West Nile virus transmission. results from the integrated surveillance system in Italy, 2008 to 2015

IIn Italy a national Plan for the surveillance of imported and autochthonous human vector-borne diseases (chikungunya, dengue, Zika virus disease and West Nile virus (WNV) disease) that integrates human and veterinary (animals and vectors) surveillance, is issued and revised annually according with the observed epidemiological changes. Here we describe results of the WNV integrated veterinary and human surveillance systems in Italy from 2008 to 2015. A real time data exchange protocol is in place between the surveillance systems to rapidly identify occurrence of human and animal cases and to define and update the map of affected areas i.e. provinces during the vector activity period from June to October. WNV continues to cause severe illnesses in Italy during every transmission season, albeit cases are sporadic and the epidemiology varies by virus lineage and geographic area. The integration of surveillance activities and a multidisciplinary approach made it possible and have been fundamental in supporting implementation of and/or strengthening preventive measures aimed at reducing the risk of transmission of WNV trough blood, tissues and organ donation and to implementing further measures for vector control

In-Building Wireless Distribution in legacy Multimode Fiber with an improved RoMMF system

A radio over multimode fiber (RoMMF) system for in-building wireless distribution employing a directly modulated Fabry-Perot (FP) transmitter and the central launch technique is presented. The worst-case spurious free dynamic range (SFDR) exceeds 105 dB×Hz2/3 up to 525 m of OM2 multimode fiber (MMF). Experimental and theoretical results are reported showing that this scheme outperforms a RoMMF system employing a distributed feed-back (DFB) laser diode (LD) and/or a mode scrambler to achieve overfilled launch (OFL). Long Term Evolution (LTE) signal transmission is achieved with high quality in terms of Adjacent Channel Leakage Ratio (ACLR) and Error Vector Magnitude (EVM)

Tipizzazione del virus della laringotracheite infettiva aviare tramite protocolli di end point PCR.

Infectious laryngotracheitis (ILT) is an acute and highly contagious respiratory disease of chickens, with a worldwide distribution and leading to severe economic losses due to decreased growth rate, reduced egg production and mortality. It is caused by Gallid Herpesvirus 1. Disease is controlled by vaccination with 2 types of live attenuated ILT vaccines are produced by sequential passages in cell cultures (tissue culture origin, TCO) or embryonated eggs (chicken embryo origin, CEO). A strong evidence exists indicating that ILTV epizootics may originate from CEO-derived strains which can regain virulence and persist in the field. Here we report the development of 3 new PCR methods able to easily differentiate vaccine and CEO related field strains, which target genes are ORFE, ORFD and UL36. Two profiles were identified: one typical of CEO vaccines (characterized by no insertion-G-R-K) and other related to field strains (insertion-A-H-R). Using this method we tested 89 ILTV PCR positive samples collected from broilers, capons, layers and broiler breeders in the period 2004-2016 and identified two distinct profiles (CEO vaccine and field profiles) without observe mixed forms

Detection and characterization of a novel reassortant mammalian orthoreovirus in bats in Europe

A renewed interest in mammalian orthoreoviruses (MRVs) has emerged since new viruses related to bat MRV type 3, detected in Europe, were identified in humans and pigs with gastroenteritis. This study reports the isolation and characterization of a novel reassortant MRV from the lesser horseshoe bat (Rhinolophus hipposideros). The isolate, here designated BatMRV1-IT2011, was first identified by electron microscopy and confirmed using PCR and virus-neutralization tests. The full genome sequence was obtained by next-generation sequencing. Molecular and antigenic characterizations revealed that BatMRV1-IT2011 belonged to serotype 1, which had not previously been identified in bats. Phylogenetic and recombination detection program analyses suggested that BatMRV1-IT2011 was a reassortant strain containing an S1 genome segment similar to those of MRV T1/bovine/Maryland/Clone23/59 and C/bovine/ Indiana/MRV00304/2014, while other segments were more similar to MRVs of different hosts, origins and serotypes. The presence of neutralizing antibodies against MRVs has also been investigated in animals (dogs, pigs, bovines and horses). Preliminary results suggested that MRVs are widespread in animals and that infections containing multiple serotypes, including MRVs of serotype 1 with an S1 gene similar to BatMRV1-IT2011, are common. This paper extends the current knowledge of MRVs and stresses the importance to continue and improve MRV surveillance in bats and other mammals through the development and standardization of specific diagnostic tools

GI-16 lineage (624/I or Q1), there and back again: The history of one of the major threats for poultry farming of our era

The genetic variability of Infectious bronchitis virus (IBV) is one of the main challenges for its control, hindering not only the development of effective vaccination strategies but also its classification and, consequently, epidemiology understanding. The 624/I and Q1 genotypes, now recognized to be part of the GI-16 lineage, represent an excellent example of the practical consequences of IBV molecular epidemiology limited knowledge. In fact, being their common origin unrecognized for a long time, independent epidemiological pictures were drawn for the two genotypes. To fix this misinterpretation, the present study reconstructs the history, population dynamics and spreading patterns of GI-16 lineage as a whole using a phylodynamic approach. A collection of worldwide available hypervariable region 1 and 2 (HVR12) and 3 (HVR3) sequences of the S1 protein was analysed together with 258 HVR3 sequences obtained from samples collected in Italy (the country where this genotype was initially identified) since 1963. The results demonstrate that after its emergence at the beginning of the XX century, GI-16 was able to persist until present days in Italy. Approximately in the late 1980s, it migrated to Asia, which became the main nucleus for further spreading to Middle East, Europe and especially South America, likely through multiple introduction events. A remarkable among-country diffusion was also demonstrated in Asia and South America. Interestingly, although most of the recent Italian GI-16 strains originated from ancestral viruses detected in the same country, a couple were closely related to Chinese ones, supporting a backward viral flow from China to Italy. Besides to the specific case-study results, this work highlights the misconceptions that originate from the lack of a unified nomenclature and poor molecular epidemiology data generation and sharing. This shortcoming appears particularly relevant since the described scenario could likely be shared by many other IBV genotypes and pathogens in general

Organic Electrochemical Transistor (OECT) for Real Time, Electrical quantification of SARS CoV 2 neutralizing antibodies

Since the outbreak of the Covid-19 pandemic, researchers have refocused their efforts on viral neutralization assays and neutralizing antibody quantifications for vaccination pre-clinical studies and long-term efficacy. Nowadays, the gold standard to assess antibody titer is the plaque reduction neutralization test (PRNT), an end-point assay which evaluates the highest serum antibody dilution that neutralizes viral replication, by inspecting the cytopathic effect (CPE) induced on cell cultures. Here, we have designed and implemented an accurate real-time technique for quantitative serum neutralization assay, employing poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (Pedot:Pss)-based Organic Electrochemical Transistors (OECT) for the automated evaluation of the CPE induced by Sars-Cov-2 on Vero E6 cells, using a customized prototype for in-vitro measurements inside the incubator. Transistors seeded with viral proliferating cell cultures reported faster time responses during the experiment, caused by the disruption of the cell layer grown onto the active area of our devices. On the contrary, when neutralizing antibodies stopped the viral infection, OECT data superimposed with standard control growth. The device reliability was proved using optical imaging (cell layer evaluation), Quantitative Reverse Transcription Polymerase Chain Reaction (investigating viral proliferation) and standard PRNT assays, obtaining robust matching. We noted that OECTs allowed to extract the neutralizing test outcome in less than 48h, earlier than the usual 72-hour required for PRNT screening, without the need of cell staining or fixing at the end of the experiment. Furthermore, the devices can be revitalized and re-used for up to three consecutive experiments, reducing plastic waste and their effective cost/experiment Our low-cost and scalable devices have the potential to speed-up large-scale viral neutralization screening without the need for cancerous staining or highly specialized operators. Finally, owing to the versatile nature of the proposed assay and the possibility to optimize the device geometry/dimensions to match the cell lines under test, the technology could be easily transferred to assess neutralizing antibody response towards different viruses in their permissive cell substrates

Hypsugopoxvirus: A Novel Poxvirus Isolated from Hypsugo savii in Italy

Interest in bat-related viruses has increased considerably during the last decade, leading to the discovery of a rising number of new viruses in several bat species. Poxviridae are a large, diverse family of DNA viruses that can infect a wide range of vertebrates and invertebrates. To date, only a few documented detections of poxviruses have been described in bat populations on three different continents (America, Africa, and Australia). These viruses are phylogenetically dissimilar and have diverse clinical impacts on their hosts. Herein, we report the isolation, nearly complete genome sequencing, and annotation of a novel poxvirus detected from an insectivorous bat (Hypsugo savii) in Northern Italy. The virus is tentatively named Hypsugopoxvirus (HYPV) after the bat species from which it was isolated. The nearly complete genome size is 166,600 nt and it encodes 161 genes. Genome analyses suggest that HYPV belongs to the Chordopoxvirinae subfamily, with the highest nucleotide identity (85%) to Eptesipoxvirus (EPTV) detected from a microbat Eptesicus fuscus in WA, USA, in 2011. To date, HYPV represents the first poxvirus detected in bats in Europe; thus, its viral ecology and disease associations should be investigated further

Organic Electrochemical Transistors as Versatile Tool for Real-Time and Automatized Viral Cytopathic Effect Evaluation

In-vitro viral studies are still fundamental for biomedical research since studying the virus kinetics on cells is crucial for the determination of the biological properties of viruses and for screening the inhibitors of infections. Moreover, testing potential viral contaminants is often mandatory for safety evaluation. Nowadays, viral cytopathic effects are mainly evaluated through end-point assays requiring dye-staining combined with optical evaluation. Recently, optical-based automatized equipment has been marketed, aimed at the real-time screening of cell-layer status and obtaining further insights, which are unavailable with end-point assays. However, these technologies present two huge limitations, namely, high costs and the possibility to study only cytopathic viruses, whose effects lead to plaque formation and layer disruption. Here, we employed poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (Pedot:Pss) organic electrochemical transistors (OECTs) for the real-time, electrical monitoring of the infection of cytolytic viruses, i.e., encephalomyocarditis virus (EMCV), and non-cytolytic viruses, i.e., bovine coronavirus (B-CoV), on cells. OECT data on EMCV were validated using a commercially-available optical-based technology, which, however, failed in the B-CoV titration analysis, as expected. The OECTs proved to be reliable, fast, and versatile devices for viral infection monitoring, which could be scaled up at low cost, reducing the operator workload and speeding up in-vitro assays in the biomedical research field

Assessment of the Costs Related to West Nile Virus Monitoring in Lombardy Region (Italy) between 2014 and 2018

In Italy, the West Nile Virus surveillance plan considers a multidisciplinary approach to identify the presence of the virus in the environment (entomological, ornithological, and equine surveillance) and to determine the risk of infections through potentially infected donors (blood and organ donors). The costs associated with the surveillance program for the Lombardy Region between 2014 and 2018 were estimated. The costs of the program were compared with a scenario in which the program was not implemented, requiring individual blood donation nucleic acid amplification tests (NAT) to detect the presence of WNV in human samples throughout the seasonal period of vector presence. Considering the five-year period, the application of the environmental/veterinary surveillance program allowed a reduction in costs incurred in the Lombardy Region of 7.7 million EUR. An integrated surveillance system, including birds, mosquito vectors, and dead-end hosts such as horses and humans, can prevent viral transmission to the human population, as well as anticipate the detection of WNV using NAT in blood and organ donors. The surveillance program within a One Health context has given the possibility to both document the expansion of the endemic area of WNV in northern Italy and avoid most of the NAT-related costs