Characterization of the immune response against Testudinid herpesvirus 3.

Numerous infectious diseases have been documented in reptiles, however minimal information isavailable concerning their immunological response. One of the most diffuse and lethal reptile pathogenis Testudinid herpesvirus 3 (TeHV3), a Alphaherpesvirinae. All species of tortoises (Testudinide) areconsidered susceptible to TeHV3, however the virus is over represented in the genus Testudo, whichincludes, among others, T. graeca, T. hermanni, T. marginata, and T. horsfieldii, that are popular pets inEurope. Incidence of TeHV3-associated disease is highest right after hibernation (Origgi, 2012).The aim of this work is to partially characterize the immunological response of T. graeca against TeHV3.A bacteriophage library composed of about 5.000 clones containing genomic DNA fragments of TeHV3was produced. Bacteriophages were amplified in a specific strain of E. coli and were screened withTeHV3-seropositive sera from T. graeca. Phagemids were excised from the positive bacteriophages,sequenced, and compare with the TeHV3 genome to identify the encoding genes. Six differentstructural and non-structural proteins have identified as immune relevant. Vero cells where transfectedwith phagemids of the positive clones, to confirm previous results. TeHV3’s proteins expression wasassessed by F.A.C.S using T. graeca seropositive sera. Of all the six selected clones, only that expressingthe partial sequence of the glycoprotein B (gB) showed a positive signal in the F.A.C.S. analysis. Thisresult is consistent with the well-known immunogenicity of gB of other herpesviruses including thoseinfecting humans and with the highly conserved role that gB plays in host-pathogen interaction acrossspecies and evolution (Beals et al., 2016)

Characterization of the immune response against Testudinid herpesvirus 3: new insight

Testudinid herpesvirus 3 (TeHV3) is one of the most lethal viral agents in tortoises worldwide. Although TeHV3 have been extensively studied, only little information is available about host-pathogen interaction. TeHV3 infections in different species of the genus Testudo correlate with various lesions profiles, disease severity and clinical outcome, suggesting the existence of a complex host-pathogen interaction. This might reflect a possible viral-host coevolution (Origgi, 2012).To study the host-pathogen interaction, we previously screened 5.000 clones from a bacteriophage library obtained from the TeHV3 genomic DNA using Testudo graeca seropositive sera. Of the six detected positive clones, only one was confirmed by F.A.C.S. Selected clone was determined to be a concatamer of different TeHV3 genomic fragments including the partial sequence of TE17, UL15, Major capsid protein (MCP), and Glycoprotein B (gB) genes. After complete sequencing of the selected clone, the MCP and the gB were antisenses compared to the phagemid promoter.In order to assess which of the gene fragments among TE17 and UL15 was encoding for the antigenic determinant that was recognized by the anti-TeHV3 tortoise sera, distinct approaches were followed.TE-17 and UL15 fragments were knock out from the original phagemid using the following approaches: a) directed-site mutagenesis, b) molecular cloning, and c) restriction enzymes cloning. All the modified constructs were cloned in two different E. coli cloning vectors (D5α and XL 1-Blue).Transformation of competent cells with the constructs described above did not yield any viable bacteria.Among the different aspects might have influenced transformation success rate, construct size was probably the most relevant (about 9Kb). Furthermore, we could not entirely exclude that genomic DNA editing might have induced mutations in the construct sequence causing toxic effects on the host bacterial cell. Cloning of TE-17 and UL15 gene fragments into different prokaryotic expression vectors is currently under way

Tularemia in the Southeastern Swiss Alps at 1,700 m above sea level.

A 37-year-old man presented with a 4-day history of nonbloody diarrhea, fever, chills, productive cough, vomiting, and more recent sore throat. He worked for the municipality in a village in the Swiss Alps near St. Moritz. Examination showed fever (40 °C), hypotension, tachycardia, tachypnea, decreased oxygen saturation (90 % at room air), and bibasilar crackles and wheezing. Chest radiography and computed tomography scan showed an infiltrate in the left upper lung lobe. He responded to empiric therapy with imipenem for 5 days. After the imipenem was stopped, the bacteriology laboratory reported that 2/2 blood cultures showed growth of Francisella tularensis. He had recurrence of fever and diarrhea. He was treated with ciprofloxacin (500 mg twice daily, oral, for 14 days) and symptoms resolved. Further testing confirmed that the isolate was F. tularensis (subspecies holarctica) belonging to the subclade B.FTNF002-00 (Western European cluster). This case may alert physicians that tularemia may occur in high-altitude regions such as the Swiss Alps

First molecular isolation of <i>Echinococcus granulosus</i> horse strain (G4) in Sardinia (Italy)

Several papers have been published in Sardinia on epidemiology and genetic characterization of Eg, although data on hydatidosis in horse are scarce and dated (Tanda S, 1960, Parassitologia, 2: 315-320). In this work we investigated the situation of Cystic Echinococcosis in horses of Sardinia through a retrospective study compared with new cases recently examined in our laboratory

Functionalized multiwalled carbon nanotubes as ultrasound contrast agents

Ultrasonography is a fundamental diagnostic imaging tool in everyday clinical practice. Here, we are unique in describing the use of functionalized multiwalled carbon nanotubes (MWCNTs) as hyperechogenic material, suggesting their potential application as ultrasound contrast agents. Initially, we carried out a thorough investigation to assess the echogenic property of the nanotubes in vitro. We demonstrated their long-lasting ultrasound contrast properties. We also showed that ultrasound signal of functionalized MWCNTs is higher than graphene oxide, pristine MWCNTs, and functionalized single-walled CNTs. Qualitatively, the ultrasound signal of CNTs was equal to that of sulfur hexafluoride (SonoVue), a commercially available contrast agent. Then, we found that MWCNTs were highly echogenic in liver and heart through ex vivo experiments using pig as an animal model. In contrast to the majority of ultrasound contrast agents, we observed in a phantom bladder that the tubes can be visualized within a wide variety of frequencies (i.e., 5.5–10 MHz) and 12.5 MHz using tissue harmonic imaging modality. Finally, we demonstrated in vivo in the pig bladder that MWCNTs can be observed at low frequencies, which are appropriate for abdominal organs. Importantly, we did not report any toxicity of CNTs after 7 d from the injection by animal autopsy, organ histology and immunostaining, blood count, and chemical profile. Our results reveal the enormous potential of CNTs as ultrasound contrast agents, giving support for their future applications as theranostic nanoparticles, combining diagnostic and therapeutic modalities

Hydrogen Refueling Stations: Prevention and Scenario Management. Large Scale Experimental Investigation of Hydrogen Jet-Fires

Hydrogen is becoming an attractive alternative for energy storage and transportation, because of the elevated energy content per unit of mass and possibility to have zero carbon-emission vehicles. For these reasons, hydrogen's share in global market is expected to grow substantially in the coming years. Today, hydrogen-fueled buses and cars are already available, and several refueling stations are operating in different countries around the world. A key role of the deployment of hydrogen fueled-vehicles is the presence of a widespread network of refueling stations, especially close to residential and industrial areas. This fact poses attention to the safety aspects related to hydrogen, with particular interest to its high flammability that can lead to catastrophic consequences for personnel and equipment. As a matter of fact, hydrogen is a comparatively less safe fuel compared to conventional fuels such as gasoline and diesel. Hydrogen infrastructures are characterized by operating pressure up to 1000 bar that, in case of an unintended loss of containments, produce a highly under expanded turbulent jet. If ignited, this hydrogen jet may give rise to very severe scenarios, mainly related to high temperatures and the oriented flows. As recently suggested by Moradi and Groth (Moradi and Groth, 2019), there is a lack of experimental and on-site data for almost all of the storage and delivery technologies relevant to the hydrogen infrastructures. Experimental data is vital to support model validation, especially in the case of the very peculiar combustion process of hydrogen. In this way, a real-scale experimental campaign is proposed to investigate the main characteristic of the hydrogen jet fire resulting from its rapid fired depressurizations. Focus of the experimental campaign is evaluation of safety distance for person and device (i.e. pressurized tanks) in order to avoid critical conditions and domino effects in real refueling station. Different initial conditions, i.e., storage pressures, are exploited, and the resulting jet across specified orifice is investigated. More specifically, temperatures at various locations are measured through an arrangement of thermocouples. Values up to 1200 °C were obtained in the core of the jet. Moreover, it was found that the recorded temperatures, especially those at the outer portion of the jet, are very sensitive to the initial conditions

Identification of Francisella tularensis Cluster in Central and Western Europe

We conducted a molecular analysis of Francisella tularensis strains isolated in Switzerland and identified a specific subpopulation belonging to a cluster of F. tularensis subsp. holarctica that is widely dispersed in central and western continental Europe. This subpopulation was present before the tularemia epidemics on the Iberian Peninsula

Long-Lasting Fever and Lymphadenitis: Think about F. tularensis

We report the case of glandular tularemia that developed in a man supposedly infected by a tick bite in Western Switzerland. Francisella tularensis (F. tularensis) was identified. In Europe tularemia most commonly manifests itself as ulcero-glandular or glandular disease; the diagnosis of tularemia may be delayed in glandular form where skin or mucous lesion is absent, particularly in areas which are assumed to have a low incidence of the disease