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Abstract Currently food fraud and authenticity of products composition are topics of great concern; ingredients quantification could allow to identify small amounts of contaminats or voluntary addition of improper components. Many molecular methods are available for species identification in foodstuffs but, for a better application, they should not be affected by the interference of other ingredients. The main purpose of this work was to verify the Real Time PCR and the Digital PCR (dPCR) quantification performances on baby food samples, specifically selected for their high miscibility to limit variability; chicken was selected as target to verify the performance of quantification of methods after having spiked the same quantity in different baby foods. The other aims were: (1) to verify a constant genome copies ratio existence between mammalian and avian species (2) to verify the dPCR performance, set up on housekeeping, to quantify mammalian and avian species in commercial products. Digital PCR showed fewer differences respect to Real Time PCR, at the same 15% w/w chicken spiking level. Despite the constant difference between mammalian and avian genome copies, in samples with the same spiking weight, the confidence intervals increasing towards the extreme values, made impossible to use genome copies ratio as a sort of correction factor between species. Finally, the dPCR system using the myostatin housekeeping gene to determine the chicken content seemed reliable to verify the labelling compliance in meat-based commercial products.

Simple Summary: Photobacterium damselae subsp. damselae (Pdd) is a marine bacterium that can infect a variety of marine animals and humans. Although this bacterium has been isolated from several stranded dolphins and whales, its pathogenic role in cetaceans is still unclear and very limited information exists on its occurrence in these animals. In this study, we report data relating to the presence of Pdd in marine mammals stranded within the Liguria Pelagos Sanctuary from 2017 to 2022. Our findings show a 41.5% (22/53) Pdd prevalence in stranded cetaceans, where 22.7% (5/22) of these were positive in at least one of the Pdd virulence factors which, in previous studies, have been related to Pdd pathogenicity. Our results also revealed that in all cases where cetaceans tested positive for Pdd, other well-known pathogens for these species were also present. This finding supports the hypothesis that Pdd is an opportunistic agent that might contribute to the worsening of health conditions in subjects already compromised by other pathogens, contributing to their death. However, further studies are necessary to investigate and deepen this hypothesis. Photobacterium damselae subsp. damselae (Pdd) is an increasingly common bacterium in post-mortem diagnostics of beached marine mammals, but little is known about its precise etiological responsibility. To estimate the prevalence of Pdd in stranded cetaceans from 2017 to 2022 on the Ligurian coast (Pelagos Sanctuary), we tested tissues from 53 stranded individuals belonging to four cetacean species. DNA extracts from cetacean tissue were screened using a polymerase chain reaction (PCR) assay targeting the Pdd ureC gene. Positive samples were screened by PCR for dly, hlyApl and hlyAch hemolysin genes, which were confirmed by sequencing. Twenty-two out of 53 (41.5%) cetaceans analyzed by PCR were confirmed for Pdd DNA in at least one tissue among those analyzed. Five of these cetaceans were positive for at least one of the hemolysin genes tested. In all Pdd-positive cetaceans, other pathogens that were considered responsible for the causa mortis of the animals were also found. The results provide new information on the spread of Pdd in cetaceans and support the thesis that Pdd might be an opportunistic agent that could contribute to worsening health conditions in subjects already compromised by other pathogens. However, further studies are needed to investigate and deepen this hypothesis. [ABSTRACT FROM AUTHOR]

Simple Summary: Oysters are sessile, filter-feeding bivalves that are vital to fisheries and ecology but are poorly understood in terms of genetic diversity and distribution. This study examines small flat oysters in the Mediterranean regions of Liguria and Sardinia (Italy) using 16S rRNA sequencing to identify species. The findings reveal new insights into the Ostrea stentina complex and introduce a new species, O. neostentina, improving knowledge of Ostrea species diversity and distribution. Oysters are sessile, filter-feeding bivalve molluscs widely distributed in estuarine and coastal habitats worldwide. They constitute a valuable resource for fisheries and extensive aquaculture and provide essential ecological services. Yet, their genetic diversity and distribution remain understudied. The variability in shell morphology complicates species classification, which is influenced by environmental and genetic factors. Although molecular phylogenetics research has refined oyster taxonomy and identified approximately 100 extant species, numerous taxonomic uncertainties persist. In the present study, we aimed to document the occurrence of small flat oysters of the genus Ostrea along the Mediterranean coastal areas of Liguria and Sardinia (Italy). Specifically, 16S rRNA sequence data were used to identify Ostrea species. Our findings offer novel insights into the O. stentina species complex and O. neostentina, a new species in the Mediterranean coastal areas of Italy. The study data further our understanding of Ostrea species diversity, distribution, and evolutionary patterns. [ABSTRACT FROM AUTHOR]

Simple Summary: The preservation of biological diversity is one of the main tasks of modern ecology. A decrease in the bee population for various reasons can negatively affect plant diversity and human economic activity. One of the most critical factors in maintaining the health of bee populations is the availability of food supplies. We compared the food preferences of bees in different ecological areas: an intensive farming area, a reserved area, and an urbanized area. The botanical composition of the honey was determined by the traditional method of melissopalynology, as well as using genetic analysis. It was found that, in the intensive farming area, the leading food for bees was monoculture plants, but proper apiary management made the diversification of the bees' diets possible. In the reserved area, the bee diet was supplemented mostly with sown plants. In urban areas, both garden and agricultural plants served as the primary food sources for bees. Bee foraging depends on the climatic and geographical conditions of the environment, as well as landscape and apiary management. Studying bee foraging is essential to understanding their use of food resources and plant preferences. This information can be used for effective measure management in order to preserve the number of bees and regulate plant pollination. The environment significantly impacts the lives of bees and their feeding. This study aimed to investigate bee foraging using melissopalynological analysis and DNA metabarcoding in intensive farming, reserved, and urbanized areas. The highest alpha diversity was observed in the reserved and intensive farming areas. The urbanized area had less diversity. In the intensive farming area, Sinapis, Helianthus, and Fagopyrum predominated; in the reserved area, Melilotus, Helianthus, and Brassica predominated. In the urbanized area, garden plants, namely radish (Raphanus sativus) and cucumber (Cucumis hystrix), and agricultural plants, namely soybean (Glycine max) and melon (Cucumis melo), were often found. The most significant agreement was between the rbcL and the melissopalynological analysis. The ITS2 revealed equal matches with both rbcL and melissopalynology, but this marker missed or underestimated some genera. Trifolium pretense and Brassica nigra were identified simultaneously by the melissopalinology method and two genetic markers in DNA metabarcoding. The species Convolvulus arvensis, Melilotus officinalis, Echium vulgare, Brassica rapa, Helianthus divaricatus, and Onobrychis viciifolia were found in all ecological areas. Imperfect databases impose some limits in the identification of some taxa using metabarcoding. The further research and expansion of plant databases is needed. Studying the food preferences of bees in different environmental conditions and landscapes is necessary to develop measures to preserve their populations. [ABSTRACT FROM AUTHOR]

The hippoboscid Lipoptena fortisetosa Maa, 1965 is a hematophagous ectoparasite of cervids that can bite humans. This fly is expanding its geographical range and is of concern for animal and human health since it can potentially harbour harmful microorganisms. This study was aimed at characterizing the bacterial communities of L. fortisetosa in its different life‐cycle stages. Pupae and wingless adults were collected from cervids hunted in Tuscan‐Emilian Apennines (central Italy) and pooled into groups of 10 by life stage (30 individual pupae; 1420 individual wingless adults). Winged flies were caught by sweep netting and separated into five pools of 10 insects. After DNA extraction, the bacterial content of each pool was analysed using 16 S metabarcoding. Results revealed that the composition and relative abundance of different taxa greatly differed in the three analysed groups. Wingless adults showed a high abundance of Bartonella (33.07%), which is almost absent in winged flies and pupae. Among the detected pathogens, four genera of concern for human health were found: Bartonella, Moraxella, Mycobacterium and Rickettsia. Interestingly reads similar to Bartonella bovis, Moraxella osloensis and Arsenophonus lipopteni Operational Taxonomic Unit (OTUs) were detected. These findings suggest the possible role of L. fortisetosa as a reservoir of pathogenic microorganisms, confirming the need for further investigation to ascertain its vectorial capacity. [ABSTRACT FROM AUTHOR]