Fluconazole Resistance and Virulence in In Vitro Induced-Fluconazole Resistant Strains and in Clinical Fluconazole Resistant Strain of Cryptococcus deuterogattii.

Neuromeningeal cryptococcosis is a life-threatening infection of the central nervous system, caused by encapsulated yeast belonging to the Cryptococcus neoformans and Cryptococcus gattii species complexes. Recent data showed that virulence and antifungal resistance are variable for yeasts belonging to the C. gattii species complex. There is an increase in resistance to fluconazole for yeasts of the C. gattii species complex and the virulence is variable according to the genotype. In the present study, (i) we explored and compared the mechanisms of resistance to fluconazole between C. deuterogattii clinically resistant strains and induced fluconazole-resistant strains by exposure to fluconazole in vitro, and (ii) we studied their virulence in the Galleria mellonella study model. We demonstrated that the fluconazole resistance mechanisms involved were different between clinically resistant strains and induced resistant strains. We also demonstrated that fluconazole-induced resistant strains are less virulent when compared to the original susceptible strains. On the contrary, the clinically resistant strain tested maintains its virulence compared to fluconazole-susceptible strains of the same sequence type.

BD BACTEC™ Mycosis IC/F culture vials for fungemia diagnosis and follow-up: a retrospective study from 2013 to 2020.

This retrospective study compared the BD BACTEC™ Mycosis IC/F with the BD BACTEC™ Plus Aerobic/F and BD BACTEC™ Lytic Anaerobic/F culture vials (i.e., standard vials) for fungemia diagnosis at Nîmes University Hospital, France. From 2013 to 2020, 57 blood samples were concomitantly collected in the 3 culture vial types. For 43.8% of these samples, all vials were positive for yeast. The mean time to positivity was shorter (32.0 hours vs 44.2 hours; -12.2 hours) and longer (89.4 hours vs 33.7 hours; +55.7 hours) with the BD BACTEC™ Mycosis IC/F culture vials than with the other culture vials in patients without and with antifungal treatment, respectively. Moreover 31.6% and 24.6% of samples were positive only with the standard vials and with the BD BACTEC™ Mycosis IC/F culture vials, respectively. The BD BACTEC™ Mycosis IC/F culture vials are useful for the initial fungemia diagnosis (before any treatment) because they provide faster results.

Epidemiological analysis of nail aspergillosis and non-dermatophyte moulds. Criteria for the involvement of Aspergillus in 102 cases of onychomycosis at Montpellier University Hospital, France (1991-2019).

BACKGROUND: Moulds are often wrongly considered contaminants, not very sensitive to conventional antifungal treatments, but they may cause ungual hyphomycosis, particularly Aspergillus. Due to the lack of precise diagnostic criteria, their real impact is underestimated. OBJECTIVES: Retrospective descriptive analysis of all ungual hyphomycosis cases diagnosed at Montpellier Hospital from 1991 to 2019 to: (i) determine the incidence of onychomycosis by pseudo-dermatophytes and moulds; (ii) perform an epidemiological analysis of nail aspergillosis; and (iii) provide simple criteria for mould involvement in onychopathy. PATIENTS/METHODS: Data concerning 4053 patients were collected: age, sex, onychomycosis location, direct examination results, species(s) identified and fungal co-infections. Moreover, clinical data of patients with nail aspergillosis were analysed to identify potential contributing factors, and the classical criteria for mould involvement in onychopathy were critically reviewed. RESULTS: A pseudo-dermatophyte or a mould was involved in nail colonisation in 17.25% of patients (men/women ratio: 0.70; mean age: 53.1 years). The identified hyphomycetes belonged mainly to the genera Fusarium (45.68%), Scopulariopsis (30.23%) and Aspergillus (16.94%). Analysis of the clinical reports of 102 patients with ungual aspergillosis (men/women ratio: 0.67; mean age: 56.3 years) identified cardiovascular (43.9%), endocrine (25.8%), cancer (19.7%) and skin (18.2%) diseases as contributing factors. CONCLUSIONS: The adoption of simple and reliable criteria (ie, characteristic filaments on direct microscopic examination after periodic acid-Schiff staining, growth at seeding points in culture) allows determining the formal involvement of a mould in chronic onychopathies and avoiding possible side effects and costs of empirical or inappropriate and repetitive treatments.

Evaluation of a new point-of-care testing for creatinine and urea measurement.

Point of care testing makes it possible to obtain results in an extremely short time. Recently, radiometer has expanded the panel of tests available on its ABL90 FLEX PLUS blood gas analyzer (ABL90) by adding urea and creatinine. The aim of this study was to verify the performance of these new parameters. This included assessment of imprecision, linearity, accuracy by comparison with central laboratory standard assays and interferences. In addition, clinical utility in a dialysis center was evaluated. Within-lab coefficients of variation were close to 2%. The mean and limits of agreement (mean ± 1.96 SD) of the difference between ABL90 and Roche enzymatic assays on cobas 8000 were 0.5 (from -1.4 to 2.3) mmol/L and -0.9 (from -19.5 to 17.8) µmol/L for urea and creatinine, respectively. The ABL90 enzymatic urea and creatinine assays met the acceptance criteria based on biological variation for imprecision and showed good agreement with central laboratory. The two assays were unaffected by hematocrit variation between 20 and 70%, hemolysis and icterus interferences. It should be noted that the relationship between lab methods and ABL90 was conserved even for high pre-dialysis values allowing easy access to dialysis adequacy parameters (Kt/V) and muscle mass evaluation (creatinine index). Rapid measurement of creatinine and urea using whole blood specimens on ABL90 appears as a fast and convenient method. Analytical performances were in accordance with our expectations without any significant interferences by hemolysis or icterus.

Rapid diagnostic tests failing to detect infections by Plasmodium falciparum encoding pfhrp2 and pfhrp3 genes in a non-endemic setting.

BACKGROUND: Rapid diagnostic tests (RDTs) detecting the histidine-rich protein 2 (PfHRP2) have a central position for the management of Plasmodium falciparum infections. Yet, variable detection of certain targeted motifs, low parasitaemia, but also deletion of pfhrp2 gene or its homologue pfhrp3, may result in false-negative RDT leading to misdiagnosis and delayed treatment. This study aimed at investigating the prevalence, and understanding the possible causes, of P. falciparum RDT-negative infections at Montpellier Academic Hospital, France. METHODS: The prevalence of falsely-negative RDT results reported before and after the introduction of a loop-mediated isothermal amplification (LAMP) assay, as part as the malaria screening strategy in January 2017, was analysed. Negative P. falciparum RDT infections were screened for pfhrp2 or pfhrp3 deletion; and exons 2 were sequenced to show a putative genetic diversity impairing PfHRP2 detection. RESULTS: The overall prevalence of P. falciparum negative RDTs from January 2006 to December 2018 was low (3/446). Whereas no cases were reported from 2006 to 2016 (0/373), period during which the malaria diagnostic screen was based on microscopy and RDT, prevalence increased up to 4.1% (3/73) between 2017 and 2018, when molecular detection was implemented for primary screening. Neither pfhrp2/3 deletion nor major variation in the frequency of repetitive epitopes could explain these false-negative RDT results. CONCLUSION: This paper demonstrates the presence of pfhrp2 and pfhrp3 genes in three P. falciparum RDT-negative infections and reviews the possible reasons for non-detection of HRP2/3 antigens in a non-endemic setting. It highlights the emergence of falsely negative rapid diagnostic tests in a non-endemic setting and draws attention on the risk of missing malaria cases with low parasitaemia infections using the RDT plus microscopy-based strategy currently recommended by French authorities. The relevance of a novel diagnostic scheme based upon a LAMP assay is discussed.

Phlebotomine sandflies (Diptera, Psychodidae) from the Ankarana tsingy of northern Madagascar: inventory and description of new taxa.

An inventory of Phlebotomine sandflies was carried out in the Ankarana tsingy located in far northern Madagascar. A total of 723 sandflies were used for morphological, morphometric, and molecular studies (sequencing of partial cytochrome B (mtDNA) and partial 28S (rDNA)). Nine species were identified: Phlebotomus fertei, Sergentomyia anka, Se. sclerosiphon, Se. goodmani, two species of the genus Grassomyia, as well as three new species described herein: Se. volfi n. sp., Se. kaltenbachi n. sp., and Se. ozbeli n. sp. The recognition of these new species is strongly supported by molecular analyses. The first two of the new species could not be classified into any existing subgenus, therefore we proposed two new subgenera (Ranavalonomyia subg. nov., and Riouxomyia subg. nov.), with combinations as: Sergentomyia (Ranavalonomyia) volfi and Sergentomyia (Riouxomyia) kaltenbachi. Our study reveals important molecular variability in Se. anka, with the recognition of a population whose taxonomic status remains below that of species. Our research confirms the need to further study the specific diversity of Malagasy sandflies, which until the start of this millennium remained mostly unknown.

TITLE: Les phlébotomes (Diptera, Psychodidae) des tsingy d'Ankarana dans le nord de Madagascar : inventaire et description de nouveaux taxons. ABSTRACT: Un inventaire des Phlébotomes a été réalisé dans les tsingy d'Ankarana à l'extrême nord de Madagascar. Au total, 723 phlébotomes ont servi à des études morphologique, morphométrique et moléculaire (séquençage d'une partie du cytochrome B (ADNmt) et d'une partie de l'ADNr 28S). Neuf espèces ont été identifiées : Phlebotomus fertei, Sergentomyia anka, Se. sclerosiphon, Se. goodmani, deux espèces du genre Grassomyia, ainsi que trois espèces nouvelles décrites dans ce travail : Se. volfi n. sp., Se. kaltenbachi n. sp., and Se. ozbeli n. sp. L'individualisation de chacune de ces espèces nouvelles est robustement soutenue par les analyses moléculaires. Les deux premières de ces espèces nouvelles ne pouvaient pas être classées dans un sous-genre existant et nous avons proposé pour elles deux sous-genres nouveaux : Ranavalonomyia subg. nov., et Riouxomyia subg. nov, avec les combinaisons Sergentomyia (Ranavalonomyia) volfi et Sergentomyia (Riouxomyia) kaltenbachi.Notre étude révèle une variabilité moléculaire marquée chez Se. anka avec l'individualisation d'une population dont le statut taxinomique demeure populationnel. Nos travaux confirment la nécessité de poursuivre l'étude de la biodiversité des Phlébotomes qui est restée méconnue dans ce pays jusqu'au début de ce millénaire.

Determining the domestic specific loads of two wastewater plants of the Paris conurbation (France) with contrasted treatments: a step for exploring the effects of the application of the European Directive.

The effluents of wastewater treatment plants (WTTP) discharged into the rivers considerably affect the biogeochemical functioning of the system. In this paper, we characterize both raw and treated domestic wastewater from two WTTPs of Parisian agglomeration using different process treatments (Achères WWTP with a secondary treatment and Colombes WWTP with a tertiary one). In addition to the classical variables, we analyse the input of bacteria, both the heterotrophs and the nitrifyers. Tertiary treatment leads to significantly decrease ammonium-specific load (< 2 g KjN inhab equ.(-1) instead of 9 g KjN inhab equ.(-1) for secondary treatment) and notably reduces the one of organic matter (approximately 2.5 g biological oxygen demand (BOD) inhab equ.(-1) instead of approximately 7.5 g BOD inhab equ.(-1) for secondary treatment); it is therefore promising to improve oxygen status of both the Seine river and its estuary. In terms of total bacterial biomass abatement (the heterotrophs mostly), bioreactors (at Colombes WWTP) eliminate 12% more bacterial biomass than the activated sludge treatment (at Achères WWTP). Regarding the nitrifying bacteria, a tertiary treatment in bioreactors eliminates reverse similar 90% of both nitrifying bacteria and nitrogen pollution of wastewater. Bacterial populations are characterized by large size bacteria (> 1 microm) with a higher growth rate, that represent in the treatment plant effluents 70% of the biomass. These large size bacteria have therefore a strong impact in the organic matter degradation and oxygen consumption. Relationships between classical physical-chemical variables routinely analysed in WWTPs laboratory and bacterial biomass (heterotrophic and nitrifying) are established, in order to quantify the ecological role of the allochthonous bacteria brought into the river system. In addition, domestic specific loads are calculated for both raw and treated effluents of the two types of WWTPs. As the application of the European Water Directive requires to upgrade the wastewater treatment at Achères WWTP as soon as 2007 for 90% nitrification and 30% denitrification and in 2015 for further denitrification (up to 70%), the results of this study can be taken as point-source constraints into the modelling approach already developed for the Seine basin, and chosen to test the implementation of the Water Frame Directive.