Detection of anti-HEV antibodies but no molecular positivity in dogs in the Campania region, southern Italy.

Paslahepevirus balayani genotypes 3 and 4 (HEV-3 and 4) have zoonotic potential and can be transmitted to humans and animals through the consumption of contaminated raw or undercooked meat. Although it has been demonstrated that dogs are susceptible to the infection and produce specific antibodies, the epidemiological role of this species is not yet well defined. This study aimed to evaluate the circulation of HEV at the serological and molecular level in the dog population of the Campania region, southern Italy. A total of 231 dogs were sampled, divided according to several variables (sex, age, origin, lifestyle, location, size, and breed), and tested for the presence of HEV antibodies using a commercial multi-species ELISA. A total of 197 blood samples and 170 stool samples were tested with two specific PCRs in order to detect viral RNA. A total of 19 out samples of 231 were seropositive, obtaining an exposure (8.2%) similar to that observed in other European countries. The univariate and multivariate analysis revealed a wide exposure to stray dogs and animals from the province of Salerno. All samples tested with molecular methods were negative. Defining the role of domestic carnivores continues to be a "one health" challenge, although it appears that they do not eliminate the virus and therefore do not pose a danger to humans. In the absence of other evidence, it is advisable to continue to carry out surveillance also for domestic animals, which, due to ethological characteristics or their position in the food chain, could be predisposed to being exposed to HEV.

Viral and Antibody Prevalence of Hepatitis E in European Wild Boars (Sus scrofa) and Hunters at Zoonotic Risk in the Latium Region.

Hepatitis E virus (HEV) is a member of the genus Hepevirus within the family Hepeviridae. Hepatitis E is recognized as a zoonosis, and swine and wild boars (Sus scrofa) are known reservoirs of HEV infection. The aim of this study was to investigate the presence of HEV in wild boars and hunters exposed to infection in central Italy (Latium region). During the hunting season, blood samples were collected from 228 wild boars and 20 hunters. The seroprevalence of HEV infection was determined using a commercial enzyme-linked immunosorbent assay, previously validated for use in man, pigs and wild boars. The estimated HEV seroprevalence in wild boars and in hunters was 40.7% (93/228; 95% confidence interval [CI] 34.4-47.1%) and 25% (5/20; 95% CI 6.1-43.9%), respectively. Liver samples were collected from the boars and HEV RNA was detected by nested reverse transcriptase polymerase chain reaction. Fifty-five of 164 tested wild boar liver samples (33.5%; 95% CI 26.2-40.7%) and three of 20 (15.0%; 95% CI 1.3-28.7%) tested human serum samples were positive for HEV RNA. Phylogenetic analysis of the nucleotide sequences obtained from PCR products indicated that the HEV strains present in wild boars and the human population all belonged to genotype 3, supporting the zoonotic role of wild boars in the spread of HEV infection.

Presence and distribution of urocortin and its receptors in the epididymis of alpaca (Vicugna pacos).

Urocortin 1 (UCN) is a 40-amino acid peptide belonging to the corticotrophin-releasing hormone (CRH) family. The biological effects of this peptide are modulated by binding two G-coupled receptors named CRH receptor 1 (CRHR1) and CRH receptor 2 (CRHR2). CRHR2 has high affinity for UCN. The aim of the present study was to investigate the presence and distribution of UCN, CRHR1 and CRHR2 in the epididymis of the South America camelid Alpaca (Vicugna pacos) by Western blotting analysis and immunohistochemistry. Tissue extracts of the organ reacted with the anti-UCN, anti-CRHR1 and anti-CRHR2 antibodies, recognizing in all the cases a single specific protein band. UCN- and CRHR2-immunoreactivities (IRs) were found in the cytoplasm of the principal cells (PCs) of the caput epididymis. A prevalent supranuclear localization of granular-shaped positive material was observed. CRHR1-IR was observed in the fibromuscular stromal cells encircling the tubules and in the smooth musculature of the blood vessels throughout the three epididymal segments. In addition, in the cauda, CRHR1-IR was observed in some apical epithelial cells (ACs) which were morphologically similar to apical mitochondria-rich cells (AMRCs). These results suggest that UCN, CRHR1 and CRHR2 are expressed in the alpaca epididymis and that CRH-related peptides might play multiple roles in maturation and storage of spermatozoa.

Presence and distribution of urocortin and corticotrophin-releasing hormone receptors in the bovine thyroid gland.

Urocortin (UCN), a 40 amino acid peptide, is a corticotrophin-releasing hormone (CRH)-related peptide. The biological actions of CRH family peptides are mediated via two types of G-protein-coupled receptors, CRH type 1 (CRHR1) and CRH type 2 (CRHR2). The aim of this study was to investigate the expression of UCN, CRHR1 and CRHR2 by immunoprecipitation, Western blot, immunohistochemistry and RT-PCR in the bovine thyroid gland. Immunoprecipitation and Western blot analysis showed that tissue extracts reacted with the anti-UCN, anti-CRHR1 and anti-CRHR2 antibodies. RT-PCR experiments demonstrated that mRNAs of UCN, CRHR1 and CRHR2 were expressed. UCN immunoreactivity (IR) and CRHR2-IR were found in the thyroid follicular and parafollicular cells and CRHR1-IR in the smooth muscle of the blood vessels. These results suggest that a regulatory system exists in the bovine thyroid gland based on UCN, CRHR1 and CRHR2 and that UCN plays a role in the regulation of thyroid physiological functions through an autocrine/paracrine mechanism.

Apocynin activity in spontaneously hypertensive rats (SHR): preliminary studies in vivo.

An elevation in angiotensin II (Ang II) levels is a common occurrence in spontaneously hypertensive rats (SHRs). Infusions of Ang II and a high salt diet increase the activity of NADPH oxidase that stimulates superoxide anion (O(-2)) generation and increases the expression of certain subunits of NADPH oxidase. Apocynin, an NADPH oxidase inhibitor with antihypertensive effects, is able to inhibit the release of superoxide anion by inhibiting NADPH oxidase activity and blocking the migration of p47 phox to the mitochondrial membrane. The aim of our study was to evaluate the antihypertensive effects of apocynin in SHRs and Wistar rats (WKYs) using a micropuncture technique. After microperfusion of both the proximal and distal tubules, we found that SHRs treated with apocynin showed a decrease in the free-flow collection of the proximal tubule (PT), which was not affected in WKYs. Moreover, significant differences were not demonstrated in the distal tubule (DT), probably due a mechanism of compensation that occurs in the loop of Henle. In conclusion, it is possible that the mechanisms of reabsorption in the PT are controlled by the interactions of O(-2) and nitric oxide (NO). These data could suggest a higher activity of NADPH oxidase and increase in reactive oxygen species (ROS) production in the PT during hypertension.

2,3,7,8-Tetrachlorodibenzo-p-dioxin increases Bovine Herpesvirus type-1 (BHV-1) replication in Madin-Darby Bovine Kidney (MDBK) cells in vitro.

Dioxin-2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a common environmental toxin of current interest. In the last years, higher levels of TCDD than those permitted in UE [European Commission. 2002. European Commission Recommendation 2002/201/CE. Official Gazette, L 67/69] were detected in milk samples from cow, water buffalo, goat, and sheep raised on some areas of Campania Region (South Italy). Dioxin often causes immunosuppression and might render the animal liable to viral infections. In addition, viral infections are able to alter the pattern of dioxin distribution in different organs of the exposed animals. Bovine Herpesvirus type-1 (BHV-1) is a widespread pathogen, which causes infectious rhinotracheitis and infectious pustular vulvovaginitis in cattle. Herein, we have studied the effects of TCDD and BHV-1 infection, in Madin-Darby Bovine Kidney (MDBK) cells, alone as well as in association, so as cellular proliferation, apoptosis, and virus replication. We have observed an increase in cell viability of confluent monolayers at low TCDD concentrations. TCDD treated cells demonstrated increased viability compared to controls as evaluated by MTT test. TCDD exposure increased cell proliferation but induced no changes on apoptosis. Cells exposed to TCDD along with BHV-1 showed a dose-dependent increase in cytopathy, represented by ample syncytia formation with the elimination of the cellular sheets and increased viral titer. These results suggest that TCDD increases viral replication in MDBK cells while BHV-1 further decreases viability of TCDD exposed cells. Since very low concentrations (0.01 pg/ml) are sufficient to augment BHV-1 titer, TCDD may contribute to reactivate BHV-1 from latency, leading to recurrent disease and increase virus transmission.