Escherichia coli, Salmonella spp., Hepatitis A Virus and Norovirus in bivalve molluscs in Southern Italy

European Legislation has fixed microbiological, chemical and biotoxicological limits for shellfish but no limits for viruses. In the present study we report the results of an investigation on Salmonella spp., Escherichia coli, Hepatitis A virus (HAV) and Norovirus (NoV) contamination in 59 bivalve shellfish collected during the years 2011-2012 in Southern Italy. All the samples of Mytilus galloprovincialis and of Solen marginatus were negative for HAV whereas 6.8% of them were positive for Norovirus GI (NoVGI) and 11.9% positive for Norovirus GII (NoVGII). Samples were also negative for Salmonella spp., while 16 of them (27%) were positive for E. coli. No correlation was found between E. coli and NoV contamination in bivalve molluscs. Moreover, the Competent Authorities are advised to take into serious consideration additional measures for the legislation in force in order to guarantee the consumer's health

First identification of porcine parvovirus 3 in a wild boar in Italy by viral metagenomics – Short communication

Metagenomic analysis revealed the presence of porcine parvovirus 3 (PPV3) in the pool of the internal organs of a wild boar found dead in Southern Italy. Phylogenetic analysis based on the complete coding sequences showed that the newly detected virus is most closely related to those found also in wild boars in Romania during 2010–2011. Even though the death could not be associated with this virus, PPV3 could have contributed to lowering the host’s immunological defences

Identification of single nucleotide polymorphisms in Toll-like receptor candidate genes associated with tuberculosis infection in water buffalo (Bubalus bubalis)

Toll-like receptors play a key role in innate immunity by recognizing pathogens and activating appropriate responses. Pathogens express several signal molecules (pathogen-associated molecular patterns, PAMPs) essential for survival and pathogenicity. Recognition of PAMPs triggers an array of anti-microbial immune responses through the induction of various inflammatory cytokines. The objective of this work was to perform a case-control study to characterize the distribution of polymorphisms in three candidate genes (toll-like receptor 2, toll-like receptor 4, toll-like receptor 9) and to test their role as potential risk factors for tuberculosis infection in water buffalo (Bubalus bubalis)

Age estimation and growth of striped dolphins Stenella coeruleoalba stranded along the coasts of south-western Italy

The knowledge of demographic traits such as longevity, growth rates and age at sexual maturity is crucial for understanding the structure of a population in its natural environment and implementing appropriate strategies for its management and conservation. Based on counts of growth layer groups in sections of decalcified teeth using the paraffin technique, we estimated the age and growth of 25 individuals of striped dolphin (Stenella coeruleoalba) found dead stranded along the coast of Campania and Calabria (south Italy, central-western Mediterranean) from 2013 to 2018. Seven individuals, with TL of 100–110 cm, were calves under 1 year old. The oldest male and female individuals were 19 and 14 years old, respectively. Growth curve estimated using the Gompertz growth model (GGM) showed that in S. coeruleoalba male growth trajectories are partly in accordance with those reported in other studies on the same species from different Mediterranean areas. The high frequency (28%) of calves strongly suggests that females of this species use the marine area all around the south-western Italian coasts to give birth to their offspring. Furthermore, a comparison with the estimated age of striped dolphins from other Mediterranean marine areas shows that the longevity of the individuals examined in this study is much lower. Our study provides information toward understanding the demographic traits of S. coeruleoalba from Mediterranean Sea. The results reported here can be useful for future research aimed at understanding population structure, mortality patterns and the effects of anthropogenic activity on the survival of this species in this marine area

Mycobacterium tuberculosis SIT42 Infection in an Abused Dog in Southern Italy

A case of Mycobacterium tuberculosis infection is described in a dead adult male dog in Southern Italy. The carcass was found by the Health Authority in a gypsy encampment. It was admitted to our forensic veterinary medicine unit, with a suspicion of cruelty to the animal. Necropsy showed beating and traumatism signs, and mistreating was confirmed. Gross lesions included multiple nodular hepatic lesions, hemorrhagic enteritis with enlarged mesenteric lymph nodes, body cavity effusions, and an adrenal neoplasm. Bacteriological and molecular analyses were carried out on the liver lesions that enabled to identify M. tuberculosis SIT42 (LAM9). Drug-resistance patterns were evaluated by screening mutations on the rpoB and katG genes that showed susceptibility to both rifampin and isoniazid, respectively. Very few studies report canine tuberculosis, and little is known about the disease in Italy. To the authors' knowledge, this is the first report of Mycobacterium tuberculosis SIT42 infection in a dog in Italy

Identification of Altered miRNAs in Cerumen of Dogs Affected by Otitis Externa

Otitis externa is one of the most common diseases in dogs. It is associated with bacteria and yeast, which are regarded as secondary causes. Cerumen is a biological substance playing an important role in the protection of ear skin. The involvement of cerumen in immune defense is poorly understood. MicroRNAs can modulate the host immune response and can provide promising biomarkers for several inflammatory and infectious disorder diagnosis. The aims of this study were to profile the cerumen miRNA signature associated with otitis externa in dogs, integrate miRNAs to their target genes related to immune functions, and investigate their potential use as biomarkers. Cerumen was collected from healthy and otitis affected dogs and the expression of miRNAs was profiled by Next Generation Sequencing; the validation of the altered miRNAs was performed using RT-qPCR. The potential ability of miRNAs to modulate immune-related genes was investigated using bioinformatics tools. The results pointed out that 32 miRNAs, of which 14 were up- and 18 down-regulated, were differentially expressed in healthy vs. otitis-affected dogs. These results were verified by RT-qPCR. To assess the diagnostic value of miRNAs, ROC analysis was carried out, highlighting that 4 miRNAs are potential biomarkers to discriminate otitis-affected dogs. Bioinformatics showed that cerumen miRNAs may be involved in the modulation of host immune response. In conclusion, we have demonstrated for the first time that miRNAs can be efficiently extracted and quantified from cerumen, that their profile changes between healthy and otitis affected dogs, and that they may serve as potential biomarkers. Further studies are necessary to confirm their diagnostic value and to investigate their interaction with immune-related genes

Comparison of automated extraction methods for the detection of hepatitis A virus and norovirus in mussels

Noroviruses and hepatitis A virus are the pathogens most frequently involved in non‑bacterial gastroenteritis and hepatitis worldwide. They are mainly transmitted via the faecal–oral route, direct person‑to‑person contact or through the consumption of contaminated water and foods. In food virology, detection methods of these viruses are currently based on real‑time reverse transcription‑polymerase chain reaction (RT‑PCR). A crucial step in this process is the acid nucleic extraction, since its performance can negatively influence viral detection and thus give false negative results. The aim of this study is to evaluate the efficiency of 2 automated extraction systems, MagMAX Express and EZ1 Advanced XL, in recovering hepatitis A virus and norovirus RNA from mussels. In the present study, we used mengovirus as a process control to assess the efficiency of the extraction process. Samples were tested for mengovirus, hepatitis A, and norovirus by real time one‑step RT‑PCR assay. Our data indicates the MagMax Express is the better system to extract hepatitis A and norovirus RNA from mussels since its extraction efficiency was higher (p < 0.05) than EZ1 Advanced XL

Escherichia coli, Salmonella spp., Virus dell'Epatite A e Norovirus in Molluschi Bivalvi nel 2011-12 in Sud Italia

La legislazione europea ha fissato limiti microbiologici, chimici e biotossicologici per i molluschi ma non ha dato indicazioni sulla contaminazione da virus a trasmissione alimentare. In questo studio sono riportati i risultati sulla contaminazione da Salmonella spp., Escherichia coli (E. coli), Virus dell'Epatite A (HAV) e Norovirus (NoV) di campioni di molluschi raccolti, nel biennio 2011-2012, in Sud Italia. Tutte le matrici analizzate di Mytilus galloprovincialis e di Solen marginatus sono risultate negative all'HAV, il 6,8% positiva a Norovirus GI (NoVGI) e l'11,9% positiva a Norovirus GII (NoVGII). Inoltre esse sono risultate negative alla contaminazione da Salmonella spp. mentre il 27% è risultata positiva a E. coli. I dati ottenuti, inoltre, hanno dimostrato l'assenza di correlazione tra la contaminazione batterica dei Molluschi Bivalvi eduli testati e quella virale. Si suggerisce alle Autorità Competenti la valutazione urgente di misure normative aggiuntive al fine di garantire la sicurezza al consumatore