Viral and Bacterial Zoonotic Agents in Dromedary Camels from Southern Tunisia: A Seroprevalence Study.

The rapid spread of SARS-CoV-2 clearly demonstrated the potential of zoonotic diseases to cause severe harm to public health. Having limited access to medical care combined with severe underreporting and a lack of active surveillance, Africa carries a high burden of neglected zoonotic diseases. Therefore, the epidemiological monitoring of pathogen circulation is essential. Recently, we found extensive Middle East respiratory syndrome coronavirus (MERS-CoV) prevalence in free-roaming dromedary camels from southern Tunisia. In this study, we aimed to investigate the seroprevalence, and thus the risk posed to public health, of two additional viral and two bacterial pathogens in Tunisian dromedaries: Rift Valley fever virus (RVFV), foot-and-mouth disease virus (FMDV), Coxiella burnetii and Brucella spp. via ELISA. With 73.6% seropositivity, most animals had previously been exposed to the causative agent of Q fever, C. burnetii. Additionally, 7.4% and 1.0% of the dromedaries had antibodies against Brucella and RVFV, respectively, while no evidence was found for the occurrence of FMDV. Our studies revealed considerable immunological evidence of various pathogens within Tunisian dromedary camels. Since these animals have intense contact with humans, they pose a high risk of transmitting serious zoonotic diseases during active infection. The identification of appropriate countermeasures is therefore highly desirable.

Prevalence of Middle East Respiratory Syndrome Coronavirus in Dromedary Camels, Tunisia.

Free-roaming camels, especially those crossing national borders, pose a high risk for spreading Middle East respiratory syndrome coronavirus (MERS-CoV). To prevent outbreaks, active surveillance is necessary. We found that a high percentage of dromedaries in Tunisia are MERS-CoV seropositive (80.4%) or actively infected (19.8%), indicating extensive MERS-CoV circulation in Northern Africa.

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.

Whole genome sequencing and phylogenetic classification of Tunisian SARS-CoV-2 strains from patients of the Military Hospital in Tunis.

In the present work, two complete genome sequences of SARS-CoV-2 were obtained from nasal swab samples of Tunisian SARS-CoV-2 PCR-positive patients using nanopore sequencing. The virus genomes of two of the patients examined, a Tunisian soldier returning from a mission in Morocco and a member of another Tunisian family, showed significant differences in analyses of the total genome and single nucleotide polymorphisms (SNPs). Phylogenetic relationships with known SARS-CoV-2 genomes in the African region, some European and Middle Eastern countries and initial epidemiological conclusions indicate that the introduction of SARS-CoV-2 into Tunisia from two independent sources was travel-related.

Screening for Carbapenemases in Ertapenem-Resistant Enterobacteriaceae Collected at a Tunisian Hospital Between 2014 and 2018.

BACKGROUND: Carbapenem-resistance is frequently detected in Enterobacteriaceae isolated from patients in Tunisia. The study was performed to identify frequent carbapenemases in Tunisian isolates. METHODS: Between May 2014 and January 2018, 197 ertapenem-resistant Enterobacteriaceae were isolated at the microbiological department of the Military Hospital of Tunis. The strains were phenotypically characterized and then subjected to in-house polymerase chain reaction (PCR) targeting the carbapenemase genes blaIMP, blaVIM, blaNDM, blaSPM, blaAIM, blaDIM,blaGIM, blaSIM, blaKPC, blaBIC , and blaOXA-48. RESULTS: The assessed 197 ertapenem-resistant Enterobacteriaceae from Tunis comprised 170 Klebsiella pneumoniae, 19 Enterobacter cloacae, 6 Escherichia coli, 1 Citrobacter sedlakii, and 1 Enterobacter asburiae. Thereby, 55 out of 197 isolates (27.9%) were from blood cultures, suggesting a systemic disease. The carbapenemase gene blaOXA-48 quantitatively dominated by far with 153 detections, followed by blaNDM with 14 detections, which were distributed about the whole study interval. In contrast, blaBIC and blaVIM were only infrequently identified in 5 and 3 cases, respectively, while the other carbapenamases were not observed. CONCLUSIONS: The carbapenemase gene blaOXA-48 was identified in the vast majority of ertapenem-resistant Tunisian Enterobacteriaceae while all other assessed carbapenemases were much less abundant. In a quantitatively relevant minority of isolates, the applied PCR-based screening approach did not identify any carbapenemases.

Two-Photon Fluorescence and Magnetic Resonance Specific Imaging of Aß Amyloid Using Hybrid Nano-GdF3 Contrast Media.

Real time in vivo detection of Amyloid ß (Aß) deposits at an early stage may lead to faster and more conclusive diagnosis of Alzheimer's disease (AD) and can facilitate the follow up of the effect of therapeutic interventions. In this work, the capability of new hybrid nanomaterials to target and detect Aß aggregates using magnetic resonance (MRI) and fluorescence imaging is demonstrated. These smart contrast agents contain paramagnetic nanoparticles surrounded by luminescent conjugated oligothiophenes (LCOs) known to selectively bind to Aß aggregates, with emission spectra strongly dependent on their conformations, opening the possibilities for several fluorescence imaging modes for AD diagnostics. Relaxivity is evaluated in vitro and ex vivo. The capability of these contrast media to link to Aß fibrils in stained sections is revealed using transmission electron microscopy and fluorescence microscopy. Preliminary in vivo experiments show the ability of the contrast agent to diffuse through the blood-brain barrier of model animals and specifically stain amyloid deposits.

Resurgence of Ebola Virus Disease in Guinea Linked to a Survivor With Virus Persistence in Seminal Fluid for More Than 500 Days.

We report on an Ebola virus disease (EVD) survivor who showed Ebola virus in seminal fluid 531 days after onset of disease. The persisting virus was sexually transmitted in February 2016, about 470 days after onset of symptoms, and caused a new cluster of EVD in Guinea and Liberia.

Impact of peripheral myeloid cells on amyloid-ß pathology in Alzheimer's disease-like mice.

Although central nervous system-resident microglia are believed to be ineffective at phagocytosing and clearing amyloid-ß (Aß), a major pathological hallmark of Alzheimer's disease (AD), it has been suggested that peripheral myeloid cells constitute a heterogeneous cell population with greater Aß-clearing capabilities. Here, we demonstrate that the conditional ablation of resident microglia in CD11b-HSVTK (TK) mice is followed by a rapid repopulation of the brain by peripherally derived myeloid cells. We used this system to directly assess the ability of peripheral macrophages to reduce Aß plaque pathology and therefore depleted and replaced the pool of resident microglia with peripherally derived myeloid cells in Aß-carrying APPPS1 mice crossed to TK mice (APPPS1;TK). Despite a nearly complete exchange of resident microglia with peripheral myeloid cells, there was no significant change in Aß burden or APP processing in APPPS1;TK mice. Importantly, however, newly recruited peripheral myeloid cells failed to cluster around Aß deposits. Even additional anti-Aß antibody treatment aimed at engaging myeloid cells with amyloid plaques neither directed peripherally derived myeloid cells to amyloid plaques nor altered Aß burden. These data demonstrate that mere recruitment of peripheral myeloid cells to the brain is insufficient in substantially clearing Aß burden and suggest that specific additional triggers appear to be required to exploit the full potential of myeloid cell-based therapies for AD.

Evidence for age-dependent in vivo conformational rearrangement within Aß amyloid deposits.

Deposition of aggregated Aß peptide in the brain is one of the major hallmarks of Alzheimer's disease. Using a combination of two structurally different, but related, hypersensitive fluorescent amyloid markers, LCOs, reporting on separate ultrastructural elements, we show that conformational rearrangement occurs within Aß plaques of transgenic mouse models as the animals age. This important mechanistic insight should aid the design and evaluation of experiments currently using plaque load as readout.