Identification of a zoonotic genotype 3 hepatitis E subtype in wildlife in north-eastern Italy.

Hepatitis E virus (HEV) is an emerging zoonosis caused by a positive RNA single stranded virus of the Hepeviridae family. In developed countries, pigs and wild boars are the main reservoir for zoonotic genotypes 3 and 4. In spring 2017, for the first time HEV was detected in wild boars captured in the Regional Park of the Euganean Hills, in north-eastern Italy. Phylogenetic analysis of two complete viral genomes and seven partial ORF1 and ORF2 sequences of HEV viruses, selected from twelve positive animals, showed that the viruses grouped together within genotype 3 but clustered separately from previously identified subtypes, thus suggesting the identification of a novel genotype 3 subtype. The phylogenetic analysis of nine partial ORF2 sequences showed the closest similarity with wild boar/human viruses identified in central-northern Italy in 2012. The circulation of HEVs in this area, characterized by a vast man-made environment, an overpopulation of wild boars and >150 swine farms, should be considered in a public health perspective. Further investigations at the wild/domestic species and human interface are therefore necessary to gain a deeper understanding of HEV dynamics.

Prevalence of vertical root fractures in teeth planned for apical surgery. A retrospective cohort study.

AIM: To evaluate retrospectively the prevalence of vertical root fractures (VRFs) in a cohort of patients during apical surgery and the factors possibly associated with VRF. METHODOLOGY: The sample consisted of 944 root filled teeth belonging to 768 patients (49.3% males and 50.7% females; mean age 43.5 ± 11.2 years, range 22-68 years), consecutively referred for endodontic surgery over a six-year period. All patients underwent a clinical assessment of their signs and symptoms. Periapical radiographs of teeth that were candidates for endodontic surgery were taken. Sixty-eight teeth with VRF were identified. Vertical root fractures were identified in pre-surgical screenings in 32 cases (47.1%), and these did not undergo surgery. Another 36 cases of VRF were noted during the intervention for root-end resection. The influence of posts, post type, tooth type, periodontal probing defects, spontaneous pain, sinus tract and follow-up duration was assessed using a logistic regression analysis. RESULTS: Vertical root fractures occurred significantly more frequently (P < 0.001) when a post was present (61 VRF out of 377 teeth with post, prevalence 16.2%) than in teeth without a post (1.2%). Threaded posts and cast posts were significantly more involved in VRF than fibre, silica or carbide posts (P < 0.001). Most fractures (80.9%) occurred 1-5 years after root canal treatment. Sinus tracts, probing defects and spontaneous pain were significantly more associated with VRF cases than with nonfractured teeth. CONCLUSIONS: In the present group of teeth, the major risk for VRF was represented by posts retained by actively engaging the canal via mechanical design (thread) or by frictional fit (cast).

Characterization of Immune System Cell Subsets in Fixed Tissues from Alpine Chamois (Rupicapra rupicapra).

Immune system cell subsets in lymph nodes and spleen from alpine chamois (Rupicapra rupicapra subspecies rupicapra) living in the Italian Alps were characterized immunohistochemically. Seven primary antibodies (against human CD3, CD79αcy, CD68, or ovine CD4, CD8, CD21 and γδ T-cell receptor [TCR] epitopes) were tested on tissues fixed either in formalin or in zinc salts (ZS) and cross-reactivity with chamois immune cell epitopes was shown. ZS fixation allowed wider identification of immune cells, without the need for antigen retrieval. CD4(+) and CD21(+) cells were labelled only in ZS-fixed tissues. Reagents specific for human CD3, CD79 and CD68 antigens successfully detected chamois immune cells, both in ZS-fixed and formalin-fixed tissues. The reactivity and distribution of immune cells in lymph nodes and spleen were similar to those described in other domestic and wild ruminants. Results from this study may allow future investigation of the immune response and pathogenesis of diseases in the chamois.

Impact of emergency oral rabies vaccination of foxes in northeastern Italy, 28 December 2009-20 January 2010: preliminary evaluation.

Fox rabies re-emerged in northeastern Italy in 2008, in an area bordering Slovenia. In 2009, the infection spread westward to Veneto region and in 2010 to the provinces of Trento and Bolzano. Aerial emergency oral fox vaccination was implemented in the winter 2009-10. Since this vaccination was performed at altitudes below the freezing level, a statistical analysis was conducted to evaluate its impact. Of the foxes sampled following the vaccination campaign, 77% showed a rabies antibody titre of >or=0.5 IU/ml.

A transcriptional cross-talk between RhoA and c-Myc inhibits the RhoA/Rock-dependent cytoskeleton.

The GTPase RhoA participates in a number of cellular processes, including cytoskeletal organization, mitogenesis and tumorigenesis. We have previously shown that the transforming activity of an oncogenic version of RhoA (Q63L mutant) was highly dependent on the transcriptional factor c-Myc. In contrast to these positive effects in the RhoA route, we show here that c-Myc affects negatively the F-actin cytoskeleton induced by RhoA(Q63L) and its downstream effector, the serine/threonine kinase Rock. This effect entails the activation of a transcriptional program that requires synergistic interactions with RhoA-derived signals and that includes the upregulation of the GTPase Cdc42 and its downstream element Pak1 as well as the repression of specific integrin subunits. The negative effects of c-Myc in the F-actin cytoskeleton are eliminated by the establishment of cell-to-cell contacts, an effect associated with the rescue of Pak1 and integrin levels at the post-transcriptional and transcriptional levels, respectively. These results reveal the presence of a hitherto unknown signaling feed-back loop between RhoA and c--Myc oncogenes that can contribute to maintain fluid cytoskeletal dynamics in cancer cells.

Genetic variability of Haemonchus contortus (Nematoda: Trichostrongyloidea) in alpine ruminant host species.

Genetic variability of the ovine parasite Haemonchus contortus from the Alpine area was investigated using mitochondrial DNA (nd4 gene), internal transcribed spacers 1 and 2 and microsatellites, in order to assess whether cross-transmission between domestic and wild ruminants occurs. The dataset was composed of 78 individual adult male H. contortus collected from chamois (Rupicapra r. rupicapra), roe deer (Capreolus capreolus), alpine ibex (Capra ibex ibex), domestic goat (Capra hircus) and sheep (Ovis aries) from different alpine areas. The data obtained show low host specificity and high genetic variation within H. contortus populations. The analyses indicate the presence of two mitochondrial haplotype clusters among host species and the absence of cryptic parasite species, confirming H. contortus as a generalist nematode and suggesting that parasite transmission between populations of domestic and wild ruminants normally occurs.

Dynamics of parasite communities and interactions between wild and domestic ruminants.

Dynamics of parasite communities and host-parasite-environment interactions can be influenced by different factors. The present note, by discussing some field experiences in wild and domestic ruminant populations, approaches eco-epidemiology of abomasal nematodes in relation with host health and dynamics. Factors possibly playing a role in the host-parasite relationship are discussed, as well as possible use of macroparasites as ecological and sanitary indicators. The above topics are approached in a management perspective, in particular regarding interactions between domestic and wild ruminant populations.

Seasonal changes in serum metabolites in free-ranging alpine marmots (Marmota marmota).

Circannual changes in serum parameters and body mass were studied in free-ranging Marmota marmota that had been shot in Switzerland (Grisons) for management reasons in May, July, and September of 1995, 1996, and 1997; and in April 1996. Markers of lipid (triglycerides, cholesterol), protein (total protein, urea-nitrogen) and mineral (calcium, inorganic phosphate) metabolism were evaluated in 111 haemolysis-free serum samples; the effects of sex, age, reproductive status, season and year were tested. Mean body mass was higher in adult males than in adult females in July and September, and serum cholesterol concentrations were lower in adult males in May. Pregnant females had lower concentrations of total protein than non-pregnant females, and triglyceride concentrations were negatively correlated with the number of uterine ampullae. Inorganic phosphate decreased and total protein increased with age. In adults, triglycerides, cholesterol and urea-nitrogen increased mainly from May to July and decreased between September and May. Total protein, calcium and phosphate did not change throughout the year. This suggests that, during the active season, ingested lipids and amino acids were utilised for metabolic needs or lipid storage, while in winter, lipids were catabolised and protein was spared. Quantitative differences between years, observed for triglycerides and total protein during the active season, were probably due to different climatic conditions in each year.

Serological study of a population of alpine chamois (Rupicapra r rupicapra) affected by an outbreak of respiratory disease.

A serological survey of respiratory virus infections was carried out from 1998 to 2001 in Lecco province, Italy, as part of a health monitoring programme in a population of alpine chamois, many of which died of pneumonia in autumn and winter 2000 to 2001; 194 carcases of all age classes were found over a short period and in a small area. Eighteen of them, which were examined postmortem, consistently showed signs of severe fibrinous lobar pneumonia or catarrhal bronchopneumonia. Samples of serum from 145 chamois collected from hunted animals and carcases were tested by a virus neutralisation test against bovine respiratory syncytial virus (BRSV), bovine viral diarrhoea virus, bovine herpesvirus type 1 and parainfluenzavirus type 3. Positive results were detected only for BRSV. The area was divided into two subunits on the basis of the distribution of deaths; in the areas where fatalities were observed there was a significant increase of BRSV titres at the beginning of the outbreak. Furthermore, during the 2000 and 2001 hunting seasons antibody titres to BRSV were significantly higher in the areas where mortality occurred. The roe deer living in the same area were not affected by pneumonia and had a low prevalence of titres to BRSV which did not vary during the period of the study.

Glycosaminoglycan-promoted muscle reinnervation and insulin-like growth factor-I levels are affected by anti-growth hormone-releasing hormone exposure.

The present study shows that exposure to antibodies to growth hormone-releasing hormone (GHRH) partially counteracted the promoting effects of treatment with glycosaminoglycans (GAGs) on muscle reinnervation. Sciatic nerve crush was performed in 2-day-old rats, and reinnervation of the extensor digitorum longus muscle was monitored. The extent of reinnervation was rather poor in saline-treated rats, whereas in GAG-treated rats the extent of muscle reinnervation, the recovery of nerve-evoked muscle twitch tension, and the number of motor neurons reinnervating the extensor digitorum longus muscle were greatly enhanced. In addition, treatment with glycosaminoglycans increased markedly insulin-like growth factor-I (IGF-I) levels in denervated muscles. Both types of stimulatory action exerted by GAGs were affected by concomitant exposure to anti-GHRH, with abolition of IGF-I muscle increase and a smaller enhancement in muscle reinnervation.

Sheep oestrosis (Oestrus ovis Linné 1761, Diptera: Oestridae) in Sardinia, Italy.

Oestrosis, the nasal myiasis of sheep and goats, is caused by the larvae of Oestrus ovis L. 1758 (Diptera, Oestridae) that develop from the first to the third stage larva in the nasal cavities and frontal sinuses of affected animals. The authors report the results of an epidemiological study of oestrosis of sheep in Sardinia, Italy. Heads of 6-month to 10-year-old Sardinian sheep (n=566) from 124 free-ranging flocks were examined for the presence and location O. ovis larvae from December 1996 to November 1997. Larvae were collected, counted, and larval stages were identified. O. ovis larvae were found in 100% of examined flocks and in 91% (514/566) of examined sheep. The monthly prevalence ranged from 69% in May to 100% in July. First stage larvae were found in 82% (463) of all heads examined, second stage larvae in 65% (367) and third stage larvae in 10% (56). The majority of sheep harboured first stage larvae, with prevalences of over 80% throughout most of the study period. The prevalence of O. ovis found in this study of Sardinian sheep is the highest reported in the Mediterranean area. The high percentage of first stage larvae found throughout the entire study period may be due to a brief period of decreased rate of larval maturation, in particular in December 1996 (96%) and January-October 1997 (94%). Third stage larvae were consistently present, often however, with extremely low prevalences compared to total larval burden.

Host specificity of abomasal nematodes in free ranging alpine ruminants.

Abomasums from 641 alpine wild ruminants representing five different species (Cervus elaphus, Capreolus capreolus, Rupicapra rupicapra, Capra ibex, Ovis musimon) and from 19 domestic sheep (Ovis aries) from alpine areas were examined in order to investigate the host-specificity of abomasal helminths. Nine out of 20 helminth species were found in at least five different host species. A discriminant analysis was able to significantly discriminate the hosts on the basis of their helminth community composition with the exception of O. musimon and O. aries. Based on the correlation between each variable represented by helminth species with the most explanatory discriminant axis, it was possible to classify helminths into specialists and generalists. Specialists are represented by the dominant species in a community of an host species or family while generalists appear in the communities of many different hosts as intermediate species. Due to the pathogenic potential of some of these generalist species (i.e. Haemonchus contortus, Trichostrongylus axei) and their ability to adapt easily to the conditions found in several different hosts, they appear to be the most important from a sanitary point of view.

Function approximation--a fast-convergence neural approach based on spectral analysis.

We propose a constructive approach to building single-hidden-layer neural networks for nonlinear function approximation using frequency domain analysis. We introduce a spectrum-based learning procedure that minimizes the difference between the spectrum of the training data and the spectrum of the network's estimates. The network is built up incrementally during training and automatically determines the appropriate number of hidden units. This technique achieves similar or better approximation with faster convergence times than traditional techniques such as backpropagation.

Impact of abomasal nematodes on roe deer and chamois body condition in an alpine environment.

Abomasal helminths were examined from 101 roe deer and 43 chamois, shot or found-dead, in the period July-November 1993-95, in the Parco Naturale Paneveggio Pale di San Martino (Italian Eastern Alps). Twelve helminth species were observed in both roe deer and chamois. For both host species, discriminant analysis, using the abundance of each parasite species in the hosts, failed to distinguish between subjects from each of the three study years. For 44 roe deer and 27 chamois data on body weight, body length, foot length, withers height, thoracic circumference and kidney fat index were also available. Principal Component Analysis was performed on the biometrical data in order to group related variables into a few compound variables. In both host species it was possible to identify two principal components, with an explained variance > 80%: the first component, led by body length, was an expression of skeletal development and the second, led by kidney fat index, of nutritional status. Parasite abundance and richness appeared to be negatively correlated with the scores of both skeletal and nutritional Components in the two host species, although only the correlation between parasite abundance and the scores of the nutritional component in roe deer was statistically significant (Spearman r = -0.411; p < 0.05). These results support the susceptibility of roe deer to abomasal helminths, which could be important considering the spatial and nutritional interspecific competition between domestic and wild ruminants often recorded in alpine environments.