Development of a real-time PCR for the specific detection of Waddlia chondrophila in clinical samples

Waddlia chondrophila is considered as an emerging human pathogen likely involved in miscarriage and lower respiratory tract infections. Given the low sensitivity of cell culture to recover such an obligate intracellular bacteria, molecular-based diagnostic approaches are warranted. We thus developed a real-time PCR that amplifies Waddlia chondrophila DNA. Specific primers and probe were selected to target the 16S rRNA gene. The PCR specifically amplified W. chondrophila but did not amplify other related-bacteria such as Parachlamydia acanthamoebae, Simkania negevensis and Chlamydia pneumoniae. The PCR exhibited a good intra-run and inter-run reproducibility and a sensitivity of less than ten copies of the positive control. This real-time PCR was then applied to 32 nasopharyngeal aspirates taken from children with bronchiolitis not due to respiratory syncytial virus (RSV). Three samples revealed to be Waddlia positive, suggesting a possible role of this Chlamydia-related bacteria in this settin

Quand l'information échappe à ses créateurs : le cas de l'artificialisation des terres agricoles en Languedoc-Roussillon

International audienceThis article aims to describe how the quantification of a phenomenon in relation to a specific worldview spreads and is distorted. This project has been supported by a regional public authority in Languedoc-Roussillon (France), and consisted in the production of information so to quantify and qualify the farmland decrease due to urban sprawl. The paper is based on Latour's actor-network theory; it characterizes the process of enrolment, and the development of dissidences. Identifying contradictory logics and dissidences help questioning the complexity of ‘closing the black-box' regarding information production and of sensemaking between heterogeneous actants involved in a supposed-to-be public debate.Cet article vise à spécifier comment la quantification d'un phénomène fondée sur une certaine vision du monde se propage et se déforme. Ce projet a été porté par une direction déconcentrée de l'Etat soucieuse de l'emprise croissante du bâti sur les terres agricoles en Languedoc-Roussillon. Le cadre théorique mobilisé, la sociologie de la traduction, permet d'identifier les grandes étapes d'intéressement, d'enrôlement, ainsi que le développement de controverses. Les difficultés rencontrées interrogent sur la complexité de clôture de processus innovants s'agissant de production d'information et de co-construction de sens associant des actants hétérogènes impliqués dans des procédures de débat public

Retrieval of total ozone quantity from high resolution infrared spectra : influence of spectroscopic and physical parameters

In this work, we present the results obtained for total ozone quantity from Jungfraujoch observatory spectrometer using 3 different spectral region

Osteocondroma intra-raquideo con afectación neurológica

Se presenta el caso de un varón de 16 años con lumbalgia consecutiva a traumatismo vertebral, sin hallazgo s radiológicos, que no mejoraba con tratamient o conservador, y que a las 5 semanas desarrolló un cuadro de paraparesia e incontinencia de esfínteres indicativo de afectación del cono medular. En el estudio mielográfico se objetivó bloqueo completo a nivel de Ll. La tomografía axial mostró una imagen sugestiva de osteocondroma intraraquídeo, que s e confirmó quirúrgicamente . Dos año s tras la intervención, se logró la total recuperación neurológica. Se destaca la rareza de la lesión, la dificultad diagnóstica, y la probable etiología traumática de la tumoración.A 16-year-old man with no improvement of low back pain after vertebral trauma conservatively treated and without radiological findings is presented. Five weeks after trauma, the patient developed paraparesia and fecal and urinary incontinence indicating compression of the medullary conus. A complet e stop of the contrast at Ll level wa s found in the myelographi c study. The CT-Scan showed an image suggesting osteochondroma whic h wa s confirmed a r surgery. Complet e neurogical recover y w a s achieved 2 year s after surgical treatment. The rare character of the lesion, the dificulty for diagnosis and the probable traumatic etiology of this tumor is discussed

Prevalence and diversity of Chlamydiales and other amoeba-resisting bacteria in domestic drinking water systems.

A growing number of human infections incriminate environmental bacteria that have evolved virulent mechanisms to resist amoebae and use them as a replicative niche. These bacteria are designated amoeba-resisting bacteria (ARB). Despite the isolation of these ARB in various human clinical samples, the possible source of infection remains undetermined in most cases. However, it is known that the ARB Legionella pneumophila, for instance, causes a respiratory infection in susceptible hosts after inhalation of contaminated water aerosols from various sources. The Chlamydiales order contains many ARB, such as Parachlamydia acanthamoebae or Simkania negevensis, previously implicated in human respiratory infections with no identified contamination sources. We thus investigated whether domestic water systems are a potential source of transmission of these Chlamydiales to humans by using amoebal culture and molecular methods. Other important ARB such as mycobacteria and Legionella were also investigated, as were their possible amoebal hosts. This work reports for the first time a very high prevalence and diversity of Chlamydiales in drinking water, being detected in 35 (72.9%) of 48 investigated domestic water systems, with members of the Parachlamydiaceae family being dominantly detected. Furthermore, various Legionella and mycobacteria species were also recovered, some species of which are known to be causal agents of human infections

On the feasibility of retrieving 16O 18O 16O ozone from high resolution ground-based FTIR spectra

We present evidence of the 16O 18O 16O ozone isotope in the 5 micron region from FTIR solar occultation spectra obtained from the Jungfraujoch Solar Observatory (47°N, 8°E, 3580 m) in Switzerland at a spectral resolution of 0,0025 cm-1 (res. = 1/2L). These spectra clearly show numerous unblended lines of the 16O 18O 16O ozone isotope. Laboratory spectra in the 5 micron region of 16O 18O 16O have been measured and have yielded line positions of the nu1 + nu3 isotopic bands which can eventually lead to their retrieval from measured ground-based solar occultation spectra

Ancestral genome estimation reveals the history of ecological diversification in Agrobacterium

Horizontal gene transfer (HGT) is considered as a major source of innovation in bacteria, and as such is expected to drive adaptation to new ecological niches. However, among the many genes acquired through HGT along the diversification history of genomes, only a fraction may have actively contributed to sustained ecological adaptation. We used a phylogenetic approach accounting for the transfer of genes (or groups of genes) to estimate the history of genomes in Agrobacterium biovar 1, a diverse group of soil and plant-dwelling bacterial species. We identified clade-specific blocks of cotransferred genes encoding coherent biochemical pathways that may have contributed to the evolutionary success of key Agrobacterium clades. This pattern of gene coevolution rejects a neutral model of transfer, in which neighboring genes would be transferred independently of their function and rather suggests purifying selection on collectively coded acquired pathways. The acquisition of these synapomorphic blocks of cofunctioning genes probably drove the ecological diversification of Agrobacterium and defined features of ancestral ecological niches, which consistently hint at a strong selective role of host plant rhizospheres

Development of a new chlamydiales-specific real-time PCR and its application to respiratory clinical samples.

Originally composed of the single family Chlamydiaceae, the Chlamydiales order has extended considerably over the last several decades. Chlamydia-related bacteria were added and classified into six different families and family-level lineages: the Criblamydiaceae, Parachlamydiaceae, Piscichlamydiaceae, Rhabdochlamydiaceae, Simkaniaceae, and Waddliaceae. While several members of the Chlamydiaceae family are known pathogens, recent studies showed diverse associations of Chlamydia-related bacteria with human and animal infections. Some of these latter bacteria might be of medical importance since, given their ability to replicate in free-living amoebae, they may also replicate efficiently in other phagocytic cells, including cells of the innate immune system. Thus, a new Chlamydiales-specific real-time PCR targeting the conserved 16S rRNA gene was developed. This new molecular tool can detect at least five DNA copies and show very high specificity without cross-amplification from other bacterial clade DNA. The new PCR was validated with 128 clinical samples positive or negative for Chlamydia trachomatis or C. pneumoniae. Of 65 positive samples, 61 (93.8%) were found to be positive with the new PCR. The four discordant samples, retested with the original test, were determined to be negative or below detection limits. Then, the new PCR was applied to 422 nasopharyngeal swabs taken from children with or without pneumonia; a total of 48 (11.4%) samples were determined to be positive, and 45 of these were successfully sequenced. The majority of the sequences corresponded to Chlamydia-related bacteria and especially to members of the Parachlamydiaceae family

Influence of age, social patterns and nasopharyngeal carriage on antibodies to three conserved pneumococcal surface proteins (PhtD, PcpA and PrtA) in healthy young children

The acquisition of specific antibodies is paramount to protect children against pneumococcal diseases, and a better understanding of how age, ethnicity and/or Streptococcus pneumoniae (Spn) nasopharyngeal carriage influence the acquisition of antibodies to pneumococcal surface proteins (PSP) is important for the development of novel serodiagnostic and immunisation strategies. IgG antibody titres against three conserved PSP (PhtD, PcpA and PrtA) in the sera of 451 healthy children aged 1 to 24months from Israel [Jewish (50.1%) and Bedouin (49.9%)] were measured by enzyme-linked immunosorbent assay (ELISA), while nasopharyngeal swabs from these children were assessed for the presence of Spn. Globally, anti-PhtD and anti-PrtA geometric mean concentrations (GMC; EU/ml) were high at <2.5months of age [PhtD: 35.3, 95% confidence interval (CI) 30.6-40.6; PrtA: 71.2, 95 % CI 60-84.5], was lower at 5-7months of age (PhtD: 10, 95 % CI 8-12.4; PrtA: 17.9, 95 % CI 14.4-22.1) and only increased after 11months of age. In contrast, an increase in anti-PcpA was observed at 5-7months of age. Anti-PcpA and anti-PrtA, but not anti-PhtD, were significantly higher in Bedouin children (PcpA: 361.6 vs. 226.3, p = 0.02; PrtA: 67.2 vs. 29.5, p < 0.001) in whom Spn nasopharyngeal carriage was identified earlier (60% vs. 38% of carriers <6months of age, p = 0.002). Spn carriage was associated with significantly higher anti-PSP concentrations in carriers than in non-carriers (p < 0.001 for each PSP). Thus, age, ethnicity and, essentially, nasopharyngeal carriage exert distinct cumulative influences on infant responses to PSP. These specific characteristics are worthwhile to include in the evaluation of pneumococcal seroresponses and the development of new PSP-based vaccine

Significance of tagI and mfd genes in the virulence of non-typeable <i>Haemophilus influenzae</i>

Non-typeable Haemophilus influenzae (NTHi) is an opportunist pathogen well adapted to the human upper respiratory tract and responsible for many respiratory diseases. In the human airway, NTHi is exposed to pollutants, such as alkylating agents, that damage its DNA. In this study, we examined the significance of genes involved in the repair of DNA alkylation damage in NTHi virulence. Two knockout mutants, tagI and mfd, encoding N(3)methyladenine-DNA glycosylase I and the key protein involved in transcription-coupled repair, respectively, were constructed and their virulence in a BALB/c mice model was examined. This work shows that N-3-methyladenine-DNA glycosylase I is constitutively expressed in NTHi and that it is relevant for its virulence.</p