Quantification of Equid herpesvirus 5 DNA in clinical and necropsy specimens collected from a horse with equine multinodular pulmonary fibrosis.

A 15-year-old Belgian gelding was referred for fever, depression, and respiratory distress. Lung biopsy revealed interstitial fibrosis consistent with chronic interstitial pneumonia. Equid herpesvirus 5 (EHV-5) DNA was detected by polymerase chain reaction (PCR) in bronchoalveolar lavage and biopsy specimens. A presumptive diagnosis of equine multinodular pulmonary fibrosis (EMPF) was made, and the horse was administered a systemic treatment with corticosteroids and antiviral drugs. Despite initial clinical improvement, 4 weeks later, the condition of the horse rapidly deteriorated, and the animal was euthanized. Postmortem examination confirmed the presumptive diagnosis of EMPF. The EHV-5 DNA load in different tissues was estimated using a quantitative real-time PCR. Lung had a remarkable viral load, higher than in other organs, especially within the pulmonary fibrotic nodules, and a linkage between high viral burden and the most severely affected tissues was observed. The results suggest that the quantitative real-time PCR is a useful tool to quantify the EHV-5 load in different organs and to understand the relationship between EHV-5 and EMPF. The bronchoalveolar lavage was determined to be a good clinical sample to estimate the EHV-5 load in lung.

Pulmonary rhodococcosis in a cat.

Feline Rhodococcus equi infection is rare, despite the bacteria is widespread in the environment. R equi infection is typically observed in equine species but the infection has also been reported in dogs, cats and other domestic animals. There are a few reports regarding pulmonary R equi infection in cats and the disease appears to be limited to the skin and the subcutaneous tissue. This report describes the pathological, microbiological and the virulence features associated with an acute necrosuppurative pneumonia in a cat. To the best of our knowledge, this is the first report of feline pulmonary R equi infection in Italy.

Molecular detection, epidemiology, and genetic characterization of novel European field isolates of equine infectious anemia virus.

The application of molecular diagnostic techniques along with nucleotide sequence determination to permit contemporary phylogenetic analysis of European field isolates of equine infectious anemia virus (EIAV) has not been widely reported. As a result, of extensive testing instigated following the 2006 outbreak of equine infectious anemia in Italy, 24 farms with a history of exposure to this disease were included in this study. New PCR-based methods were developed, which, especially in the case of DNA preparations from peripheral blood cells, showed excellent correlation with OIE-approved agar gel immunodiffusion (AGID) tests for identifying EIAV-infected animals. In contrast, the OIE-recommended oligonucleotide primers for EIAV failed to react with any of the Italian isolates. Similar results were also obtained with samples from four Romanian farms. In addition, for the first time complete characterization of gag genes from five Italian isolates and one Romanian isolate has been achieved, along with acquisition of extensive sequence information (86% of the total gag gene) from four additional EIAV isolates (one Italian and three Romanian). Furthermore, in another 23 cases we accomplished partial characterization of gag gene sequences in the region encoding the viral matrix protein. Analysis of this information suggested that most Italian isolates were geographically restricted, somewhat reminiscent of the "clades" described for human immunodeficiency virus type 1 (HIV-1). Collectively this represents the most comprehensive genetic study of European EIAV isolates conducted to date.

Age-dependent prevalence of equid herpesvirus 5 infection.

Equid herpesvirus 5 (EHV-5) infection was detected in a farm in Italy by the use of a semi-nested polymerase chain reaction (PCR) targeting glycoprotein B of EHV-5 on nasal swabs and blood samples of clinically healthy and randomly selected Lipizzaner horses (n = 55). Twenty-five horses at the age of 4-17 years and 30 at an age of 1-3 years were sampled once. The association of the infection with these age-groups and the gender of the horses was investigated. The apparent prevalence of EHV-5 infection was significantly different between age-cohorts: it was higher in the younger group of horses with 73,3% and 80% positives in nasal swabs and blood respectively, compared to 40% of nasal swabs and 20% of blood in the older horses. An age-dependence therefore was observed: the young age is more frequently associated with EHV-5 infection.

Characterization of genes encoding virulence determinants and toxins in Staphylococcus aureus from bovine milk in Central Italy.

The aim of this study was to evaluate the genotypic characteristics of Staphylococcus aureus isolates (n=170) from bovine milk collected from seven dairy farms in Italy. On the basis of cultural and biochemical properties and by amplification of the 23S rRNA specific to S. aureus, all isolates were identified as S. aureus. To genotypically characterize S. aureus isolates, genes encoding virulence determinants (nuc, clfA, spa-IgG-binding, spa-X-region, fnbA and fnbB, cap5 and cap8) and staphylococcal enterotoxins (sea, seb, sec, sed, see, seg, seh, sei, sej) were investigated using a PCR technique. The results showed that the isolates of S. aureus in each farm had the same genotypic characteristics, while the isolates genotipically differed between the different farms. The present study might help to understand the distribution of prevalent S. aureus strains in dairy farms.

Anaplasma phagocytophilum in horses and ticks: a preliminary survey of Central Italy.

Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, affects several species of wild and domesticated mammals, including horses. In this work we compared direct and indirect methods to evaluate A. phagocytophilum presence in Central Italy: 135 sera were screened by IFA for A. phagocytophilum and other haemopathogens (Theileria equi and Babesia caballi). Each horse was also tested for A. phagocytophilum 16S rRNA with a nested-PCR technique. In order to examine the risk of A. phagocytophilum transmission, 114 ticks were examined for the presence of A. phagocytophilum by PCR targeting the 16S rRNA. The seroprevalence against A. phagocytophilum was 17.03% and 11 horses (8.14%) showed positive PCR results. The concordance rate of A. phagocytophilum detection between IFAT and PCR had a K value of 0.34.