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Fournier, P.-E.

Publications and source records attributed to Fournier, P.-E..

5 recordsLinked to original sources

Description of Rickettsia senegalensis sp. nov.: a new Rickettsia species detected worldwide

Members of the genus Rickettsia are widespread obligate intracellular bacteria with high medical and veterinary relevance. While Rickettsia senegalensis was previously isolated from cat fleas, Ctenocephalides felis, in Senegal, a comprehensive polyphasic characterization is required for its formal taxonomic validation. This study aims to provide the first detailed phenotypic, ultrastructural, and genomic characterization of the type strain PU01-02. Cultivation assays demonstrated that strain PU01-02 grows aerobically in XTC-2, SF9, and LD652 cell lines at 28 {degrees}C in a CO2-free atmosphere. Ultrastructural examination via scanning electron microscopy using a backscattered electron detector confirmed its free cytoplasmic localization and a typical rickettsial morphology characterized by rod-shaped cells approximately 1.5 {micro}m in length and 0.3 {micro}m in width. Whole-genome sequencing yielded a 1.62-Mb genome with a G+C content of 33.2%, and three plasmids were detected. Genomic analyses confirmed its status as a distinct species, placing it within the transitional Rickettsia group, specifically inside the Rickettsia felis cluster. A review of epidemiological data revealed that rickettsial genes identical to those of R. senegalensis had already been identified in several hematophagous arthropods, including fleas and ticks parasitizing various hosts such as cats, dogs, opossums, and rodents worldwide. It has also been detected in cat tissues, suggesting a potential host-pathogen association. Based on these comprehensive characterization data, we formally propose Rickettsia senegalensis sp. nov. as a new species, with PU01-02 (= CSUR R184 = DSM 28250) as the type strain. RepositoriesThe genome sequence of Rickettsia senegalensis sp. nov. strain PU01-02T has been deposited in GenBank under accession number JBVYTQ000000000, and the rrs, gltA, ompB and sca4 gene sequences under accession numbers KF666476, KF666472, KF666470, KF666474, respectively. The plasmid accession numbers are PZ272915, PZ272916, and PZ272917, for pRS01, pRS02 and pRS03, respectively.

microbiology↗

Dental pulp metagenomics reveals ancient human Herpesviridae.

Following a recent report on ancient human Herpesviridae (HHV), we adapted a standard protocol used for the high-throughput sequencing of modern DNA to recover ancient DNA (aDNA) from dental pulp specimens collected at two historical burial sites. Investigation of 36 soldiers buried in Sevastopol, Crimea, during the 1853-1856 Crimean War yielded genomic sequences covering 8.5% of the herpes simplex virus (HSV)-1 genome, 15.7% of the HSV-2 genome, 1.9% of the Epstein-Barr virus (EBV) genome and 3.9% of the cytomegalovirus genome. Further investigation of 16 civilians buried in the 17th century in Amiens, France, yielded genomic sequences covering 9.8% of the HHV-1 genome and 2.2% of the HHV-6A genome; all data obtained with parallel negative controls was appropriately negative. Specific qPCRs confirmed the presence of four HSV-1, two HSV-2 and one EBV sequences in Sevastopol and two HSV-1 and one HHV-6 sequences in Amiens. HHV-6 was further detected by fluorescent in situ hybridization. These results demonstrated the usefulness of modern metagenomic protocols for high-throughput sequencing of aDNA in dental pulp.

microbiology↗

Genomic diversity and evolution of coronavirus (SARS-CoV-2) in France from 309 COVID-19-infected patients

The novel coronavirus (SARS-CoV-2) causes pandemic of viral pneumonia. The evolution and mutational events of the SARS-CoV-2 genomes are critical for controlling virulence, transmissibility, infectivity, severity of symptoms and mortality associated to this infectious disease. We collected and investigated 309 SARS-CoV-2 genomes from patients infected in France. Detailed genome cartography of all mutational events (SNPs, indels) was reported and correlated to clinical features of patients. A comparative analysis between our 309 SARS-CoV-2 genomes from French patients and the reference Wuhan coronavirus genome revealed 315 substitution mutations and six deletion events: ten were in 5/3 UTR, 178 were nonsynonymous, 126 were synonymous and one generated a stop codon. Six different deleted areas were also identified in nine viral variants. In particular, 30 substitution mutations (18 nonsynonymous) and one deletion ({Delta}21765-21770) concerned the spike S glycoprotein. An average of 7.8 mutational events (+/- 1.7 SD) and a median of 8 (range, 7-9) were reported per viral isolate. Comparative analyses and clustering of specific mutational signatures in 309 genomes disclose several divisions in groups and subgroups combining their geographical and phylogenetic origin. Clinical outcomes of the 309 COVID-19-infected patients were investigated according to the mutational signatures of viral variants. These findings highlight the genome dynamics of the coronavirus 2019-20 and shed light on the mutational landscape and evolution of this virus. Inclusion of the French cohort enabled us to identify 161 novel mutations never reported in SARS-CoV-2 genomes collected worldwide. These results support a global and continuing surveillance of the emerging variants of the coronavirus SARS-CoV-2.

genomics↗

High Content Screening, a reliable system for Coxiella burnetii isolation from clinical samples

Q fever, caused by Coxiella burnetii, is a worldwide zoonotic disease that may cause severe forms in humans and requires a specific and prolonged antibiotic treatment. Although the current serological and molecular detection tools enable a reliable diagnosis of the disease, culture of C. burnetii strains is mandatory to evaluate their antibiotic susceptibility and sequence their genome in order to optimize patient management and epidemiological studies. However, cultivating this fastidious microorganism is difficult and restricted to reference centers as it requires biosafety-level 3 laboratories and relies on cell culture performed by experienced technicians. In addition, the culture yield is low, which results in a small number of isolates being available. In this work, we developed a novel high content screening (HCS) isolation strategy based on optimized high-throughput cell culture and automated microscopic detection of infected cells with specifically-designed algorithms targeting cytopathic effects. This method was more efficient than the shell-vial assay when applied to both frozen specimens (7 isolates recovered by HCS only, sensitivity 91% vs 78% for shell-vial) and fresh samples (1 additional isolate using HCS, sensitivity 7% vs 5% for shell-vial). In addition, detecting positive cultures by an automated microscope reduced the need for expertise and saved 24% of technician working time. Application of HCS to antibiotic susceptibility testing of 12 strains demonstrated that it was as efficient as the standard procedure that combines shell-vial culture and quantitative PCR. Overall, this high-throughput HCS system paves the way to the development of improved cell culture isolation of human viruses.

microbiology↗

The dynamics and interactions of respiratory pathogen carriage among French pilgrims during the 2018 Hajj

Respiratory tract infections are frequent among Hajj pilgrims. We conducted this study to describe the dynamics of the acquisition of respiratory pathogens, their potential interactions and risk factors for possible lower respiratory tract infection symptoms (LRTI) among French pilgrims during the 2018 Hajj. Pilgrims from Marseille who were participating in the Hajj were recruited. Each participant underwent four successive systematic nasopharyngeal swabs before and during their stay in Saudi Arabia. Carriage of the main respiratory pathogens was assessed by PCR. 121 pilgrims were included and 93.4% reported respiratory symptoms during the study period. Polymicrobial carriage was observed in 73.8% samples. The acquisition of rhinovirus, coronaviruses and Staphylococcus aureus occurred soon after arrival in Saudi Arabia and rates decreased gradually after days 5 and 6. In contrast, Streptococcus pneumoniae and Klebsiella pneumoniae carriage increased progressively until the end of the stay in Saudi Arabia. Haemophilus influenzae and Moraxella catarrhalis carriage increased starting around days 12 and 13, following an initial clearance. Influenza viruses were rarely isolated. We observed an independent positive mutual association between S. aureus and rhinovirus carriage and between H. influenzae and M. catarrhalis carriage. Dual carriage of H. influenzae and M. catarrhalis was strongly associated with S. pneumoniae carriage (OR = 6.22, 95%CI [2.04-19.01]). Finally, our model showed that M. catarrhalis carriage was negatively associated with K. pneumoniae carriage. Chronic respiratory disease was associated with symptoms of LRTI. K. pneumoniae, M. catarrhalis-S. aureus and H. influenzae-rhinovirus dual carriage was associated with LRTI symptoms. Our data suggest that RTIs at the Hajj are a result of complex interactions between a number of respiratory viruses and bacteria.\n\nAuthor summaryDespite the recommendation to take individual preventive measures to prevent respiratory tract infections, these infections remain common among Hajj pilgrims. Respiratory pathogens acquired during the Hajj are usually studied individually, although in their natural environment they often compete or coexist with multiple microbial species. A better understanding of polymicrobial interactions in the nasopharynx among Hajj pilgrims is important. Our study describes the dynamics of the acquisition of respiratory pathogens and their potential interactions among pilgrims during the Hajj. We found that polymicrobial carriage was observed in most individuals and that some pathogens associated positively while other did not. Some pathogen associations correlated with symptoms of lower respiratory tract infections.

microbiology↗