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 responsabile di una sindrome emorragica altamente letale. In Italia è stato isolato da bovini che erano stati trattati con vaccini anti-IBR contaminati. Recentemente è stata anche ipotizzata la presenza in Italia di una terza variante, il BVDV-III. L’infezione da BVDV spesso è associata con disordini a livello riproduttivo e nelle bovine gravide può esitare in aborto, malformazioni fetali o nascita di vitelli persistentemente infetti (PI). Scopo del presente lavoro è stato quello di genotipizzare i ceppi di BVDV isolati, al fine di approfondire la conoscenza circa la diffusione delle diverse varianti virali in Sicilia. Sono stati sequenziati ed analizzati filogeneticamente 18 ceppi, isolati su linee cellulari MDBK, da campioni di animali che erano risultati positivi in RT-PCR specifica per una porzione della regione 5’ UTR e in ELISA per la ricerca dell’antigene virale. Per il sequenziamento, i ceppi virali sono stati sottoposti ad estrazione dell’RNA ed amplificazione di una porzione della regione 5’ UTR, secondo quanto descritto da Vilcek et al. (1994). Le sequenze ottenute sono state successivamente allineate e comparate con altre sequenze di riferimento disponibili in GeneBank. L’analisi filogenetica condotta attraverso il metodo Neighbour-Joining, ha permesso di evidenziare la presenza di due sottotipi virali appartenenti al genotipo BVDV-I: BVDV-Ib e BVDV-Ie. Nessun ceppo virale è risultato appartenere al genotipo BVDV-II. Una successiva analisi di restrizione condotta sugli amplificati utilizzando l’enzima di restrizione AvaI, ha confermato infine l’origine bovina di ciascun isolato. L’analisi filogenetica dei ceppi virali isolati in un determinato territorio, rappresenta un valido strumento per il monitoraggio di una malattia infettiva, permettendo l’evidenziazione di nuove eventuali varianti causa di emergenze sanitarie, la realizzazione di test diagnostici specifici e la realizzazione di una profilassi vaccinale mirata

Cardiac filariosis in migratory Mute swans (Cygnus olor) in Sicily

Sarconema eurycerca is a common parasitic disease of North America swans and geese. The infection has been correlated with severe heart lesions, often resulting in cardiac failure and death of the animals. Heartworms infections have been previously reported in European swans, and specifically in the United Kingdom and Nederland. Both the countries are characterized by a cold temperate weather, similar to the one that can be found in swan wintering areas of U.S.A. and Canada. The first record of cardiac filariasis associated with Sarconema eurycerca infection in four swans in Italy. Twelve mute swans were examined during avian influenza surveillance activities on migratory birds. Birds were collected in the year 2006, in wintering areas of Eastern Sicily (Italy). Four of the twelve swans showed necrotic-haemorrhagic myocarditis with intra-lesional nematodes. Morphological characteristics identified the parasite as a filarial nematode. Birds lungs samples were used for parasites DNA extraction. The latter was used as template for polymerase chain reaction (PCR) amplification and sequencing of part of the 12S rDNA gene. Comparison of genomic DNA extracted from a reference S. eurycerca isolate confirmed parasite identity and provided the first sequence resources for this species of value to future diagnostic and epidemiological studies

Microbial diversity of traditional Sicilian cheeses

Traditional Sicilian cheeses are manufactured in small size farms with raw milk from animals of indigenous breeds and without the addition of starter cultures. In order to transform milk into cheese, the presence of lactic acid bacteria (LAB) is required. The main sources of desirable LAB are generally the milk, the rennet, the equipment used during processing and the dairy environment. In the last years, the microbial characterisation of traditional Sicilian cheeses, such as Caciocavallo Palermitano, Protected Designation of Origin (PDO) Pecorino Siciliano and PDO Vastedda della valle del Belìce have been the object of different studies conducted by our research group. To this purpose, the aim of the present study was to describe the microbial population of traditional Sicilian cheeses. The analysis of the microbial diversity of these cheeses revealed the presence of several dairy LAB at dominant levels, while the undesired microorganisms, including coliforms and coagulase-positive staphylococci (CPS), were at very low densities. Overall, the presence of the pathogenic bacteria Listeria monocytogenes and Salmonella spp. was never detected. This may be due to the quality of milk, the optimal maintenance of wooden vats and on following good production practices. Furthermore, the technological characterisation of the LAB found in these cheeses and in raw materials showed interesting dairy properties, including acidification capacity, diacetyl formation, autolytic properties and the ability to inhibit undesired bacteria

Investigation of disease hazards in cattle in South of Italy (Sicily)

Istituto Zooprofilattico Sperimentale, Palermo, Italy. Objective: Infectious diseases represent a serious limitation of bovine production. The etiology of these diseases is diverse and comprises a variety of viral, bacterial, protozoan and chlamydial agents, some of which are zoonotic [1]. Infectious- parasitic agents associated with reproductive disorders in ruminants include Neospora caninum, Coxiella burnetii, Chlamydia abortus and Toxoplasma gondii; they cause the greatest economic losses for the livestock industry. This is a cross-sectional study to assess the presence of antibodies in ruminants against selected pathogens including the zoonotic agents C. burnetii, T gondii, N. caninum, Clamydia spp. and Theileria annulata in cattle in Sicily region and to determine the molecular status for T. gondii in order to determine the serological and molecular status of bovine in the Sicily region

Viral Encephalopathy and Retinopathy (VER) in Mediterranean wild and farmed fish species : the experience of the ‘Istituto Zooprofilattico Sperimentale’ in Sicily

Betanodavirus infection is widespread in a broad spectrum of fish species worldwide. In Italy, it is responsible for outbreaks of Viral Encephalo-Retinopathy (VER) that causes mortality and economic losses in sea fish farming. The infection is also widespread in the wild and not only in managed systems, where there are generally no observed clinical manifestations.peer-reviewe

Molecular characterization of human enteric viruses in food, water samples, and surface swabs in Sicily

Objectives: Enteric viruses are responsible for foodborne and waterborne infections affecting a large number of people. Data on food and water viral contamination in the south of Italy (Sicily) are scarce and fragmentary. The aim of this study was to evaluate the presence of viral contamination in food, water samples, and surface swabs collected in Sicily Methods: The survey was conducted on 108 shellfish, 23 water samples (seawater, pipe water, and torrent water), 52 vegetables, one peach and 17 berries, 11 gastronomic preparations containing fish products and/or raw vegetables, and 28 surface swabs. Hepatitis A virus (HAV), genogroup GI, GII, and GIV norovirus (NoV), enterovirus (EV), rotavirus (RoV), hepatitis E virus (HEV), adenovirus (AdV), and bocavirus (BoV) were detected by nested (RT) PCR, real-time PCR, and sequence analysis. Results: The most frequently detected viruses in shellfish were HAV (13%), NoV (18.5%), and EV (7.4%). Bocavirus was found in 3.7%, HEV in 0.9%, and AdV in 1.9% of the molluscs. Of the 23 water samples, 21.7% were positive for GII NoV and 4.3% for RoV and HEV genotype 3. Of the 70 vegetable samples, 2.9% were positive for NoV GI (GI.5 and GI.6), 2.9% for EV, and 1.4% for HEV. In the gastronomic preparations, only one EV (9%) was detected. No enteric viruses were detected in the berries, fruit, or swabs analyzed. Conclusions: Molecular surveillance of water and food samples clearly demonstrated that human pathogenic viruses are widely found in aquatic environments and on vegetables, and confirmed the role of vegetables and bivalve molluscs as the main reservoirs. Keywords: Enteric virus, PCR, Genotyping, Shellfish, Waters, Vegetable

The Biobank of the ‘Istituto Zooprofilattico Sperimentale’ of Sicily: an important resource in medical research for safe and quality storage of biological specimens

The Mediterraean Biobank has been recently launched at the ‘Istituto Zooprofilattico Sperimentale’ (IZS) of Sicily in Palermo. The main aim of the Biobank is to collect and store, under standardised conditions and in a centralised system, several types of certified animal and zoonotic biological resources such as bacterial and viral strains, parasites, nucleic acids, positive/negative sera, cell cultures and tissues, in accordance to quality System procedures as regulated by UNI CEI EN ISO/IEC 17025:2005.peer-reviewe

Microbial diversity of traditional Sicilian cheeses

In the last years, the microbial characterisation of traditional Sicilian cheeses, such as Caciocavallo Palermitano, Protected Designation of Origin (PDO) Pecorino Siciliano and PDO Vastedda della valle del Belìce have been the object of different studies conducted by our research group. To this purpose, the aim of the present study was to describe the microbial population of traditional Sicilian cheeses.peer-reviewe

Detection of human enteric viruses in water and shellfish samples collected in Sicily

Historical flora and/or vegetation surveys highlight how the small islands of the Ionian coast of Sicily (Bella, Ciclopi islands, Vendicari, Capo Passero), currently protected as Nature Reserves and/or part of the Natura 2000 network have undergone major landscape changes following alterations in anthropogenic pressure. Landscape changes have been more prominent for the islands located in the northern part of the Ionian coast, with the consequence of increasing populations of invasive alien species. In contrast, landscape changes have decreased in the southernmost islands as indicated by the recovery of natural wood vegetation.peer-reviewe