Computer-Assisted Molecular Traceability for Dairy Farming Products

Food integrity and food safety have received much attention in recent years due to the dramatic increasing number of food frauds. In this article we analyze dairy products for which one of the crucial issues is traditional cheese traceability. In this paper we propose a computer- assisted molecular traceability system able to certify a traditional dairy product. We investigate the use of short tandem repeat analysis data processed by a Covariance Matrix Adaptation Evolution Strategy algo- rithm in order to predict the traceability between dairy products and the corresponding producer and to highlight possible adulterations and/or inconsistencies. Preliminary results collected from two farms are pre- sented in this study to show the capability of the proposed algorithm in a real setup

Efficient isolation on Vero.DogSLAMtag cells and full genome characterization of Dolphin Morbillivirus (DMV) by next generation sequencing

The Dolphin Morbillivirus (DMV) genome from the frst Mediterranean epidemic (1990-\u201992) is the only cetacean Morbillivirus that has been completely sequenced. Here, we report the frst application of next generation sequencing (NGS) to morbillivirus infection of aquatic mammals. A viral isolate, representative of the 2006-\u201908 Mediterranean epidemic (DMV_IZSPLV_2008), efciently grew on Vero.DogSLAMtag cells and was submitted to whole genome characterization by NGS. The fnal genome length was 15,673 nucleotides, covering 99.82% of the DMV reference genome. Comparison of DMV_IZSPLV_2008 and 1990-\u201992 DMV strain sequences revealed 157 nucleotide mutations and 47 amino acid changes. The sequence similarity was 98.7% at the full genome level. Whole-genome phylogeny suggested that the DMV strain circulating during the 2006-\u201908 epidemics emerged from the 1990-\u201992 DMV strain. Viral isolation is considered the \u201cgold standard\u201d for morbillivirus diagnostics but efcient propagation of infectious virus is difcult to achieve. The successful cell replication of this strain allowed performing NGS directly from the viral RNA, without prior PCR amplifcation. We therefore provide to the scientifc community a second DMV genome, representative of another major outbreak. Interestingly, genome comparison revealed that the neglected L gene encompasses 74% of the genetic diversity and might serve as \u201chypervariable\u201d target for strain characterization

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

Differentiation between Fresh and Thawed Cephalopods Using NIR Spectroscopy and Multivariate Data Analysis

The sale of frozen–thawed fish and fish products, labeled as fresh, is currently one of the most common and insidious commercial food frauds. For this reason, the demand of reliable tools to identify the storage conditions is increasing. The present study was performed on two species, commonly sold in large-scale distribution: Cuttlefish (Sepia officinalis) and musky octopus (Eledone spp.). Fifty fresh cephalopod specimens were analyzed at refrigeration temperature (2 ± 2°C), then frozen at −20°C for 10 days and finally thawed and analyzed again. The performance of three near-infrared (NIR) instruments in identifying storage conditions were compared: The benchtop NIR Multi Purpose Analyzer (MPA) by Bruker, the portable MicroNIR by VIAVI and the handheld NIR SCiO by Consumer Physics. All collected spectra were processed and analyzed with chemometric methods. The SCiO data were also analyzed using the analytical tools available in the online application provided by the manufacturer to evaluate its performance. NIR spectroscopy, coupled with chemometrics, allowed discriminating between fresh and thawed samples with high accuracy: Cuttlefish between 82.3–94.1%, musky octopus between 91.2–97.1%, global model between 86.8–95.6%. Results show how food frauds could be detected directly in the marketplace, through small, ultra-fast and simplified handheld devices, whereas official control laboratories could use benchtop analytical instruments, coupled with chemometric approaches, to develop accurate and validated methods, suitable for regulatory purposes

A New Approach Against Food Frauds: The Portable Near-Infrared Device for Fish Fillets Identification

The demand for analytical methods for fish product authenticity has increased dramatically, particularly in rapid and non- destructive food authentication. Near-infrared (NIR) handheld devices can meet this need for fast, reliable, non-destructive and in situ analysis. Aim of this study was to verify fish fillets species by using a pocket-sized NIR sensor, called SCiO, a handheld NIR spectrometer that can easily scan solid and liquid samples. The species were chosen from among the most commonly sold on the market as fresh. Samples were divided in two groups, one for calibration and one for validation. The first was performed to set up the instrument and the second to cross-validate model performance. The fish species were correctly identified with a global accuracy between 93.97% and 96.58% and were all confirmed by a validated method based on genetic marker. The samples correspond to the declaration, suggesting this method as a good screening approach to avoid fish frauds with good accuracy

Molecular characterization of Pseudomonas fluorescens isolates involved in the Italian "blue mozzarella" event.

Between June and September 2010, widespread Italian consumer reports of unusual blue spoilage on fresh dairy products were publicized, resulting in the so-called blue mozzarella event. An inordinately high number of samples from mozzarella and whey cheese products of Italian and German production subsequently tested positive for Pseudomonas fluorescens. The aim of this study was to verify whether a selected P. fluorescens strain was responsible for this apparently unusual event. Molecular characterization of 181 isolated P. fluorescens strains was conducted using a newly optimized pulsed-field gel electrophoresis protocol. Although a high number of pulsotypes was found (132), only four pulsotypes were associated with more than one production plant, and only one German isolate had the same pulsotype as was detected in two Italian plants. This is the only evidence of possible cross-contamination among cheeses from the two countries. The overall results did not support the spread of contamination from German to Italian plants or the presence of one environmental strain that spread in both countries

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)

Detection of Invasive Borrelia burgdorferi Strains in North-Eastern Piedmont, Italy.

SummaryFollowing reports of human cases of Lyme borreliosis from the Ossola Valley, a mountainous area of Piemonte, north‐western Italy, the abundance and altitudinal distribution of ticks, and infection of these vectors with Borrelia burgdorferi sensu lato were evaluated. A total of 1662 host‐seeking Ixodes ricinus were collected by dragging from April to September 2011 at locations between 400 and 1450 m above sea level. Additional 104 I. ricinus were collected from 35 hunted wild animals (4 chamois, 8 roe deer, 23 red deer). Tick density, expressed as the number of ticks per 100 m2, resulted highly variable among different areas, ranging from 0 to 105 larvae and from 0 to 22 nymphs. A sample of 352 ticks (327 from dragging and 25 from wild animals) was screened by a PCR assay targeting a fragment of the 16S rRNA gene of B. burgdorferi s.l. Positive samples were confirmed with a PCR assay specific for the 5S‐23S rRNA intergenic spacer region and sequenced. Four genospecies were found: B. afzelii (prevalence 4.0%), B. lusitaniae (4.0%), B. garinii (1.5%) and B. valaisiana (0.3%). Phylogenetic analysis based on the ospC gene showed that most of the Borrelia strains from pathogenic genospecies had the potential for human infection and for invasion of secondary body sites