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Dalipi R.,University of Brescia | Borgese L.,University of Brescia | Zacco A.,University of Brescia | Tsuji K.,Osaka City University | And 4 more authors.
International Journal of Environmental Analytical Chemistry

This work was performed to highlight the advantages of total reflection X-ray fluorescence spectroscopy (TXRF) for multi-elemental qualitative and quantitative analysis of wine. Indeed the International Organization of Vine and Wine (OIV) selected some potentially toxic elements and proposed limit values for their concentration in wines. Direct TXRF analysis of nine wine samples from Emilia Romagna region of Italy was performed in two different laboratories: Italy and Japan. Wine dehydration was also evaluated as sample conservation mean. Traces of Fe, Cu, Zn and Pb are present in all the analysed samples, with concentrations lower than the limits established by the OIV. The target hazard quotients (THQs) were also calculated for seven elements (Cl, Mn, Fe, Ni, Cu, Zn and Sr) to determine their potential detrimental effects. The results show that TXRF is a fast, simple and accurate analytical technique for trace element analysis of wine. Moreover, dehydration is an effective way to store wine samples for further elemental analysis. © 2015 Taylor & Francis. Source

Gomez-Morales M.A.,Parasitic and Immunomediated Diseases | Ludovisi A.,Parasitic and Immunomediated Diseases | Amati M.,Parasitic and Immunomediated Diseases | Bandino E.,Istituto Zooprofilattico della Sardegna | And 8 more authors.
Parasites and Vectors

Background: Trichinella spp. infections in wild boar (Sus scrofa), one of the main sources of human trichinellosis, continue to represent a public health problem. The detection of Trichinella spp. larvae in muscles of wild boar by digestion can prevent the occurrence of clinical trichinellosis in humans. However, the analytical sensitivity of digestion in the detection process is dependent on the quantity of tested muscle. Consequently, large quantities of muscle have to be digested to warrant surveillance programs, or more sensitive tests need to be employed. The use of indirect detection methods, such as the ELISA to detect Trichinella spp. infections in wild boar has limitations due to its low specificity. The aim of the study was to implement serological detection of anti-Trichinella spp. antibodies in meat juices from hunted wild boar for the surveillance of Trichinella spp. infections. Methods. Two tests were used, ELISA for the initial screening test, and a specific and sensitive Western blot (Wb) as a confirmatory test. The circulation of anti-Trichinella IgG was determined in hunted wild boar muscle juice samples in 9 provinces of 5 Italian regions. Results: From 1,462 muscle fluid samples, 315 (21.5%, 95% C.I. 19.51-23.73) were tested positive by ELISA. The 315 ELISA-positive muscle fluid samples were further tested by Wb and 32 (10.1%, 95% C.I. 7.29-13.99) of these were positive with a final seroprevalence of 2.2% (95% C.I 1.55-3.07; 32/1,462). Trichinella britovi larvae were detected by artificial digestion in muscle tissues of one (0.07%, 95%C.I. 0.01-0.39) out of the 1,462 hunted wild boars. No Trichinella spp. larvae were detected in Wb-negative wild boar. From 2006 to 2012, a prevalence of 0.017% was detected by muscle digestion in wild boar hunted in the whole Italian territory. Conclusions: The combined use of both serological methods had a sensitivity 31.4 times higher than that of the digestion (32/1,462 versus 1/1,462), suggesting their potential use for the surveillance of the Trichinella spp. infection in wild boar populations. © 2014 Gómez-Morales et al.; licensee BioMed Central Ltd. Source

Canelli E.,Istituto Zooprofilattico della Lombardia e dellEmilia Romagna | Cordioli P.,Istituto Zooprofilattico della Lombardia e dellEmilia Romagna | Barbieri I.,Istituto Zooprofilattico della Lombardia e dellEmilia Romagna | Catella A.,Istituto Zooprofilattico della Lombardia e dellEmilia Romagna | And 4 more authors.
Avian Diseases

Astroviruses (AstVs) are nonenveloped RNA small round viruses (SRVs) with a genome of 6.87.9 kb. Known avian AstVs are spread worldwide; they have been associated with poult enteritis and mortality syndrome in the United States and reported in Italy in intensive turkey and guinea fowl flocks. Nevertheless, their real prevalence and their pathogenic role in avian enteritis affecting Italian flocks is far from clear. Negative staining electron microscopy (nsEM) is used for the routine diagnosis of avian enteric SRVs, although it cannot distinguish morphologically similar particles. Enzyme-linked immunosorbent assay (ELISA), reverse-transcription PCR (RT-PCR), and genomic sequencing are now used for this specific purpose. We analyzed 329 samples of chicken, turkey, and guinea fowl intestinal contents from Italian poultry flocks. Most samples were from enteritis outbreaks, but we also included samples from three longitudinal studies (one on 11 broiler flocks and the other two on a guinea fowl flock). We first examined the samples with nsEM. SRVs, including AstVs, are often associated with rotaviruses and were the most commonly detected morphotypes in avian enteric diseases. We then analyzed 124 of the samples with an RT-PCR targeting the open reading frame (ORF)-1b of AstV. This gene codes for an RNA-dependent polymerase. We then sequenced and genetically analyzed the RT-PCR positive samples. Phylogenetic analysis distinguished three defined clusters: the first included guinea fowl AstVs and turkey AstVs-2; the second, chicken AstVs; and the third was formed by avian nephritis viruses (ANVs). No strains clustered with turkey AstVs-1. The results indicate that ORF-1b presents certain genetic variability, even among AstVs from the same species. In longitudinal studies, samples retrieved from the same shed were homogeneous, with some exceptions suggesting possible coexistence of different genetic types in the same unit. The finding of ANV-like viruses in commercial guinea fowls underlines the genetic variability of AstVs and strengthens the hypothesis of a varied intraherd situation. © 2012 American Association of Avian Pathologists. Source

Nogarol C.,University of Turin | Bertolotti L.,University of Turin | De Carlo E.,Centro Of Referenza Nazionalesulligiene E Le Tecnologie Dellallevamento E Delle Produzioni Bufaline | Masoero L.,Istituto Zooprofilattico Sperimentale del Piemonte | And 9 more authors.
Journal of Virological Methods

Bubaline herpesvirus 1 (BuHV1) is a member of ruminant alphaherpesviruses antigenically related to bovine herpesvirus 1 (BoHV1). The impact of BuHV1 infection in infectious bovine rhinotracheitis control program is difficult to establish, due to the lack of specific diagnostic test. The ectodomain of glycoprotein E of BuHV1 was expressed as recombinant secreted protein and used in indirect ELISA as well as in a discriminatory test using the BoHV1 counterpart. A panel of monoclonal antibodies was produced against BuHV1; 6 out of 7 anti-gE monoclonal antibodies specifically recognized the BuHV1 gE. Results indicated BuHV1 gE as a sensitive marker of infection compared to seroneutralization (SN) test or blocking ELISA. When BoHV1 and BuHV1 gEs were immobilized in different wells of the same ELISA microplate, bovine and water buffalo sera were more reactive against the respective infecting virus. About one third of seropositive buffaloes with no history of contact with cattle and having higher SN titres, reacted in BoHV1 gE blocking ELISA, possibly because of steric hindrance. Since in two occasions BuHV1 was also isolated from water buffalo scoring gB+/gE+ BoHV1 blocking ELISA, we conclude that the combination of the two blocking ELISAs is not suitable to differentiate between BoHV1 and BuHV1. © 2014 Elsevier B.V. Source

Riva A.,University of Milan | Borghi E.,University of Milan | Cirasola D.,University of Milan | Colmegna S.,Istituto Zooprofilattico della Lombardia e dellEmilia Romagna | And 4 more authors.
Journal of Food Protection

Staphylococcus aureus is a known major cause of foodborne illnesses, and raw milk and dairy products are often contaminated by enterotoxigenic and antimicrobial-resistant S. aureus strains. In the present study, 35 S. aureus strains were isolated from 383 raw milk samples collected from various dairy herds in the province of Milan (northern Italy). The isolates were characterized based on their antimicrobial susceptibility patterns and the presence of genes encoding staphylococcal enterotoxins (sea, seb, sec, sed, and see). About half (45.7%) of the strains were enterotoxigenic, and 37.1% were resistant to at least one of the antimicrobial drugs tested. Seven (20%) of 35 isolates were identified as methicillin-resistant S. aureus (MRSA), and SCCmec typing performed with a multiplex PCR assay revealed the presence of gene cassettes IV and V, typical of communityacquired MRSA, and I and II, characteristic of health care-associated MRSA. The MRSA strains were evaluated for the presence of the Panton-Valentine leukocidin gene, but this gene was not found. The results of the present study revealed the presence of toxin-producing S. aureus and MRSA strains in raw milk. MRSA and enterotoxigenic S. aureus in dairy farms are an important risk factor for the spread of staphylococcal infections; therefore, further studies are needed to find strategies for monitoring and controlling the presence of S. aureus, especially MRSA, in dairy products. Copyright ©, International Association for Food Protection. Source

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