Ontogenic and ecological control of metamorphosis onset in a carapid fish, <i>Carapus homei</i>: experimental evidence from vertebra and otolith comparisons

In Carapus homei, reef colonisation is associated with a penetration inside a sea cucumber followed by heavy transformations during which the length of the fish is reduced by 60%. By comparing vertebral axis to otolith ontogenetic changes, this study aimed (i) to specify the events linked to metamorphosis, and (ii) to establish to what extent these fish have the ability to delay it. Different larvae of C. homei were caught when settling on the reef and kept in different experimental conditions for at least 7 days and up to 21 days: darkness or natural light conditions, presence of sea cucumber or not, and food deprivation or not. Whatever the nutritional condition, a period of darkness seems sufficient to initiate metamorphosis. Twenty-one days in natural light conditions delayed metamorphosis, whereas the whole metamorphosis process is the fastest (15 days) for larvae living in sea cucumbers. Whether the metamorphosis was initiated or not, otoliths were modified with the formation of a transition zone, whose structure varied depending on the experimental conditions. At day 21, larvae maintained in darkness had an otolith transition zone with more increments (around 80), albeit wider than those (more or less 21) of individuals kept under natural lighting. These differences in otolith growth could indicate an increased incorporation rate of released metabolites by metamorphosing larvae. However, the presence of a transition zone in delayed-metamorphosis larvae suggests that these otolith changes record the endogenously-induced onset of metamorphosis, whereas body transformations seem to be modulated by the environmental conditions of settlement

Molecular Diagnosis of Invasive Aspergillosis and Detection of Azole Resistance by a Newly Commercialized PCR Kit

Item does not contain fulltextAspergillus fumigatus is the main species responsible for aspergillosis in humans. The diagnosis of aspergillosis remains difficult, and the rapid emergence of azole resistance in A. fumigatus is worrisome. The aim of this study was to validate the new MycoGENIE A. fumigatus real-time PCR kit and to evaluate its performance on clinical samples for the detection of A. fumigatus and its azole resistance. This multiplex assay detects DNA from the A. fumigatus species complex by targeting the multicopy 28S rRNA gene and specific TR34 and L98H mutations in the single-copy-number cyp51A gene of A. fumigatus The specificity of cyp51A mutation detection was assessed by testing DNA samples from 25 wild-type or mutated clinical A. fumigatus isolates. Clinical validation was performed on 88 respiratory samples obtained from 62 patients and on 69 serum samples obtained from 16 patients with proven or probable aspergillosis and 13 patients without aspergillosis. The limit of detection was <1 copy for the Aspergillus 28S rRNA gene and 6 copies for the cyp51A gene harboring the TR34 and L98H alterations. No cross-reactivity was detected with various fungi and bacteria. All isolates harboring the TR34 and L98H mutations were accurately detected by quantitative PCR (qPCR) analysis. With respiratory samples, qPCR results showed a sensitivity and specificity of 92.9% and 90.1%, respectively, while with serum samples, the sensitivity and specificity were 100% and 84.6%, respectively. Our study demonstrated that this new real-time PCR kit enables sensitive and rapid detection of A. fumigatus DNA and azole resistance due to TR34 and L98H mutations in clinical samples