Search

Objectives: The emergence of Enterobacteriaceae isolates resistant to the last-resort antibiotic fosfomycin outside of Asia is a public-health issue. Here we report the draft genome of an Escherichia coli isolate presenting both an extended-spectrum β-lactamase (ESBL) and the fosA3 gene in a healthy cow in Brazil., Methods: Whole genomic DNA from E. coli E12 was extracted and 2×150-bp paired-end reads were generated using Illumina sequencing technology. De novo genome assembly was performed using SPAdes v.3.11 and the draft genome was annotated by the NCBI Prokaryotic Genome Annotation Pipeline. Further analyses were performed using the Center for Genomic Epidemiology databases., Results: The 5045934-bp genome displayed several resistance genes, including the fosA3 and bla CTX-M-8 genes. Southern blot experiments showed that they were co-located on an IncI1/ST113 plasmid., Conclusion: Presence of the fosA3 gene on the same common plasmid as bla CTX-M-8 will have to be monitored. This draft genome provides data that will help in tracing the dissemination of this gene and the evolution of its plasmidic support., (Copyright © 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Crimean-Congo hemorrhagic fever virus (CCHFV) circulates in many countries of Asia, Africa, and Europe. CCHFV can cause a severe hemorrhagic fever in humans with case-fatality rates of up to 80%. CCHF is considered to be one of the major emerging diseases spreading to and within Europe. Ticks of the genus Hyalomma function as vector as well as natural reservoir of CCHFV. Ticks feed on various domestic animals (e.g. cattle, sheep, goats) and on wildlife (e.g. hares, hedgehogs). Those animal species play an important role in the life cycle of the ticks as well as in amplification of CCHFV. Here we present a competitive ELISA (cELISA) for the species-independent detection of CCHFV-specific antibodies. For this purpose nucleocapsid (N) protein specific monoclonal antibodies (mAbs) were generated against an Escherichia coli (E. coli) expressed CCHFV N-protein. Thirty-three mAbs reacted with homologous and heterologous recombinant CCHFV antigens in ELISA and Western blot test and 20 of those 33 mAbs reacted additionally in an immunofluorescence assay with eukaryotic cells expressing the N-protein. Ten mAbs were further characterized in a prototype of the cELISA and nine of them competed with positive control sera of bovine origin. The cELISA was established by using the mAb with the strongest competition. For the validation, 833 sera from 12 animal species and from humans were used. The diagnostic sensitivity and specificity of the cELISA was determined to be 95% and 99%, respectively, and 2% of the sera gave inconclusive results. This cELISA offers the possibility for future large-scale screening approaches in various animal species to evaluate their susceptibility to CCHFV infection and to identify and monitor the occurrence of CCHFV., (Copyright © 2016 Elsevier B.V. All rights reserved.)