PUBLICATIONS FROM 2006 TO 2010

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1 Istituto Zooprofilattico Sperimentale della Lombardia e dell Emilia Romagna Bruno Ubertini PUBLICATIONS FROM 2006 TO 2010 ID nr Brocchi E, Bergmann IE, Dekker A, Paton DJ, Sammin DJ, Greiner M, Grazioli S, De_Simone F, Yadin H, Haas B, Bulut N, Malirat V, Neitzert E, Goris N, Parida S, Sorensen K, De_Clercq K Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus Vaccine. - Vol. 24 no ( 2006). - p bib ref Impact factor: 2,822 Project SSPE-CT Improvement of foot and mouth disease control by ethically acceptable methots based on scientifically validated assays and new knowledge on FMD vaccines, including the impact of vaccination (FMD-IMPROCON). - De Clercq Dr. Kris - UO - CE ,00 EUR - 01/01/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I To validate the use of serology in substantiating freedom from infection after foot-and-mouth disease (FMD) outbreaks have been controlled by measures that include vaccination, 3551 sera were tested with six assays that detect antibodies lo the non-structural proteins of FMD virus. The sera came from naïve, vaccinated, infected and vaccinated-and-infected animals two-thirds from cattle, the remainder from sheep and pigs. The assays were covariant for sensitivity, but not necessarily for specificity. A commercial kit from Cedidiagnostica and an in-house assay from 1ZS-Brescia were comparable to the NCPanaftosa-screening index method described in the Diagnostic Manual of the World Animal Health Organisation. Using these three tests the specifìcity and sensitivity. for the detection of carriers in vaccinated cattle approaches or exceeds 99% and 90% respectively. ID nr Stacchiotti A, Rodella LF, Ricci F, Rezzani R, Lavazza A, Bianchi R Stress proteins expression in rat kidney and liver chronically exposed to aluminium suiphate Histol Histopathol. - Vol. 21 ( 2006). - p bib ref Impact factor: 2,023 Aluminium (Al) is the third most widespread metal in the environment. It is toxic for the brain, bone and haematological system but unfortunately very little data exist for other organs. Stress proteins are induced or enhanced against metal toxicity with an essential role in the recovery of organules and other cellular proteins. This immunohistochemical study was performed to analyze the distribution of three stress proteins (HSP25, HSP72, GRP75) in rat kidney and liver orally exposed to Al sulphate daily for 3 and 6 months. A1-induced alterations were further studied by histopathology (H&E, PAS, Perl's, Masson) and ultrastructural morphometry. In the kidney: HSP25 was enhanced in proximal tubules after 6 months A1-exposure when abnormal brush borders were observed; HSP72 was induced in proximal tubules only after long Al-treatment; GRP75 was raised in midcortical area sometimes within nuclei. Furthermore, lysosomal and lipofuscins densities increased in the juxtamedullary tubules after 3 months Al exposure with respect to controls. In the liver: Perl'spositive deposits and fibrosis became evident after Al treatment. HSP25 was very weak; HSP72 focal in pericentral hepatocytes at 3 months and induced also in Kupffer cells at 6 months; GRP75 diffuse in periportal hepatocytes and non parenchymal cells at 6 months. Prolonged Al exposure stimulated stress proteins strictly organ-dependently in the rat. Their distribution in kidney and liver seems related to cumulative sublethal effects induced by metal and could be a sensitive index of Al susceptibility of these organs. Pag. 1 di /08/2011

2 ID nr Paton DJ, De_Clercq K, Greiner M, Dekker A, Brocchi E, Bergmann I, Sammin JD, Gubbins S, Parida S Application of non-structural protein antibody tests in substantiating freedom from foot-and-mouth disease virus infection after emergency vaccination of cattle Vaccine. - Vol. 24 no ( 2006). - p bib ref Impact factor: 2,822 Project SSPE-CT Improvement of foot and mouth disease control by ethically acceptable methots based on scientifically validated assays and new knowledge on FMD vaccines, including the impact of vaccination (FMD-IMPROCON). - De Clercq Dr. Kris - UO - CE ,00 EUR - 01/01/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I There has been rnuch debate about the use of the so-called "vaccinate-to-live policy for the control of footand-mouth disease (FMD) in Europe. according to which, spread of the FMD virus (FMDV) from future outbreaks could be controlled by a short period of "emergency)," vaccination of surrounding herds, reducing the need for large-scale pre-emptive culling of at-risk animals. Since vaccinated animals may become subclinically infected with FA9DV following challenge exposure, it is necessary to either remove all vaccinates (vaccinate-to-kill) or to detect and remove vaccinates in which virus is circulating or has established persistent infections (vaccinate-to-live), in order to rapidly regain the most favoured trading status of FMD-free without vaccination. The latter approach can be supported by testing vaccinated animals for the present of antibodies to certain non-structural proteins NSP) of FMDV, which are induced by infection with the virus, but not by vaccination wíth purified FMD vaccines. Using test sensitivity and specificity data established at a recent workshop on NSP assays [Brocchi E. Bergmann I. Dekker A. Paton DJ. Sammin DJ. Greiner M. et al. Comparative performance of six ELISAs for antibodies to the non-structural proteins of foot-and-mouth disease. Vaccine. in press, this paper examines the ways in which serological testing with NSP ELISAs can he used and interpreted and the effect that this will have un the confidence with which freedom from infection can be demonstrated within guidelines specified by the World Animal Health Organisation and the European Commission. ID nr Pedretti E, Passeri B, Amadori M, Isola P, Di_Pede P, Telera A, Vescovini R, Quintavalla F, Pistello M Low-dose interferon-alfa treatment for feline immunodeficiency virus infection Vet Immunol Immunopathol. - Vol. 109 ( 2006). - p bib ref Impact factor: 1,626 Feline immunodeficiency virus sustains an AIDS-like syndrome in cats, which is considered a relevant model for human AIDS. Under precise enrolment requirements, 30 naturally infected cats showing overt disease were included in a trial of lowdose, oral human interferon-a treatment. Twenty-four of them received 10 IU/Kg of human interferon-alfa and 6 placebo only on a daily basis under veterinary supervision. The low-dose human interferon-alfa treatment significantly prolonged the survival of virus-infected cats (p < 0.01) and brought to a rapid improvement of disease conditions in the infected hosts. Amelioration of clinical conditions was neither correlated with plasma viremia, nor with proviral load in leukocytes. A good survival of CD4+T cells and a slow increase of CD8+T cells were also observed in human interferon-alfa-treated cats. Interestingly, the improvement of the total leukocyte counts showed a much stronger correlation with the recovery from serious opportunistic infections. As shown in other models of low-dose interferon-alfa treatment, there was a rapid regression of overt immunopathological conditions in virus-infected cats. This hints at a major role of interferon-a in the control circuits of inflammatory cytokines, which was probably the very foundation of the improved clinical.score and survival despite the unabated persistence of virus and Pag. 2 di /08/2011

3 virusinfected cells. ID nr Nunez JI, Fusi P, Borrego B, Brocchi E, Pacciarini ML, Sobrino F Genomic and antigenic characterization of viruses from the 1993 Italian foot-and-mouth disease outbreak Arch Virol. - Vol. 151 ( 2006). - p bib ref Impact factor: 1,819 Project SSPE-CT Improvement of foot and mouth disease control by ethically acceptable methots based on scientifically validated assays and new knowledge on FMD vaccines, including the impact of vaccination (FMD-IMPROCON). - De Clercq Dr. Kris - UO - CE ,00 EUR - 01/01/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I PRC Update of reagents and procedures for diagnosis and control of Foot-and-mouth disease toward recognition of newly arrived viral strains - Brocchi Dr.ssa Emiliana - Solo - MINSAN /10/ /09/ ,00 LIT 1 - H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I The origin and evolution of the type O foot-and-mouth disease viruses (FMDV) that caused the outbreak occurrence in Italy in 1993, the first episode of the disease in the EU after adoption of a non-vaccination policy in 1991, have been studied by the analysis of sequences encoding three main antigenic sites on the viral capsid proteins. The phylogenetic tree derived from sequences spanning the carboxyterminal end of VP 1 showed that these Italian viruses were grouped in the ME-SA topotype, closely related to viruses that circulated previously in the Middle East. The analysis of the nucleotide sequences in VP1, VP2 and VP3 showed a co-circulation during the epizootic of genetic variants, including viruses with amino acid replacements in VP3. For some of the isolates analyzed, values of fixation of nucleotide substitutions per year were observed in the three regions analyzed, ranging from 1.5 to 5.1 x The use of a panel of new monoclonal antibodies raised against an isolate from this outbreak, as well as monoclonal antibodies to FMDV 01-Switzerland 1965, showed differences in the reactivity pattern among some of the Italian isolates analyzed, which were consistent with the co-circulation of antigenic variants. These results support the potential for FMDV diversification in a limited period of time and under epidemiological conditions in which no vaccination campaigns were being implemented. ID nr Gelmetti D, Meroni A, Brocchi E, Koenen F, Cammarata G Pathogenesis of encephalomyocarditis experimental infection in young piglets: a potential animal model to study viral myocarditis Vet Res. - Vol. 37 ( 2006). - p bib ref Impact factor: 2,324 Project PL Molecular characterisation and epidemiology of Encephalomyocarditis virus: a model for emerging diseases - Koenen Dr. F. - UO - CE ,00 EUR - 01/02/ /01/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I The pathogenesis of encephalomyocarditis (ENIC) due to the ENIC virus (EMCV) was studied in 24 piglets oro-nasally infected with the field isolate B279/95. Two pigs were kept as negative controls and were euthanised at hour 0. The remaining 24 were euthanised every 6 h up to 78-h post infection (hpi). Virus isolation, histological examination and EMCV immunodetection were performed on the spleen, intestine, pancreas, liver, kidneys, heart, lungs, lymph nodes, tonsils and brain. EMCV was isolated at 6-hpl from the intestine and lymph nodes and at 12-hpi from the heart. From 6 to 12-hpi, scattered degenerate myocardiocytes were immunolabelled. Subsequently, myocarditis developed and progressively worsened. Immunopositive reaction in tonsil macrophages, observed in the early stage of infection (6-hpi), suggests that Pag. 3 di /08/2011

4 tonsils are the portal of entry, and by mean of wandering macrophages the ENIC virus is then distributed through the body. Afterwards, EMCV-B279/95 replicates intensively in the cytoplasm of myocardiocytes and the acute myocarditis is strictly related to the tropism of these cells. Four pigs died spontaneously. In three animals no post mortem lesions or virus were isolated/detected, although all of them showed mild myocarditis. The experimental infection with EMCV B279,95 indicates: (i) the experimental protocol mimics the individual variability observed in natural disease, (ii) tonsils are the portal of entry of infection and the heart is the target organ, (iii) EMCV provides a valuable animal model for comparative studies on progressive viral myocarditis. ID nr Bolelli L, Bobrovovà Z, Ferri E, Fini F, Menotta S, Scandurra S, Fedrizzi G, Girotti S Bioluminescent bacteria assay of veterinary drugs in excreta of food-producing animals J Pharm Biomed Anal. - Vol. 42 ( 2006). - p bib ref Impact factor: 1,889 The residues of pharmacological treatments on food-producing animals, present in the manure dispersed on agricultural land, can impact environmental and human health through toxic, genotoxic, and drug-resistance development effects. Biotoxicity assays can easily reveal the presence of noxious substances and those based on bioluminescent bacteria (BLB) are particularly simple and rapid. A BLB assay was developed as microplate format by using various strains of Vibrio sp. and was employed to evaluate their response to pure antibiotic solutions and to residues extracted from excreta of antibiotic treated pigs and turkeys. The residues were quantified by HPLC analysis. The BLB assay can be proposed as an easy-to-perform screening tool to assess the presence of residues due to undeclared current, or recently ended, pharmacological treatments, as well as to evaluate their permanence in manure. ID nr Petracci M, Bianchi M, Cavani C, Gaspari P, Lavazza A Preslaughter mortality in broiler chickens, turkeys, and spent hens under commercial slaughtering Poult Sci. - Vol. 85 no 9 ( 2006). - p bib ref Impact factor: 1,747 The incidence of dead on arrival (DOA) birds was surveyed over 33 broiler, 11 turkey, and 19 spent hen abattoirs representing the majority (around 70%) of the Italian poultry slaughter plants. Data were recorded monthly during a 4-yr period (August 2001 to July 2005), considering a total of 1,266 million chicken broilers, 118 million turkeys, and 54 million spent hens, which represent 67.7, 84.0, and 28.4% of the national production, respectively. The overall average incidence of DOA was found to be 0.35, 0.38, and 1.22% in broilers, turkeys, and spent hens, respectively. The season significantly (P S 0.01) influenced the mortality of all considered poultry categories, with higher incidence being observed during the summer (0.47, 0.52, and 1.62% for broilers, turkeys, and spent layers, respectively). The incidence of DOA broilers was found to be lower in small slaughter plants compared with medium and large slaughter plants (0.28 vs and 0.35%, P <_ 0.01). The data obtained in this study might be used for establishing limit values of DOA as a welfare indicator during the preslaughter time of birds, including catching, loading, transportation, and lairage. ID nr Alborali L Climatic variations related to fish diseases and production Vet Res Commun. - Vol. 30 suppl 1 ( 2006). - p bib ref Impact factor: 0,328 First author Alborali, Giovanni Loris Pag. 4 di /08/2011

5 ID nr Petrini S, Ferrari M, Vincenzetti S, Vita A, Amici A, Ramadori G An immunoenzyme linked assay (ELISA) for the detection of antibodies to Truncated Glycoprotein D (tgd) of bovine Herpesvirus-1 Vet Res Commun. - Vol. 30 suppl 1 ( 2006). - p bib ref Impact factor: 0,328 Project PRC New strategies for control of Influenza virus infection in swine and horses. - Barigazzi Dr. Giuseppe - Capofila - MINSAN ,00 LIT - 02/05/ /08/ N5 - Parma - Foni Dr.ssa Emanuela - I 2 - N1 - Bologna - Vecchi Dr. Giovanni - I 3 - ISS - Donatelli Dr.ssa Isabella - E 4 - ISS 2 - Campitelli Dr.ssa Laura - E 5 - L1 - Centro Substrati Cellulari - Ferrari Dr.ssa Maura - I ID nr Ferris NP, King DP, Reid SM, Hutchings GH, Shaw AE, Paton DJ, Goris N, Haas B, Hoffmann B, Brocchi E, Bugnetti M, Dekker A, De Clercq K Foot-and-mouth disease virus : a first inter-laboratory comparison trial to evaluate virus isolation and RT-PCR detection methods Vet Microbiol. - Vol. 117 ( 2006). - p bib ref Impact factor: 2,175 Project 2003-SSP New and emerging technologies: improved laboratory and on-site detection of oie list a viruses in animals and animal products (LAB-ON.SITE) - Belak Dr. Sandor - UO - CE ,00 EUR - 01/11/ /07/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I PRC Development and validation of innovating serological methods for the differentiation of vaccinated from FMD-virus infected animals based on the use of multiple non-structural antigens - Brocchi Dr.ssa Emiliana - Solo - MINSAN /08/ /08/ ,00 LIT 1 - H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I Five European reference laboratories participated in an exercise to evaluate the sensitivity and specificity of their routinely employed RT-PCR tests and cell cultures for the detection and isolation of foot-and-mouth disease (Fk1D) virus. Five identical sets of 20 coded samples were prepared from 10 vesicular epithelia, which were derived from submissions from suspect cases of FNID or swine vesicular disease (SVD). Sixteen samples were derived from six FMD virus positive epithelia representing four different serotypes (two each of types O and A and one each of types Asia 1 and SAT 2), two from samples which had been found to be negative by antigen ELISA and virus isolation (VI) in cell culture and two from SVD virus positive epithelia. Some of the FNID virus positive samples were prepared from 10-fold serial dilutions of three of the initial suspensions. Each laboratory tested the samples by one or more of its available RT-PCR procedures and inoculated cell cultures that it routinely uses for FMD diagnosis in attempts to isolate virus, the specificity of which was confirmed by antigen ELISA. The best of the RT-PCR assays used in each laboratory gave comparable results while the sensitivity of cell cultures was variable from high in one laboratory. moderate in two and low in two others. This prototype panel of samples would appear suitable for external quality assurance of these tests but would benefit from the inclusion of more negative samples and an extension in the serial dilution range of one or more of the FMD positive sample titration series. ID nr Stacchiotti A, Ricci F, Rezzani R, Li_Volti G, Borsani E, Lavazza A, Bianchi R, Rodella LF Tubular stress proteins and nitric oxide synthase expression in rat kidney exposed to mercuric Pag. 5 di /08/2011

6 chioride and melatonin J Histochem Cytochem. - Vol. 54 no 10 ( 2006). - p bib ref Impact factor: 2,208 Stress proteins such as HSP70 members (HSP72 and GRP75) and metallothionein (MT) protect the Kidney against oxidative damage and harmful metals, whereas inducible nitric oxide synthase (inos) regulates tubular functions. A single dose of mercuric chloride (HgCI2) can cause acute renal failure in rats, its main target being the proximal tubule. Oxidative damage has been proposed as one of its pathogenic mechanisms. In this study we tested whether melatonin (MEL), a powerful antioxidant compound, is effective against HgCI2 nephrotoxicity. Rats were treated with saline, HgCI2 (3.5 mg/kg), MEL (5 mg/i<9), and MEL + HgCl2 and examined after 24 hr for HSP72, GRP75, MT, and inos by immunohistochemistry and immunoblotting. Tubular effects of the treatment were then characterized by ultrastructure. In the HgCI2 group, all markers were overexpressed in convoluted proximal tubules and sometimes in distal tubules. In the MEL + HgCI2 group, GRP75 and inos decreased in convoluted and straight proximal tubules, whereas HSP72 and MT persisted more than the saline and MEL-only groups. Tubular damage and mitochondrial morphometry were improved by MEL pretreatment. In conclusion, the beneficial effect of MEL against nephrotoxicity HgCI2 nephrotoxicity was outlined morphologically and by the reduction of the tubular melatonin expression of stress proteins and inos. These markers could represent sensitive recovery stress proteins index against mercury. ID nr Scala A, Garippa G, Varcasia A,Tranquillo VM, Genchi C Cystic echinococcosis in slaughtered sheep in Sardinia (Italy) Vet Parasitol. - Vol. 135 nr. 1 ( 2006). - p bib ref Impact factor: 1,686 Of 771 regularly slaughtered Sardinian breed sheep, 580 (75%) were found infected with Echinococcus granulosus hydatid cysts. Seventy-nine sheep (10.3%) had at least 1 fertile cyst. The prevalence of sheep infected with purulent/caseous cysts, calcified cysts and sterile cysts was 13, 59 and 28%, respectively. The age of sheep was positively associated with the probability of infection that increased 1.15 fold for each further year of age. Fertile cysts were found in the lungs of 46 sheep (6%) and in the liver of 13 sheep (1.7%), and in the lung and the liver of 20 sheep (2.6%). Most fertile cysts were found in the lungs (314) and most sheep were infected with less than 10 cysts. When analyzed by a mixed-effect logistic model, the probability to find fertile cysts in the lungs was three times higher compared to the liver and it increased with the age of the sheep (p = 0.70, p < 0.001). Of 4072 collected cysts, 532 were fertile, 178 purulent/caseous calcified and 1023 sterile. ID nr Furlanello T, Tasca S, Caldin M, Carli E, Patron C, Tranquillo M, Lubas G, Solano_Gallego L Artifactual changes in canine blood following storage, detected using the ADVIA 120 hematology analyzer Vet Clin Pathol. - Vol. 35 no 1 ( 2006). - p bib ref Impact factor: 0,795 Artifactual changes in blood may occur as a consequence of delayed analysis and may complicate interpretation of CBC data. Objective: The aim of this study was to characterize artifactual changes in canine blood, due to storage, using the ADVIA 120 hematology analyzer. Methods: Blood samples were collected into EDTA from 5 clinically healthy dogs. Within 1 hour alter blood sample collection and at 12, 24, 36 and 48 hours alter storage of the samples at either 4'C or room temperature (-24'C), a CBC was clone using the ADVIA 120 and multispecies software. A linear mixed model was used to statistically evaluate significant differences in values over time, compared with initial values. Results: The HCT and MCV were increased Pag. 6 di /08/2011

7 significantly after 12 hours of collection at both 4'C and 24'C, and continued to increase through 48 hours. The MCHC initially decreased significantly at hours and then continued to decrease through 48 hours at both temperatures. Changes in HCT, MCV, and MCHC were greater at 24'C than at 4'C at all time points. A significant increase in MPV and a decrease in mean platelet component concentration were observed at all time points at 24'C. Samples stored at 24'C for 48 hours had significantly higher percentages of normocytichypochromic RBCs, and macrocytic-normochromic RBCs, and lower platelet and total WBC counts. Conclusions: Delayed analysis of canine blood samples produces artifactual changes in CBC results, mainly in RBC morphology and platelet parameters, that are readily detected using the ADVIA 120. Refrigeration of specimen, even after 24 hours of storage at room temperature, is recommended to improve the accuracy of CBC results for canine blood samples. ID nr Furianello T, Caldin M, Stocco A, Tudone E, Tranquillo M, Lubas G, Solano-Gallego L Stability of stored canine plasma for hemostasis testing Vet Clin Pathol. - Vol. 35 no 2 ( 2006). - p bib ref Impact factor: 0,795 ID nr De_Lorenzi D, Mandara MT, Tranquillo M, Baroni M, Gasparinetti N, Gandini G, Masserdotti C, Bonfanti U, Bertolini G, Vian P, Bernardini M Squash prep cytology in the diagnosis of canine and feline nervous system lesions: a study of 42 cases Vet Clin Pathol. - Vol. 35 no 2 ( 2006). - p bib ref Impact factor: 0,795 ID nr Croci L, Losio MN, Suffredini E, Pavoni E, Di_Pasquale E, Fallacara F, Arcangeli G Assessment of human enteric viruses in shellfish from the northern Adriatic sea Int J Food Microbiol. - Vol. 114 no 2 ( 2007). - p bib ref Impact factor: 2,608 Incidence and circulation of different strains of hepatitis A and Norovirus in shellfish were studied on 235 samples (Tapes philippinarum. mytilus galloprovincialis. Ostrea spp. and Chlamys spp.) obtained from different sites, representing the shellfish production areas of the northem Adriatic sea. Shellfish were harvested in the period of one year and, after depuration, were examined for bacterial (Escherichia coli and Salmonella) and viral (HAV and NoV) contamination. Viral contamination was present on average in 22% of samples: specifically, 6% of samples tested positive for HAV. 14% for NoV and 2% for both viruses. None of the samples revealed the presence of Salmonella, and in most of them (93%) the number of E. coli was below the European legislation limit of 230 MPN/100 g. T. philippinarum was the species most often contaminated, as well as bein the only species in which the legal limit for E. coli was, in some cases, exceeded. Both HAV and NoV contamination were detected throughout the year; NoV detection was slightly more frequent during winter months, but positive samples were also present in summer. The sequencing of the PCR products showed the circulation of only one HAV genotype (LA) and four different NoV genotypes (Hawaii, Melksham. Lordsdale and GGIIb) with a prevalence of the GGIIb genotype in the sceond period of the monitoring. ID nr Ostanello F, Dottori M, Gusmara G, Leotti G, Sala V Pneumonia disease assessment using a slaughterhouse lung-scoring method Pag. 7 di /08/2011

8 J Vet Med A. - Vol. 54 no 2 ( 2007). - p bib ref Impact factor: 0,627 The aims of this study were the evaluation of a quantitative method for the assessment of pneumonia lesions applied lo heavy-weight slaughtered pies. the identification of risk factors connected with the increase in the prevalence and severity of the lesions and the evaluation of a possible correlation between the presence of pneumonia lesions and the decrease in the carcass quality. The lungs of pigs (109 slaughtered batches) coming from 91 farms located in Northern Italy were examined. Lung lesions were scored using the method developed by Madec and Kobisch (Juurn. Rech. Porc. Fr ). Before the scoring. anamnestic information regarding the farm of origin of each batch were collected. For 41 batches (3603 pigs). information about carcass quality were also collected. Pneumonia lesions were found in 59.6 '0 of the lungs (range 3-91%), and the average batch score was 2.11 (range ). We identified as farm risk factors those related to an increase in the severity of the lung lesions. the presence of breeders within the herd. the starting of a growing cycle during the winter season and the lack of vaccination programmes to Mycoplasma hyopneumoniae. Moreover, we also found a statistically significant association between the increase in the mean lung score of the batch and the decrease of the carcass quality. ID nr Capobianco R, Casalone C, Suardi S, Mangieri M, Miccolo C, Limido L, Catania M, Rossi G, Di_Fede G, Giaccone G, Bruzzone MG, Minati L, Corona C, Acutis P, Gelmetti D, Lombardi G, Groschup MH, Buschmann A, Zanusso G, Monaco S, Caramelli M, Tagliavini Conversion of the base prion strain into the BSE strain: the origin of BSE? PLoS Pathog. - Vol. 3 no 3 ( 2007). - p bib ref Impact factor: 6,056 Project PRC Preliminary assessment of the presence of hepatitis E infection in swine of Lombardy, set-up of an ELISA serological method and PCR - Lombardi Dr. Guerino - Solo - MINSAN ,00 EUR - 01/07/ /06/ G3 - Reparto benessere animale, immunoprofilassi, allevamenti e sperim. Animale - Lombardi Dr. Guerino - I Atypical neuropathological and molecular phenotypes of bovine spongiform encephalopathy (BSE) have recently been identified in different countries. One of these phenotypes, named bovine "amyloidotic" spongiform encephalopathy (BASE), differs from classical BSE for the occurrence of a distinct type of the disease-associated prion protein (PrP), termed PrPSc, and the presence of PrP amyloid plaques. Here, we show that the agents responsible for BSE and BASE possess different biological properties upon transmission to transgenic mice expressing bovine PrP and inbred lines of nontransgenic mice. Strikingly, serial passages of the BASE strain to nontransgenic mice induted a neuropathological and molecular disease phenotype indistinguishable from that of BSE-infected mice. The existence of more than one agent associated with prion disease in cattle and the ability of the BASE strain to convert into the BSE strain may have important implications with respect to the origin of BSE and spongiform encephalopathies in other species, including humans. ID nr Amadori M The role of IFN-a as homeostatic agent in the inflammatory response: a balance between danger and response? J Interferon Cytokine Res. - Vol. 27 ( 2007). - p bib ref Impact factor: 2,472 Project PRC Assessment and modulation of the inflammatory effects of early weaning in the piglet - Amadori Dr. Massimo - Solo - MINSAN ,00 EUR - 15/05/ Pag. 8 di /08/2011

9 14/11/ G3 - Reparto benessere animale, immunoprofilassi, allevamenti e sperim. Animale - Amadori Dr. Massimo - I Interferon-a (IFN- a) is thought of by most immunologists as a fundamental component of the antiviral innate immune response, with other, accessory properties. Several lines of evidence point to a fundamental reappraisal of this conceptual framework because it may neglect other crucial functions of this cytokine under both health and disease conditions. Among these, a regulatory role in the inflammatory response is probably of paramount importance, as shown by in vivo and in vitro studies on humans, pets, and farm and laboratory animals. This role would not conflict with the main functions of IFN-a in the innate immune system and would complement these in line with major, evolution-based needs of the host. A hinge role of IFN-a between innate and adaptive immunity was recognized in the past on the basis of compelling evidence. This concept should now be widened; not only does IFN-a act to trigger, amplify, and sustain the different phases of the immune response, but it could also promote a substantial balance between danger and inflammatory response when an infectious challenge is either declining or completely over. ID nr Duan L, Campitelli L, Fan HX, Leung, YHC, Vijaykrishna D, Zhang JX, Donatelli I, Delogu M, Li KS, Foni E, Chiapponi C, Wu WL, Kai H, Webster RG, Shortridge KF, Peiris JSM, Smith GJD, Chen H, Guan Y Characterization of low-pathogenic H5 subtype influenza viruses from Eurasia: implications for the origin of highly pathogenic H5N1 viruses J Virol. - Vol. 81 no 14 ( 2007). - p bib ref Impact factor: 5,341 Highly pathogenic avian influenza (HPAI) HSNI viruses are now endemiec in many Asian countries, resulting in repeated outbreaks in poultry and increased cases of human infection. The immediate precursor of these HPAI viruses is believed to be Algoose/Guangdong/1/96 (Gs/GD)-like H5NI HPAI viruses first detected in Guangdong, China, in From 2000 onwards, many novel reassortant H5N1 influenza viruses or genotypes have emerged in southern China. However, precursore of the Gs/GD-like viruses and their subsequent reassortants have not been fully determined. Here we characterize low-pathogenic avian influenza (LPAI) HS subtype viruses isolated from poultry and migratory birds in southern China and Europe from the 1970s to the 2000s. Phylogenetíc analyses revealed that Gs/GD-like virus was likely derived from an LPAI H5 virus in migratory birds. However, its variants arose from multiple reassortments between Gs/GD-like virus and viruses from migratory birds or with those Eurasian viruses isolated in the 1970s. It is of note that unlíke HPAI H5N1 viruses, those recent LPAI H5 viruses have not become established in aquatie or terrestrial poultry. Phylogenetic analyses revealed the dynamic nature of the influenza virus gene pool in Eurasia with repeated transmissions between the eastern and western extremities of the continent. The data allo show reassortment between influenza viruses from domestie and migratory birds in thìs region that has contributed to the expanded diversity of the influenza virus gene pool among poultry in Eurasia. ID nr Gasparini M, Assini W, Bozzoni E, Tognoli N, Dusi G Development and validation of a liquid chromatography tandem mass spectrometry method for the separation of conjugated and unconjugated 17(alfa).- and 17 ß-boldenone in urine sample Anal Chim Acta. - Vol. 586 ( 2007). - p bib ref Impact factor: 2,894 Natural occurrence or illegal treatment of boldenone (BOLD) presence in cattle urine is under debate within the European Union. Separation of conjugated and unconjugated forms of 17a-boldenone (a-bold) and 17(3-boldenone ((3-BOLD) and presence of related molecules as androsta-1,4-diene-3,17-dione (ADD) appear critical points for the decision of an illegal use. The aim of this study is a new analytical approach of Pag. 9 di /08/2011

10 BOLD and ADD confirmation in cattle urine. The separation between conjugated and unconjugated forms of BOLD was obtained by a preliminary urine liquid liquid extraction step with ethyl acetate. In this step the organic phase extracts only unconjugated BOLD and ADD, while BOLD in conjugated form remain in urine phase.. Afterwards the urine phase, contains conjugated BOLD, was suhjected to an enzymatic deconjugation. Solid-phase extraction (OASIS-HLB Waters) was used for the purification and concentration of analytes in organic and urine phases and liquid chromatography ion electrospray tandem mass spectrometry (LC MS MS) was applied for the confirmation of BOLD and ADD, using deuteriumlabelled 17(3-boldenone (BOLD-d3) as internal standard. The method was validated as a quantitative confirmatory method according to the Commission Decision 2002/657/CE. The results obtained demonstrate that the developed method show very high specificity, pecision, trueness and ruggedness. Decision limits (CCoc) smaller than 0.5 ng ml-' were obtained for each analyte. ID nr Di_Francesco A, Baldelli R, Cevenini R, Magnino S, Pignanelli S, Salvatore D, Galuppi R, Donati M Seroprevalence to chlamydiae in pigs in Italy Vet Rec. - Vol. 159 no 25 ( 2006). - p bib ref Impact factor: 1,017 ID nr Lanzi G, de_miranda JR, Boniotti MB, Cameron GE, Lavazza A, Capucci L, Camazine SM, Rossi C Molecular and biological characterization of deformed wing virus of honeybees (Apis mellifera L.) J Virol. - Vol. 80 no 10 ( 2006). - p bib ref Impact factor: 5,178 Project AMA - An investigation into Varroa-associated viruses of bees. - Sabatini Dr.ssa Anna Gloria - UO - MIPA ,00 LIT - 06/12/ /01/ H2 - Reparto di Biologia Molecolare - Rossi Dr. Cesare - I Deformed wing virus (DWV) of honeybees (Apis mellifera) is closely associated with characteristic wing deformities, abdominal bloating, paralysis, and rapid mortality of emerging adult bees. The virus was purified from diseased insects, and its genome was cloned and sequenced. The genomic RNA of DWV is 10,140 nucleotides in length and contains a single large open reading frame encoding a 328-kDa polyprotein. The coding sequence is flanked by a 1,144-nucleotide 5' nontranslated leader sequence and a 317-nucleotide 3' nontranslated region, followed by a poly(a) tail. The three major structural proteins, VP1 (44 kda), VP2 (32 kda), and VP3 (28 kda), were identified, and their genes were mapped to the N-terminal section of the polyprotein. The C-terminal part of the polyprotein contains sequence motifs typical of well-characterized picornavirus nonstructural proteins: an RNA helicase, a chymotrypsin-like 3C protease, and an RNAdependent RNA polymerase. The genome organization, capsid morphology, and sequence comparison data indicate that DWV is a member of the recently established genus I flavirus. ID nr Stacchiotti A, Borsani E, Riccia F, Lavazza A, Rezzani R, Bianchi R, Rodella LF Bimoclomol ameliorates mercuric chloride nephrotoxicity through recruitment of stress proteins Toxicol Lett. - Vol. 166 ( 2006). - p bib ref Impact factor: 2,43 Bimoclomol (BIM), is a stress proteins coinducer, that acts synergistically with a mild stressor to activate cytoprotective stress proteins. BIM has been successfully utilized in animal models for the treatment of Pag. 10 di /08/2011

11 various nervous, cardiac and cerebrovascular diseases. Mercuric chloride (HgC12) induces acute renal failure in rats by a single dosage. The present in vivo study was conduc to assess the efficacy of BIM against acute HgC12 nephrotoxicity. At different times after BIM and/or HgC12 exposure we evaluated renal morphology and the localization/abundance of three stress proteins (HSP72, GRP75, HSP60) by electron microscopy immunohistochemistry and Western blot analysis. BIM delivery to rats 6 h before mercury, ameliorated damage to renal ultrastructure, with recovery of tubular and mitochondrial membranes 24 h after mercury treatment. In rats pretreated with BIM prior HgC12 exposure, HSP72 was significantly overexpressed in proximal tubules in a time-dependent manner. In contrast, the amou of GRP75 and HSP60 after BIM pretreatment were comparable to the group treated with mercury alone, but these stress proteins had translocated to the nuclei at 14 and 24 h, respectively. These novel findings suggest that BIM mitigates H902 nephrotoxicity in rats through the early recruitment of stress proteina midcortical proximal tubules that are the main renal mercury-targets. ID nr Martella V, Bànyai K, Ciarlet M, Iturriza_Gómara M, Lorusso E, De_Grazia S, Arista S, Decaro N, Elia G, Cavalli A, Corrente M, Lavazza A, Baselga R, Buonavoglia C Relationships among porcine and human P[6] rotaviruses : evidence that the different human P[6] lineages have originated from multiple interspecies transmission events Virology. - Vol. 344 ( 2006). - p bib ref Impact factor: 3,08 Project PRF Genetic and antigenic diversity of rotaviruses study of the evolutionary mechanisms and implications for diagnosis and vaccination - Gentile Dr. Mattia - UO - MINSAN ,00 EUR - 01/03/ /10/ G1 - Reparto di Virologia e Sierologia Specializzata - Lavazza Dr. Antonio - I Porcine rotavirus strains (PoRVs) bearing human-like VP4 P[6] gene alleles were identified. Genetic characterization with either PCR genotyping or sequence analysis allowed to determine the VP7 specificity of the PoRVs as G3, G4, G5 and G9, and the VP6 as genogroup I, that is predictive of a subgroup I specificity. Sequence analysis of the VP8* trypsin-cleavage product of VP4 allowed PoRVs to be characterized further into genetic lineages within the P[6] genotype. Unexpectedly, the strains displayed significantly higher similarity (up to 94.6% and 92.5% at aa and nt level, respectively) to human M37-like P[6] strains (lineage 1), serologically classifiable as P2A, or to the atypical Hungarian P[6] human strains (HRVs), designated as lineage V (up to 97.0% aa and 96.1% nt), than to the porcine P[6] strain Gottfried, lineage II «85.1% aa and 82.2 nt), which is serologically classified as P213. Interestingly, no P[6] PoRV resembling the original prototype porcine strain, Gottfried, was detected, while Japanase P[6] PoRV clustered with the atypical Japanasc GI human strain AU 19. By analysis of the l Oth and 11th genome segments, all the strains revealed a NSP413 genogroup (Wa-like) and a NSP5/6 gene of porcine origin. These findings strongly suggest interspecies transmission of rotavirus strains and/or genes, and may indicate the occurrence of at least 3 separate rotavirus transmission events between pigs and human, providing convincing evidence that evolution of human rotaviruses is tightly intermingled with the evolution of animal rotaviruses. ID nr Martella V, Ciarlet M, Bànyai K, Lorusso E, Cavalli A, Corrente M, Elia G, Arista S, Camero M, Desario C, Decaro N, Lavazza A, Buonavoglia C Identification of a novel VP4 genotype carried by a serotype G5 porcine rotavirus strain Virology. - Vol. 346 ( 2006). - p bib ref Impact factor: 3,08 Project PRF Genetic and antigenic diversity of rotaviruses study of the evolutionary mechanisms and implications for diagnosis and vaccination - Gentile Dr. Mattia - UO - MINSAN ,00 EUR - 01/03/ /10/ G1 - Reparto di Virologia e Sierologia Specializzata - Lavazza Dr. Antonio - I Pag. 11 di /08/2011

12 Rotavitus genome segment 4, encoding the spike outer capsid VP4 protein, of a porcine rotavirus (PoRV) strain, , identified in Italy was sequenced, and the predicted amino acid (aa) sequence was compared to those of all known VP4 (P) genotypes. The aa sequence of the fullength VP4 protein of the PoRV strain 134.'04-15 showed aa identity values ranging from 59.7% (bovine strain KK3, P8[11]) to 86.09% (porcine strain A46, P[13]) with those of the remaining 25 P genotypes. Moreover, aa sequence analysis of the corresponding VP8* trypsin cleavage fragment revealed that the PoRV strain 134/ 4-15 shared low identity, ranging from 37.52'% (bovine strain 993'83, P[t 7]) to 73.6 % (porcine strain MDR-13, P[13]), with those of the remaining 25 P genotypes. Phylogenetic relationships showed that the VP4 of the PoRV strain 134.;04-15 shares a common evolutionary origin with porcine P[13] and tapine P[22] rotavirus strains. Additional sequence analyses of the VP7, VP6, and NSP4 genes of the PoRV strain 134/04-15 revealed the highest VP7 aa identity (95.9%r) to G5 porcine strains, a porcine-like VP6 within VP6 genogroup I, and a Walike (genotype B) NSP4, respectively. Altogether, these results indicate that the PoRV strain 134/04-15 should be considered as prototype of a new VP4 genotype, P[26], and provide further evidence for the vast genetic and antigenic diversity of group A rotavinises. ID nr Ferrari M Differentiative potential of cardiomyocyte satellite cells and possible utilization in cardiac tissue repair Eur J Histochemistry. - Vol. 3 ( 2006). - p 210? Workshop "Skeletal Muscle Fiber: Morpho-Functional Patterns and Technical Approaches " : Urbino, Italy : October 3-6th, 2006) Impact factor: 0,99 Project PRF Comparative study of the efficacy of the immunizing systems towards type 1 herpetic viruses in humans and animals - Guercio Dr.ssa Annalisa - UO - MINSAN ,00 EUR - 01/02/ /09/ L1 - Centro Substrati Cellulari - Ferrari Dr.ssa Maura - I One possible strategy in the utilization of cell precursors in cardiac tissue damage repair concerns satellite cells. These are localised between mature myofiber basal lamina and cell membrane, on fiber surface. There are responsible of muscle tissue maintaining, as well as of its repair and regeneration. There are generally quiescent ce progenitors, but, alter activation, they generate a population of myoblasts which proliferate and differentiate, to form plurinucleate myotubes. Satellite cells are different, if compared to myoblasts, for biochemical and biological characteristics. Skeletal muscle satellite cells have been also reported to have a certain plasticity and consequent differentiation capabilites directed to different cell lineages, including myocytes. These observations, together with autologous cell utilization, addressed the interest to the possible use of differentiated satellite cells in altered cardiac tissue repair. The experiments have been carried out on ovine cells, which, after in vitro amplification, have been inoculated in the same animal, undergoing an experimentally induced cardiac lesion. The results demonstrated a ce scarce capability to repair the altered tissue, probably because of an impaired integration with cardiomyocytes. ID nr Sassera D, Beninati T, Bandi C, Bouman EAP, Sacchi L, Fabbi M, Lo N Candidatus Midichloria mitochondrii', an endosymbiont of the tick Ixodes ricinus with a unique intramitochondriai lifestyle Int J Syst Evol Microbiol. - Vol. 56 ( 2006). - p bib ref Impact factor: 2,744 Project PRF Diagnosis of avian influenza and investigation into gene exchange between animal and human influenza virus strains. - Capua Dr.ssa Ilaria - UO - MINSAN ,00 EUR - 02/05/ /09/ G1 - Reparto di Virologia e Sierologia Pag. 12 di /08/2011

13 Specializzata - Cordioli Dr. Paolo - I An intracellular bacterium with the unique ability to enter mitochondria exists in the European vector of Lyme disease, the hard tick Ixodes ricinus. Previous phylogenetic analyses based on 16S rrna gene sequences suggested that the bacterium formed a divergent lineage within the Rickettsiales (Alphaproteobacteria). Here, we present additional phylogenetic evidence, based on the gyrb gene sequence, that confirms the phylogenetic position of the bacterium. Based on these data, as well as electron microscopy (EM), in situ hybridization and other observations, we propose the name Candidatus Midichloria mitochondrii' for this bacterium. The symbiont appears to be ubiquitous in females of 1. ricinus across the tick's distribution, while lower prevalence is observed in males (44%). Based on EM and in situ hybridization studies, the presence of 'Candidatus M. mitochondrii' in females appears to be restricted to ovarian cells. The bacterium was found to be localized both in the cytoplasm and in the intermembrane space of the mitochondria of ovarian cells. 'Candidatus M. mitochondrii' is the first bacterium to be identified that resides within animal mitochondria. ID nr Bochkov YA, Tosi P, Massi P, Drygin VV Phylogenetic analysis of partial S1 and N gene sequences of infectious bronchitis virus isolates from Italy revealed genetic diversity and recombination Virus Gen. - Vol. 35 ( 2007). - p bib ref Impact factor: 1,102 Project PRC Molecular characterization of viral variants responsible for diffusive infections of domestic animals - Pacciarini Dr.ssa Maria - Solo - MINSAN ,00 EUR - 01/07/ /12/ H2 - Reparto di Biologia Molecolare - Fallacara Dr.ssa Francesca - I 2 - G1 - Reparto di Virologia e Sierologia Specializzata - Cordioli Dr. Paolo - I 3 - N8 - Reggio Emilia - Dottori Dr. Michele - I 4 - N3 - Forlì - Massi Dr.ssa Paola - I A total of ten infectious bronchitis virus (IBV) isolates collected from commercial chickens in Italy in 1999 were characterized by RT-PCR and sequencing of the S1 and N genes. Phylogenetic analysis based on partial S1 gene sequences showed that five field viruses clustered together with 793/B-type strains, having % nucleotide identity within the group, and one isolate had very close sequence relationship (94.6% identity) with 624/I strain. These two IBV types have been identified in Italy previously. The other three variant isolates formed novel genotype detected recently in many countries of Western Europe. For one of these variant viruses, Italy-02, which afterwards became the prototype strain, the entire S1 gene was sequenced to confirm its originality. In contrast, phylogenetic analysis of more conserved partial N gene sequences, comprising nucleotides, revealed different clustering. Thus, three variant IBVs of novel Italy-02 genotype, which had % S1 gene nucleotide identity with each other, belonged to three separate subgroups based on N gene sequences. 624/I-type isolate Italy-06 together with Italy-03, which was undetectable using S1 gene primers, shared 97.7% and 99.3% identity, respectively, in N gene region with vaccine strain H120. Only one of the 793/B-type isolates, Italy-10, clustered with the 793/B strain sharing 99.3% partial N gene identity, whereas the other four isolates were genetically distant from them (only % identity) and formed separate homogenous subgroup. The results demonstrated that both mutations and recombination events could contribute to the genetic diversity of the Italian isolates. ID nr Cattoli G, De_Battisti C, Toffan A, Salviato A, Lavazza A, Cerioli M, Capua I Co-circulation of distinct genetic lineages of astroviruses in turkeys and guinea fowl Arch Virol. - Vol. 152 ( 2007). - p bib ref Impact factor: 1,85 Pag. 13 di /08/2011

14 Viruses belonging to the genus Astrovirus have been increasingly associated with enteritis in mammalian and avian species, including turkeys. More recently, astroviruses have also been detected in diseased guinea fowl. In turkeys, two genetically distinct types of astrovirus have been reported, namely turkey astrovirus 1 and 2 (TAstV1 and TAstV2). The prevalence and the pathogenesis of astrovirus infections in this species is currently unknown, with the exception of data generated in the USA. In the present report, we have demonstrated the co-circulation of distinct genetic lineages of astroviruses infecting turkeys and guinea fowl. The predominant lineage infecting turkeys is genetically related to the American TAstV2, but genetic variability within this lineage was demonstrated. Other isolates appeared to be either TastV1-related or unrelated to any other known isolate. Astroviruses infecting guinea fowl were more closely related to TAstV2, and interspecies transmissions between turkeys and guinea fowl is suggested as a possible mechanism by which these viruses may have evolved, based on the genetic data available. This investigation provides genetic and epidemiological information contributing to a better understanding of enteric viral infections in turkeys and guinea fowl. ID nr Paltrinieri S, Spagnolo V, Giordano A, Moreno_Martin A, Luppi A Influenza virus tye A serosurvey in cats Emerg Inf Dis. - Vol. 13 no 4 ( 2007). - p bib ref Impact factor: 5,094 ID nr Parida S, Fleming L, Gibson D, Hamblin PA, Grazioli S, Brocchi E, Paton DJ Bovine serum panel for evaluating foot-and-mouth disease virus nonstructural protein antibody tests J Vet Diagn Investig. - Vol. 19 no 5 ( 2007). - p bib ref Impact factor: 1,145 Project SSPE-CT Improvement of foot and mouth disease control by ethically acceptable methots based on scientifically validated assays and new knowledge on FMD vaccines, including the impact of vaccination (FMD-IMPROCON). - De Clercq Dr. Kris - UO - CE ,00 EUR - 01/01/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I A panel of 36 sera has been assembled from experimental cattle that had been infected by inoculation or contact exposure with 4 serotypes of foot-and-mouth disease virus (FMDV) with or without prior vaccination. Virus replication and persistente had been characterized in all of the animals. The proportion of the sera scored positive by 5 tests for ant"bodies to the nonstructural proteins of FMDV varied, suggesting that the panel can discriminate between the sensitivity with which such tests are able to identify infected cattle. Use of this panel will help in assessment of new tests and quality control of existing methods. ID nr Bonilauri P, Merialdi G, Dottori M, Barbieri I Presence of PRRSV in wild boar in Italy Vet Rec. - Vol. 158 no 3 ( 2006). - p bib ref Impact factor: 1,017 ID nr Circella E, Camarda A, Martella V, Bruni G, Lavazza A, Buonavagoglia G Coronavirus associated with an enteric syndrome on a quail farm Avian Pathol. - Vol. 36 n 3 ( 2007). - p bib ref Pag. 14 di /08/2011

15 Impact factor: 1,809 An enteric syndrome was observed in quail (Coturnix coturnix) semi-intensively reared for restocking in Apulia southern Italy ). The birds showed depression, severe diarrhoea, dehydration and reduced growth. mortality occurred particularly in young birds. At necropsy the prominent lesion was enteritis. A coronavirus was detected by electron microscopy and reverse transcriptase-polymerase chain reaction in the faeces and in the intestinal content of the dead quails. The virus could not be cultivated in chicken embryos. By sequence analyses of a fragment (409 nucleotides) of region lb of the polymerase gene, the quail coronavirus displayed =93% nucleotide identity to avian coronaviruses (group 3 coronaviruses) whereas by analysis of the Sl portion of the spike protein-encoding gene, the quail coronavirus displayed 16 % to 18% amino acid identity with infectious bronchitis virus. and 79% to 81% identity with turkey coronavirus. Altogether, the findings suggest the existence of a novel coronavirus genetically related to turkey coronavirus. ID nr Terregino C, De_Nardi R, Guberti V, Scremin M, Raffini E, Moreno_Martin A, Cattoli G, Bonfanti l, Capua I Active surveillance for avian influenza viruses in wild birds and backyard flocks in Northern Italy during 2004 to 2006 Avian Pathol. - Vol. 36 n 4 ( 2007). - p bib ref Impact factor: 1,809 Following the avian influenza epidemics that occurred in Italy between 1997 and 2003, the Italian Ministry of Health in collaboration with veterinary authorities promoted, funded and implemented a national surveillance programme. The main objectives of the surveillance effort were to identify avian influenza viruses circulating in wild birds and to investigate the role of backyard poultry flocks in the dynamics of infection in a densely populated poultry area. Over 2 years (2004 to 2006), 164 backyard flocks and 4083 wild birds (mainly migratory Anseriformes and Charadriiformes) were sampled in three regions in the North of Italy. Samples collected were screened by means of real-time reverse transcriptase-polymerase chain reaction and the positive samples were processed for attempted virus isolation in embryonated fowl s specific pathogen free eggs. At the end of the study period, 27 low-pathogenic avian influenza viruses had been isolated from backyard flocks and 49 strains obtained from wild birds. Of these, 26 belonged to the H5 or H7 subtype and were closely related to contemporary low-pathogenic strains of Eurasian lineage. The findings confirm that backyard free-range farming is at high risk for avian influenza virus introduction, and confirm the role of wild waterfowl in the introduction and perpetuation of low-pathogenic avian influenza viruses during the winter season in Southern Europe. ID nr Recordati C, Gualdi V, Tosi S, Vailati_Facchini R, Pengo G, Luini M, Simpson KW, Scanziani E Detection of Helicobacter spp. DNA in the oral cavity of dogs Vet Microbiol. - Vol. 119 ( 2007). - p bib ref Impact factor: 2,073 The mode of acquisition of gastric Helicobacter spp. infection in dogs has not been determined. It is suspected that oral oral and faecal oral transmission may be involved. The present study sought to determine if Helicobacter spp. DNA is present in the oral cavity of healthy and vomiting dogs. Thirty-eight pet dogs (27 vomiting and 11 clinically healthy) were studied. The presente of Helicobacter spp. was determined by single and nested PCR evaluation of DNA extracted from saliva, dental plaque and gastric biopsy samples. Helicobacter spp. DNA was detected by nested PCR in 36-(94.7%) gastric biopsies, 17 (44.7%) dental plaque and 19 (50%) saliva samples out of the 38 dogs examined. Overall 27 (71.1 %) dogs screened by nested PCR were found to harbour Helicobacter spp. DNA in the oral cavity (dental plaque and/or saliva). There was Pag. 15 di /08/2011

16 no significant differente in the prevalence of Helicobacter spp. DNA in the oral cavity of vomiting and healthy dogs, and the time from vomiting to oral sampling did not have significant impact. This study confirms the high prevalence of gastric Helicobacter spp. infection in dogs, and reveals that Helicobacter spp. DNA is detectable in the oral cavity of over 70% of dogs. These findings support the possibility of oral oral transmission between dogs and that the canine oral cavity may act as source of non-pylori Helicobacter spp. infection for humans. ID nr Moroni P, Pisoni G, Varisco G, Boettcher P Effect of intramammary infection in Bergamasca meat sheep on milk parameters and lamb growth J Dairy Res. - Vol. 74 no 3 ( 2007). - p bib ref Impact factor: 1,407 Pooled milk samples from 115 Bergamasca meat sheep were collected aseptically five times from lambing Io weaning Io determine the prevalence of intramammary infection, somatic cell counts and milk quality parameters (protein, fat and lactose), and effects of infection on lamb weight gain. The global prevalence of subclinical intramammary infection was 51.2%. The Staphylococcus genus was responsible (or the greatest prevalence (53-3% among infected udders). Staphylococcus aureus was isolated in 8.4% of infected milk samples. Infection status had significant effects on fat and protein percentage and on somatic cell count. Lamb growth was greatest for lambs of ewes with no infection and decreased as the number of infected samples increased. No significant differences were detected in the growth of lambs with dams infected by different bacterial species. ID nr Cattaneo D, Dell'Orto V, Varisco G, Agazzi A, Savoini G Enrichment in n 3 fatty acids of goat's colostrum and milk by maternal fish oil supplementation Small Rumin Res. - Vol. 64 ( 2006). - p bib ref Impact factor: 0,777 Long-chain n 3 polyunsaturated fatty acids (PUFAs), particularly docosahexaenoie acid (DHA), have been shown to be essential during perinatal growth and development of mammals. The study was conducted to determine whether the inclusion of fish oil in dairy goats diet during late gestation and early lactation would increase the proportion of n 3 PUFAs in colostral and mature mille From 3 weeks before kidding throughout 42 days of lactation, 14 dairy goats were fed either a control diet or a diet containing 1.1 % fish oil (FO). Dietary treatment did not affect main components of colostrum, but lowered percentages of fat and lactose in mature mille. In colostrum, FO supplementation lowered the proportions of C18:0, C18:2, C18:3 and elevated those of C16:1 and very long-chain n 3 PUFAs, mainly EPA (eicosapentaenoic acid) and DHA. In mature milk, FO supplementation lowered the proportion of C 18:O and raised C 16:1 and n 3 PUFAs (C18:3,EPA and DHA). Estimated transfer efficiencies for EPA and DHA into mature milk were 14 and 7%, respectively. Results suggest that goat's colostrum and milk can be enriched in n-3 PUFAs by maternal dietary fish oil supplementation during late gestation and early lactation. ID nr Paci G, Lavazza A, Ferretti M, Bagliacca M Relationship between habitat, densities and metabolic profile in brow hares (Lepus europaeus Pallas) Ital J Anim Sci. - Vol. 6 no 3 ( 2007). - p bib ref Impact factor: 0,172 Pag. 16 di /08/2011

17 È stato effettuato uno studio per conoscere le relazioni tra alcune caratteristiche ambientali, la densità di popolazione ed i parametri ematici nella lepre. A tale scopo, 33 aree protette (Zone di Ripopolamento e Cattura) della provincia di Firenze sono state sottoposte a monitoraggio ambientale e censimento delle lepri. Per ogni area sono stati effettuati i seguenti rilievi ambientali: altitudine; rapporto tra aree aperte e area totale; esposizione prevalente; natura del suolo; dotazioni idriche naturali; acclività; antropizzazione; presenza predatori; adiacenze boschive perimetrali; presenza di alberi e cespugli; controllo caccia di frodo; controllo predatori; coltivazioni; colture a perdere. Dopo il censimento tutte le lepri catturate sono state traslocate in zone libere di caccia. Un totale di 172 soggetti è stato sottoposto ai seguenti rilievi: prelievo ematico dalla vena auricolare per le analisi del glucosio, colesterolo, urea, AST, ALT, Ca, P, Mg, Na, K e Cl, determinazione dell'età, del sesso e del peso vivo. Le relazioni fra densità di popolazione e caratteristiche ambientali sono state analizzate singolarmente, quindi tutte le caratteristiche ambientali sono state sottoposte al processo di selezione "stepwise" in funzione del peso corporeo rilevato nelle lepri. 1 dati ematici sono stati analizzati mediante il metodo dei minimi quadrati considerando le caratteristiche ambientali, la densità censita, l'età ed il sesso degli animali come fonti di variazione categoriche, l'interazione sesso * età e lo stato fisiologico delle femmine "nested' nel sesso femminile. I risultati hanno evidenziato la presenza di una più alta densità di popolazione e le migliori condizioni corporee nelle zone collinari caratterizzate da aree aperte con scarsi boschetti e confini boscosi, con suolo di medio impasto, scarsa presenza di predatori, limitata antropizzazione e abbondante disponibilità idrica naturale e zone ricche di cespugli adatti al rifugio. Lo studio dei valori assoluti del profilo metabolico, indicatori dello stato fisiologico e nutritivo degli animali allevati, non ha evidenziato alcuna carenza nutrizionale negli animali selvatici nel periodo invernale considerato. Presumibilmente, come i dati dei censi-menti, devono essere ripetuti per più anni dal momento che è la variazione ad essere utile come indicatore di stato e non il valore assoluto. ID nr Knowles NJ, Wilsden G, Reid SM, Ferris NP, King DP, Paton DJ, Fevereiro M, Brocchi E Reappearance of swine vesicular disease virus in Portogal Vet Rec. - Vol. 161 no 2 ( 2007). - p 71 Impact factor: 1,168 Project PRC Improvement of SVD diagnosis: identification and neutralization of swine enteroviruses affecting SVD virus isolation in tissue cultures. - Brocchi Dr.ssa Emiliana - Capofila - MINSAN ,00 EUR - 15/04/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I 2 - G1 - Reparto di Virologia e Sierologia Specializzata - Cordioli Dr. Paolo - I 3 - IZSVE - Nardelli Dr. Stefano - E 4 - ISS - Tollis Dr.ssa Maria - E ID nr Ferrari M, Corradi A, Lazzaretti M, De'Cillà M, Losi CG, Villa R, Lanfranchi A Adult stem cells : perspectives for therapeutic applications Vet Res Commun. - Vol. 31 suppl 1 ( 2007). - p bib ref Impact factor: 0,377 The use of adult stem cells in tissue regeneration appears to be a powerful research tool, due to the intrinsic characteristics of these cells, i.e., self-renewal and unlimited capacity for proliferation. In particular, mesenchymal stem cells (MSCs) obtained from bone marrow or peripheral blood can be easily isolated, cultivated, propagated and can be differentiated into several specialized celi types thanks to their plasticity. Among these cells, MSCs can evolve into cardiac cell lineages. Since heart damage leads to the irreversible loss of cardiac function, cell transplantation could be a potential therapy for heart injury. Our laboratory has focused on the purification and expansion of rat and sheep MSCs, their differentiation into cardiomyocytes and their characterisation. Numerous results indicate that MSCs could be promising for therapy, however we need to better understand the biology of stem cells to improve methods for delivery and/or pharmacological activation. These techniques can indeed track engrafted cells and systems to guarantee their safe use. Pag. 17 di /08/2011

18 ID nr Goris N, Praet N, Sammin D, Yadin H, Paton D, Brocchi E, Berkvens D, De_Clercq K Foot-and-mouth disease non-structural protein serology in cattle: use of a Bayesian framework to estimate diagnostic sensitivity and specificity of six ELISA tests and true prevalence in the field Vaccine. - Vol. 25 no 41 ( 2007). - p bib ref Impact factor: 3,159 Project SSPE-CT Improvement of foot and mouth disease control by ethically acceptable methots based on scientifically validated assays and new knowledge on FMD vaccines, including the impact of vaccination (FMD-IMPROCON). - De Clercq Dr. Kris - UO - CE ,00 EUR - 01/01/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I The diagnostic performance of six foot-and-mouth disease (FMD) assays for detection of antibodies to the non-structural proteins (NSP) of the FMD virus (FMDV) was estimated using a Bayesian analysis on field sera from cattle of unknown infection status originating from post-fmdv outbreak situations in Israel and Zimbabwe. Estimations of the disease prevalence in both populations were also obtained. The diagnostic sensitivity estimates did not differ between both field studies, although overall Bayesian estimates were markedly higher than those previously reported based on sera from comparable experimentally infected (vaccinated) cattle populations. All NSP-based assays demonstrated a lower diagnostic specificity when applied to the Zimbabwean sera compared to both published specificities and similar Bayesian specificity estimates derived for the Israeli dataset. In Israel, the disease prevalence was estimated at 23.9% (95% credibility interval: %), whereas 65.4% ( %) was found in Zimbabwe. The need for reliable diagnostic test performance estimates and the benefits of Bayesian analysis in obtaining them are also addressed. ID nr Roncada P, Begni B, Amadori M, Cristoni S, Archetti IL, Boldetti C, Fortin R, Deriu F, Greppi GF Blood serum proteome for welfare evaluation in pigs Vet Res Commun. - Vol. 31 suppl 1 ( 2007). - p bib ref Impact factor: 0,377 ID nr Bonardi S, Paris A, Bacci C, D'Incau M, Ferroni L, Brindani F Detection and characterization of Yersinia enterocolitica from pigs and cattle Vet Res Commun. - Vol. 31 suppl 1 ( 2007). - p bib ref Impact factor: 0,377 ID nr Ghidini S, Zanardi E, Conter M, Varisco G, Bolzoni G, Ianieri A Residues of aminoglycosides in milk : confirmatory analysis Vet Res Commun. - Vol. 31 suppl 1 ( 2007). - p bib ref Impact factor: 0,377 Project PRC Design, development and production of reference samples with known content for the inhibiting-substances assay in milk and validation of a test procedure for cheese - Bolzoni Dr. Giuseppe - Solo - MINSAN ,00 EUR - 01/07/ /12/ I4 - Reparto produzioni zootecniche e sorveglianza epidemiologica degli allevamenti - Varisco Dr. Giorgio - I Pag. 18 di /08/2011

19 2 - I4 - Reparto produzioni zootecniche e sorveglianza epidemiologica degli allevamenti - Bertocchi Dr. Luigi - I 3 - I4 - Reparto produzioni zootecniche e sorveglianza epidemiologica degli allevamenti - Bolzoni Dr. Giuseppe - I ID nr Bonardi S, Foni E, Chiapponi C, Salsi A, Brindani F Detection of verocytotoxin-producing Escherichia coli serogroups O157 and O26 in the cecal content and lymphatic tissue of cattle at slaughter in Italy J Food Prot. - Vol. 70 no 6 ( 2007). - p bib ref Impact factor: 1,921 Verocytotoxin-producing Escherichia coli (VTEC) has emerged as a foodborne pathogen that can cause severe and potentially fatal illnesses, such as hemorrhagic colitis or the hemolvtie uremie syndrome. In this study, 182 cattle at slaughter (119 dairy cows and 63 feedlot cattle) were randomly selected and tested for the presence of VTEC serogroups 026, 0103, 0111, 0145, and 0157 in their cecal content and lymphatic tissue (tonsils or mesenteric lymph nodes). A total of 364 samples were evaluated with an immunomagnetic separation technique followed by slide agglutination. Presumptive VTEC , 0111, 0145, and 0157 isolates were tested by Vero cell assay for verocvtotoxin production and by multiplex PCR assay for the detection of vtx1, vtx2 eae, and E-hlyA genes. VTEC 0157 was detected in 6 (3.3%o) of 182 animals, and VTEC 026 was detected in 1 (0.5%) of 182 animals. No VTEC VTEC 0111, or VTEC 0145 isolates were found in cattle feces, but one VTEC 091:H- vtx2(+in apice) eae(- in apice), E-hlyA(-in apice) strain nonspecifically cross-reacted with the VTEC 0103 type. The prevalence of VTEC 0157 in the lymphatic tissue of cattle was in both tonsils (1 of 93 samples) and mesenteric lymph nodes (1 of 89 samples). Lymphatic tissue contamination was observed only in VTEC 0157 intestinal carriers: two (33.3%) of six fecal carriers were simultaneously VTEC 0157 lymphatic carriers. This finding suggests that VTEC 0157 contamination of meat does not necessarily come from feces or the environment. No other VTEC serogroups were detected in the lymphatic tissue of slaughtered cattle. ID nr Campitelli L, Di_Martino A, Spagnolo D, Gavin JDS, Di_Trani L, Facchini M, De_Marco MA, Foni E, Chiapponi C, Moreno_Martin A, Chen H, Guan Y, Delogu M, Donatelli I Molecular analysis of avian H7 influenza viruses circulating in Eurasia in : detection of multiple reassortant virus genotypes J Gen Virol. - Vol. 89 no 1 ( 2008). - p bib ref Impact factor: 3,120 Project CE ISS FLUPAN - Preparing for an influenza Pandemic (Flupan) - Donatelli Dr.ssa Isabella - UO - CE ,00 EUR - 23/04/ /06/ N5 - Parma - Barigazzi Dr. Giuseppe - I 3 - G1 - Reparto di Virologia e Sierologia Specializzata - Cordioli Dr. Paolo - I PRC Epidemiological surveillance of influenza virus re-circulation in domestic and wild animals: an investigation into antigenic characterization of circulating strains in susceptible domestic animal species (pig, horse, avian species), intensification of surveillance in wild avian species and from rural flocks, new immunizing strategies for swine - Barigazzi Dr. Giuseppe - Capofila - MINSAN /03/ /08/ ,00 LIT 1 - N5 - Parma - Foni Dr.ssa Emanuela - I 2 - G1 - Reparto di Virologia e Sierologia Specializzata - Moreno Martin Dr.ssa Ana - I 3 - N3 - Forlì - Massi Dr.ssa Paola - I 4 - ISS - Donatelli Dr.ssa Isabella - E 5 - INFS - De Marco Dr.ssa Maria Alessandra - E 6 - ISS 2 - Castrucci Dr.ssa Maria Rita - E Avian influenza infections by high and low pathogenicity H7 influenza viruses have caused several outbreaks Pag. 19 di /08/2011

20 in European poultry in recent years, also resulting in human infections. Although in some cases the source of H7 strains from domestic poultry was shown to be the viruses circulating in the wild bird reservoir, a thorough characterization of the entire genome of H7 viruses from both wild and domestic Eurasian birds, and their evolutionary relationships, has not been conducted. In our study, we have analysed low pathogenicity H7 influenza strains isolated from wild and domestic ducks in Italy and southern China and compared them with those from reared terrestrial poultry such as chicken and turkey. Phylogenetic analysis demonstrated that the H7 haemaggiutinin genes were all closely related to each other, whereas the remaining genes couid be divided into two or more phylogenetic groups. Almost each year different H7 reassortant viruses were identified and in at least two different years more than one H7 genotype co-circulated. A recent precursor in wild waterfowl was identified for most of the gene segments of terrestrial poultry viruses. Our data suggest that reassortment allows avian influenza viruses, in their natural reservoir, to increase their genetic diversity. In turn this might help avian influenza viruses colonize a wider range of hosts, including domestic poultry. ID nr Decaro N, Mari M, Desario C, Campolo M, Elia G, Martella V, Greco G, Cirone F, Colaianni ML, Cordioli P, Buonavoglia C Severe outbreak of bovine coronavirus infection in dairy cattle during the warmer season Vet Microbiol. - Vol. 126 ( 2008). - p bib ref Impact factor: 2,010 Project PRF Mammal coronaviruses: molecular epidemiology, vaccine development And implications for animal and human health - Cordioli Dr. Paolo - Capofila - MINSAN ,00 EUR - 01/09/ /08/ U1 - REPARTO VIROLOGIA - Cordioli Dr. Paolo - I 2 - Università di Bari - Dipartimento di Sanità Pubblica e Zootecnica - Decaro Prof. Nicola - E 3 - IstitutoZooprofilattico della Sardegna - Oggiano Dr.ssa Annalisa - E 4 - Istituto Zooprofilattico della Sicilia - Guercio Dr.ssa Annalisa - E 5 - Istituo Zooprofilattico delle Venezie - Nardelli Dr. Stefano - E A severe outbreak of enteric and respiratory disease associated with bovine coronavirus (BCoV) infection is described. The outbreak occurred in a dairy herd of southern Italy in the first decade of September 2006, when summer temperatures were still recorded, affecting calves, heifers and adult cows, with a marked decrease in milk production. By virus isolation and RT-PCR targeting the S gene, BCoV was identified as the etiological agent of the outbreak, whereas bacteriological, parasitological and toxicological investigations failed to detect other causes of disease. BCoV strains with % nucleotide identity in the S gene were isolated from nasal, ocular and rectal swabs, thus proving the absence of separate clusters of virus on the basis of tissue tropism. Sequence analysis of the haemagglutination-esterase and spike proteins of the strain detected in one rectal sample ( ) showed a high genetic relatedness with recent BCoV isolates (98-99% amino acid identity), with several unique amino acid substitutions in the S protein. The BCoV outbreak described in this paper presents interesting aspects: (i) the occurrence of a severe form of disease in the warmer season; (ii) the simultaneous presente of respiratory and enteric disease; (iii) the involvement of young as well as adult cattle. ID nr Bellini S, Santucci U, Zanardi G, Brocchi E, Marabelli R Swine vesicular disease surveillance and eradication activities in Italy Rev Sci Tech OIE. - Vol. 26 no 3 ( 2007). - p bib ref Impact factor: 0,744 Project PRC Improvement of SVD diagnosis: identification and neutralization of swine enteroviruses affecting SVD virus isolation in tissue cultures. - Brocchi Dr.ssa Emiliana - Capofila - MINSAN ,00 EUR - 15/04/ /12/ H1 - Reparto di Biotecnologie - Brocchi Dr.ssa Emiliana - I Pag. 20 di /08/2011

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