Woody encroachment and soil carbon stocks in subalpine areas in the Central Spanish Pyrenees

Woody encroachment has been an ongoing process in the subalpine belt of Mediterranean mountains, after land abandonment, the disappearance of the transhumant system and the decrease of the livestock number. The main objectives of this study were: (i) to identify land use/land cover (LULC) changes from 1956 to 2015, and (ii) to investigate the effects of LULC changes in physical and chemical soil properties and soil organic carbon (SOC) and nitrogen (N) stocks. It is hypothesized that woody encroachment in the subalpine belt may lead to significant changes in soil properties, and will generate an increase in the SOC stocks. A land use gradient was identified in the subalpine belt of the Central Spanish Pyrenees: (i) subalpine grasslands, (ii) shrublands, (iii) young forests, and (iv) old forests. Mineral soil samples were collected every 10 cm, down to 40 cm, at three points per each LULC and a total of 48 samples were analyzed. The results showed that (i) woody encroachment has occurred from 1956 to 2015 due to the expansion of coniferous forests and shrublands (at the expense of grasslands), (ii) land cover and soil depth had significant effects on soil properties (except for pH), being larger in the uppermost 0–10 cm depth, (iii) SOC and N contents and stocks were higher in the grassland sites, and (iv) the woody encroachment process initially produced a decrease in the SOC stocks (shrublands), but no differences were observed considering the complete soil profile between grasslands and young and old forests. Further studies, describing SOC stabilization and quantifying above-ground carbon (shrub and tree biomass) are required

In Vivo IS6110 profile changes in a Mycobacterium tuberculosis strain as determined by tracking over 14 years

Transposition and homologous recombination of IS6110 appear in Mycobacterium tuberculosis along in vivo sequential infec- tions. These events were checked in different clones of a successful strain, M. tuberculosis Zaragoza, with the focus on a variant in which integration of a copy of IS6110 in the origin of replication (oriC) region occurred

Therapeutic efficacy of pulmonary live tuberculosis vaccines against established asthma by subverting local immune environment

Background: Substantial recent advances in the comprehension of the molecular and cellular mechanisms behind asthma have evidenced the importance of the lung immune environment for disease outcome, making modulation of local immune responses an attractive therapeutic target against this pathology. Live attenuated mycobacteria, such as the tuberculosis vaccine BCG, have been classically linked with a type 1 response, and proposed as possible modulators of the type 2 response usually associated with asthma. Methods: In this study we used different acute and chronic murine models of asthma to investigate the therapeutic efficacy of intranasal delivery of the live tuberculosis vaccines BCG and MTBVAC by regulating the lung immune environment associated with airway hyperresponsiveness (AHR). Findings: Intranasal administration of BCG, or the novel tuberculosis vaccine candidate MTBVAC, abrogated AHR-associated hallmarks, including eosinophilia and lung remodeling. This correlated with the re-polarization of allergen-induced M2 macrophages towards an M1 phenotype, as well as with the induction of a strong allergen-specific Th1 response. Importantly, vaccine treatment was effective in a scenario of established chronic asthma where a strong eosinophil infiltration was already present prior to immunization. We finally compared the nebulization efficiency of clinical formulations of MTBVAC and BCG using a standard commercial nebulizer for potential aerosol application. Interpretation: Our results demonstrate that pulmonary live tuberculosis vaccines efficiently revert established asthma in mice. These data support the further exploration of this approach as potential therapy against asthma

Detection of a putative TetR-like gene related to Mycobacterium bovis BCG growth in cholesterol using a gfp-transposon mutagenesis system

In vitro transposition is a powerful genetic tool for identifying mycobacterial virulence genes and studying virulence factors in relation to the host. Transposon shuttle mutagenesis is a method for constructing stable insertions in the genome of different microorganisms including mycobacteria. Using an IS1096 derivative, we have constructed the Tngfp, a transposon containing a promoterless green fluorescent protein (gfp) gene. This transposon was able to transpose randomly in Mycobacterium bovis BCG. Bacteria with a single copy of the gfp gene per chromosome from an M. bovis BCG::Tngfp library were analyzed and cells exhibiting high levels of fluorescence were detected by flow cytometry. Application of this approach allowed for the selection of a mutant, BCG_2177c::Tngfp (BCG-Tn), on the basis of high level of long-standing fluorescence at stationary phase. This BCG-Tn mutant showed some particular phenotypic features compared to the wild type strain, mainly during stationary phase, when cholesterol was used as a sole carbon source, thus supporting the relationships of the targeted gene with the regulation of cholesterol metabolism in this bacteria. This approach showed that Tngfp is a potentially useful tool for studying the involvement of the targeted loci in metabolic pathways of mycobacteria

Global study of is6110 in a successful mycobacterium tuberculosis strain: Clues for deciphering its behavior and for its rapid detection

The Mycobacterium tuberculosis insertion sequence IS6110, besides being a very useful tool in molecular epidemiology, seems to have an impact on the biology of bacilli. In the present work, we mapped the 12 points of insertion of IS6110 in the genome of a successful strain named M. tuberculosis Zaragoza (which has been referred to as the MTZ strain). This strain, belonging to principal genetic group 3, caused a large unsuspected tuberculosis outbreak involving 85 patients in Zaragoza, Spain, in 2001 to 2004. The mapping of the points of insertion of IS6110 in the genome of the Zaragoza strain offers clues for a better understanding of the adaptability and virulence of M. tuberculosis. Surprisingly, the presence of one copy of IS6110 was found in Rv2286c, as was recently described for a successful Beijing sublineage. As a result of this analysis, a rapid method for detecting this particular M. tuberculosis strain has been designed

Minimal surfaces and particles in 3-manifolds

We use minimal (or CMC) surfaces to describe 3-dimensional hyperbolic, anti-de Sitter, de Sitter or Minkowski manifolds. We consider whether these manifolds admit ``nice'' foliations and explicit metrics, and whether the space of these metrics has a simple description in terms of Teichm\"uller theory. In the hyperbolic settings both questions have positive answers for a certain subset of the quasi-Fuchsian manifolds: those containing a closed surface with principal curvatures at most 1. We show that this subset is parameterized by an open domain of the cotangent bundle of Teichm\"uller space. These results are extended to ``quasi-Fuchsian'' manifolds with conical singularities along infinite lines, known in the physics literature as ``massive, spin-less particles''. Things work better for globally hyperbolic anti-de Sitter manifolds: the parameterization by the cotangent of Teichm\"uller space works for all manifolds. There is another description of this moduli space as the product two copies of Teichm\"uller space due to Mess. Using the maximal surface description, we propose a new parameterization by two copies of Teichm\"uller space, alternative to that of Mess, and extend all the results to manifolds with conical singularities along time-like lines. Similar results are obtained for de Sitter or Minkowski manifolds. Finally, for all four settings, we show that the symplectic form on the moduli space of 3-manifolds that comes from parameterization by the cotangent bundle of Teichm\"uller space is the same as the 3-dimensional gravity one.Comment: 53 pages, no figure. v2: typos corrected and refs adde

Respiratory Immunization With a Whole Cell Inactivated Vaccine Induces Functional Mucosal Immunoglobulins Against Tuberculosis in Mice and Non-human Primates

Vaccination through the natural route of infection represents an attractive immunization strategy in vaccinology. In the case of tuberculosis, vaccine delivery by the respiratory route has regained interest in recent years, showing efficacy in different animal models. In this context, respiratory vaccination triggers lung immunological mechanisms which are omitted when vaccines are administered by parenteral route. However, contribution of mucosal antibodies to vaccine- induced protection has been poorly studied. In the present study, we evaluated in mice and non-human primates (NHP) a novel whole cell inactivated vaccine (MTBVAC HK), by mucosal administration. MTBVAC HK given by intranasal route to BCG-primed mice substantially improved the protective efficacy conferred by subcutaneous BCG only. Interestingly, this improved protection was absent in mice lacking polymeric Ig receptor (pIgR), suggesting a crucial role of mucosal secretory immunoglobulins in protective immunity. Our study in NHP confirmed the ability of MTBVAC HK to trigger mucosal immunoglobulins. Importantly, in vitro assays demonstrated the functionality of these immunoglobulins to induce M. tuberculosis opsonization in the presence of human macrophages. Altogether, our results suggest that mucosal immunoglobulins can be induced by vaccination to improve protection against tuberculosis and therefore, they represent a promising target for next generation tuberculosis vaccines

Analysis of Mycobacterium africanum in the last 17 years in Aragon identifies a specific location of IS6110 in Lineage 6

The purpose of this study was to increase our knowledge about Mycobacterium africanum and report the incidence and characteristics of tuberculosis (TB) due to their lineages in Aragon, Spain, over the period 2003–2019. The study includes all the cases in our region, where all the M. tuberculosis complex isolates are systematically characterised. We detected 31 cases of M. africanum among 2598 cases of TB in the period studied. TB caused by M. africanum is rare (1.19%) in our population, and it affects mainly men of economically productive age coming from West African countries. Among the isolates, Lineage (L) 6 was more frequent than L5. The genotyping of these strains identified five clusters and 13 strains with a unique pattern. The isolates’ characterisation identified a copy of IS6110 within the moaX gene, which turned out to be specific for L6. It will allow the differentiation of this lineage from the rest of MTBC with a simple PCR reaction. It remains to be established whether this polymorphism may limit M. africanum transmission. Furthermore, a mutation in the mutT2 promoter was found as specific for L6 strains, which could be related to the high variability found for L6 compared to L5. © 2021, The Author(s)