytoseiidae. However, many protocols for DNA extraction of such small specimens require crushing the entire sample, precluding deposition of the carcass as a museum voucher. This study aimed to determine the efficiency of a modified Qiagen DNeasy tissue kit extraction method to both extract enough DNA for performing PCR and DNA sequencing, and recover intact the adult female specimens for identification. Two types of biological material were studied for two species: 100 % alcohol preserved females and females mounted in lactic acid for four hours. All specimens were retrieved after DNA extraction and were mounted on slides. All the structures used for species identification were visible with a phase microscope. DNA was successfully extracted from all the specimens considered and fragments of CytB mtDNA of 426 bp were amplified and corresponded to those of the two species considered. The Qiagen DNeasy tissue kit extraction protocol thus permits to retrieve, prepare and identify mites for which DNA has been successfully extracted, whatever the material considered: alcohol preserved or lactic acid treated mites. This result is of major interest for future taxonomic studies of the family Phytoseiidae, as it will allow the assignment of morphological and molecular attributes to a same specimen.

Antibiotic resistance is a serious concern for public health. Farm environments are relevant reservoirs of antibiotic resistant bacteria and antibiotic resistance genes ( ARGs ), thus strategies to limit the spread of ARGs from farms to the environment are needed. In this study a broiler farm, where antibiotics have never been used for any purpose, was selected to evaluate if this measure is effective in reducing the ARGs load in farm environment ( FE ) and in meat processing environment ( MPE ). Faecal samples from FE and MPE were processed for DNA extraction. Detection and quantification of the 16S rRNA gene and selected ARGs ( bla TEM , qnr S, sul 2, and tet A) were carried out by PCR and digital droplet PCR ( ddPCR ), respectively. Generally, the relative abundance of the quantified ARGs in FE was similar or higher than that measured in intensive farms. Furthermore, apart for tet A, no differences in relative abundances of the other ARGs between FE and MPE were determined. These results suggest that the choice to not use antibiotics in broiler farming is not so effective to limit the ARGs spread in MPE and that further sources of ARGs should be considered including the preceding production phase with particular reference to the breeding stage.

Tetrodotoxins (TTXs) are a group of potent neurotoxins named after the Tetraodontidae fish family (pufferfish). TTXs have been reported in several animal taxa, both terrestrial and marine. The ingestion of TTX‐contaminated flesh can cause serious neurotoxic symptomatology and can eventually lead to death. Traditionally, TTXs have been associated with Asian countries, in particular with pufferfish consumption. However, they have also been reported in bivalve mollusks farmed in the Pacific area and, recently, in European seas. In Europe, different countries have reported TTXs, especially those bordering the Mediterranean Sea. As a consequence, in 2017 the European Food Safety Authority (EFSA) released an opinion with reference to TTX present in marine gastropods and bivalves, proposing a safety limit of 44 µg/kg TTXs in shellfish meat, below which no adverse effects should be observed in humans. Nevertheless, this limit has been exceeded on many occasions in European shellfish and, while for bivalves there have been no registered human intoxications, that is not the case for marine gastropods. However, TTXs have not yet been included in the list of marine biotoxins officially monitored in live bivalve mollusks within the European Union (EU). Thus, the aims of this manuscript are to discuss the increasing occurrence of TTXs in live bivalve mollusks from European sea waters, to acknowledge the still ongoing knowledge gaps that should be covered and to stimulate constructive debate on the eventuality of adopting a shared regulatory context, at least in the EU, for monitoring and managing this potential threat to food safety.

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A side effect of antibiotic usage is the emergence and dissemination of antibiotic resistance genes (ARGs) within microbial communities. The spread of ARGs among pathogens has emerged as a public health concern. While the distribution of ARGs is documented on a global level, their routes of transmission have not been clarified yet; for example, it is not clear whether and to what extent the emergence of ARGs originates in farms, following the selective pressure exerted by antibiotic usage in animal husbandry, and if they can spread into the environment. Here we address this cutting edge issue by combining data regarding antimicrobial usage and quantitative data from selected ARGs (blaTEM, blaCTXM, ermB, vanA, qnrS, tetA, sul2, and mcr-1) encoding for resistance to penicillins, macrolides-lincosamides-streptogramins, glycopeptides, quinolones, tetracyclines, sulfonamides, and colistin at the farm level. Results suggest that dairy farms could be considered a hotspot of ARGs, comprising those classified as the highest risk for human health and that a correlation existed between the usage of penicillins and blaTEM abundances, meaning that, although the antibiotic administration is not exclusive, it remains a certain cause of the ARGs' selection and spread in farms. Furthermore, this study identified the role of calves as the main source of ARGs spread in dairy farms, claiming the need for targeted actions in this productive category to decrease the load of ARGs along the production chain.