Isolation and characterization of lytic phage TUN1 specific for Klebsiella pneumoniae K64 clinical isolates from Tunisia.

BACKGROUND: Multidrug-resistant Klebsiella pneumoniae spp. (kp) are emerging agents of severe infections of the respiratory, urinary tract and wounds that can progress to fatal septicemia. The use of bacteriophages is currently being considered as an effective alternative or adjuvant to antibiotic therapy. RESULTS: In this study, we report capsule (K)-typing of 163 carbapenem-resistant Kp (CRKP) isolated 2014-2018 at the Military Hospital of Instruction of Tunis (MHT), Tunisia, by partial amplification and sequencing of the Kp wzi gene. The most prevalent K-type overall was K64 with 50.3% followed by K17 and K27 (22.7 and 11.0%, respectively). K64 Kp strains were most common and associated with increased case/fatality rates, especially at the intensive care unit (ICU). Using a K64 Kp strain we isolated and characterized a lytic Kp phage, vB_KpP_TUN1 (phage TUN1), from wastewater samples of the ICU at the MHT. TUN1 belongs to the Autographiviridae family and specifically digests K64 Kp capsules most probably via a depolymerase encoded by gp47. Furthermore, we successfully assembled phage TUN1 in a non-replicative host (E. coli) raising the possibility of in vitro assembly in the absence of live bacterial hosts. We propose that phage TUN1 is a promising candidate to be used as an adjuvant or an alternative to antibiotic therapy in CRKP infections, facilitating regulatory approval of phage therapy. CONCLUSIONS: K64, K17 and K27 are the most common wzi capsule types in this geographical location in Northern Africa. The lytic phage TUN1 efficiently lyses K64 Kp strains associated with increased case/fatality rates at body temperature. Together with its ability to be rescued in a non-replicative host these features enhance the utility of this phage as an antibacterial agent.

Novel Nucleoside Analogues as Effective Antiviral Agents for Zika Virus Infections.

Previously considered a neglected flavivirus, Zika virus has recently emerged as a public health concern due to its ability to spread rapidly and cause severe neurological disorders, such as microcephaly in newborn babies from infected mothers, and Guillain-Barré syndrome in adults. Despite extensive efforts towards the identification of effective therapies, specific antivirals are still not available. As part of ongoing medicinal chemistry studies to identify new antiviral agents, we screened against Zika virus replication in vitro in a targeted internal library of small-molecule agents, comprising both nucleoside and non-nucleoside agents. Among the compounds evaluated, novel aryloxyphosphoramidate prodrugs of the nucleosides 2'-C-methyl-adenosine, 2-CMA, and 7-deaza-2'C-methyl-adenosine, 7-DMA, were found to significantly inhibit the virus-induced cytopathic effect in multiple relevant cell lines. In addition, one of these prodrugs exhibits a synergistic antiviral effect against Zika virus when applied in combination with an indirect antiviral agent, a l-dideoxy bicyclic pyrimidine nucleoside analogue, which potently inhibits vaccinia and measles viruses in vitro by targeting a host pathway. Our findings provide a solid basis for further development of an antiviral therapy for Zika virus infections, possibly exploiting a dual approach combining two different agents, one targeting the viral polymerase (direct-acting antiviral), the second targeting a host-directed autophagy mechanism.

Genetic analysis of Rhipicephalus sanguineus sensu lato ticks parasites of dogs in Africa north of the Sahara based on mitochondrial DNA sequences.

The aim of this work was to determine the evolutionary relationship among tick populations of Rhipicephalus sanguineus sensu lato distributed in Africa north of the Sahara and different lineages of R. sanguineus s.l. distributed in different regions of Sub-Saharan Africa, America and Europe through the analysis of DNA sequences of two mitochondrial genes. One hundred and thirty six 16S rRNA gene sequences and twenty-seven 12S rRNA gene sequences of R. sanguineus s.l. were analyzed. Phylogenetic analyses were performed including different lineages of R. sanguineus s.l. from America, Europe and Africa, and species belonging to the R. sanguineus group as Rhipicephalus camicasi, Rhipicephalus guilhoni, Rhipicephalus sulcatus, Rhipicephalus rossicus, Rhipicephalus pusillus, Rhipicephalus turanicus and Rhipicephalus leporis. At least two different lineages of R. sanguineus s.l. are living in sympatry in Africa north of the Sahara. One of these mitochondrial lineages belongs to the same evolutionary entity that R. sanguineus s.l. from tropical areas of America, R. sanguineus s.l. from Sub-Saharan Africa, R. camicasi and R. guilhoni. The other mitochondrial lineage of R. sanguineus s.l. present in Africa north of the Sahara is phylogenetically associated to R. sanguineus s.l. ticks from southeastern Europe (Romania, Turkey and Greece). Both evolutionary entities are clearly different to the evolutionary entity formed by R. sanguineus s.l. from western Europe and temperate areas of America. Thus, the name R. sanguineus s.s. cannot be assigned to any of the two evolutionary entities present in Africa north of the Sahara. The taxonomic status of these taxa will remain unresolved until new lines of evidence become available to complement the current results based on mitochondrial DNA.