Ecotoxicological evaluation of an aqueous phytoextract of Melia azedarach L.

Melia azedarach L. is a Meliaceae that has shown important insecticidal activities. However, few researchers have extensively studied the toxicology of aqueous extracts of M. azedarach (MAE). Therefore, the main objective of this study was to characterize the phyto-eco-toxicological profile of MAE. First, a botanical and phytochemical characterization of MAE was performed using a histological, and metabolomic multi-analytical approach. Second, the toxicological effects on pollinating insects (Apis mellifera ligustica) and soil collembola (Folsomia candida) were evaluated. In addition, acute toxicity was evaluated in zebrafish (Danio rerio) to assess effects on aquatic fauna, and toxicity was determined in human neuroblastoma (SH-SY5Y) and fibroblast (FB-21) cell models. Finally, phytotoxic effects on germination of Cucumis sativus L., Brassica rapa L. and Sorghum vulgare L. were considered. Metabolomic analyses revealed the presence of not only limonoids but also numerous alkaloids, flavonoids and terpenoids in MAE. Histological analyses allowed us to better localize the areas of leaf deposition of the identified secondary metabolites. Regarding the ecotoxicological data, no significant toxicity was observed in bees and collembola at all doses tested. In contrast, severe cardiac abnormalities were observed in zebrafish embryos at concentrations as low as 25 µg/mL. In addition, MAE showed toxicity at 1.6 µg/mL and 6.25 µg/mL in FB-21 and SH-SY5Y cells, respectively. Finally, MAE inhibited seed germination with inhibitory concentrations starting from 5.50 µg/mL in B. rapa, 20 µg/mL in S. vulgare, and 31 µg/mL in C. sativus. Although M. azedarach extracts are considered valuable natural insecticides, their ecological impact cannot be underestimated. Even the use of an environmentally friendly solvent (an aqueous solution), for the first time, is not without side effects. Therefore, the data collected in this study show the importance of evaluating the dosages, modes of administration and production methods of M. azedarach phytoextracts in agricultural settings.

Beyond the border: comparative ethnobotany in Valmalenco (SO, Italy) and Valposchiavo (Canton of Grisons, Switzerland).

BACKGROUND: The ethnobotanical analysis of two bordering areas allows for the in-depth understanding of the dynamics of Local Ecological Knowledge, which mirrors the naturalistic, historical, and sociopolitical features of each area. As part of the Interreg Italy-Switzerland B-ICE&Heritage and GEMME projects, this work is an ethnobotanical comparative study of two neighboring Alpine territories: Valmalenco (Italy) and Valposchiavo (Switzerland). METHODS: A total of 471 informants were interviewed on different fields of use (medicinal, food, veterinary, etc.). All data were organized in Excel™ spreadsheets. Informant Consensus Factor was calculated for the pathologies reported. Jaccard's similarity indices were calculated to compare the Valmalenco and Valposchiavo areas. Subsequently, another comparison between Valmalenco/Valposchiavo and Italian/Swiss Alpine neighboring areas was carried out. RESULTS: The number of taxa for Valmalenco was 227 (77 families) and 226 in Valposchiavo (65). Out of the 10 most cited species, 7 were mentioned in both. Arnica montana L. was the most cited in Valmalenco, and Sambucus nigra L. in Valposchiavo. The 5 most cited families were the same. Regarding the medicinal and food fields, the similarity indices were fairly low (0.31 and 0.34 for the species; 0.22 and 0.31 for the uses). Concerning the comparison with Italian and Swiss Alps, similarity values were slightly higher with Italy (Valmalenco food species: 0.38 with Italy and 0.26 with Switzerland, medicinal: 0.26 IT and 0.14 SW; Valposchiavo food species: 0.36 with IT and 0.26 with SW, medicinal: 0.21 IT and 0.14 SW). CONCLUSION: Although Valmalenco and Valposchiavo partly share natural environment, language, history, and culture, they had low similarity indices. They both seemed to have more similarities with Italy than Switzerland, maintaining low values with the surrounding territories too. They showed a common core of Local Ecological Knowledge with several divergent branches possibly due to pivotal historical happenings, as well as more modern external influences.

Catha edulis Leaves: Morphological Characterization and Anti-Inflammatory Properties in an In Vitro Model of Gastritis.

Khat leaves, indigenous to eastern Africa, have been chewed for centuries for their stimulant effects, attributed to alkaloids such as cathinone and cathine. Although associated with gastric disorders like gastritis and gastro-oesophageal reflux disease, the underlying molecular mechanisms remain unclear. This study aimed to examine the morpho-anatomy of khat leaves using light microscopy and histochemistry and to assess the effects of leaf extracts and alkaloids on human gastric epithelial cells (GES-1). The study identified specific cells in the palisade-spongy transition zone as storage sites for psychoactive alkaloids. Leaf extracts were prepared by mimicking the chewing process, including a prolonged salivary phase followed by a gastric phase. Cytotoxicity and cell viability were evaluated using LDH and MTT assays, respectively. Additionally, the impact on IL-8 secretion, a key chemokine in gastric inflammation, was analysed under normal and TNF-α-stimulated conditions. The results showed no increase in cytotoxicity up to 250 µg/mL. However, there was a significant decrease in cell metabolism and a reduction in both basal and TNF-α-induced IL-8 secretion, but cathinone and cathine were inactive. These findings suggest that khat may not directly cause the gastric issues reported in the literature, which would rather be attributed to other confounding factors, highlighting the need for further research to clarify its biological impacts.

From Primary Data to Ethnopharmacological Investigations on Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson as a Remedy against Gastric Ailments in Valmalenco (Italy).

(1) Background: Within the framework of the European Interreg Italy-Switzerland B-ICE & Heritage project (2018-2022), this study originated from a three-year ethnobotanical survey in Valmalenco (Sondrio, Italy). Following a preliminary work published by our group, this research further explored the folk therapeutic use of Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson (Asteraceae) for dyspepsia disorders, specifically its anti-inflammatory potential at a gastrointestinal level. (2) Methods: Semi-structured interviews were performed. The bitter taste was investigated through molecular docking software (PLANTS, GOLD), while the anti-inflammatory activity of the hydroethanolic extract, infusion, and decoction was evaluated based on the release of IL-8 and IL-6 after treatment with TNFα or Helicobacter pylori. The minimum inhibitory concentration and bacterial adhesion on the gastric epithelium were evaluated. (3) Results: In total, 401 respondents were interviewed. Molecular docking highlighted di-caffeoylquinic acids as the main compounds responsible for the interaction with bitter taste receptors. The moderate inhibition of IL-6 and IL-8 release was recorded, while, in the co-culture with H. pylori, stronger anti-inflammatory potential was expressed (29-45 µg/mL). The concentration-dependent inhibition of H. pylori growth was recorded (MIC = 100 µg/mL), with a significant anti-adhesive effect. (4) Conclusions: Confirming the folk tradition, the study emphasizes the species' potentiality for dyspepsia disorders. Future studies are needed to identify the components mostly responsible for the biological effects.

An Integrative Approach to Selected Species of Tanacetum L. (Asteraceae): Insights into Morphology and Phytochemistry.

In this work, we studied Tanacetum vulgare, Tanacetum parthenium, and Tanacetum corymbosum (Asteraceae) cultivated at the Ghirardi Botanic Garden (Toscolano Maderno, Brescia, Northern Italy) of the University of Milan. An integrative research approach was adopted: microscopic and histochemical, with special focus on the secretory structures responsible for the productivity of secondary metabolites; phytochemical, with the analysis of the essential oil (EO) profiles from the air-dried, flowered aerial parts collected in June 2021; bio-ecological, with emphasis, based on literature data, on the ecology and biological activity of the main EO components. In all three species, two basic trichome morphotypes (flagellar non-glandular and biseriate glandular) occurred with different distribution patterns. The glandular ones produced terpenes, along with flavonoids. A high level of chemical variability in the EO compositions emerged, specifically for qualitative data. T. vulgare profile was more complex and heterogeneous than those obtained from T. parthenium and T. corymbosum, with camphor as the predominant compound, followed by farnesol and α-santalone, respectively. Finally, the obtained scientific findings were made available to the visitors of the botanic garden through new dissemination labeling that highlights the "invisible", microscopic features of the plants, from an Open Science perspective ("Botanic Garden, factories of molecules…work in progress"-Lombardy Region Project Lr. 25/2016, year 2021).

A urokinase-associated outbreak of Ralstonia mannitolilytica bloodstream infections in haemodialysis patients in north-eastern Italy, January to April 2023.

An outbreak of Ralstonia mannitolilytica bloodstream infections occurred in four hospitals in north-eastern Italy, involving 20 haemodialysis patients with tunnelled central vascular catheter access. We identified as the outbreak source a batch of urokinase vials imported from India contaminated with R. mannitolilytica. Whole genome sequences of the clinical and urokinase strains were highly related, and only urokinase-treated patients were reported with R. mannitolilytica infections (attack rate = 34%; 95% confidence interval: 22.1-47.4). Discontinuation of the contaminated urokinase terminated the outbreak.

Remnants from the Past: From an 18th Century Manuscript to 21st Century Ethnobotany in Valle Imagna (Bergamo, Italy).

BACKGROUND: This project originated from the study of an 18th century manuscript found in Valle Imagna (Bergamo, Italy) which contains 200 plant-based medicinal remedies. A first comparison with published books concerning 20th century folk medicine in the Valley led to the designing of an ethnobotanical investigation, aimed at making a thorough comparison between past and current phytotherapy knowledge in this territory. METHODS: The field investigation was conducted through semi-structured interviews. All data collected was entered in a database and subsequently processed. A diachronic comparison between the field results, the manuscript, and a 20th century book was then performed. RESULTS: A total of 109 interviews were conducted and the use of 103 medicinal plants, belonging to 46 families, was noted. A decrease in number of plant taxa and uses was observed over time, with only 42 taxa and 34 uses reported in the manuscript being currently known by the people of the valley. A thorough comparison with the remedies in the manuscript highlighted similar recipes for 12 species. Specifically, the use of agrimony in Valle Imagna for the treatment of deep wounds calls back to an ancient remedy against leg ulcers based on this species. CONCLUSIONS: The preliminary results of this study allow us to outline the partial passage through time fragments of ancient plant-based remedies once used in the investigated area.

Alpine Diet in Valmalenco (Lombardy, Italy): Nutritional Features of Spontaneous Plants and Traditional Dishes.

BACKGROUND: Along the Alps, the Alpine diet is considered to be one of the most common nutritional models. Next to traditional animal-based products, spontaneous plants of the territory are collected and eaten. AIM: The aim of this study is to evaluate the nutritional features of selected autochthonous plants of the territory and the typical recipe of green gnocchi. METHODS: The analyses of proximate composition, carotenoid, total phenol, and mineral contents in raw and cooked plant samples and the chemical composition and in vitro starch digestibility in green and control gnocchi were performed. RESULTS: Except for Aruncus dioicus, all the wild plants contained high levels of carotenoids (15-20 mg/100 g FW), mainly as xanthophylls. Rumex acetosa showed the highest levels of total phenols (554 mg GAE/100 g FW), and Urtica dioica can be considered to be a good dietary source of iron, calcium, and magnesium (4.9, 410, and 72 mg/100 g FW). Cooking significantly decreased the potassium and magnesium contents in all wild species, and total phenols and carotenoids in Aruncus dioicus, Blitum bonus-henricus, and Silene vulgaris (p < 0.05). The slowly digestible fraction of starch (%SDS/available starch), which is inversely correlated to insulin demand, was significantly increased in green gnocchi compared to matched control gnocchi (p < 0.05). CONCLUSIONS: Traditional consumption of spontaneous plants in the Alpine regions might increase the dietary intakes of several bioactive substances and contribute to cover the nutritional needs of micronutrients.

Teucrium fruticans L., a Multi-Scale Study: From Trichomes to Essential Oil.

This work represents the first multi-scale study on Teucrium fruticans L. cultivated at the Ghirardi Botanic Garden (Lombardy, Northern Italy), combining a micromorphological and a phytochemical survey on the plant's aerial parts. Micromorphological investigations, performed by Light Microscopy, Fluorescence Microscopy and Scanning Electron Microscopy, highlighted the presence of five trichomes morphotypes, distinguished by a different distribution pattern: peltates, short-stalked and ball-like medium-stalked capitates, ubiquitous on the whole plant, medium-stalked and long-stalked capitates, exclusive to the floral whorls. Both peltates and medium-stalked capitates were recognized as the main terpene production sites. Phytochemical characterization focused on the essential oils (EOs), obtained by Clevenger-type hydrodistillation in February and April 2022 and characterized by Gas Chromatography-Mass Spectrometry (GC/MS), which resulted mainly formed by sesquiterpene hydrocarbons. The February EO profile was characterized by ß-caryophyllene (28.30 %) and germacrene D (19.16 %) as main compounds, while in April ß-myrcene was detected at high percentage (13.77 %), in addition to the previous two components (15.72 % and 11.55 %, respectively). Literature data, dealing with the biological activities of the main oil constituents, highlighted an anti-microbial, anti-inflammatory, and anti-tumor potential, due to the high content in sesquiterpenes and, particularly, of ß-caryophyllene and germacrene D.

The Leaf Essential Oil of Myrtus communis subsp. tarentina (L.) Nyman: From Phytochemical Characterization to Cytotoxic and Antimigratory Activity in Human Prostate Cancer Cells.

The aim of this study was to investigate the chemical profile and the cytotoxic activity in two castration-resistant prostate cancer (CRPC) cell lines of the leaf essential oil in Myrtus communis subsp. tarentina (L.) Nyman (EO MT), which was cultivated at the Ghirardi Botanical Garden (Toscolano Maderno, Brescia, Italy). The leaves were air-dried and extracted by hydrodistillation with a Clevenger-type apparatus, and the EO profile was characterized by GC/MS. For the cytotoxic activity investigation, we analyzed the cell viability by MTT assay, and the apoptosis induction by Annexin V/propidium iodide assay/Western blot analysis of cleaved caspase 3 and cleaved PARP proteins. Moreover, the cellular migration was analyzed by Boyden's chamber assay and the distribution of actin cytoskeleton filaments by immunofluorescence. We identified 29 total compounds; the main compound classes were oxygenated monoterpenes, monoterpene hydrocarbons, and sesquiterpenes. The main constituents were α-pinene, α-humulene, α-terpineol, durohydroquinon, linalool, geranyl acetate, and ß-caryophyllene. We found that EO MT was able to reduce cellular viability, activating an apoptotic process, and to decrease the migratory capacity of CRPC cells. These results suggest that it might be interesting to further investigate the effects of single compounds present in EO MT for their possible use in prostate cancer treatment.

Achillea moschata Wulfen: From Ethnobotany to Phytochemistry, Morphology, and Biological Activity.

A multidisciplinary investigation on Achillea moschata Wulfen (Asteraceae) is outlined herein. This work, part of the European Interreg Italy-Switzerland B-ICE project, originated from an ethnobotanical survey performed in Chiesa in Valmalenco (Sondrio, Lombardy, Northern Italy) in 2019-2021 which highlighted this species' relevance of use in folk medicine to treat gastrointestinal diseases. In addition, this contribution included analyses of the: (a) phytochemical profile of the aqueous and methanolic extracts of the dried flower heads using LC-MS/MS; (b) morpho-anatomy and histochemistry of the vegetative and reproductive organs through Light, Fluorescence, and Scanning Electron Microscopy; (c) biological activity of the aqueous extract concerning the antioxidant and anti-inflammatory potential through cell-based in vitro models. A total of 31 compounds (5 phenolic acids, 13 flavonols, and 13 flavones) were detected, 28 of which included in both extracts. Covering and secreting trichomes were observed: the biseriate 10-celled glandular trichomes prevailing on the inflorescences represented the main sites of synthesis of the polyphenols and flavonoids detected in the extracts, along with volatile terpenoids. Finally, significant antioxidant and anti-inflammatory activities of the aqueous extract were documented, even at very low concentrations; for the first time, the in vitro tests allowed us to formulate hypotheses about the mechanism of action. This work brings an element of novelty due to the faithful reproduction of the traditional aqueous preparation and the combination of phytochemical and micromorphological research approaches.

Resilience of Stevia rebaudiana (Bertoni) Bertoni in the Underwater Biospheres of Nemo's Garden®: Adaptation to New Cultivation Systems.

The Nemo's Garden® project is an alternative production system for areas with scarce cultivable land but significant presence of water; thus, it is an interesting intervention to address the climate crisis. This work aimed to evaluate the micromorphological, biochemical, and phytochemical characteristics of Stevia rebaudiana (Bertoni) Bertoni grown underwater compared to the terrestrial specimens. The micromorphological analyses, performed on the leaves using light microscopy, fluorescence microscopy, and scanning electron microscopy, evidenced a general uniformity of the trichome morphotype and distribution pattern. The histochemical investigation indicated the simultaneous presence of terpenes and polyphenols in the trichome secreted material from the underwater samples and a prevailing polyphenolic content in the terrestrial specimens; this was also confirmed by biochemical analyses (26.6 mg GAE/g DW). The characterization of non-volatile components, performed using HPLC-MS, showed similar chemical profiles in all the samples, which were characterized by phenolic compounds and steviol glycosides. The volatile compounds, evaluated using HS-SPME coupled with GC-MS, showed sesquiterpene hydrocarbons as the main class in all the analyzed samples (80.1-93.9%). However, the control plants were characterized by a higher content of monoterpene hydrocarbons (12.1%). The underwater biosphere environment did not alter S. rebaudiana micro-morphological characters, although slight qualitative changes were evidenced for the compounds produced as a response to the growth conditions.

Terpene Profiles Composition and Micromorphological Analysis on Two Wild Populations of Helichrysum spp. from the Tuscan Archipelago (Central Italy).

Two wild populations of Helichrysum (Mill.) located at Elba Island (Tuscan Archipelago, Central Italy) were morphologically and phytochemically analyzed to taxonomically identify H. litoreum (population A) and H. italicum subsp. italicum (population B). Micromorphological and histochemical analyses were performed on the indumentum using Scanning Electron Microscope (SEM) and Light Microscope (LM). Morphometric analyses on vegetative and reproductive traits were also conducted. Finally, a chemotaxonomic analysis was carried out on the terpene profile of flowers, leaves and bark tissues using gas chromatography (GC-MS). Results suggested that morphological discriminant traits were mainly in leaves and cypselae glandular tissues. Phytochemical analysis indicated that a high relative content of α-pinene and ß-caryophyllene were the main markers for population A, while a high relative content of neryl-acetate, α-curcumene, isoitalicene and italicene, especially in the terpene profile of bark tissue, were the main compounds for discriminating population B. The analysis suggested that the wild population A could be mainly ascribed to H. litoreum, whilst population B is defined by H. italicum.

A Comprehensive Phytochemical Analysis of Terpenes, Polyphenols and Cannabinoids, and Micromorphological Characterization of 9 Commercial Varieties of Cannabis sativa L.

New hemp (Cannabis sativa L.) strains developed by crossbreeding selected varieties represent a novel research topic worthy of attention and investigation. This study focused on the phytochemical characterization of nine hemp commercial cultivars. Hydrodistillation was performed in order to collect the essential oils (EO), and also the residual water and deterpenated biomass. The volatile fraction was analyzed by GC-FID, GC-MS, and SPME-GC-MS, revealing three main chemotypes. The polyphenolic profile was studied in the residual water and deterpenated biomass by spectrophotometric assays, and HPLC-DAD-MSn and 1H-NMR analyses. The latter were employed for quali-quantitative determination of cannabinoids in the deterpenated material in comparison with the one not subjected to hydrodistillation. In addition, the glandular and non-glandular indumentum of the nine commercial varieties was studied by means of light microscopy and scanning electron microscopy in the attempt to find a possible correlation with the phytochemical and morphological traits. The EO and residual water were found to be rich in monoterpene and sesquiterpene hydrocarbons, and flavonol glycosides, respectively, while the deterpenated material was found to be a source of neutral cannabinoids. The micromorphological survey allowed us to partly associate the phytochemistry of these varieties with the hair morphotypes. This research sheds light on the valorization of different products from the hydrodistillation of hemp varieties, namely, essential oil, residual water, and deterpenated biomass, which proved to be worthy of exploitation in industrial and health applications.

Botanic Garden as a Factory of Molecules: Myrtus communis L. subsp. communis as a Case Study.

A novel perception of botanic gardens as complex "factories of molecules" (Lombardy Region Project-Lr. 25/2016, year 2021), that mediate plant-environment interactions, and are the basis of their utility for humans, is presented. The core-topic is the medicinal plant heritage of the Ghirardi Botanic Garden (Toscolano Maderno, Brescia, Italy) of the University of Milan. In this work, we studied Myrtus communis L. subsp. communis (Myrtaceae) at multiple scale levels: macro- and micromorphological, with special emphasis on the secretory structures responsible for the production of secondary metabolites; phytochemical, with the analysis of the essential oil (EO) composition from leaves (fresh, dried, stored at -20 °C and at -80 °C) and fruits over two consecutive years (2018 and 2019); bio-ecological, with a focus, based on literature data, on the ecology and biological activity of the main EO components. The occurrence of secretory cavities producing terpenes, along with flavonoids, was proven. A high level of chemical variability across the obtained EO profiles emerged, especially that concerning quantitative data. However, regardless of the different conservation procedures, the examined plant part, or the phenological stage, we detected the presence of three ubiquitous compounds: α-pinene, 1,8-cineole, and linalool. The overall results will serve to enrich the Ghirardi Botanic Garden with novel labeling showing accurate and updated scientific information in an Open science perspective.

Scutellaria brevibracteata subsp. subvelutina (Rech.f.) Greuter & Burdet: morphological and phytochemical characterization.

A micromorphological and phytochemical survey was performed on Scutellaria brevibracteata subsp. subvelutina cultivated in Italy. The indumentum of the vegetative and reproductive organs was investigated: peltate, short-, medium- and long-stalked capitates were described. Histochemistry evidenced similar results for peltates and long-stalked capitates, differences for short and medium capitates. For the first time, this work reported the characterization of volatile organic compounds from leaves and flowers, along with the first analysis of the essential oil obtained from the aerial parts of Italian samples. The floral profile resulted more complex than the foliar one, due to the higher number of the total compounds (40 vs 27) and of the exclusive constituents (24 vs 11). 16 common compounds were detected, with ß-caryophyllene as the most abundant. The essential oil was characterized by 23 compounds, with ß-caryophyllene dominating. The peltates, the medium and the long-stalked capitates resulted the producers of the investigated compounds.

Ca' Granda, Hortus simplicium: Restoring an Ancient Medicinal Garden of XV-XIX Century in Milan (Italy).

This work is based on the study of 150 majolica vases dated back to the mid XVII century that once preserved medicinal remedies prepared in the ancient Pharmacy annexed to the Ospedale Maggiore Ca' Granda in Milan (Lombardy, Italy). The Hortus simplicium was created in 1641 as a source of plant-based ingredients for those remedies. The main objective of the present work is to lay the knowledge base for the restoration of the ancient Garden for educational and informative purposes. Therefore, the following complementary phases were carried out: (i) the analysis of the inscriptions on the jars, along with the survey on historical medical texts, allowing for the positive identification of the plant ingredients of the remedies and their ancient use as medicines; (ii) the bibliographic research in modern pharmacological literature in order to validate or refute the historical uses; (iii) the realization of the checklist of plants potentially present in cultivation at the ancient Garden, concurrently with the comparison with the results of a previous in situ archaeobotanical study concerning pollen grains. For the species selection, considerations were made also regarding drug amounts in the remedies and pedoclimatic conditions of the study area. Out of the 150 vases, 108 contained plant-based remedies, corresponding to 148 taxa. The remedies mainly treated gastrointestinal and respiratory disorders. At least one of the medicinal uses was validated in scientific literature for 112 out of the 148 examined species. Finally, a checklist of 40 taxa, presumably hosted in the Hortus simplicium, was assembled.

Altered feeding behavior and immune competence in paper wasps: A case of parasite manipulation?

Paper wasps (Polistes dominula), parasitized by the strepsipteran Xenos vesparum, are castrated and desert the colony to gather on plants where the parasite mates and releases primary larvae, thus completing its lifecycle. One of these plants is the trumpet creeper Campsis radicans: in a previous study the majority of all wasps collected from this plant were parasitized and focused their foraging activity on C. radicans buds. The unexpected prevalence and unusual feeding strategy prompted us to investigate the influence of this plant on wasp behavior and physiology through a multidisciplinary approach. First, in a series of laboratory bioassays, we observed that parasitized wasps spent more time than non-parasitized ones on fresh C. radicans buds, rich of extra-floral nectaries (EFNs), while the same wasps ignored treated buds that lacked nectar drops. Then, we described the structure and ultra-structure of EFNs secreting cells, compatible with the synthesis of phenolic compounds. Subsequently, we analysed extracts from different bud tissues by HPLC-DAD-MS and found that verbascoside was the most abundant bioactive molecule in those tissues rich in EFNs. Finally, we tested the immune-stimulant properties of verbascoside, as the biochemical nature of this compound indicates it might function as an antibacterial and antioxidant. We measured bacterial clearance in wasps, as a proxy for overall immune competence, and observed that it was enhanced after administration of verbascoside-even more so if the wasp was parasitized. We hypothesize that the parasite manipulates wasp behavior to preferentially feed on C. radicans EFNs, since the bioactive properties of verbascoside likely increase host survival and thus the parasite own fitness.

Tools to Tie: Flower Characteristics, VOC Emission Profile, and Glandular Trichomes of Two Mexican Salvia Species to Attract Bees.

A plant can combine physical and chemical tools to interact with other organisms. Some are designed for pollinator attraction (i.e., colors and volatile organic compounds-VOCs); others can act to discourage herbivores (i.e., non-glandular trichomes). Few studies fully address available tools in a single species; notwithstanding, this information can be pivotal in understanding new interactions out of the home range. We characterized flower traits, emission profiles of constitutive compounds from flowers and leaves, micro-morphology of the glandular trichomes, and listed flower visitors of two Mexican bird-pollinated Salvia species (S. blepharophylla and S. greggii), growing in an Italian botanical garden. Flowers were highly variable in their morphometric characteristics. In both species, four trichome morphotypes with similar histochemistry and distribution were documented for leaves and flowers except the calyx abaxial side. The vegetative emission profiles were qualitatively more complex than the floral ones; however, common compounds occurring in high relative percentages were ß-caryophyllene and germacrene D. Floral bouquets were dominated by limonene and ß-pinene in S. greggii and by 1,8-cineole in S. blepharophylla. Two potential (non-bird) pollinators were especially abundant: small bees belonging to the genus Lasioglossum and large bees belonging to the species Xylocopa violacea. Our study highlights the plasticity of these plants, as well as tools that can be conveniently used to establish novel interactions.

Anatomical Investigation and GC/MS Analysis of 'Coco de Mer', Lodoicea maldivica (Arecaceae).

Lodoicea maldivica (J.F.Gmel.) Pers. (Arecaceae), 'Coco de Mer', is a palm, growing as endemic in the Seychelles islands. Its fruit weighs up to 20 kg and is characterized by a fleshy and fibrous envelope surrounding the nutlike portion. The present work combines a morpho-anatomical and a phytochemical analysis of the fruit exocarp and mesocarp. The exocarp is composed by a layer of palisade cells. The mesocarp is characterized by vascular bundles and by sclereids. In the aerenchyma, the internal zone of the mesocarp, cells aggregates were positive to phenols, while idioblasts were positive to terpenes. We performed a GC/MS analysis with a semi-quantitative relative amount calculation of the recorded compounds. The GC/MS essential oil profile revealed the dominance of acyclic sesquiterpenoids (53.95 %), followed by bicyclic sesquiterpenoids (31.69 %), monoterpenes (11.89 %) and monocyclic sesquiterpenoids (2.44 %). The terpenes detected in higher amounts, ß-caryophyllene and bicyclogermacrene, are known for activity against insect larvae, but have been proposed as antiviral candidates against SARS-CoV-2. The third compound in amount, aromadendrene, is active against bacteria and, again, known to possess insecticidal properties.