BUSCADOR RECOLECTA

Growth (A), biofilm formation (B), EPS production (C) and motility (D) of cavA and vfr related strains demonstrating that the observed phenotypes were not attributed to growth alternations of the strains. Moreover, the empty miniTn7 system (EV) with Tetracycline (Tc) or Tellurate (Tel) resistance markers, used for the complementations and genes expression in different backgrounds, did not have an effect on any of the tested phenotypes. Growth and biofilm formation were assessed after 24 h at 37°C shaking. EPS production was assessed on Congo agar after 5 days incubation of the strains at 37°C and motility was tested after 48 h on soft agar at 37°C. ns p>0.05, *p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001—One-Way ANOVA with Tukey post-hoc test., Peer reviewed

A–Twitching motility of ΔcavA related strains showing that miniTn7 EV has no impact on A. baumannii motility. B–Natural transformation of ΔcavA mutant was bellow the detection limit (< d. l.) and was significantly decreased compared to the WT AB5075. ns p>0.05, ** p<0.01, ***p<0.001, **** p<0.0001—One-Way ANOVA with Tukey post-hoc test (A) and Unpaired t-test (B)., Peer reviewed

A–Representative images of Congo agar plates showing that the empty miniTn7 vector does not affect A. baumannii EPS production. B–Representative image of Congo red plates demonstrating the effect of pgaA disruption (pgaA::Tn) on Congo red dye binding to A. baumannii EPS and on the increased EPS production in the ΔcavA mutant. This shows that the effect of cavA on EPS production is mainly due to changes in the production of poly-β-1,6-N-acetylglucosamine (PNAG) and pgaABCD operon expression. C & D–Growth (C) and biofilm formation (D) of AB5075 WT and its derivative ΔabaI, csuC::Tn, pgaA::Tn and ΔcavA single and double mutants after being incubated at 37°C shaking for 24 h. Deletion of the autoinducer synthase gene abaI in ΔcavA did not affect the increased ΔcavA biofilm. In contrast, disruption of csuC or pgaA in the ΔcavA mutant significantly decreased the high biofilm levels caused by the deletion of cavA but did not completely reversed the ΔcavA phenotype. This data demonstrates that the regulation of A. baumannii biofilm formation by CavA is dependent on Csu pili and EPS production and is the additive effect of the global simultaneous regulation of multiple genes. ns p>0.05, **p<0.01, **** p<0.0001—One-Way ANOVA with Tukey post-hoc test., Peer reviewed

A—Heatmap of the protein profile of CavA, CavB and Vfr in the A. baumannii pangenome. The genomes are clustered (rows) by the variants of the proteins they present. The most frequent variant of the protein (ref) is depicted in orange and the other variants are shown in different shades of purple. The left side of the heatmap shows the metadata: Multilocus sequence typing (MLST) (the 5 most frequent, highlighting ST2), and host source (predominantly human but also hospital environment, environment and other). Highlighted are A. baumannii strains commonly used as reference: ATCC17978 (GCF_902728005.1), ATCC19606 (GCF_014116795.1), ACICU (GCF_000018445.1), AYE (GCF_000069245.1), AB5075 (GCF_000770605.1), AB0057 (GCF_000021245.2). B—Proportion of different sequence variants (v) of CavA, CavB and Vfr proteins in the A. baumannii pangenome. The vref variant represents the most frequent, and "not found" appears when the protein has not been found. The number of genomes per group is as follows: Human (6589), Hospital environment (131), Environmental (140), Other (2836)., Peer reviewed

A–PCR validating T26 insertion in the ABUW_2208 gene in five strains annotated as ABUW_2208::T26 transposon mutants, labelled with their corresponding number from the Manoil transposon mutant library. C.ABUW_2208 fw/rv (annealing at the beginning and end of the ABUW_2208 gene) were used. Wild-type ABUW_2208 gene is 1.5-kb in size (C+), while the cavA gene with T26 transposon (1.8-kb) is ~3.2-kb. Genomic DNA (20 ng) from AB5075 was used as positive control (C+) and water was used as a negative one (C-). Only AB05781, AB05784 and AB05783 were confirmed to be ABUW_2208::T26 mutants. The other two strains (AB05780 and AB05782) did not harbour the T26 transposon in the ABUW_2208 gene and thus, were incorrectly assigned as ABUW_2208::T26 mutants. B–Schematic representation of CavA protein (489 aa) showing its predicted transmembrane (TM) regions and the adenylate/guanylate cyclase domain (PF00211). C & D–Growth measured as optical density at 600 nm (OD600) (C) and biofilm formation measured as optical density at 570 nm (OD570) (D) after 24 h at 37°C shaking, demonstrating that both phenotypes remained unchanged in the controls bearing chromosomal insertion of the empty miniTn7 (EV) in the wild-type (WT) and ΔcavA backgrounds. E—Intracellular cAMP concentrations presented as μmol per milligram protein of wild-type (WT), ΔcavA and ΔcavB single and ΔcavAΔcavB double mutants and their derivatives with EV showing that the empty miniTn7 did not alter cAMP production in these strains. ns p>0.05, *p<0.05, **p<0.01, ****p<0.0001 One-Way ANOVA with Tukey post-hoc test. F–Representation of CavB protein (487 aa) with its CYTH (PF01928) and CHAD (PF05235) domains. T26 transposon insertion positions in the transposon mutants from the Manoil mutant library [24] are presented by red triangles. G–Growth (OD600) of ΔcavB mutant and WT after 24 h period incubation at 37°C shaking. ns p>0.05—Unpaired t-test., Peer reviewed

Current global change scenarios demand knowledge on how anthropogenic impacts affect ecosystem functioning through changes in food web structure. Frugivorous mesocarnivores are a key link in trophic cascades because, while their abundance and behaviour are usually controlled by apex predators, they can provide high-quality seed-dispersal services to plant communities. Thus, the recent rewilding of large carnivores worldwide can trigger cascading effects for plants. We investigated the top-down effects of an apex predator (Iberian lynx Lynx pardinus) on seed-dispersal services mediated by two mesocarnivore species (red fox Vulpes vulpes and stone marten Martes foina) at the plant community level by comparing areas with and without lynx in a Mediterranean mountain range in Southern Spain. We collected scats of mesocarnivores (n = 1575) to assess frugivory and seed dispersal of 15 plant species over two consecutive fruiting seasons and two habitat types (open and forest). Specifically, we assessed the effect of lynx presence on (i) seed occurrence and fleshy-fruit biomass per scat, (ii) the number of scats containing seeds, and (iii) the diversity of dispersed seeds. The quantity and diversity of dispersed seeds drastically decreased under predation risk for both mesocarnivore species. Seed dispersal by stone martens was negatively affected by the presence of lynx, with a marked reduction in the number of scats with seeds (93%) and the diversity of dispersed seeds (46%). Foxes dispersed 68% fewer seeds in open habitats when coexisting with lynx, probably leading to differential contributions to seed-dispersal effectiveness among habitats. Our study reveals a novel trophic cascade from apex predators to plant communities. The behavioural responses of frugivorous mesocarnivores to predation risk and the reduction in the intensity of their faecal deposition pattern are probably related to their lower abundance when co-occurring with apex predators. While rewilding apex predators is a successful conservation tool, attention should be paid to cascading effects across food webs, particularly where frugivore megafauna are missing and mesocarnivores provide unique services to plant communities., Ministerio de Ciencia e Innovación: CGL2017-84633-P, Generación de conocimiento Ministerio de Ciencia e Innovación: FPU17/04375, Formación del profesorado universitario, Peer reviewed

Riera-Ferrer et al 2024_tables_figures_ESM.pdf, Peer reviewed

Peer reviewed

[EN] Data collection from the UV-vis spectra and corresponding plot, powder X-ray diffraction data, data collection of cell viability, values of Z-potential and data collection of XPS core level analysis of Ag 3d; Au 4f; C 1s and O 1s with fittings and plots., Programa Bilateral CSIC-TEC Monterrey (ref. BILTC22001); Spanish Ministry of Science and Innovation to grant PID2021-126524NB-100 funded by MCIN/AEI/10.13039/501100011033 and by "ERDF A way of making Europe. BILTC22001; PID2021-126524NB-100 SCALAR; RYC2018-024561-I; CEX2023-001286-S, Au_AgNPs UV-Vis.mppz Au_Ag DRX magic plot.mppz Cell viability.xlsx potencial z.xlsx XPS.pxp, No

Description of the project Context: In the face of anthropogenic global change, Mediterranean oak-dominated ecosystems confront increased biotic (ungulate herbivory) and abiotic (drought) stressors, compromising forest regeneration. Restoration measures are imperative to address this scenario. Aims: This study assesses the impact of different mitigation measures on the survival and biochemical traits of two widespread Mediterranean oak species. 4. Description of the dataset The dataset encompasses information from the planting of 300 Quercus seedlings in Cabañeros (Quercus ilex; N=200) and Doñana (Quercus suber; N=100), subjected to various treatments: cotyledon/acorn removal, seedling age (1 vs. 2-year-old), and herbivore protection (fenced vs. non-fenced). Monthly measurements of the seedlings were conducted from February to September 2021. Each visit consisted of recording: the seedling survival status (alive vs. dead), the damage caused by biotic and abiotic stress agents and the identification of the main stress agent, namely wild boar, deer, rabbit, fungi, insect or drought. We considered the main stress agent the one leading to the plant death, or, in the case of surviving seedlings, the one identified with the highest intensity score. To compare seedling physiological leaf biochemical responses to different treatments and plant ages, several non-destructive measures of chlorophyll, anthocyanin and flavonoid concentrations and the nitrogen balance index (NBI) were taken using the leaf clamp fluorescence sensor Dualex ® TMScientific+, Force One. The Dualex measures were taken on 5 leaves previously marked, from top to bottom, only in alive seedlings., Peer reviewed