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BACKGROUND: Invasive aspergillosis(IA) by a triazole resistant Aspergillus fumigatus is associated with a high mortality. Real-time resistance detection will result in earlier initiation of appropriate therapy. METHODS: In a prospective study in the Netherlands and Belgium, we evaluated the clinical value of the multiplex AsperGenius®PCR in hematology patients from 12 centers. This PCR detects the most frequent cyp51A mutations in A. fumigatus conferring azole-resistance. Patients were included when a CT-scan showed a pulmonary infiltrate and bronchoalveolar lavage(BALf) sampling was performed. The primary endpoint was antifungal treatment failure in patients with azole-resistant IA. Patients with mixed azole-susceptible/resistant infections were excluded. RESULTS: Of 323 patients enrolled, complete mycological and radiological information was available in 276/323(94%) and probable IA diagnosed in 99/276(36%). Sufficient BALf for PCR testing was available in 293/323(91%). Aspergillus DNA was detected in 116/293(40%) and A.fumigatus DNA in 89/293(30%). The resistance PCR was conclusive in 58/89(65%) and resistance detected in 8/58(14%). Two had a mixed azole-susceptible/resistant infection. In the 6 remaining patients, treatment failure was observed in one. Galactomannan positivity was associated with higher mortality(p=0.004). In contrast, mortality of patients with an isolated positive Aspergillus PCR was comparable to those with a negative PCR(p=0.83). CONCLUSIONS: Real-time PCR-based resistance testing may help to limit the clinical impact of triazole resistance. In contrast, the clinical impact of an isolated positive Aspergillus PCR on BALf seems limited. The interpretation of the EORTC/MSGERC PCR criterion for BALf may need further specification (e.g. minimum Ct-value and/or PCR positive on >1 BALf sample).

Background Surveillance of human leishmaniasis in Europe is mostly limited to country-specific information from autochthonous infections in the southern part. As at the end of 2021, no integrated analysis has been performed for cases seen across centres in different European countries. Aim To provide a broad perspective on autochthonous and imported leishmaniasis cases in endemic and non-endemic countries in Europe. Methods We retrospectively collected records from cutaneous, mucosal and visceral leishmaniasis cases diagnosed in 15 centres between 2014 and 2019. Centres were located in 11 countries: Belgium, France, Germany, Italy, the Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. Data on country of infection, reason for travelling, infecting species, age and sex were analysed. Results We obtained diagnostic files from 1,142 cases, of which 76%, 21% and 3% had cutaneous, visceral, and mucosal disease, respectively. Of these, 68% were men, and 32% women, with the median age of 37 years (range: 0–90) at diagnosis. Visceral leishmaniasis was mainly acquired in Europe (88%; 167/190), while cutaneous leishmaniasis was primarily imported from outside Europe (77%; 575/749). Sixty-two percent of cutaneous leishmaniasis cases from outside Europe were from the Old World, and 38% from the New World. Geographic species distribution largely confirmed known epidemiology, with notable exceptions. Conclusions Our study confirms previous reports regarding geographic origin, species, and traveller subgroups importing leishmaniasis into Europe. We demonstrate the importance of pooling species typing data from many centres, even from areas where the aetiology is presumably known, to monitor changing epidemiology.

Background: Limited prospective data are available on the acquisition of viral, bacterial and parasitic diarrhoeagenic agents by healthy individuals during travel.Methods: To determine the frequency of travel associated acquisition of 19 pathogens in 98 intercontinental travellers, qPCR was used to detect 8 viral pathogens, 6 bacterial enteric pathogens and 5 parasite species in faecal samples collected immediately before and after travel.Results: We found high pre-travel carriage rates of Blastocystis spp. and Dientamoeba fragilis of 32% and 19% respectively. Pre-travel prevalences of all other tested pathogens were below 3%. Blastocystis spp. (10%), Plesiomonas shigelloides (7%), D. fragilis (6%) and Shigella spp. (5%) were the most frequently acquired pathogens and acquisition of enteral viruses and hepatitis E virus in this relatively small group of travellers was rare or non-existent.Conclusions: Our findings suggest that the role of viruses as the cause of persisting traveller's diarrhoea is limited and bacterial pathogens are more likely as a cause of traveller's diarrhoea. The substantial proportion of travellers carrying Blastocystis spp. and D. fragilis before travel warrants cautious interpretation of positive samples in returning travellers with gastrointestinal complaints. (C) 2017 Elsevier Ltd. All rights reserved.

International audience

Introduction: A lack of prospective and longitudinal data on pre- and post-travel carriage of Blastocystis spp. complicates interpretation of a positive WA post-travel. Therefore we studied dynamics of Blastocystis carriage in a cohort of Dutch travellers.Methods: From the prospective, multicentre COMBAT study among 2001 Dutch travellers, a subset of 491 travellers was selected based on travel destination to 7 subregions (70 or 71 travellers each). Faecal samples taken directly before and after travel were screened for Blastocystis with qPCR, followed, when positive, by sequence analysis to determine subtypes.Results: After exclusion of 12 samples with missing samples or inhibited qPCR-reactions, stool samples of 479 travellers were analysed. Before travel, 174 of them (36.3%) carried Blastocystis and in most of these, the same subtype was persistently carried. However, in 48/174 of those travellers (27.6%; CI95 20.8-36.6%) no Blastocystis or a different subtype was detected in the post-travel sample, indicating loss of Blastocystis during travel. Only 26 (5.4%; CI95 3.7%-8.0%) of all travellers acquired Blastocystis, including two individuals that were already positive for Blastocystis before travel but acquired a different subtype during travel.Discussion: This study shows that Blastocystis carriage in travellers is highly dynamic. The observed acquisition and loss of Blastocystis could either be travel-related or reflect the natural course of Blastocystis carriage. We demonstrate that the majority of Blastocystis detected in post-travel samples were already carried before travel.

International audience; Blastocystis is a highly prevalent anaerobic eukaryotic intestinal parasite found in the intestinal tract of human and various animals. Although the role of Blastocystis as human pathogen remains unclear, it can cause acute or chronic digestive disorders and some studies have suggested an association with irritable bowel syndrome. Seventeen subtypes (ST1-ST17), among which the first nine are found in human, have been identified based on the gene coding for the small-subunit ribosomal RNA. We have previously sequenced the first whole genome of Blastocystis ST7 (Denoeud et al., 2011). It consists of a 18.8 Mb nuclear genome with more than 6000 genes and a circular genome of 29 Kbp located within mitochondria-like organelles (MLO). Here we report the sequencing and annotation of the genome of a Blastocystis ST4 isolate. Genome sequencing was done with the Illumina HiSeq 2000 system generating more than 43 millions of 100-bp paired-end reads. The sequence reads were de novo assembled using the IDBA-ud algorithm. In total, 3996 scaffolds higher than 200 bp were obtained, with a scaffold N50 determined to be 20,431 bp. The draft genome sequence of Blastocystis ST4 has a total of 13.36 Mbp. As expected, assembly also provided a circular genome of ~27 kb in size corresponding to the whole MLO genome sequence. Genes were predicted using the Maker gene annotation pipeline. A first run of Maker was performed using the ab initio predictor Augustus trained with 413 genes manually designed from the ST4 scaffolds, the ~6000 annotated genes of the ST7 genome and available ESTs data from both ST7 and ST1. Genes determined from this first run were then used to train another ab initio gene prediction program called SNAP. A second run of Maker similar to the first run and including the newly trained gene predictor SNAP was finally performed. This led to significant improvements in gene prediction accuracy with a final annotation set of 6046 genes. The same pipeline was also used to complete and correct the previous annotation of the ST7 genome (Denoeud et al., 2011). Gene functions (for ST4 predicted genes) were annotated by Blast2GO and blastP analyses with NCBI, Swissprot/Uniprot and KEGG databases. Finally OrthoMCL was applied to compare both ST4 and ST7 genomes. This led to the identification of new candidate genes, in particular some potential virulence factors that may be involved in the physiopathology of this parasite. The sequencing of other ST genomes is under progress and should be very helpful for a better understanding of the genetic diversity, pathogenesis, metabolic potential and genome evolution of this neglected human parasite.

A total of 8-10 million persons are infected worldwide with Trypanosoma cruzi, the causative parasite of Chagas disease, most of whom are inhabitants of Latin America. Due to the increased migration of peoples, Chagas disease has been on the uprise outside Latin America, including in Europe. The course of Chagas, also called American trypanosomiasis, runs in 2 phases: an acute phase lasting approximately 2 months, and a chronic phase in which symptoms may appear years after infection. Without treatment, the patient will remain infected for life. The acute phase is usually asymptomatic; in the chronic phase of American trypanosomiasis, severe gastro-intestinal and cardiac abnormalities may develop, finally with fatal course. In the Netherlands, the number of immigrants who would serologically test positive for American trypanosomiasis is estimated to be between 726 and 2929. Healthcare providers in the Netherlands may encounter patients who have Chagas disease more and more frequently. The screening of pregnant women and blood donors at risk for American trypanosomiasis should be considered

A 28-year-old woman presented with extensive erythematous lesions on her back after visiting Malawi. Skin biopsies showed ova, which could belong to Schistosoma spp. Sequencing of the Schistosoma 28S rRNA gene, extracted and amplified from paraffin biopsies, identified DNA of Schistosoma haematobium. Cutaneous ectopic schistosomiasis can present with extensive lesions and should be considered in the differential diagnosis of skin lesions in returning travelers. Microscopy and serology are the classical methods to obtain a diagnosis. Alternatively, molecular methods can be a valuable new tool for diagnosis and species determination.