Setting the stage for new ecological indicator species: A holistic case study on the Iberian dolphin freshwater mussel Unio delphinus Spengler, 1793

Supplementary data to this article can be found online at https://doi.org/10.1016/j.ecolind.2019.105987.Due to their sensitivity and dramatic declines, freshwater mussels are prime targets for conservation and environmental monitoring. For this, however, information is needed on life history and ecological traits, which is lacking in many taxa, including threatened species. Species recently described or recognized as valid are of particular concern, due to the shortage of even basic knowledge. A case in point is the recently recognized and Near Threatened dolphin freshwater mussel Unio delphinus Spengler, 1793, which is endemic to the western Iberian Peninsula and has suffered marked population declines. To overcome information gaps for U. delphinus, we carried out a holistic biological study across the species range, aiming to: i) estimate the area of occupancy (AOO) and extent of occurrence (EOO) based on updated distribution data taken from the literature and recent surveys; ii) estimate growth patterns from biometrical (shell dimensions and growth annuli) measurements taken on specimens from seven populations; iii) estimate sex ratios from gonad tissue biopsies collected on specimens from eight populations; iv) estimate gametogenesis and sex ratio through histological examination of gonad and gill tissues collected monthly for a year, from a single population; and v) determine host species from infestation trials of glochidia with co-occurring fish species. We estimated an EOO of 706 km2 and an AOO of 61 km2, which together with data on declines assigns the species to the Endangered category using IUCN criteria. Unio delphinus was found to grow faster and to be shorter-lived (up to 11 years, maturity at around 2 years old) than other European freshwater mussels. Growth and life span are similar across the range in lotic habitats, but different from that in lentic habitats. The larvae of U. delphinus may attach to most co-occurring fish species, but only native species were effective hosts. Native cyprinids, especially those from the genus Squalius, seem to be the primary hosts. Overall, the information provided contributes to a better conservation status assessment, selection of conservation and rehabilitation areas, guidance for the establishment of propagation programs and better timing for specimens’ manipulation including monitoring and possible translocations. The framework presented here highlights the importance of basic biological studies to define good ecological and physiological status.This work was supported by Portuguese FCT – Foundation for Science and Technology, Projects FRESHCO: Multiple implications of invasive species on Freshwater Mussel co-extinction processes (PTDC/AGR-FOR/1627/2014-04/SAICT/2015), MUSSELFLOW: Host-depen dent evolution, ecology and conservation of freshwater mussels under varying hydrological conditions: consequences of climate change (PTDC/BIA-EVL/29199/2017), and ConBiomics: The missing approach for the Conservation of freshwater Bivalves Project N° NORTE-01-0145- FEDER-030286, co-financed by COMPETE 2020, Portugal 2020 and the European Union through the ERDF. FCT also supported MLL under grant (SFRH/BD/115728/2016), CM under grant (SFRH/BD/111133/ 2015) and also through Strategic Funding UID/Multi/04423/2019. PB was supported by EDP Biodiversity Chair

Catalytic Mechanism for the Conversion of Salicylate Into Catechol by the Flavin-Dependent Monooxygenase Salicylate Hydroxylase

Salicylate hydroxylase (NahG) is a flavin-dependent monooxygenase that catalyzes the decarboxylative hydroxylation of salicylate into catechol in the naphthalene degradation pathway in Pseudomonas putida G7. We explored the mechanism of action of this enzyme in detail using a combination of structural and biophysical methods. NahG shares many structural and mechanistic features with other versatile flavin-dependent monooxygenases, with potential biocatalytic applications. The crystal structure at 2.0 Å resolution for the apo form of NahG adds a new snapshot preceding the FAD binding in flavin-dependent monooxygenases. The kcat/Km for the salicylate reaction catalyzed by the holo form is \u3e105 M−1 s−1 at pH 8.5 and 25 °C. Hammett plots for Km and kcat using substituted salicylates indicate change in rate-limiting step. Electron-donating groups favor the hydroxylation of salicylate by a peroxyflavin to yield a Wheland-like intermediate, whereas the decarboxylation of this intermediate is faster for electron-withdrawing groups. The mechanism is supported by structural data and kinetic studies at different pHs. The salicylate carboxyl group lies near a hydrophobic region that aids decarboxylation. A conserved histidine residue is proposed to assist the reaction by general base/general acid catalysis

Changes in Glucose and Glutamine Lymphocyte Metabolisms Induced by Type I Interferon α

In lymphocytes (LY), the well-documented antiproliferative effects of IFN-α are associated with inhibition of protein synthesis, decreased amino acid incorporation, and cell cycle arrest. However, the effects of this cytokine on the metabolism of glucose and glutamine in these cells have not been well investigated. Thus, mesenteric and spleen LY of male Wistar rats were cultured in the presence or absence of IFN-α, and the changes on glucose and glutamine metabolisms were investigated. The reduced proliferation of mesenteric LY was accompanied by a reduction in glucose total consumption (35%), aerobic glucose metabolism (55%), maximal activity of glucose-6-phosphate dehydrogenase (49%), citrate synthase activity (34%), total glutamine consumption (30%), aerobic glutamine consumption (20.3%) and glutaminase activity (56%). In LY isolated from spleen, IFNα also reduced the proliferation and impaired metabolism. These data demonstrate that in LY, the antiproliferative effects of IFNα are associated with a reduction in glucose and glutamine metabolisms