Patrizia Bassi

Evidence for bovine besnoitiosis being endemic in Italy - first in vitro isolation of Besnoitia besnoiti from cattle born in Italy

Until 2009, bovine besnoitiosis had never been considered endemic in Italy and the only report on the disease in this country referred to animals imported from France shortly before. However, recently, an autochthonous outbreak of bovine besnoitiosis was reported in four herds located at the intersection of the borders between Emilia-Romagna, Toscana and Marche (Northern Apennine Mountains), which has led to an increased awareness concerning this disease. The present study describes a further outbreak of bovine besnoitiosis in Italy. The afflicted herd was a dairy herd with no evidence for contact with cattle from regions known to be endemic for bovine besnoitiosis. The farm investigation was initiated after a three-year old Holstein Friesian dairy cow with generalized thickening and lichenification of the skin was diagnosed with bovine besnoitiosis. The clinical diagnosis was confirmed by gross pathology, histopathology, serology and PCR. Bradyzoites released from tissue cysts obtained from the skin of this animal enabled the first in vitro isolation of Besnoitia besnoiti in Italy. This isolate was named Bb-Italy1. Sequencing of a 2118 bp spanning region including the complete internal transcribed spacer 1 and parts of the 18S and the 5.8S rRNA gene from DNA extracted from skin-derived zoites revealed a 99.9% identity to sequences known for other B. besnoiti isolated from cattle in Europe. Two GKO mice which had been inoculated intraperitoneally with bovine skin-derived bradyzoites became ill 7 days post inoculation. Parasitophorous vacuoles with multiplying zoites were observed in the cell culture inoculated with peritoneal fluids of these mice and a B. besnoiti infection in the mice and in the cell culture could be confirmed by real-time PCR. A serological investigation in the afflicted herd using immunoblots and an immunofluorescent antibody test (IFAT) revealed an overall herd seroprevalence of 9.7% (31/321), whereas within the female animals older than 2 years 17.0% (29/171) of the dams were tested positive. With one exception, an imported cow from Germany, all the seropositive animals were born in Italy. In connection with previously described autochthonous cases of bovine besnoitiosis the case described herein suggests that bovine besnoitiosis should be considered endemic in Italy.

Helicobacter apri sp. nov., isolated from wild boars

Three isolates (A19T, C21 and F12) with spiral-shaped cells and one bipolar sheathed flagellum were obtained from gastric mucosa and caecal contents of three different wild boars (Sus scrofa) and subjected to a polyphasic taxonomic study. A genus-specific PCR showed that these isolates belonged to the genus Helicobacter. Phylogenetic analysis based on 16S rRNA, 60-kDa heat-shock protein (HSP60) and atpA genes demonstrated they formed a novel lineage within this genus. Pairwise 16S rRNA, HSP60 and atpA gene sequence comparisons of the three isolates revealed 99.7, 99.4 and 99.9 % similarity, respectively, among the three isolates; the 16S rRNA gene of isolate A19T shared 98.5 % sequence similarity with its nearest validly named neighbouring species, Helicobacter mastomyrinus (to the type strain MIT 97-5577T). The taxonomic uniqueness of the wild boar isolates was confirmed by protein analysis performed by matrix-assisted laser desorption/ionization time-of-flight MS and by a distinctive biochemical profile. These data indicated that isolates A19T, C21 and F12 represent a novel taxon, for which the name Helicobacter apri sp. nov. is proposed, with isolate A19T (=DSM 28990T=LMG 28471T) as the type strain.

Retroperitoneal Pulmonary Choristoma in a Newborn Calf

A newborn male Romagnola calf, who had died a few hours after birth, was submitted for postmortem examination. Necropsy revealed a 23 cm × 22 cm × 5 cm, pale pink, lobulated, elastic, partially fluctuant mass that protruded from the dorsal retroperitoneal space into the abdominal cavity, extending from the diaphragm to the left kidney. The mass consisted of mature pulmonary tissue and was consistent with a pulmonary choristoma. The gross and microscopic appearance of this rare tumor-like congenital lesion and the possible pathogenesis are discussed.

p16 Immunostaining of Canine Squamous Cell Carcinomas Is Not Associated with Papillomaviral DNA.

While papillomavirus (PVs) are an established cause of human cancer, few reports have supported a relationship between PV and canine squamous cell carcinomas (SCCs). Human oncogenic PVs lead to an increased expression of the p16 tumor suppressor protein, and the latter can be demonstrated immunohistochemically to support a likely causal relationship between tumor and PV infection. In the present study, archive samples of canine SCC from different anatomical locations were tested by polymerase chain reaction for the presence of PV DNA and by p16 immunohistochemistry. The aims were to investigate the relationship between p16 expression and presence of PV DNA, in order to assess the utility of p16 overexpression as a biomarker of PV infection in canine SCC. A total of 52 SCCs were included. Nine cases (17.3%) showed moderate p16 immunoreactivity, with no association with tumor degree of differentiation, histotype or mitotic activity. The canPVf/FAP64 primers amplified Canis familiaris PV-1 DNA from 3 out of 52 tumors (5.8%), one cutaneous, one oral and one tonsillar SCC. There was no association between PV presence and p16 immunostaining. These results do not support a significant role of PVs in the development of canine SCCs. Additionally, PV infection was apparently not the cause of the p16 immunostaining observed in a subset of canine SCCs. A better awareness of p16 level of expression and cellular function in canine cancer may help to define its diagnostic and prognostic role.

Histopathological findings and proliferative activity of canine sebaceous gland tumours with a predominant reserve cell population.

Sebaceous gland tumours represent the third most common skin tumours in dogs, but diagnostic criteria for tumours with basal differentiation (i.e. sebaceous epithelioma) are poorly defined and there is lack of correlation with biological behaviour. The aim of this study was to identify the main histological criteria associated with malignancy in 30 canine sebaceous gland tumours with a predominant reserve cell population. For each case, tumour proliferative activity was assessed by determining mitotic index and the Ki67/MIB-1 index. Additional histological features included endophytic or exophytic growth, proportion of reserve/intermediate/mature cells, connection to the epidermis, nuclear characteristics, peripheral invasion, neoplastic emboli and necrosis. Mitotic and Ki67 indexes were variable, but correlated (R = 0.66; P < 0.001), and both were significantly higher in infiltrative tumours (P = 0.018 and P < 0.001, respectively). No significant difference in histological features was observed between tumours comprised of more or less than 90% reserve cells, nor among tumours showing proliferative activity in sebocytes. This study suggests that high proliferative activity and peripheral invasion should be considered the most significant parameters for the differentiation between benign and malignant sebaceous gland tumours. Furthermore, the incidence of circumanal gland and testicular tumours in these dogs was significantly higher compared with an age-matched control population, suggesting a potential androgen-related pathway for the tumourigenesis of canine sebaceous gland neoplasms.

Enrichment tools for undocked heavy pigs: effects on body and gastric lesions and carcase and meat quality parameters

Abstract Two independent trials were carried out to assess the effect of two enrichment tools on the prevalence of skin and tail lesions recorded prior of slaughtering, lesions of the pars oesophagea (OGL) of the stomach, and on carcase and meat quality traits in Italian heavy pigs (body weight range: 25–160 kg). Eighty undocked barrows (Landrace × Large White) were used in two trials (20 pigs/experimental group, 5 pigs/pen). In Trial 1, the control group received a hanging metal chain (C1), while the other group received wood logs (WL) placed inside a metal rack. In Trial 2, the control group was provided with hanging chain (C2), while the pen of the other group was enriched with a vegetal edible block (EB) placed inside the metal rack. In both trials, no differences were observed in the prevalence and severity of skin, tail and gastric lesions (p > .05). In Trial 1, WL pigs presented lower backfat (p = .01), higher lean meat percentage (p = .03) and higher drip loss in the loin muscle (p = .02) than C1 pigs. Tail score and gastric lesions showed a moderate correlation (r = 0.42; p = .01) in Trial 1. Treatments had no effect on carcase or meat quality traits in Trial 2 (p > .05). In conclusion, the two enrichments provided did not affect body and gastric lesions, carcase and meat quality of Italian heavy pigs, if compared to the metal chains.

Evidence for bovine parafilariosis in Italy: First isolation of Parafilaria bovicola (Tubangui, 1934) from autochthonous cattle

In May and July 2010 and in March 2011, the presence of multiple bleeding nodules at the level of the neck, shoulders, withers, back and rump in five cattle from three herds in north-central Italy were observed. A diagnosis of parafilariosis was made. Fragments of adult females of Parafilaria bovicola could be identified from the derma of three to five bioptic extirpates. Larvated eggs and free larvae were observed in serohemorrhagic exudates. This paper is the first report of the occurrence of parafilariosis in Italy and subsequent to the various outbreaks reported, it can now be said that bovine parafilariosis is also currently present in Italy.