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The recent reinforcement of CoV surveillance in animals fuelled by the COVID-19 pandemic provided increasing evidence that mammals other than bats might hide further diversity and play critical roles in human infectious diseases. This work describes the results of a two-year survey carried out in Italy with the double objective of uncovering CoV diversity associated with wildlife and of excluding the establishment of a reservoir for SARS-CoV-2 in particularly susceptible or exposed species. The survey targeted hosts from five different orders and was harmonised across the country in terms of sample size, target tissues, and molecular test. Results showed the circulation of 8 CoV species in 13 hosts out of the 42 screened. Coronaviruses were either typical of the host species/genus or normally associated with their domestic counterpart. Two novel viruses likely belonging to a novel CoV genus were found in mustelids. All samples were negative for SARS-CoV-2, with minimum detectable prevalence ranging between 0.49% and 4.78% in the 13 species reaching our threshold sample size of 59 individuals. Considering that within-species transmission in white-tailed deer resulted in raising the prevalence from 5% to 81% within a few months, this result would exclude a sustained cycle after spillback in the tested species.

The genus Pestivirus comprises globally distributed members of the family Flaviviridae, which cause severe losses in livestock. The most common species of the genus are bovine viral diarrhoea virus type 1 (BVDV-1) and type 2 (BVDV-2), classical swine fever virus (CSFV) and border disease virus (BDV). Recently, a novel ovine pestivirus was repeatedly detected in aborted lamb foetuses on a farm located in the Brescia Province (Italy). Complete genome characterization of this isolate showed that it was highly divergent from known pestivirus species and that it was genetically closely related to CSFV. The aim of this study was to determine the serological relatedness between the identified novel pestivirus and BVDV, BDV and CSFV selected strains for which homologous serum was available, by antigenic characterization performed using cross-neutralization assays. The serological relatedness was expressed as the coefficient of antigenic similarity (R). Both field and specific antisera raised against the ovine pestivirus neutralized the CSFV reference strain Diepholz with titres significantly higher than those specific for the BDV and BVDV strains. Furthermore, the calculated R values clearly indicated that the novel ovine pestivirus is antigenically more related to CSFV than to ruminant pestiviruses, in agreement with the results of the genomic analysis. This would have severe consequences on CSFV serology in the event of a switch to porcine hosts with implications for CSFV surveillance and porcine health management., (© 2020 Blackwell Verlag GmbH.)

Although  the  eradication  of  Pseudorabies  virus  (PrV)  in  domestic  pigs  is  ongoing,  the circulation  of  this  virus  in  wild  boars  remains  a  threat  in  the  currently  unprotected, 'low prevalence', pig population. In this study, we reported PrV prevalence data and the influence of  possible  risk  factors  in  2  North-West  Italian  wild  boar  populations  (free  and  enclosed) with the goal of supporting the implementation of a risk-based AD surveillance system. Sera from  1,425  wild  boars  were  collected  between  2011  and  2015  and  tested  by  ELISA  for  the presence  of  PrV  antibodies;  the  overall  raw  seroprevalence  was  30.39%  (433/1,425;  CI  95% 28.01-32.85%).  A  significant  difference  was  however  observed  between  the  prevalence rates  of  the  free  range  population  (9.98%;  90/902;  CI  95%;  8.10-12.12%)  and  the  enclosed  population of La Mandria park (Piedmont, Italy) (65.58%; 343/523; CI 95%; 61.51-69.65%). In both populations a significantly higher number of adults and females were found positive to PrV  ELISA.  Specific  territorial  data  on  PrV  circulation  in  wild  boars  should  be  acquired  from other regions for guiding risk-based measures in order to reduce the threat of AD re-infection in a more cost-effective manner.

Sixteen haemagglutinin (HA) subtypes of avian influenza viruses (AIV) have been described to date. Rapid subtype identification of any AIV is of major interest because of the possible serious consequences for the poultry industry and even public health. Molecular techniques currently allow immediate accurate subtype characterisation prior to virus isolation. In this study, a set of fourteen specific real-time RT-PCR methods were developed and evaluated for AIV HA subtyping (H1-H4, H6-H8, H10-H16), H5 and H9 being excluded on the basis of the current validity of the European Union (EU) recommended specific assays. Specific primers and probes sets for each HA-subtype were designed to hybridise the largest isolates range within each single subtype, considering the Eurasian lineage as a major target. The robustness and general application of the 14 HA-subtype methods were verified by the analysis of 110 AIV isolates belonging to all 16 HA-subtypes, performed in different laboratories. The developed real-time RT-PCR assays proved to be highly specific and revealed suitable sensitivity, allowing direct HA-subtyping of clinical material. In summary, this study provides for the first time a panel of molecular tests using specific hydrolysis probes for rapid and complete AIV HA-subtype identification., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Five chicken lines were experimentally infected with a HPAI H7N1 virus, to examine the variation in susceptibility to infection. Three lines showed high susceptibility to the virus, while two showed some resistance, with 7 out of 20, and 11 out of 15 birds, respectively, remaining healthy and surviving the experimental infection. Genotyping for the G/A polymorphism at position 2032 of Mx cDNA showed that one line was fixed for the G allele, and two were segregating for A and G alleles. Birds in the other two lines were selected to be fixed for the A allele. Statistical analyses indicated that the Mx genotype did not affect the clinical status or the time course of infection after viral inoculation.