Analisi filogenetica condotta su ceppi di Bovine Diarrhea Virus (BVDV) isolati in Sicilia

La Diarrea Virale del Bovino-Malattia delle Mucose, è una malattia infettiva che colpisce i bovini, ampiamente diffusa a livello mondiale. L’agente infettivo responsabile è un virus provvisto di envelope e con un genoma ad RNA a singolo filamento e a polarità positiva, appartenente al genere Pestivirus, famiglia Flaviviridae. A causa dell’elevata capacità di andare in contro a mutazioni genetiche, esistono numerose varianti di BVDV con diverso assetto antigenico e differente patogenicità. Ad oggi sono noti due genotipi: il BVDV-I e il BVDV-II. Il BVDV-I comprende almeno 13 sottotipi, mentre ne sono stati descritti solo 2 per il BVDV-II. Quest’ultimo, poco diffuso nel nostro territorio, è re…

Quality controls for cell cultures: identification of interspecies cross-contamination by PCR-RFLP analysis of the cytochrome b gene

Cross-contaminations of a cell line with cells of different species represent a potential risk in laboratories handling human and animal cells. Therefore, it is necessary to control such contaminations. Tests based on mitochondrial DNA (mtDNA) are used in forensic analysis, phylogenetic studies and in food authentication. However, the use of mtDNA in quality controls of cell cultures is recent. Mitochondrial sequence differences of closely related animal species are five- to tenfold higher than those of nuclear genes. On the contrary, intraspecies variation in mitochondrial sequences is low in most animal species. Moreover, each cell contains 100–10.000 mitochondrial genomes. The amount of …

Surveillance of Municipal drinking-water supply after a Norovirus outbreak in Italy

BACKGROUND Norovirus (NoV), in the family Caliciviridae, is the major causative agent of non-bacterial acute gastroenteritis, whose possible routes of transmission include drinking water. During March 2011 an outbreak of gastroenteritis was identified by the local public health service in S. Stefano di Quisquina, Sicily, Italy. Over two weeks, 156 cases were identified and NoV genotype GII.4 v2010 was detected in stool samples from eleven out of twelve patients. The finding of identical genomic sequences between patients suggested a common infection source and an epidemiological investigation indicated a possible correlation to municipal drinking water consumption. One of the wells supplyin…

Analisi biomolecolare di ceppi di Canine Distemper Virus (CDV) in furetti domestici (Mustela putorius furo)

L’agente eziologico del cimurro (CDV) appartiene alla famiglia Paramyxoviridae, genere Morbillivirus ed è causa di una patologia infettiva e contagiosa in canidi, mustelidi e procionidi. Gli Autori, descrivono un caso clinico di cimurro in due furetti domestici vaccinati con un ceppo Onderstepoort avianizzato. e la caratterizzazione biomolecolare del CDV isolato. I furetti presentavano un quadro clinico caratterizzato da dermatite pruriginosa e squamo-purulenta, alla regione del mento, peri-labiale, peri-vulvare ed al condotto uditivo esterno. I soggetti venivano a morte tre settimane dopo l’esordio dei sintomi. I campioni di croste prelevati sono stati sottoposti ad estrazione degli acidi …

Diagnosi di enteriti infettive di origine virale nel cane

Canine Viral Enteritis are widespread in farms and kennels. Responsible viruses are Parvovirus (CPV), Coronavirus (CCoV), Rotavirus (CRV) and Distemper Virus (CDV). Aim of this study was to assess their prevalence in Sicily and to characterize the strains isolated during 2009-2012. For this purpose, samples (stools, rectal swabs, intestine, liver, spleen, heart, lung, brain) collected from dogs were analyzed by PCR, RT-PCR and Real Time RT-PCR. Positive samples were processed for virus isolation on cell lines. Viruses isolated were analyzed by RFLP and sequencing for molecular characterization. Results show an high prevalence of CPV infection in dogs, followed by CCoV, CRV and CDV. CPV prev…

Evidenza di un caso di epatite infettiva del cane in Sicilia

L’Adenovirus Canino di tipo 1 (CAV-1), appartenente al genere Mastadenovirus, famiglia Adenoviridae, è l’agente causale dell’epatite infettiva del cane (ICH). Il virus replica negli endoteli vascolari e negli epatociti, causando epatite necrotico emorragica acuta. I sintomi neurologici sono rari e sono causati da una vasculite del SNC. Negli ultimi anni, la diffusa vaccinazione ha ridotto la circolazione di CAV-1 ed i casi clinici segnalati sono diventati rari ed isolati. In questo lavoro, gli Autori descrivono un caso clinico in un cucciolo di 2 mesi, non vaccinato, con febbre, prostrazione, convulsioni, vomito, diarrea ed esito infausto in una settimana. A seguito dell’esame autoptico, so…

In vitro production of differentiated Endotelual Cells from Mesenchimal Stem Cells, collected from adult rat Adipose Tissue (AD-MSCs): preliminary results

Mesenchymal Stem Cells (MSCs) are multipotent cells that reside within various tissues, including adipose tissue and bone marrow. Research concerning the potential therapeutic role of MSCs has advanced significantly over the last decade. Many studies have demonstrated that adipose tissue is an excellent source of MSCs (AD-MSCs), easily accessible in experimental animals. AD-MSCs can be expanded in vitro over the short term and they can differentiate toward a variety of cell types of mesodermal lineage. Therefore, they are thought an attractive tool for cell therapy. Selection of appropriate animal models for preclinical evaluations is crucial for optimization and validation of therapeutic p…

Artrite-Encefalite Caprina (CAEV) una patologia emergente nel territorio siciliano: programma volontario di eradicazione

L’Artrite-Encefalite Virale Caprina è una malattia infettiva segnalata per la prima volta nel 1974; l’agente causale è stato isolato solamente nel 1979: si tratta di un virus della famiglia Retroviridae, genere Lentivirus. Il virus si diffonde principalmente attraverso colostro, latte (durante la mungitura) e, in minor misura, attraverso sangue, saliva, espettorato, secrezioni vaginali. La malattia può evolvere in forma subdola e tende ad essere sottostimata negli allevamenti infetti. Le forme cliniche negli animali adulti si presentano sotto forma di artrite e/o mastite, mentre negli animali giovani, nei quali l’evidenza è ancora più rara, si osserva sintomatologia clinica riferibile ad en…

Synthetic scaffolds may be used for the orientation of cardiac stem cells properties, differentiation and extracellular matrix interactions in cardiac tissue engineering: in vitro and in vivo studies.

GIV noroviruses and other enteric viruses in bivalves: a preliminary study.

We evaluated the presence of the enteric viruses: norovirus, adenovirus, enterovirus, astrovirus, hepatitis A virus, and hepatitis E virus in bivalves using nested PCR methods and cell culture assays. Noroviruses GII.4 and GIV.1, adenoviruses types 1 and 2, hepatitis A, and echovirus type 7 were detected in the shellfish tested, which were often co-infected. This is the first study to detect such a high level of viral contamination in Italian mussels (up to four different viral groups in a single sample), and the first to document the presence of GIV NoV in shellfish.

Canine Mesenchymal Stem Cells from visceral and subcutaneuous adipose tissue for cell-based therapy

This study compared some characteristics of canine Adipose tissue-Derived Mesenchymal Stem Cells (cAD-MSCs) from subcutaneous and visceral fat. These findings were directed to obtain high quantity and quality cAD-MSCs for clinical cell-based therapy.

An innovative method for the detection of contaminant viral genome in cell cultures

The use of cell cultures involves different fields of biology, from diagnosis to research. Moreover, technologies based on animal cells represent a useful tool to the development of biological products for the prophylaxis and therapy in humans and animals. Therefore, it is necessary to perform quality controls, including virological tests. Several tests performed in research laboratories are able to discriminate one or more viral species, but it is not possible to demonstrate the presence of contaminant viral genome with one non-specific method. The aim of this work consisted on the realization of a biomolecular method able to detect and to identify by sequencing extraneous viral genome in …

Detection of bovine Papillomavirus and efficacy of treatment using saponinDetection of bovine Papillomavirus and efficacy of treatment using saponin

Papillomas of teats and udders in cows may cause skin lesions, thelitis, mastitis and poor milking resulting in economic losses. We describe an outbreak of bovine papillomavirus (BPV) and successful treatment with a purified extract of Quillaja saponaria bark (Quil-A).

GIV noroviruses and other enteric viruses in bivalves

The impact of sewage discharge on the microbiological quality of shellfish is well known. Bivalves filter large volumes of water as part of their feeding activities, and thus accumulate and concentrate different types of pathogens from human fecal pollution. The aim of the study was to evaluate the presence, in these molluscs, of six different groups of enteric viruses: norovirus (genogroups I, II, and GIV), adenovirus, enterovirus, astrovirus, hepatitis A virus, and hepatitis E virus. The present study is the first to document the presence of GIV noroviruses in shellfish.

Ovine Catarrhal fever (bluetongue): Analysis of Culicoides species in seropositive farms

Bluetongue (BT) is an orbiviral disease of wild and domestic ruminants, mainly sheep. In Sicily, the first Bluetongue outbreak occurred in October 2000; there have been 76 recorded outbreaks so far. The National Surveillance Plan, based on European Union Commission Decision 138/2001/CE, establishes serological and entomological surveys. This plan consists of controls of seronegative cattle, called 'sentry' as indicators for the presence and circulation of virus in defined areas. To check the seroconversions, the regional territory has been subdivided in 400 km2 areas including 58 seronegative cattle, periodically checked by serological tests. All positive sera have been tested to detect the…

Risultati di un test RFLP su ceppi vaccinali di Canine Distemper Virus in Italia

Canine Distemper (CD) is a highly contagious and multisystemic viral disease of domestic and wild carnivores. A published Restriction Fragment Length Polymorphism (RFLP) test, based on the presence of a PsiI cleavage site on hemagglutinin (H) gene, allows a rapid differentiation of all currently used vaccine strains by virulent field strains. The present study describes the results of this test carried out on different CD vaccines available in Italy in 2010. RFLP has also revealed that the CD strain present in the Vanguard (Pfizer Animal Health) vaccine reacts as a wild-type strain. Moreover, genetic analysis of H gene sequence has showed that Vanguard vaccine strain does not cluster in the…

Haemorragic syndrome in cattle by subgenotype BVDV-1h

Bovine viral diarrhoeaemucosal disease (BVD-MD) is caused by a Pestivirus. Two genotypes are recognized: BVDV-1 and BVDV-2. BVDV-1 causes diarrhoea, respiratory disorders and reproductive disorders. BVDV-2 causes a haemorrhagic syndrome. We describe several hypervirulent outbreaks characterized by severe haemorrhagic signs caused by BVDV-1 in Sicily.

Virus influenzali H1N1pdm e riassortanti isolati dal suino

Sin dai primi giorni della circolazione del virus pandemico H1N1-2009 nella popolazione, è stata segnalata la trasmissione del virus dall’uomo a specie animali diverse (suino, tacchino, cane, gatto). La trasmissione di virus influenzali dall’uomo agli animali non rappresenta solo un problema limitato al settore zootecnico, ma un problema di sanità pubblica per il ruolo di “serbatoio” dei virus influenzali rappresentato da tali specie. Il suino non ha sinora dimostrato un ruolo epidemiologico importante nella diffusione di (H1N1)pdm all’uomo; tuttavia ciò non esclude, che a seguito dell’evoluzione virale, o di fenomeni di riassortimento, da tale specie emergano e si trasmettano all’uomo viru…

Messa a punto di un protocollo sperimentale per la ricerca di Enterovirus in sedimenti marini

In riferimento al Decreto Ministeriale del 24/01/1996, il controllo dei fondali marini utilizzati per il ripascimento dei litorali, costituisce un passo importante per la valutazione della salubrità dell’ambiente marino e dei prodotti ittici da allevamento, con particolare riferimento alla coltivazione di molluschi eduli lamellibranchi. Diversi studi, infatti, hanno dimostrato come i sedimenti, per le loro caratteristiche chimico-fisiche, rappresentano dei “serbatoi” di agenti microbici di varia natura che, da qui possono essere risospesi nelle acque, compromettendo la salubrità dell’ambiente. Poiché i molluschi eduli lamellibranchi sono organismi filtratori e trattengono agenti patogeni ca…

Rapid sequencing and genetic analysis of the pandemic (H1N1)V influenza virus circulating in pigs in Italy

will continue to pose challenge to public health and scientifi c community. General concern exists about possible mutation or reassortment between the H1N1pdm and infl uenza viruses circulating in human and animal, giving rise to more transmissible or pathogenic viruses. Emergence of resistance during antiviral treatment is a wellrecognised phenomenon in infl uenza viruses; surveillance for emergence of resistant viruses is of importance for monitoring this potential public health problem in the context of the H1N1 pandemic. Thus preparedness to identify new strains would require fast sequencing of the full genome of virus. Here we present an optimised workfl ow for rapid sequencing of the …

Preliminary study on the production of canine mesenchymal stem cells from adipose tissue and possibile applications in dogs with orthopaedic lesions.

Caratterizzazione molecolare di norovirus GIV ed altri virus enterici in molluschi bivalvi : uno studio preliminare

I virus enterici si replicano nell’intestino umano e possono essere eliminati dagli individui infetti ad elevate concentrazioni; pertanto i reflui urbani possono trasportare una grande quantità di patogeni responsabili di infezioni oro-fecali e contaminare le acque superficiali, incluse quelle marine. I molluschi bivalvi eduli sono spesso implicati nella trasmissione di malattie gastroenteriche di origine virale in quanto, essendo organismi filtratori, rappresentano un potenziale serbatoio di agenti patogeni e sostanze tossiche. Nonostante i miglioramenti delle tecniche per il trattamento delle acque reflue e la classificazione delle aree di allevamento e raccolta dei molluschi, il rischio …

Foreign body response to subcutaneously implanted scaffolds for cardiac tissue engineering.

Genotyping of bovine diarrhea virus (BVDV) isolated from cattle in Sicily

Bovine Viral Diarrhoea - Mucosal Disease (BVD-MD) is a widely spread infectious disease that cause economically important losses in farms. Several epidemiological studies indicate a high genetic heterogeneity among Bovine Viral Diarrhoea Virus (BVDV) strains circulating in Italy. The aim of this study was to investigate the genotypes of BVDV in Sicily, a region in the South of Italy. For this purpose, 17 BVDV strains collected from cattle breed in Sicily between 2005 and 2008 were genetically typed by sequencing of 5'-untraslated region (5’-UTR) of the viral genome. In this study, phylogenetic analysis showed that all 17 examined strains were clustered within the BVDV genotype 1. Particular…

Fibroin and poly-lactic acid scaffolds may be used in cardiac tissue engineering to drive the differentiation of cardiac progenitor cells: in vitro and in vivo studies.

Nuove varianti di Bronchite Infettiva negli allevamenti avicoli siciliani

Avian Infectious Bronchitis (IB) is a disease caused by a Coronavirus, included in Coronaviridae family. The disease, endemic in Italy, affects both broilers and laying hens. It represents one of the main health issue in sicilian poultry farms. The presence of new antigenic variants makes problematic the implementation of an adequate prophylaxis through the use of appropriate vaccines. The present work aims to study the spread of IB strains in Sicily by serological and biomolecular tests in order to investigate the presence of "historical" strains as well as new strains and to carry out the genotyping of viruses isolated. The serological results show that the used vaccination protocols are …