Bovine papillomatosis: First detection of bovine papillomavirus types 6, 7, 8, 10 and 12 in Italian cattle herds.

Limited information about the distribution of different bovine papillomavirus (BPV) types in Italy is available; therefore, this study aimed to investigate the presence of BPVs in bovine lesions in the Emilia Romagna region. Sixty-four proliferative lesions were collected between december 2011 and december 2014, and subsequently analysed by qualitative PCR with genus- and type-specific primer pairs, as well as rolling circle amplification (RCA). The results demonstrated, for the first time in Italy, the presence of BPV 6, 7, 8, 10 and 12 and also types previously described elsewhere. In addition, the high prevalence of viral co-infections in this sample set provides new information about viral tropism.

Equus asinus papillomavirus (EaPV1) provides new insights into equine papillomavirus diversity.

We detected a novel papillomavirus (EaPV1) from healthy skin and from sun associated cutaneous lesions of an Asinara (Sardinia, Italy) white donkey reared in captivity in a wildlife recovery centre. The entire genome of EaPV1 was cloned, sequenced, and characterised. Genome is 7467 bp long, and shows some characteristic elements of horse papillomaviruses, including a small untranslated region between the early and late regions and the lack of the retinoblastoma tumour suppressor binding domain LXCXE in E7. Additionally, a typical E6 ORF is missing. EaPV1 DNA was detected in low copies in normal skin of white and grey donkeys of the Asinara Island, and does not transform rodent fibroblasts in standard transformation assays. Pairwise nucleotide alignments and phylogenetic analyses based on concatenated E1-E2-L1 amino acid sequences revealed the highest similarity with the Equine papillomavirus type 1. The discovery of EaPV1, the prototype of a novel genus and the first papillomavirus isolated in donkeys, confirms a broad diversity in Equidae papillomaviruses. Taken together, data suggest that EaPV1 is a non-malignant papillomavirus adapted to healthy skin of donkeys.

Cervus elaphus papillomavirus (CePV1): new insights on viral evolution in deer.

We identified a novel papillomavirus (CePV1) in a fibropapilloma of a 1.5 year old male red deer (Cervus elaphus) shot in the Italian Alps in Brescia province. PV particles were first observed by electron microscopy and PV DNA was then identified by PCR using degenerate primers. Subsequently we cloned the entire genome and determined its complete sequence. CePV1 genome is 8009 bp long and contains all 9 ORFs and the long untranslated regulatory region characteristic for Delta-papillomaviruses. Pairwise nucleotide alignments and phylogenetic analyses based on concatenated E1-E2-L1 ORFs allowed to determine the highest similarity with the Capreolus caprelus papillomavirus CcaPV1. The analysis of the host-parasite phylogenetic tree interactions suggest the co-divergence of CePV1 and C. elaphus while the identified topological incongruences leading us to speculate that CcaPV1 could eventually be the result of an earlier host switch event.

Original findings associated with two cases of bovine papular stomatitis.

Bovine papular stomatitis virus was isolated from two calves in an animal house with biosafety level 3 confinement. The hypotheses on the origin of the infection, the interesting features of the partial amino acid sequences of the major envelope viral protein, and the importance of diagnostic tools available for animal diseases that are not listed by the World Organization for Animal Health (OIE) are discussed.

Antiviral activity of HPMPC (cidofovir) against orf virus infected lambs.

(S)-1-[3-hydroxy-2-(phosphonomethoxy)propyl]cytosine [corrected] (HPMPC, cidofovir, CDV, Vistide) is an acyclic nucleoside analogue with a potent and selective activity against a broad spectrum of DNA viruses including the poxviruses. In this study we present the results of different treatment regimens in lambs experimentally infected with orf virus with different cidofovir formulations prepared in Beeler basis and Unguentum M. Our results show that choice of excipient, concentration of codofovir [corrected] and treatment regimen were all important to the clinical outcome of the therapy. Whilst one particular regimen appeared to exacerbate the lesion, treatment with 1% (w/v) cidofovir cream, prepared in Beeler basis, for 4 consecutive days did result in milder lesions that resolved in milder lesions that resolved [corrected] more quickly than untreated lesions. Furthermore the scabs of the treated animals contained significantly lower amounts of viable virus meaning there should be less contamination of the environment with virus than would normally occur.

A real time PCR assay for the detection and quantification of orf virus.

A real time quantitative PCR assay based on TaqMan technology was developed for orf virus (ORFV) DNA quantification in clinical samples, infected cells and organotypic cultures. This method was based on the amplification of a 70 bp fragment from the ORFV B2L gene (orthologue of the Vaccinia virus Copenhagen F13L gene) that encodes the major envelope protein. Both intra- and inter-assay variability were well within +/-0.25 log(10) S.D. showing the high efficiency and reproducibility of the assay. The TaqMan PCR was subsequently used to determine the titre of several batches of the ORFV strain NZ-2, with it being possible to quantify virus solutions in the range of 1 x 10(1) to 1 x 10(6) TCID(50)/ml. A good correlation between the titre determined by the TaqMan PCR and by conventional endpoint dilution was found. The PCR assay is reproducible and can be used for a rapid quantification of ORFV in vitro and ex vivo, being readily achievable within 1h.

Heparin binding activity of orf virus F1L protein.

The orf virus is the type species of the Parapoxvirus genus and is the causative agent of contagious echtyma, a debilitating skin disease of sheep and goats, which can also affect man. The virus exhibits a restricted host range, even if it has been shown to bind to a wide range of tissues of non-permissive species. This ability is an argument for its potential use as an expression vector. Since most mammalian cell types express heparan sulfate (HS) surface receptors, we assumed that HS could serve as receptors to mediate orf virus binding. In this study, we showed that orf virus is inhibited by the addition of soluble heparin in cell cultures. Affinity chomatography using heparin agarose demonstrated that orf virus F1L is the major heparin binding protein. Furthermore, the recombinant F1L protein was visualised on the cell surface by confocal microscopy, and rabbits immunised with recombinant F1L protein produced virus neutralising antibodies. These results confirm that the F1L immunodominant protein is also involved in virus binding to cells as for the vaccinia homologue H3L protein. Heparin also inhibited the binding of the F1L protein to cells showing that this protein has a role in the early stages of infection.

Diagnosis of orf virus infection in humans by the polymerase chain reaction.

The orf virus is the causal agent of contagious ecthyma in goats and sheep. The infection can be transmitted to humans and represents a typical example of occupational zoonosis. In Italy, the incidence of human infection remains uncertain because the disease is rarely reported or diagnosed. In this paper, we report a case of human orf virus infection and the laboratory methods of diagnosis. We demonstrated a genomic identity between the conserved and the variable regions of the genome of the viral strains isolated from the human patient and from the infected sheep confirming that there is no specific clone infecting humans rather than animals.

[Surgical treatment of thyroid diseases in geriatric patients]. / Il trattamento delle tireopatie chirurgiche nel paziente in età geriatrica.

Thyroid diseases have a characteristic evolution in geriatric age, whether for the symptomatology frequently mingled with typical manifestations of again, or for glandular involution. Moreover, in the aged patient, the particular aspect of the epidemiology, physiopathology, clinic and therapy are to know and interpret. In most cases, the presence of a uni- or multi-nodular goiter does not cause compression problems or cancerization risk. In the presence of these problems and in multi-nodular goiter, we prefer total thyroidectomy because, at the present time, it is possible to put at zero the risks of this operation, neither we fear hypothyroidism which all the same also appears in less extensive operations. Between thyroid diseases, cancer has a typical biological behaviour and prognosis in geriatric patients. While most tumors have a better curability in geriatric age, these have a worse prognosis. So therapeutic indications very as a function of age. About this the authors present preliminary data of a prospective trial started in 1992.