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Background Hyalomma marginatum is a hard tick vector of various pathogens, including Crimean-Congo Hemorrhagic fever virus, recently detected in French specimens. This species has a wide distribution from North Africa to Eastern Europe and has only recently been considered established in Southern France. These changes in species distribution led us to explore the genetic structure of tick populations in the Mediterranean basin and attempt to infer the origin of French populations. Methods We used two mitochondrial markers (12S rRNA and Cytochrome Oxidase 1) and genotyped ticks from nine Mediterranean countries. We compared genetic indices and haplotypic composition between these countries and the various French geographical populations. Results Across all countries, we showed significant genetic differentiation, with a certain proximity between neighboring countries. We found very different genetic compositions among the French geographic populations: some exhibited signs of recent expansion, while others suggested the presence of ancient populations. Conclusions It is possible that small populations of H. marginatum were already present in France and are now more abundant. This recent change in population structure could be owing to increased human activity and climate change. These factors, combined with a potentially high level of phenotypic plasticity, could facilitate H. marginatum conquest of more northerly latitudes in France and other European countries. Graphical Abstract

Hyalomma marginatum is an invasive tick species reported in regions bordering the Mediterranean Sea. It has recently become established in mainland France in 2016. H. marginatum hosts a diverse range of human and animal pathogens such as Rickettsia aeschlimannii, Theileria equi, Anaplasma phagocytophilum, Anaplasma marginale, Ehrlichia minasensis and West Nile virus. The objective of this study was to characterize the current spatial distribution of H. marginatum-borne pathogens in the region Occitanie, southern France and the influence of other factors (tick sex, engorgement status). A large-scale tick collection program was organized in the region Occitanie to answer these questions. A total of 510 ticks from 27 sites of four departments were analysed for pathogens presences and loads using both a high-throughput microfluidic real time PCR system and quantitative PCR. In the analysed ticks, we identified the tick-borne pathogens and their prevalence: Theileria equi (9.2%), Theileria orientalis (0.2%), Anaplasma phagocytophilum (1.6%), Anaplasma marginale (0.8%) and Rickettsia aeschlimannii (87.3%). Interestingly, we highlighted a spatial clustered distribution for the pathogen R. aeschlimannii between two geographically isolated areas with infection rates and bacterial loads significantly lower in Herault/Gard departments (infection rate 78.6% in average) compared to Aude/Pyrenees-Orientales (infection rate 92.3% in average). R. aeschlimannii infection rates varied from one site to another, ranging from 29% to 100%, highlighting the importance of the sampling effort to better characterize the circulation of pathogens. Such high infection rate (87.3% in average) and the effective maternal transmission of R. aeschlimannii suggest a role as a tick symbiont in H. marginatum. On the other hand, currently identified as a human pathogen, its pathogenic status in humans is questioned given the low number of human cases. Further studies are thus needed to understand both the status and the role of R. aeschlimannii in H. marginatum ticks.Competing Interest StatementThe authors have declared no competing interest.

Wildflower honeys produced in mountain grasslands are an expression of the biodiversity of these fragile habitats. Despite its importance, the botanical origin of honey is often defined without performing formal analysis. The aim of the study was to characterize six wildflower mountain honeys produced in the Italian Alps with different analytic techniques (SPME–GC–MS, HPLC-Orbitrap, cicatrizing and antioxidant activity) alongside melissopalynological analysis and botanical definition of the production area. Even though the apiaries were in mountain grasslands rich in Alpine herbaceous species, the honey could be defined as rhododendron/raspberry unifloral or raspberry and rhododendron bifloral while the honey produced at the lowest altitude differed due to the presence of linden, heather and chestnut. The non-compliance of the honey could be due to habitat (meadows and pastures) fragmentation, but also to specific compounds involved in the plant–insect relationship, such as kynurenic acid, present in a high quantity in the sample rich in chestnut pollen. 255 volatile compounds were detected as well as some well-known markers of specific botanic essences, in particular chestnut, linden and heather, also responsible for most of the differences in aroma profiling. A high correlation between nicotinaldehyde content and percentage of raspberry pollen (r = 0.853, p  < 0.05) was found. Phenolic acid and hydroxy-fatty acid were predominant in the chestnut pollen dominant honey, which presented the highest antioxidant activity and the lowest cicatrizing activity, while the flavonoid fraction was accentuated in one sample (rhododendron pollen prevalent), that was also the one with the highest effect on wound closure, although all samples had similar cicatrizing effects apart from the chestnut pollen dominant honey (lowest cicatrizing activity). Our study highlighted the difficulty of producing mountain wildflower honey and the importance of a thorough characterization of this product, also to encourage its production and valorisation.

Honey is a variegate matrix depending significantly on the floral origin, and it could become an important agri-food product to valorise specific territories. Being so diverse, different analytical techniques are necessary for its description. Herein we characterized the honey produced in one of the Italian sand dunes systems hosting beekeeping activities. In terms of floristic origin, phytochemical characterization, and sensory and colour analysis, honey collected in 2021 and 2022 was comparable. Honey was polyfloral, with several pollens from dune habitat plants classified as minor. The presence of the allochthonous Amorpha fruticosa L. and the ruderal Rubus fruticosus L. pollens in the category of the secondary pollens testifies the alteration of the park vegetation. The phytochemical profile was rich in polyphenols. Other interesting compounds were coumarine derivatives, likely attributable to resin-laden plants as rockroses, long chain hydroxyacids typical of royal jelly and nicotinic acid and its analogues (2-hydroxynicotinic acid and 2-hydroxyquinoline). The above-mentioned honey showed interesting features and was a good representation of the vegetation of this area. Our study pointed out the importance of relying on multiple analytical techniques for the characterization of honey and the advisability of a technical support toward beekeepers to correctly describe and valorise their product.

Sanguisorba dodecandra Moretti is endemic to the Italian Alps. It grows in very different habitats in contrast to most other endemics that tend to occupy specific niches. It is a resource for honeybees, but it is not clear if they or other insects act as the main pollinator. The volatile profile of S. dodecandra flowers has not been studied, nor its impact on the aroma of their respective honey. We investigated the volatile profiles of S. dodecandra flowers and honey, the insects visiting its inflorescences, and the seed set resulting from open pollination compared to insect exclusion treatments. S. dodecandra was mainly visited by Diptera, especially hoverflies, both in mid mountain (52.46%) and high mountain (41.12%) habitats. Open pollination resulted in a higher seed set (4.42 ± 3.14 seeds/cm) than from bagged inflorescences (0.96 ± 1.50 seeds/cm). Although bees were rarely recorded visiting S. dodecandra , pollen spectra of honey produced in the area where the endemic species grows showed its pollen and that of other plants increased when flowering of more attractive species, such as linden, declined. Terpenes were well represented in honey and flowers. Out of 104 compounds identified in honey containing S. dodecandra as an important component and 116 in the inflorescences only 15 were common to both. Some of these (e.g., rose oxide or jasmone) are characterized by a geranium-jasmine smell. Our study highlighted new aspects of the reproductive strategy of this endemic species, that is different from its more common relative, Sanguisorba officinalis L. Despite a floral morphology aligned with a wind pollination syndrome it may be an important resource for bees where it occurs.

Sanguisorba dodecandra Moretti is an endemic plant of the Alps of the Lombardy region (Northern Italy). Differently from most endemic species, this plant grows in diverse environments, and it is often very abundant and a distinctive element of some mountain and sub-alpine agro-ecosystems. The ecological features and the role of this species in some mountain agricultural activities are poorly investigated. This article shows the results of a synecological analysis of S. dodecandra and the evaluation of its functional strategy. Furthermore, its forage value was investigated and melissopalynological analysis was used to characterize the honey produced in an area where this species grows. The ecological analysis defined this plant as euriecious and ruderal/competitive-ruderal strategist. Bromatological analysis showed a good forage value, confirming the ethnobotanical knowledge concerning this species. In fact, it has good protein content (12.92 ± 1.89%) and non-fiber carbohydrates (47.12 ± 3.62%) in pre-flowering. S. dodecandra pollen was identified as a “frequent pollen” in the honey, showing that this plant is attractive to honeybees. This research allowed a deeper knowledge of S. dodecandra ecology and showed that this species is a resource for traditional and sustainable agricultural activities of the Lombardy Alps such as pastoralism and beekeeping.

Sanguisorba dodecandra Moretti is an endemic plant of the Alps of the Lombardy region (Northern Italy). Differently from most endemic species, this plant grows in diverse environments, and it is often very abundant and a distinctive element of some mountain and sub-alpine agro-ecosystems. The ecological features and the role of this species in some mountain agricultural activities are poorly investigated. This article shows the results of a synecological analysis of S. dodecandra and the evaluation of its functional strategy. Furthermore, its forage value was investigated and melissopalynological analysis was used to characterize the honey produced in an area where this species grows. The ecological analysis defined this plant as euriecious and ruderal/competitive-ruderal strategist. Bromatological analysis showed a good forage value, confirming the ethnobotanical knowledge concerning this species. In fact, it has good protein content (12.92 ± 1.89%) and non-fiber carbohydrates (47.12 ± 3.62%) in pre-flowering. S. dodecandra pollen was identified as a “frequent pollen” in the honey, showing that this plant is attractive to honeybees. This research allowed a deeper knowledge of S. dodecandra ecology and showed that this species is a resource for traditional and sustainable agricultural activities of the Lombardy Alps such as pastoralism and beekeeping.