Heavy metals in impacted ecosystem : a pressure favoring the selection of antibiotic resistant opportunistic pathogens ?

Pseudomonas aeruginosa and Stenotrophomonas maltophilia, two major opportunistic pathogens, could acquire antibiotic resistance in the environment under heavy metal pressure that co-selects both resistances. We first investigated the distribution and abundance of these species in a wide range of soils of different geographical origin (France and Africa) and evaluated the influence of human activities that may expose soils to metallic elements on this distribution. While the presence of P. aeruginosa is rather sporadic and could be linked to exogenous intake, S. maltophilia is present in all studied soils, that suggests its endemicity. Evaluating resistance capacities of strains isolated from these soils also showed differences between the two species. Environmental strains of P. aeruginosa are mostly characterized by a wild type phenotype, whereas those of S. maltophilia present a wide diversity of phenotypes depending on the site, sometimes similar to those of clinical strains. This diversity could be attributed to a deep adaptation to the very different environmental conditions encountered in the original niche but it is difficult to attribute specifically to metals a role in coselection of resistance. The study conducted on the bacterial community present in a contaminated soil has also highlighted a high proportion of bacteria resistant to different antibiotics represented by species qualified as opportunistic pathogens and the presence of the gene blaIMP, enabling resistance to imipenem, used in the hospital to treat infections due to multidrug-resistant clones.Pseudomonas aeruginosa et Stenotrophomonas maltophilia, pathogènes opportunistes majeurs, pourraient acquérir leur résistance aux antibiotiques dans l’environnement, sous la pression exercée par les métaux lourds par co-sélection de résistance. Nous avons tout d’abord évalué la distribution et l’abondance de ces espèces dans un large panel de sols d’origine géographique différente (France et Afrique) et évalué l’influence d’activités anthropiques susceptibles d’exposer les sols en éléments métalliques sur cette distribution. Alors que la présence de P. aeruginosa est sporadique et plutôt liée à un apport exogène, S. maltophilia est présente dans tous les sols étudiés, suggérant son endémicité. L’évaluation des résistances des souches isolées de ces sols a également montré des différences entre les deux espèces. Les souches environnementales de P. aeruginosa sont pour la plupart caractérisées par un phénotype sauvage alors que celles de S. maltophilia présentent une grande diversité de phénotypes en fonction des sites, parfois similaires à ceux de souches cliniques. Cette diversité peut être attribuée à l’adaptation aux conditions environnementales très différentes rencontrées mais il est difficile d‘attribuer précisément aux métaux un rôle dans la co-sélection de ces résistances. L’étude menée sur la communauté bactérienne d’un sol contaminé a également permis de mettre en évidence une forte proportion de bactéries résistantes à différents antibiotiques représentée par des espèces qualifiées de pathogènes opportunistes ainsi que la présence du gène blaIMP, permettant la résistance à l’imipénème, utilisé en milieu clinique pour le traitement de clones multi-résistants

Les métaux lourds dans les écosystèmes anthropisés : une pression favorisant la sélection de pathogènes opportunistes résistants à des antibiotiques ?

Pseudomonas aeruginosa and Stenotrophomonas maltophilia, two major opportunistic pathogens, could acquire antibiotic resistance in the environment under heavy metal pressure that co-selects both resistances. We first investigated the distribution and abundance of these species in a wide range of soils of different geographical origin (France and Africa) and evaluated the influence of human activities that may expose soils to metallic elements on this distribution. While the presence of P. aeruginosa is rather sporadic and could be linked to exogenous intake, S. maltophilia is present in all studied soils, that suggests its endemicity. Evaluating resistance capacities of strains isolated from these soils also showed differences between the two species. Environmental strains of P. aeruginosa are mostly characterized by a wild type phenotype, whereas those of S. maltophilia present a wide diversity of phenotypes depending on the site, sometimes similar to those of clinical strains. This diversity could be attributed to a deep adaptation to the very different environmental conditions encountered in the original niche but it is difficult to attribute specifically to metals a role in coselection of resistance. The study conducted on the bacterial community present in a contaminated soil has also highlighted a high proportion of bacteria resistant to different antibiotics represented by species qualified as opportunistic pathogens and the presence of the gene blaIMP, enabling resistance to imipenem, used in the hospital to treat infections due to multidrug-resistant clones.Pseudomonas aeruginosa et Stenotrophomonas maltophilia, pathogènes opportunistes majeurs, pourraient acquérir leur résistance aux antibiotiques dans l’environnement, sous la pression exercée par les métaux lourds par co-sélection de résistance. Nous avons tout d’abord évalué la distribution et l’abondance de ces espèces dans un large panel de sols d’origine géographique différente (France et Afrique) et évalué l’influence d’activités anthropiques susceptibles d’exposer les sols en éléments métalliques sur cette distribution. Alors que la présence de P. aeruginosa est sporadique et plutôt liée à un apport exogène, S. maltophilia est présente dans tous les sols étudiés, suggérant son endémicité. L’évaluation des résistances des souches isolées de ces sols a également montré des différences entre les deux espèces. Les souches environnementales de P. aeruginosa sont pour la plupart caractérisées par un phénotype sauvage alors que celles de S. maltophilia présentent une grande diversité de phénotypes en fonction des sites, parfois similaires à ceux de souches cliniques. Cette diversité peut être attribuée à l’adaptation aux conditions environnementales très différentes rencontrées mais il est difficile d‘attribuer précisément aux métaux un rôle dans la co-sélection de ces résistances. L’étude menée sur la communauté bactérienne d’un sol contaminé a également permis de mettre en évidence une forte proportion de bactéries résistantes à différents antibiotiques représentée par des espèces qualifiées de pathogènes opportunistes ainsi que la présence du gène blaIMP, permettant la résistance à l’imipénème, utilisé en milieu clinique pour le traitement de clones multi-résistants

Impact of untreated urban waste on the prevalence and antibiotic resistance profiles of human opportunistic pathogens in agricultural soils from Burkina Faso

This study examined the long-term effects of the landfill disposal of untreated urban waste for soil fertilization on the prevalence and antibiotic resistance profiles of various human opportunistic pathogens in soils from Burkina Faso. Samples were collected at three sites in the periphery of Ouagadougou during two campaigns in 2008 and 2011. At each site, amendment led to changes in physico-chemical characteristics as shown by the increase in pH, CEC, total C, total N, and metal contents. Similarly, the numbers of total heterotrophic bacteria were higher in the amended fields than in the control ones. No sanitation indicators, i.e., coliforms, Staphylococci, and Enterococci, were detected. Pseudomonas aeruginosa and Burkholderia cepacia complex (Bcc) were detected at a low level in one amended field. Stenotrophomonas maltophilia was detected from both campaigns at the three sites in the amended fields and only once in an unamended field. Diversity analysis showed some opportunistic pathogen isolates to be closely related to reference clinical strains responsible for nosocomial- or community-acquired infections in Northern countries. Antibiotic resistance tests showed that P. aeruginosa and Bcc isolates had a wild-type phenotype and that most S. maltophilia isolates had a multi-drug resistance profile with resistance to 7 to 15 antibiotics. Then we were able to show that amendment led to an increase of some human opportunistic pathogens including multi-drug resistant isolates. Although the application of untreated urban waste increases both soil organic matter content and therefore soil fertility, the consequences of this practice on human health should be considered

RegA, the transcriptional regulator of the two-component system RegB/RegA of Brucella suis is a controller of both oxidative respiration and denitrification, required for chronic infection in mice.

International audienc

RegA, the transcriptional regulator of the two-component system RegB/RegA of Brucella suis is a controller of both oxidative respiration and denitrification, required for chronic infection in mice.

International audienc

Identification of Stenotrophomonas maltophilia strains isolated from environmental and clinical samples: a rapid and efficient procedure

International audienceAims: Aim of the study is to identify accurately Stenotrophomonas maltophilia isolates recovered from environmental and clinical samples. Methods and Results: Recovery of Sten. maltophilia-like isolates from soil samples using the vancomycin, imipenem, amphotericin B (VIA) selective agar medium enabled distinction of various morphotype colonies. A set of soil and clinical isolates was tested for species identification using different methods. 16S rDNA analyses showed the dark green with a blue halo morphotype to be typical Sten. maltophilia strains. The API-20NE, Vitek-2 and Biolog phenotypic analyses typically used for the identification of clinical isolates did not perform well on these soil isolates. The species-specific PCR screening targeting Sten. maltophilia 23S rDNA and the multiplex smeD/ggpS PCR, differentiating Sten. maltophilia from Stenotrophomonas rhizophila, were tested for improvement of these identification schemes. The latter multiplex PCR identified all isolates tested in this study, whatever be their origin. Conclusions: Isolation on VIA medium and confirmation of Sten. maltophilia species membership by smeD PCR is proposed to identify environmental and clinical isolates of Sten. maltophilia. Significance and Impact of the Study: The proposed approach enables isolation and identification of Sten. maltophilia from different environments in an easy and rapid way. This approach will be useful to accurately manage studies on the abundance and distribution of Sten. maltophilia in hospital and nonhospital environments

RegA, the Regulator of the Two-Component System RegB/RegA of Brucella suis, Is a Controller of Both Oxidative Respiration and Denitrification Required for Chronic Infection in Mice

International audienc

Detection and enumeration of Pseudomonas aeruginosa in soil and manure assessed by an ecfX qPCR assay

International audienceAimsTo develop a qPCR approach for the detection of Pseudomonas aeruginosa in soil and manure and explore its efficacy and limitations compared with that of a classical culture-dependent approach.Methods and ResultsA Ps. aeruginosa ecfX qPCR assay was developed. This assay was optimized for soils of contrasting physico-chemical properties and evidenced a three-log dynamic range of detection [5 × 104 – 5 × 106 cells (g drywt soil)−1] in inoculated microcosms. Sensitivity was determined to be around 5 × 104 cells (g drywt soil)−1. In parallel, the minimum detection limit was estimated in the range of 10–100 CFU (g drywt soil)−1 using a culture-dependent approach based on the use of a selective medium (cetrimide agar base medium supplemented with nalidixic acid), coupled to ecfX gene amplification to confirm isolate identity. These soil samples led to the growth of abundant non-Ps. aeruginosa colonies mainly belonging to other Pseudomonas species but also some beta-Proteobacteria. These bacteria strongly impacted the detection threshold of this approach. Efficacy of these approaches was compared for Ps. aeruginosa enumeration among manure and agricultural soil samples from various sites in France, Tunisia and Burkina Faso.ConclusionsThe developed qPCR assay enabled a specific detection of Ps. aeruginosa in soil and manure samples. The culture-based approach was usually found more sensitive than the qPCR assay. However, abundance of non-Ps. aeruginosa species among the indigenous communities able to grow on the selective medium affected the sensitivity of this latter approach.Significance and Impact of the StudyThis study describes the first specific and sensitive qPCR assay for the detection and enumeration of Ps. aeruginosa in soil and manure and shows its complementarity with a culture-based approach

Soils as reservoir of opportunistic human material pathogens and impact of agricultural practices

National audienc