Parallel declines in species and genetic diversity driven by anthropogenic disturbance: a multispecies approach in a French Atlantic dune system.

Numerous studies assess the correlation between genetic and species diversities, but the processes underlying the observed patterns have only received limited attention. For instance, varying levels of habitat disturbance across a region may locally reduce both diversities due to extinctions, and increased genetic drift during population bottlenecks and founder events. We investigated the regional distribution of genetic and species diversities of a coastal sand dune plant community along 240 kilometers of coastline with the aim to test for a correlation between the two diversity levels. We further quantify and tease apart the respective contributions of natural and anthropogenic disturbance factors to the observed patterns. We detected significant positive correlation between both variables. We further revealed a negative impact of urbanization: Sites with a high amount of recreational infrastructure within 10 km coastline had significantly lowered genetic and species diversities. On the other hand, a measure of natural habitat disturbance had no effect. This study shows that parallel variation of genetic and species diversities across a region can be traced back to human landscape alteration, provides arguments for a more resolute dune protection, and may help to design priority conservation areas

Epidemiology of airway colonization by Scedosporium apiospermum during cystic fibrosis

With a frequency of about 10%, species of the Scedosporium apiospermum complex (which comprises at least five distinct species with different antifungal susceptibility patterns) rank the second among the filamentous fungi colonizing the airways in cystic fibrosis (CF). Additionally, it is clearly established that these fungi may disseminate in case of immunodeficiency and that a chronic colonization of the airways by these pathogens may hinder the success of lung transplantation. In this study, we develop a new genotyping method to investigate the epidemiology of the airway colonization by these fungi. 63 multiple and sequential isolates of S. apiospermum collected from 9 CF patients, and selected among those previously studied by random amplification of polymorphic DNA (RAPD), were analyzed using the automated typing system DiversiLab (bioMérieux) based on PCR amplification of repetitive sequences. The DiversiLab Aspergillus rep-PCR kit which uses specific primers designed for Aspergillus fumigatus, was compared with the pan-fungus DiversiLab Fungal kit. Amplification products were separated by capillary electrophoresis on Agilent B2100 bioanalyzer, leading to single profiles for each isolate which were then compared using the DiversiLab software. In addition, species identification of these isolates was clarified by sequencing the betatubulin gene. Results obtained with both kits were comparable. Nevertheless, differentiation was easier using the DiversiLab Fungal kit. Additionally, rep-PCR usually confirmed the colonization patterns described by RAPD. Only two patients showed distinct genotypes. For Patient 2, two isolates were analyzed which were undistinguishable by RAPD, but rep-PCR revealed that they belonged to distinct genotypes, suggesting a transient colonization. For Patient 8 which showed by RAPD two distinct genotypes, 5 genotypes were found by rep-PCR with a dominant one represented by 5 isolates and two very close genotypes (corresponding to 3 isolates), while 4 other isolates belonged to two distant genotypes. In conclusion, the automated typing system DiversiLab proved to be an easy and efficient method to investigate the molecular epidemiology of the airway colonization by S. apiospermum in CF. Our results also confirm the capacity of the different species from the S. apiospermum complex to chronically colonize the airways of CF patients

Geosmithia argillacea: an emerging agent of airway colonization in patients with cystic fibrosis?

Date du colloque&nbsp;: 06/2009</p

Rasamsonia argillacea species complex: taxonomy, pathogenesis and clinical relevance.

Since 2010, colonizations/infections by Rasamsonia argillacea species complex, previously known as Geosmithia argillacea, have been regularly reported in literature. We reviewed all available cases focusing on pathogenesis and clinical relevance. The number of cases may be underestimated, as these fungi are frequently misidentified as Penicillium or Paecilomyces species. Major underlying conditions that predispose for infections by the R. argillacea species complex include cystic fibrosis (CF) and chronic granulomatous disease (CGD). While the pathogenic role of the colonization of CF lungs is still under debate, these molds are the causative agent of pneumonia and/or invasive infections in CGD patients. Given their thermotolerance and their resistance to various antifungals, especially the azole drugs, a special attention should be paid to the chronic colonization of the airways by these fungi in CF and CGD patients

Human cryptosporidiosis in immunodeficient patients in France (2015-2017)

Cryptosporidiosis is a common disease in children and immunodeficient individuals. In 2006, a national network was set up on the surveillance of human cryptosporidiosis in France. Since January 2015, the 41 tertiary care hospitals and the 3 private laboratories of the French National Network on the surveillance of human cryptosporidiosis have been able to declare confirmed cases of cryptosporidiosis online. Between 2015 and 2017, 210 cases of cryptosporidiosis were declared in immunodeficient patients in France; Cryptosporidium parvum and Cryptosporidium hominis represented 66% and 22% of cases, respectively. A peak was observed in autumn. Cryptosporidiosis occurred mainly in a context of solid organ transplantation (SOT) (49%) and of HIV infection (30%). In SOT recipients, cryptosporidiosis appeared more frequently in the first 6 months post transplantation. Regarding cases declared in SOT recipients, mycophenolate mofetil was used in 68%. A mortality rate of 6% was observed. Present results underline the importance of screening for cryptosporidiosis in immunocompromised patients suffering from diarrhea, especially in the course of major cell mediated immunodeficiency or even systematic screening before SOT. Exclusive Cryptosporidium free water feeding could be suggested during major cell mediated immunodeficiency

Clinical and microbiological efficacy of micafungin on Geosmithia argillacea infection in a cystic fibrosis patient

Cystic fibrosis are at risk of colonization by a number of fungi, including Geosmithiaargillacea which appears to be an emerging pathogen in these patients. This pathogen has been recently reported as a cause of invasive/systemic mycosis in immunocompromized patients such as colonized patients who are immunosuppressed for lung transplantation. In this context, we report here a case of clinical and microbiological efficacy of micafungin in a French cystic fibrosis patient chronically colonized with G. argillacea. O.D., a female F508Del-CFTR homozygous patient was diagnosed at birth with cystic fibrosis in January 1996. She was found chronically colonised with multi-resistant Staphylococcus aureus (MRSA) from 1997 to 2011, and with Aspergillus fumigatus from 2001 to 2006. She was treated alternatively with oral voriconazole and itraconazole from 2004 to 2008, and with posaconazole since february 2008. Geosmithia argillacea was first diagnosed in May 2007, and chronic colonisation was persistent from this date to August 2010 with 23/28 fungus positive sputum samples, in spite of posaconazole therapy. For an isolate obtained in October 2008, minimal inhibitory/effective concentrations (MIC/MEC, mg/ml) determined using the Eucast method were 2.0, 2.0, 16.0, 2,0, 0.25 and 0.015 for amphotericin B, itraconazole, voriconazole, posaconazole, caspofungin and micafungin, respectively. The FEV1 predicted value was 73% at the time of first fungus isolation and was decreased to 47% in October 2009. She then was given caspofungin for 21 days ((70 mg/day, later reduced to 50 mg) which resulted in clinical improvement (FEV1 = 64% in January 2010) without eradication of G. argillacea. In June 2010, treatment with micafungin (75 mg, 21days) was realized owing to deterioration of the respiratory function (FEV1 = 56%),without clinical improvement ( FEV1 = 47% in August 2010). O.D. was then treated from September, 23 to November 3, 2010 with micafungin (100mg bid for 21 days and 100mg/day for the following 21 days) which resulted in clinical and microbiological improvement. FEV1 predicted ranged 67-68% in October and December 2010, and February and May 2011, and from the end of treatment to December 2010, 5/6 sputum samples were found negative for G. argillacea. The positive sample contained fungus of the same genotype as previous isolates. The present case is to our knowledge the first description of G. argillacea eradication in a chronically colonized cystic fibrosis patient. Similar to previous studies, G. argillacea colonization was detected in the presence of chronic MRSA after A. fumigatus eradication. Since no change in bacterial colonization was observed before, during, and after G. argillacea colonization, the present case is consistent with a pathogenic role of the fungus in cystic fibrosis patients. In vitro antifungal susceptibility assays suggested that echinocandins are most effective agents against this fungus with a lowest MEC for micafungin (7 isolates studied, MEC range: 0.015-0.03), although eradication could only be obtained with high dose micafungin for a long time (6 weeks)

Fungal colonization in Cystic Fibrosis (CF): Epidemiology and antifungal resistance in a French cohort of CF patients – Focused on Aspergillus fumigatus colonization

Introduction: Cystic fibrosis (CF) is the major genetic inherited disease in the European Caucasian population, with an average of 1 in 3000 living births in France. Prognostic depend essentially on the lung impairments. While considerable attention therefore has been paid over recent decades to prevent and treat bacterial respiratory infections, we observed emergence of fungi colonization in CF respiratory tract. In particular, Aspergillus fumigatus represents the most common causative agent colonizing the airways of CF patients; it can be responsible for Allergic Bronchopulmonary Aspergillosis (ABPA). Since oral corticosteroids and itraconazole represent the mainstay of ABPA treatment, long-term therapy may increase the risk of acquired resistance to azoles that is mainly associated with amino acid substitutions in the CYP51A gene of A. fumigatus. Objective: First, we managed to have exhaustive epidemiological data on species of filamentous fungi able to colonize the airway tract of 300 CF patients followed-up in our national prospective study ("MucoFong" study – PHRC1902). Second, CF patients being chronically exposed to azole (especially to itraconazole), our study aimed to evaluate the prevalence of azole resistance in isolates prospectively collected from CF patients followed-up in seven French hospitals involved in our national prospective study. Third, we focused on the most prevalent species: Aspergillus fumigatus, studying the azole resistance at molecular level. To our knowledge, it is the first multicenter study focused on azole resistance of A. fumigatus in CF. Methods: A total of 243 sputa were analyzed using the same protocol in each centre. The MICs of antifungal drugs were evaluated for each isolate using the E-test ® strips. Focusing on A. fumigatus, a total of 87 isolates was collected in 85 patients. These isolates were characterized at the molecular level by targeting ITS, ß-tubulin and MAT-A/α genes. The CYP51A gene as well as its promoter was sequenced; a 3D Cyp51A protein homology model was built. Results and discussion: 300 patients were enrolled in this study. At inclusion time, most of them were adults colonized with A. fumigatus (about 35% of the patients). Scedosporium was isolated in 5%, and Exophiala in about 2%. Regarding antifungal susceptibility, isolates of Scedosporium and Exophiala exhibited antifungal resistance comparable with published data. Regarding A. fumigatus, a majority of isolates (88.1%) were found sensitive to itraconazole (MIC≤ 2μg/ml), and 2 new mutations were identified and localized within 3-dimensional Cyp51A protein model. To obtain insight into azole resistance of A. fumigatus, the results are analyzed taking into account clinical data, itraconazole exposition, and the potential correlation between the identified CYP5IA mutations and azole resistance is discussed based on the Cyp51A protein homology model

Drug Discovery for Schistosomiasis: Hit and Lead Compounds Identified in a Library of Known Drugs by Medium-Throughput Phenotypic Screening

The flatworm disease schistosomiasis infects over 200 million people with just one drug (praziquantel) available—a concern should drug resistance develop. Present drug discovery approaches for schistosomiasis are slow and not conducive to automation in a high-throughput format. Therefore, we designed a three-component screen workflow that positions the larval (schistosomulum) stage of S. mansoni at its apex followed by screens of adults in culture and, finally, efficacy tests in infected mice. Schistosomula are small enough and available in sufficient numbers to interface with automated liquid handling systems and prosecute thousands of compounds in short time frames. We inaugurated the workflow with a 2,160 compound library that includes known drugs in order to cost effectively ‘re-position’ drugs as new therapies for schistosomiasis and/or identify compounds that could be modified to that end. We identify a variety of ‘hit’ compounds (antibiotics, psychoactives, antiparasitics, etc.) that produce behavioral responses (phenotypes) in schistosomula and adults. Tests in infected mice of the most promising hits identified a number of ‘leads,’ one of which compares reasonably well with praziquantel in killing worms, decreasing egg production by the parasite, and ameliorating disease pathology. Efforts continue to more fully automate the workflow. All screen data are posted online as a drug discovery resource

Evaluation and mangement of fungal risk in cystic fibrosis: first results of a national French study

Date du colloque&nbsp;: 06/2009</p