Burkholderia cepacia complex species differ in the frequency of variation of the lipopolysaccharide O-antigen expression during cystic fibrosis chronic respiratory infection

Burkholderia cepacia complex (Bcc) bacteria can adapt to the lung environment of cystic fibrosis (CF) patients resulting in the emergence of a very difficult to eradicate heterogeneous population leading to chronic infections associated with rapid lung function loss and increased mortality. Among the important phenotypic modifications is the variation of the lipopolysaccharide (LPS) structure at level of the 0-antigen (OAg) presence, influencing adherence, colonization and the ability to evade the host defense mechanisms. The present study was performed to understand whether the loss of OAg expression during CF infection can be considered a general phenomenon in different Bcc species favoring its chronicity. In fact, it is still not clear why different Bcc species/strains differ in their ability to persist in the CF lung and pathogenic potential. The systematic two-decade-retrospective-longitudinal-screening conducted covered 357 isolates retrieved from 19 chronically infected patients receiving care at a central hospital in Lisbon. The study involved 21 Bcc strains of six/seven Bcc species/lineages, frequently or rarely isolated from CF patients worldwide. Different strains/clonal variants obtained during infection gave rise to characteristic OAg-banding patterns. The two most prevalent and feared species, B. cenocepacia and B. multivorans, showed a tendency to lose the OAg along chronic infection. B. cenocepacia recA lineage IIIA strains known to lead to particularly destructive infections exhibit the most frequent OAg loss, compared with lineage IIIB. The switch frequency increased with the duration of infection and the level of lung function deterioration. For the first time, it is shown that the rarely found B. cepacia and B. contaminans, whose representation in the cohort of patients examined is abnormally high, keep the OAg even during 10- or 15-year infections. Data from co-infections with different Bcc species reinforced these conclusions. Concerning the two other rarely found species examined, B. stabilis exhibited a stable OAg expression phenotype over the infection period while for the single clone of the more distantly related B. dolosa species, the OAg-chain was absent from the beginning of the 5.5-year infection until the patient dead. This work reinforces the relevance attributed to the OAg-expression switch suggesting marked differences in the various Bcc species

Selective sequential recovery of zinc and copper from acid mine drainage

The observed imbalance between the unsustainable consumption of available natural metal resources and finite deposits makes the recovery and recycling of metals from metal-containing wastes an imperative. Here, ionic-liquid-based aqueous biphasic systems (IL-based ABSs) are proposed as an efficient alternative for selective metal recovery from real copper acid mine drainage (AMD) effluents. ABSs composed of different ILs and Na2SO4 were evaluated for Zn, Al, Cu, Co, and Ni extraction from both model solutions and AMD samples. It is shown that IL composed of thiocyanate anion ([SCN]−) presented a remarkable ability to extract metals from AMD through the formation of stable metal complexes. The addition of NaSCN to ABSs composed of tetrabutylammonium chloride ([N4444]Cl) allowed to mimic the use of [SCN]-based IL with additional advantages: tunable metal selectivity by the concentration of [SCN]− added to the ABS and a reduction in system cost and environmental impact. Furthermore, at the [SCN]− concentration range studied here, the formation of a hydrophobic salt composed of IL cations and metal complex anions is observed, which allows the selective extraction and recovery of transition metals in a single step. The IL-rich phase recyclability in three extraction cycles is demonstrated, showing the possibility to recover two times more Zn than with a single extraction cycle while using the same amount of IL and thiocyanate. Salt-rich phases were also recycled in a new IL-based ABS for the subsequent Cu extraction and recovery. These results allow the development of a sustainable process for the selective sequential recovery of transition metals from AMD.publishe

Adaptation and survival of Burkholderia cepacia and B. contaminans during long-term Incubation in saline solutions containing benzalkonium chloride

TheBurkholderia cepaciacomplex (Bcc) is a group of opportunistic pathogenic bacteria with a remarkable metabolic capacity and broad genotypic/phenotypic plasticity, allowing their adaptation to hostile conditions, including nutrient depleted solutions containing antimicrobial agents. Bcc bacteria are feared contaminants in pharmaceutical industries and cause nosocomial outbreaks, posing health threats to immunocompromised individuals and cystic fibrosis (CF) patients. In this study, the adaptation and survival ofB. cepaciaandB. contaminansisolates was investigated after long-term incubation in nutrient depleted saline solutions supplemented with increasing concentrations of the biocidal preservative benzalkonium chloride (BZK), recreating the storage conditions of pharmaceutical products. These epidemiologically related isolates were recovered from intrinsically contaminated saline solutions for nasal application and from two CF patients. Long-term incubation in saline solutions containing BZK led to the development of bacterial sub-populations that survived for at least 16 months, despite an initial 2-3 log decrease in viability, displaying a progressive dose-dependent decrease of colony and cell size, including the appearance of small colony variants (SCVs). Bacterial colonies lost pigmentation, changed the morphotype from rough to smooth and produced more spherical cells during extended incubation with BZK. The development of macroscopically visible cellular aggregates, rich in polysaccharide and harboring viable cells in their interior was triggered by BZK. The existence of a metabolic pathway for BZK degradation was confirmed through genome analysis. This study reveals mechanisms underlying the prevalence of Bcc bacteria as contaminants of pharmaceutical products containing BZK, which often lead to false-negative results during quality control and routine testing

Proteomic profiling of Burkholderia cenocepacia clonal isolates with different virulence potential retrieved from a cystic fibrosis patient during chronic lung infection

Respiratory infections with Burkholderia cepacia complex (Bcc) bacteria in cystic fibrosis (CF) are associated with a worse prognosis and increased risk of death. In this work, we assessed the virulence potential of three B. cenocepacia clonal isolates obtained from a CF patient between the onset of infection (isolate IST439) and before death with cepacia syndrome 3.5 years later (isolate IST4113 followed by IST4134), based on their ability to invade epithelial cells and compromise epithelial monolayer integrity. The two clonal isolates retrieved during late-stage disease were significantly more virulent than IST439. Proteomic profiling by 2-D DIGE of the last isolate recovered before the patient's death, IST4134, and clonal isolate IST439, was performed and compared with a prior analysis of IST4113 vs. IST439. The cytoplasmic and membrane-associated enriched fractions were examined and 52 proteins were found to be similarly altered in the two last isolates compared with IST439. These proteins are involved in metabolic functions, nucleotide synthesis, translation and protein folding, cell envelope biogenesis and iron homeostasis. Results are suggestive of the important role played by metabolic reprogramming in the virulence potential and persistence of B. cenocepacia, in particular regarding bacterial adaptation to microaerophilic conditions. Also, the content of the virulence determinant AidA was higher in the last 2 isolates. Significant levels of siderophores were found to be secreted by the three clonal isolates in an iron-depleted environment, but the two late isolates were more tolerant to low iron concentrations than IST439, consistent with the relative abundance of proteins involved in iron uptake.This work was supported by FEDER and FCT – Fundação para a Ciência e a Tecnologia (contract PEst-OE/EQB/LA0023/2011_ research line: Systems and Synthetic Biology; PhD grant to A.M. – SFRH/BD/37012/2007, and PD grants to S.S. – SFRH/BPD/75483/2010 and C.C. – SFRH/BPD/ 81220/2011. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.info:eu-repo/semantics/publishedVersio

Towards an applied metaecology

The complexity of ecological systems is a major challenge for practitioners and decision-makers who work to avoid, mitigate and manage environmental change. Here, we illustrate how metaecology – the study of spatial interdependencies among ecological systems through fluxes of organisms, energy, and matter – can enhance understanding and improve managing environmental change at multiple spatial scales. We present several case studies illustrating how the framework has leveraged decision-making in conservation, restoration and risk management. Nevertheless, an explicit incorporation of metaecology is still uncommon in the applied ecology literature, and in action guidelines addressing environmental change. This is unfortunate because the many facets of environmental change can be framed as modifying spatial context, connectedness and dominant regulating processes - the defining features of metaecological systems. Narrowing the gap between theory and practice will require incorporating system-specific realism in otherwise predominantly conceptual studies, as well as deliberately studying scenarios of environmental change.We thank FAPESP (grants 2014/10470-7 to AM, 2013/04585-3 to DL, 2013/50424-1 to TS and 2015/18790-3to LS), CNPq (Productivity Fellowships 301656/2011-8 to JAFDF,308205/2014-6 to RP, 306183/2014-5 to PIP and 307689/2014-0 to VDP), the National Science Foundation (DEB 1645137 toJGH), the Natural Sciences and Engineering Council of Canada (SJL,PPN), and the Academy of Finland (grants 257686 and 292765 toMC) for support. This work contributes to the Labex OT-Med (no.ANR-11-LABX-0061), funded by the French government throughthe A*MIDEX project (no. ANR-11-IDEX-0001-02)

Towards an applied metaecology

The complexity of ecological systems is a major challenge for practitioners and decision-makers who work to avoid, mitigate and manage environmental change. Here, we illustrate how metaecology - the study of spatial interdependencies among ecological systems through fluxes of organisms, energy, and matter - can enhance understanding and improve managing environmental change at multiple spatial scales. We present several case studies illustrating how the framework has leveraged decision-making in conservation, restoration and risk management. Nevertheless, an explicit incorporation of metaecology is still uncommon in the applied ecology literature, and in action guidelines addressing environmental change. This is unfortunate because the many facets of environmental change can be framed as modifying spatial context, connectedness and dominant regulating processes - the defining features of metaecological systems. Narrowing the gap between theory and practice will require incorporating system-specific realism in otherwise predominantly conceptual studies, as well as deliberately studying scenarios of environmental change. (C) 2019 Associacao Brasileira de Ciencia Ecologica e Conservacao. Published by Elsevier Editora Ltda.Peer reviewe

Survey NEN / Inquérito NEN

Introduction: Although new treatments and diagnostic methods for Neuroendocrine Neoplasms (NEN) have been introduced, the level of access to them needs somehow to be further investigated. Objectives: to understand the aspects that influence the access to the diagnosis, follow-up and treatment of patients with NEN in Brazil. Methods: This is a cross-sectional electronic survey conducted by the Brazilian Group of Gastrointestinal Tumors (GTG) and containing sixteen questions sent to Brazilian Oncologists via messaging app, aiming to identify access profiles to diagnostic and follow-up tests among patients with NEN, in addition to proven effective treatments in the Public and Private Brazilian Health Care System. Descriptive analysis was used to report the outcomes.Results: The survey was carried out with 201 Oncologists. Since (31.8%) of the Oncologists responded that they have been trained for more than 15 years and have been working with clinical practice within the scope of the Brazilian National Health System and/or the Private Health Care. For follow-up, the most requested marker was Chromogranin A (39%). Regarding diagnosis, 35.8% of the 201 participants claim to ask for a slide review in their clinical practice, and their access to tomography and resonance (58%). When contextualizing the performance of PET Scan with Gallium 68, it is available in (15%), but a significant percentage did not have access in the Brazilian National Health System Service, corresponding to (95%). Subgroup analysis revealed statistically significant differences in the availability of Somatostatin Analogue, being offered in 56% in the Brazilian National Health System and 84% in the Private Health Care (p < 0.05). Conclusion: This is the first Brazilian research that evaluated access to diagnostic and therapeutic tools in the Brazilian scenario, showing important access limitations, especially in the public sector. In view of the results presented, these restrictions can lead to unfavorable clinical outcomes in patients with NEN in Brazil

Genomic Expression Analysis Reveals Strategies of Burkholderia cenocepacia to Adapt to Cystic Fibrosis Patients' Airways and Antimicrobial Therapy

Pulmonary colonization of cystic fibrosis (CF) patients with Burkholderia cenocepacia or other bacteria of the Burkholderia cepacia complex (Bcc) is associated with worse prognosis and increased risk of death. During colonization, the bacteria may evolve under the stressing selection pressures exerted in the CF lung, in particular, those resulting from challenges of the host immune defenses, antimicrobial therapy, nutrient availability and oxygen limitation. Understanding the adaptive mechanisms that promote successful colonization and long-term survival of B. cenocepacia in the CF lung is essential for an improved therapeutic outcome of chronic infections. To get mechanistic insights into these adaptive strategies a transcriptomic analysis, based on DNA microarrays, was explored in this study. The genomic expression levels in two clonal variants isolated during long-term colonization of a CF patient who died from the cepacia syndrome were compared. One of the isolates examined, IST439, is the first B. cenocepacia isolate retrieved from the patient and the other isolate, IST4113, was obtained three years later and is more resistant to different classes of antimicrobials. Approximately 1000 genes were found to be differently expressed in the two clonal variants reflecting a marked reprogramming of genomic expression. The up-regulated genes in IST4113 include those involved in translation, iron uptake (in particular, in ornibactin biosynthesis), efflux of drugs and in adhesion to epithelial lung tissue and to mucin. Alterations related with adaptation to the nutritional environment of the CF lung and to an oxygen-limited environment are also suggested to be a key feature of transcriptional reprogramming occurring during long-term colonization, antibiotic therapy and the progression of the disease