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…

A comprehensive one-year correlation analysis between prevalence and number of sars-cov-2 rna copies detected in wastewater from 8 sicilian cities in italy

Background and objectives: Several authors have reported positive correlations between wastewater SARS-CoV-2 concentrations and the community’s burden of infection, providing information about the disease’s epidemiological trend. This pre-post study evaluated – within a year of active surveillance, from October 1st,2021, to September 30th, 2022 – the relation between daily SARSCoV-2 prevalence and the number of genome copies/L detected in West Sicily wastewater samples. This study is part of a national framework coordinated by Istituto Superiore di Sanità. Methods: Quantitative data from analysing 486 wastewater samples collected from 9 purification plants in 8 Sicilian cities (Agrigento, B…

Investigation and control of a Norovirus outbreak of probable waterborne transmission through a municipal groundwater system

During March 2011 an outbreak of gastroenteritis occurred in Santo Stefano di Quisquina, Agrigento, Sicily, Italy. Within two weeks 156 cases were identified among the 4,965 people living in the municipality. An epidemiological investigation was conducted to characterize the outbreak and target the control measures. A case was defined as a person developing diarrhea or vomiting during February 27–March 13, 2011. Stool specimens were collected from 12 cases. Norovirus (NoV) genotype GII.4 variant New Orleans 2009 was identified in stool samples from 11 of 12 cases tested (91.7%). Epidemiological investigations suggested a possible association with municipal drinking water consumption. Water …

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

Valutazione di alcune performance diagnostiche di kit ELISA per la diagnosi sierologica di Anemia Infettiva Equina (AIE)

Data on evaluation of some diagnostic parameters of all ELISA kits available in Italy for the serodiagnosis of AIE are presented and discussed. Ten laboratories were involved using a panel of 30 sera ran with 4 commercial kits and 2 in-house kits. Kits were also evaluated using a panel of sera from vaccinated animals at different days post vaccination (p.v.). All sera were also tested in agar gel immunodiffusion (AGID). The parameters evaluated were: diagnostic sensitivity (DSe) and specificity (DSp), Cohen K, weighted Cohen K, coefficient of variation (CV), accordance, concordance. Analysis of these parameters indicates that all kits have a higher sensitivity than AGID, even if a complete …

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…

Differentiation and characterization of rat adipose tissue mesenchymal stem cells into endothelial-like cells

In this study, mesenchymal stem cells were isolated from rat adipose tissue (AD-MSCs) to characterize and differentiate them into endothelial-like cells. AD-MSCs were isolated by mechanical and enzymatic treatments, and their identity was verified by colony-forming units (CFU) test and by differentiation into cells of mesodermal lineages. The endothelial differentiation was induced by plating another aliquot of cells in EGM-2 medium, enriched with specific endothelial growth factors. Five subcultures were performed. The expression of stemness genes (OCT4, SOX2 and NANOG) was investigated. The presence of CD90 and the absence of the CD45 were evaluated by flow cytometry. The endothelial-like…

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…

Waterborne Norovirus outbreak at a seaside resort likely originating from municipal water distribution system failure

AbstractIn May 2016 a Norovirus (NoV) gastroenteritis outbreak involved a high school class visiting a seaside resort near Taormina (Mascali, Sicily). Twenty-four students and a teacher were affected and 17 of them showed symptoms on the second day of the journey, while the others got ill within the following 2 days. Symptoms included vomiting, diarrhoea and fever, and 12 students required hospitalisation. Stool samples tested positive for NoV genome by Real-Time polymerase chain reaction assay in all 25 symptomatic subjects. The GII.P2/GII.2 NoV genotype was linked to the outbreak by ORF1/ORF2 sequence analysis. The epidemiological features of the outbreak were consistent with food/waterbo…

Surveillance of bovine tuberculosis in Sicily

Bovine Tuberculosis (bTB) by M. bovis is a chronic infectious disease of cattle that can represent a risk to human health. bTB European Eradication Plans are based on tuberculin testing and slaughtering of reactive subjects. Despite control measures have been applied in Italy since 1964, illness is still present in Sicily. On 2012, regional official data reported an herd infection prevalence of 3.09% and an herd incidence of 2.37%. In the same year, head prevalence was of 0.70%. This study has analyzed the results of the control activity performed on cattle farm in Sicily between 2009-2012, with the aim to evaluate the bTB prevalence and incidence.

OBTAINING MESENCHYMAL STEM CELLS FROM ADIPOSE TISSUE OR MURIN ORIGIN: EXPERIMENTAL STUDY.

The aim of this study was to isolate and characterize rat adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order to evaluate their proliferative potential and their ability to different cell types. AD-MSCs and Derived Mesenchymal Stem Cells (BM-MSCs) have the same characteristic in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtained from subcutaneous adipose tissue of Wistar rats. first of all microbiological controls were made to exclude the presence of bacteria of fungi in then tissue. Adipose tissue was mechanically and enzimatically fragmented and st…

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…

Obtaining Mesenchymal Stem Cells From Adipose Tissue Of Murin Origin: Experimental Study

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…

Further spread of West Nile virus in Italy.

Following two consecutive years of West Nile virus (WNV) circulation in Italy, new foci of infection were observed in August 2010 in Sicily and Molise in southern and central Italy, respectively. These incidents were far from the previous infected area in northern Italy, thereby confirming the ability of WNV to spread to new areas and affect new host populations. © Istituto G. Caporale 2010.

VALUTAZIONE DELLA PREVALENZA DI NOROVIRUS E VIRUS DELL’EPATITE A (HAV) NEI MOLLUSCHI EDULI LAMELLIBRANCHI IN SICILIA, STRATEGIE DI CONTROLLO E COMUNICAZIONE DEL RISCHIO

PRODUCTION OF CANINE MESENCHYMAL STEM CELLS FROM ADIPOSE TISSUE AND APPLICATION IN DOGS WITH CHRONIC OSTEOARTHRITIS OF THE HUMERORADIAL JOINTS

Genomic Analysis and Lineage Identification of SARS-CoV-2 Strains in Migrants Accessing Europe Through the Libyan Route

Many African countries, representing the origin of the majority of refugees, asylum-seekers, and other migrants, toward regions bordering on the Mediterranean area, are experiencing sustained local transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Sicily is one of the main entry gates of migrants crossing into Europe. We conducted a pilot study, based on the full-genome sequencing of SARS-CoV-2 strains isolated from migrants coming to Sicily by crossing the Mediterranean Sea, with the aim to investigate the viral genome polymorphism and to describe their genetic variations and the phylogenetic relationships. On June 21, a nongovernmental organization vessel rescu…

West Nile Disease (WND) in Sicily

WND is an emerging vector-borne zoonosis caused by an RNA virus included in the Japanese encephalitis group within the Flavivirus genus. In Italy, after the first appearance in Tuscany in 1998, West Nile Virus (WNV) has been continuously circulating since 2008. A National Surveillance Program consisting of monitoring horses, “sentinel poultries”, wild bird mortality and mosquitoes is active in the entire national territory since 2002. Authors describe the outbreaks which occurred in Sicily in 2010 and 2011. Case study: Between September-October 2010, seven horses from the western part of the island, in the Trapani province, showed neurological symptoms. In October 2011, another two horses s…

Importance of dogs as sentinels of West Nile Virus activity in urban and suburban areas

Background: West Nile Virus (WNV) is a virus included in the Japanese encephalitis sero-complex within the genus Flavivirus. In August 2010, cases of West Nile disease were reported for the first time in Sicily. Neurological symptomswere observed in native horses resident in the rural areas around the province of Trapani, in the western part of the island. During the epidemic, important critical questions onwhen the viruswas introduced in the area and aboutwhether the virus had circulated/was circulating in theurban area, emerged and needed to be answered. A retrospective study using dog serum samples was designed to answer these questions. Methods: Between January 2009 and September 2010, …

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.

Ricerca di Enterovirus in sedimenti marini delle coste siciliane

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 …

CUTANEOUS NEOFORMATIONS ASSOCIATED WITH COINFECTION BY ORF VIRUS AND ORTHOPOXVIRUS IN GOATS IN SICILY

Parapoxvirus - ORF virus (ORFV) is the etiological agent of contagious ecthyma, a severe zoonotic exanthematic disease that affects small ruminants. It is usually characterized by highly infectious pustules and papillomatous lesions on the skin of the lips, tongue, and around the mouth. In more severe cases, the skin of the eyes, feet, vulva or udder may also be affected. Orthopoxvirus has been sometimes associated with naturally spreading disease in domestic and wild animals. Parapoxvirus infection can be complicate by opportunistic agents, mainly Fusiformis necrophorus, Dermatophilus congolensis or be invaded by the larvae of flies. Little is reported on co-infection with other viruses. T…

Wastewater-Based Epidemiology as a Tool to Detect SARS-CoV-2 Circulation at the Community Level: Findings from a One-Year Wastewater Investigation Conducted in Sicily, Italy

Wastewater-based epidemiology is a well-established tool for detecting and monitoring the spread of enteric pathogens and the use of illegal drugs in communities in real time. Since only a few studies in Italy have investigated the correlation between SARS-CoV-2 in wastewater and the prevalence of COVID-19 cases from clinical testing, we conducted a one-year wastewater surveillance study in Sicily to correlate the load of SARS-CoV-2 RNA in wastewater and the reported cumulative prevalence of COVID-19 in 14 cities from October 2021 to September 2022. Furthermore, we investigated the role of SARS-CoV-2 variants and subvariants in the increase in the number of SARS CoV-2 infections. Our findin…

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…

Wastewater surveillance of SARS-CoV-2 variants in October-November 2022 in Italy: detection of XBB.1, BA.2.75 and rapid spread of the BQ.1 lineage

This study adds insight regarding the occurrence and spread of SARS-CoV-2 Variants of Concern (VOCs) and Var-iants of Interest (VOIs) in Italy in October and November 2022, by testing urban wastewater collected through-out the country. A total of 332 wastewater samples were collected from 20 Italian Regions/Autonomous Provinces (APs) within the framework of national SARS-CoV-2 environmental surveillance. Of these, 164 were collected in the first week of October and 168 in the first week of November. A similar to 1600 bp fragment of the spike protein was sequenced by Sanger (for individual samples) and long-read nanopore sequencing (for pooled Region/AP samples).In October, mutations charact…

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 …