Piggybacking on Niche Adaptation Improves the Maintenance of Multidrug-Resistance Plasmids.

The persistence of plasmids in bacterial populations represents a puzzling evolutionary problem with serious clinical implications due to their role in the ongoing antibiotic resistance crisis. Recently, major advancements have been made toward resolving this "plasmid paradox" but mainly in a nonclinical context. Here, we propose an additional explanation for the maintenance of multidrug-resistance plasmids in clinical Escherichia coli strains. After coevolving two multidrug-resistance plasmids encoding resistance to last resort carbapenems with an extraintestinal pathogenic E. coli strain, we observed that chromosomal media adaptive mutations in the global regulatory systems CCR (carbon catabolite repression) and ArcAB (aerobic respiration control) pleiotropically improved the maintenance of both plasmids. Mechanistically, a net downregulation of plasmid gene expression reduced the fitness cost. Our results suggest that global chromosomal transcriptional rewiring during bacterial niche adaptation may facilitate plasmid maintenance.

Tn1 transposition in the course of natural transformation enables horizontal antibiotic resistance spread in Acinetobacter baylyi.

Transposons are genetic elements that change their intracellular genomic position by transposition and are spread horizontally between bacteria when located on plasmids. It was recently discovered that transposition from fully heterologous DNA also occurs in the course of natural transformation. Here, we characterize the molecular details and constraints of this process using the replicative transposon Tn1 and the naturally competent bacterium Acinetobacter baylyi. We find that chromosomal insertion of Tn1 by transposition occurs at low but detectable frequencies and preferably around the A. baylyi terminus of replication. We show that Tn1 transposition is facilitated by transient expression of the transposase and resolvase encoded by the donor DNA. RecA protein is essential for the formation of a circular, double-stranded cytoplasmic intermediate from incoming donor DNA, and RecO is beneficial but not essential in this process. Absence of the recipient RecBCD nuclease stabilizes the double-stranded intermediate. Based on these results, we suggest a mechanistic model for transposition during natural transformation.

Host dependent maintenance of a blaNDM-1-encoding plasmid in clinical Escherichia coli isolates.

Dissemination of bacterial clones carrying plasmid-mediated resistance genes is a major factor contributing to the increasing prevalence of antibiotic resistance. Understanding the evolution of successful clones and the association to mobile resistance elements are therefore crucial. In this study, we determined the sequence of a 145 kb IncC multi-drug resistance plasmid (pK71-77-1-NDM), harbouring resistance genes to last-resort antibiotics including carbapenems. We show that the plasmid is able to transfer into a range of genetically diverse clinical Escherichia coli strains and that the fitness cost imposed on the host is often low. Moreover, the plasmid is stably maintained under non-selective conditions across different genetic backgrounds. However, we also observed a lower conjugation frequency and higher fitness cost in the E. coli sequence type (ST) 73 background, which could partially explain why this clone is associated with a lower level of antibiotic resistance than other E. coli clones. This is supported by a bioinformatical analysis showing that the ST73 background harbours plasmids less frequently than the other studied E. coli STs. Studying the evolution of antibiotic resistance in a clinical context and in diverse genetic backgrounds improves our understanding of the variability in plasmid-host associations.

Conserved collateral antibiotic susceptibility networks in diverse clinical strains of Escherichia coli.

There is urgent need to develop novel treatment strategies to reduce antimicrobial resistance. Collateral sensitivity (CS), where resistance to one antimicrobial increases susceptibility to other drugs, might enable selection against resistance during treatment. However, the success of this approach would depend on the conservation of CS networks across genetically diverse bacterial strains. Here, we examine CS conservation across diverse Escherichia coli strains isolated from urinary tract infections. We determine collateral susceptibilities of mutants resistant to relevant antimicrobials against 16 antibiotics. Multivariate statistical analyses show that resistance mechanisms, in particular efflux-related mutations, as well as the relative fitness of resistant strains, are principal contributors to collateral responses. Moreover, collateral responses shift the mutant selection window, suggesting that CS-informed therapies may affect evolutionary trajectories of antimicrobial resistance. Our data allow optimism for CS-informed therapy and further suggest that rapid detection of resistance mechanisms is important to accurately predict collateral responses.

Development and validation of risk profiles of West African rural communities facing multiple natural hazards.

West Africa has been described as a hotspot of climate change. The reliance on rain-fed agriculture by over 65% of the population means that vulnerability to climatic hazards such as droughts, rainstorms and floods will continue. Yet, the vulnerability and risk levels faced by different rural social-ecological systems (SES) affected by multiple hazards are poorly understood. To fill this gap, this study quantifies risk and vulnerability of rural communities to drought and floods. Risk is assessed using an indicator-based approach. A stepwise methodology is followed that combines participatory approaches with statistical, remote sensing and Geographic Information System techniques to develop community level vulnerability indices in three watersheds (Dano, Burkina Faso; Dassari, Benin; Vea, Ghana). The results show varying levels of risk profiles across the three watersheds. Statistically significant high levels of mean risk in the Dano area of Burkina Faso are found whilst communities in the Dassari area of Benin show low mean risk. The high risk in the Dano area results from, among other factors, underlying high exposure to droughts and rainstorms, longer dry season duration, low caloric intake per capita, and poor local institutions. The study introduces the concept of community impact score (CIS) to validate the indicator-based risk and vulnerability modelling. The CIS measures the cumulative impact of the occurrence of multiple hazards over five years. 65.3% of the variance in observed impact of hazards/CIS was explained by the risk models and communities with high simulated disaster risk generally follow areas with high observed disaster impacts. Results from this study will help disaster managers to better understand disaster risk and develop appropriate, inclusive and well integrated mitigation and adaptation plans at the local level. It fulfills the increasing need to balance global/regional assessments with community level assessments where major decisions against risk are actually taken and implemented.

Low biological cost of carbapenemase-encoding plasmids following transfer from Klebsiella pneumoniae to Escherichia coli.

OBJECTIVES: The objective of this study was to determine the biological cost, stability and sequence of two carbapenemase-encoding plasmids containing blaKPC-2 (pG12-KPC-2) and blaVIM-1 (pG06-VIM-1) isolated from Klebsiella pneumoniae when newly acquired by uropathogenic Escherichia coli clinical isolates of different genetic backgrounds. METHODS: The two plasmids were transferred into plasmid-free E. coli clinical isolates by transformation. The fitness effect of newly acquired plasmids on the host cell was assessed in head-to-head competitions with the corresponding isogenic strain. Plasmid stability was estimated by propagating monocultures for ∼312 generations. Plasmid nucleotide sequences were determined using next-generation sequencing technology. Assembly, gap closure, annotation and comparative analyses were performed. RESULTS: Both plasmids were stably maintained in three of four E. coli backgrounds and resulted in low to moderate reductions in host fitness ranging from 1.1% to 3.6%. A difference in fitness cost was observed for pG12-KPC-2 between two different genetic backgrounds, while no difference was detected for pG06-VIM-1 between three different genetic backgrounds. In addition, a difference was observed between pG12-KPC-2 and pG06-VIM-1 in the same genetic background. In general, the magnitude of biological cost of plasmid carriage was both host and plasmid dependent. The sequences of the two plasmids showed high backbone similarity to previously circulating plasmids in K. pneumoniae. CONCLUSIONS: The low to modest fitness cost of newly acquired and stably maintained carbapenemase-encoding plasmids in E. coli indicates a potential for establishment and further dissemination into other Enterobacteriaceae species. We also show that the fitness cost is both plasmid and host specific.

Growth phase-specific evolutionary benefits of natural transformation in Acinetobacter baylyi.

Natural transformation in bacteria facilitates the uptake and genomic integration of exogenous DNA. This allows horizontal exchange of adaptive traits not easily achieved by point mutations, and has a major role in the acquisition of adaptive traits exemplified by antibiotic resistance determinants and vaccination escape. Mechanisms of DNA uptake and genomic integration are well described for several naturally transformable bacterial species; however, the selective forces responsible for its evolution and maintenance are still controversial. In this study we evolved transformation-proficient and -deficient Acinetobacter baylyi for 175 days in serial transfer cultures where stress was included. We found that natural transformation-proficient populations adapted better to active growth and early stationary phase. This advantage was offset by the reduced performance in the late stationary/death phase. We demonstrate fitness trade-offs between adaptation to active growth and survival in stationary/death phase caused by antagonistic pleiotropy. The presented data suggest that the widely held assumption that recombination speeds up adaptation by rapid accumulation of multiple adaptive mutations in the same genetic background is not sufficient to fully account for the maintenance of natural transformation in bacteria.

Interdisciplinary assessment of sea-level rise and climate change impacts on the lower Nile delta, Egypt.

CLImate-induced changes on WAter and SECurity (CLIWASEC) was a cluster of three complementary EC-FP7 projects assessing climate-change impacts throughout the Mediterranean on: hydrological cycles (CLIMB - CLimate-Induced changes on the hydrology of Mediterranean Basins); water security (WASSERMed - Water Availability and Security in Southern EuRope and the Mediterranean) and human security connected with possible hydro-climatic conflicts (CLICO - CLImate change hydro-COnflicts and human security). The Nile delta case study was common between the projects. CLIWASEC created an integrated forum for modelling and monitoring to understand potential impacts across sectors. This paper summarises key results from an integrated assessment of potential challenges to water-related security issues, focusing on expected sea-level rise impacts by the middle of the century. We use this common focus to illustrate the added value of project clustering. CLIWASEC pursued multidisciplinary research by adopting a single research objective: sea-level rise related water security threats, resulting in a more holistic view of problems and potential solutions. In fragmenting research, policy-makers can fail to understand how multiple issues can materialize from one driver. By combining efforts, an integrated assessment of water security threats in the lower Nile is formulated, offering policy-makers a clearer picture of inter-related issues to society and environment. The main issues identified by each project (land subsidence, saline intrusion - CLIMB; water supply overexploitation, land loss - WASSERMed; employment and housing security - CLICO), are in fact related. Water overexploitation is exacerbating land subsidence and saline intrusion, impacting on employment and placing additional pressure on remaining agricultural land and the underdeveloped housing market. All these have wider implications for regional development. This richer understanding could be critical in making better policy decisions when attempting to mitigate climate and social change impacts. The CLIWASEC clustering offers an encouraging path for the new European Commission Horizon 2020 programme to follow.

Seasonal water demand in Benin's agriculture.

This paper describes and analyzes agricultural water demands for Benin, West Africa. Official statistical data regarding water quantities as well as knowledge on factors influencing the demand for water are extremely rare and often reveal national trends without considering regional or local differences. Thus policy makers usually work with this estimated and aggregated data, which make it very difficult to adequately address regional and local development goals. In the framework of an interdisciplinary analysis the following paper provides insight into water quantification and detects water problems under seasonal aspects for agriculture according to regional differences. Following the definition of the Food and Agriculture Organization [FAO, 1995. Water Report 7. Irrigation in Africa in Figures. Rome] agriculture is divided into irrigation and livestock watering, which were analyzed using different field methods. The study reveals that although water supply in absolute terms seems to be sufficient in Benin, seasonal water problems occur both in irrigation and in livestock management. Thus arising seasonal water problems are not the consequence of general water scarcity but more linked to three major problems. These problems emerge from difficulties in technical equipment and financial means of farmers, from the specific local conditions influencing the access to water sources and the extraction of groundwater, and third from the overall low organizational structure of water management. Therefore regional differences as well as a general improvement of knowledge on better management structures, technical know how, and access to credits for farmers need to be considered in national strategies in order to improve the agricultural water usage in Benin.