Francesca Vaccari

TaqMan Based Real Time PCR for the Quantification of Canine Distemper Virus

Scagliarini, A., Dal Pozzo, F., Gallina, L., Vaccari, F. and Morganti, L., 2007. TaqMan based real time PCR for the quantification of canine distemper virus. Veterinary Research Communications, 31(Suppl. 1), 261–263

Antiviral efficacy of EICAR against canine distemper virus (CDV) in vitro.

Canine distemper virus (CDV) is a highly contagious pathogen of carnivores. In dogs, the disease is characterized by high lethality rates and no specific antiviral therapy is available. The aim of this study was to verify the in vitro antiviral activity of the 5-ethynyl-1-beta-d-ribofuranosylimidazole-4-carboxamide (EICAR) and to compare it with the 1-(beta-d-ribofuranosyl)-1,2,4-triazole-3-carboxamide (ribavirin, RBV). EICAR was more active than RBV against CDV replication, while both molecules exhibited low selectivity indexes. A reversal of their antiviral activity was observed after addition of guanosine, suggesting their involvement in the inhibition of the inosine monophosphate dehydrogenase enzyme (IMPDH). RBV and EICAR had a time- and concentration-dependent anti-CDV activity, mainly displayed during the first 10h post-infection. The involvement of the inhibition of the viral RNA-dependent RNA polymerase (vRdRp) is discussed, as well as the role of CDV as a model to study more potent and selective antiviral molecules active against other Paramyxoviridae.

Analysis of deletion within the reindeer pseudocowpoxvirus genome.

Cases of contagious pustular stomatitis have been reported in Finnish reindeer for many years. Two species of the genus Parapoxvirus of the family Poxviridae have been identified as the causative agent of the disease; orf virus (ORFV) was found during the 1992-1993 epidemic and pseudocowpoxvirus (PCPV) was connected to the 1999-2000 epidemic. The genome of reindeer parapoxvirus from the latter outbreak, isolate F00.120R, was recently sequenced and confirmed as PCPV. The six gene deletion of the right terminus of the F00.120R genome, in comparison to ORFV, was investigated in an attempt to use it in differentiating viruses causing pustular stomatitis in reindeer. The present study describes discovery and analysis of genes 116-121 in reindeer PCPV and in an Italian field isolate of bovine PCPV. The results show that a 5431 bp sequence containing genes 116-121 was likely to have been deleted from the F00.120R genome between the 6th and 7th passage in cell culture, and that these genes are present in other isolates of reindeer and bovine PCPV isolated in Finland during the years 2005-2010. The data presented here extends our knowledge of the PCPV genome, confirming that it contains homologues of all known ORFV genes and further reinforces their close genetic relationship. The similarity between the EEV envelope and GM-CSF inhibitory factor genes from reindeer PCPV and ORFV isolates, Finnish sheep ORFV and cattle PCPV isolates indicate that these viruses have been circulating among Finnish reindeer, cattle and sheep over a long period of time.

Parapoxvirus Infections of Red Deer, Italy

To characterize parapoxviruses causing severe disease in wild ruminants in Stelvio Park, Italy, we sequenced and compared the DNA of several isolates. Results demonstrated that the red deer isolates are closely related to the parapox of red deer in New Zealand virus.

Co-infection with multiple variants of canine parvovirus type 2 (CPV-2).

Battilani, M., Gallina, L., Vaccari, F. and Morganti, L., 2007. Co-infection with multiple variants of canine parvovirus type 2 (CPV-2). Veterinary Research Communications, 31(Suppl. 1), 209–212

Virulent feline calicivirus disease in a shelter in Italy: a case description.

Abstract Feline calicivirus (FCV) is a common pathogen of cats that is particularly widespread in multi-cat environments such as shelters and catteries. FCV infections are usually associated with acute, mild and self-limiting upper respiratory tract disease characterized by oral vesicles/ulcers. Recently, virulent systemic disease (VSD) associated with FCV infection has been reported in the USA and Europe. This paper describes a case of VSD affecting one adult, FIV infected cat (“Oscar”) living in a shelter located in Northern Italy; the clinical, post-mortem and laboratory findings indicate that this is the first case of suspected FCV-VSD in this country. Similar to a previous report (Meyer et al., 2011), the disease affected only one cat, while others remained asymptomatic, despite their direct contact with “Oscar”. Phylogenetic analysis identified unique features in the “Oscar” FCV isolate. The FIV infection of the patient might have favoured the generation of the virulent FCV strains in this cat.

In Vitro Evaluation of Antiviral Activity of Ribavirin Against Canine Distemper Virus

Canine distemper virus (CDV) a member of the Morbillivirus genus of the family Paramyxoviridae, is a highly infectious and an acknowledged lethal pathogen of many carnivores. The disease is controlled by vaccination but an increasing number of distemper cases has been recorded even in vaccinated dogs all over the world and the affected animals can display catharral and or neurological symptoms. At present, there is no specific therapy, even if there is an increasing demand for one from the dog owners. For this reason the research on antiviral compounds against distemper is aimed at finding active molecules to treat the infection. Ribavirin (1-β-D-ribofuranosyl-1,2,4-triazole-3-carbox-amide, RBV) is a broad spectrum antiviral agent (Garcia et al., 2001; Crance et al., 2003; Poynard et al., 1998) that has been licensed for clinical use (as an aerosol) in the treatment of respiratory syncytial virus and for the therapy of measles pneumonia (Forni et al., 1994) and subacute slerosing panencephalitis (Hara et.al, 2003; Hosoya et al., 2004). The aim of the present study was to evaluate the in vitro antiviral activity of ribavirin against CDV.

Detection and quantification of Betanodavirus by real time PCR

Betanodavirus infection is now widespread in several fish species worldwide and it causes severe economic losses in European sea bass (Dicentrarchus labrax) farming in Italy. Betanodavirus are small ssRNA non-envelope viruses. Based on phylogenetic analysis of the coat protein gene, Betanodavirus can be classified into four genotypes: SJNNV, TPNNV, BFNNV, RGNNV (Nishizawa et al., 1997). The virus is widespread both in farmed and wild animals (Gagné et al., 2004; Gomez et al., 2004) and it has high resistance to environmental and chemico-physical disinfectant treatment, so control by direct prophylaxis is particularly difficult. Furthermore, fattening-farm is often conducted in sea cages or in extensive brackish ponds with no clear-cut separation between wild and farmed fish (Munday et al., 2002). Despite a recent work (Thiéry et al., 2004) reporting the presence of a SJNNV-like virus in a sole farmed in the Mediterranean, the main genotype widespread in this region and responsible for the outbreaks on sea bass farms is the RGNNV. Unfortunately, there is no vaccine available for this disease, due to a limited knowledge about the pathogenesis of Betanodavirus infection, so control is based on early diagnosis and broodstock control. In this study we set up a new Real Time PCR diagnostic method. This technique was found to be rapid, practical and sensitive. Several Real Time PCR methods have recently been set up to detect numerous pathogens significantly contributing to studies on pathogenesis (Gilad et al., 2004), diagnosis and antiviral activity (Yun et al., 2003).