Food additives are redefined in European legislation (EC Regulation No. 1333/2008). Sulphur dioxide (E220) and sulphite (E221- E228) are widely used in food processing as preservatives because they slow down bacteri- al growth on foods and prevent oxidation or browning developing on shrimp and lobster. Shellfish processors, farmers and fishermen have long used sulfiting agents in a variety of species of warm and coldwater crustaceans as a treatment to prevent prawns and shrimps melanosis (blackspot), which is a natural process that makes the shell black after har- vesting caused by Polyphenoloxidase enzyme systems which remain active during refrigera- tion or ice storage. Sulfite-induced hypersensi- tivity is the most well-established adverse response in humans to this food additive. In the present study the presence of sulfites in different frozen and thawed shrimp and prawn species belonging to Penaeoidea superfamily has been evaluated by the Monier-Williams procedure, in order to carry out a risk assess- ment and evaluate the levels of consumer exposure to this class of additives from these fish products. In addition to assessing and monitoring the correct use of the additive, according to the limits imposed by the European regulations, the correct consumer information on labels was also evaluated. Analysis were performed on both whole shrimp (shell on) and inedible parts (head and peeled shell). Sulphites concentration in frozen sam- ples (expressed as SO2 mg/kg mean value±S.D.) was 214±17.43 for head on shell on shrimps; 170.73±14.99 for shell on headless shrimps; 112.90±27.55 for peeled and deveined shrimps. Thawed shrimps were purchased at mass retailers channel and local fish markets and local seafood retailers and purveyors: for these samples, all head on shell on, the sul- phites concentration (expressed as SO2 mg/kg mean value±S.D.) was 160.05±26.15 and 292.54±146.04, respectively. Non-edible parts showed, in all samples, much higher concen- trations.

Cinquanta cavalli da Tiro Pesante Rapido, alleva- ti in modo semibrado, presentavano eccessivo dimagrimento; di questi 4 erano deceduti e 10 si presentavano letargici. Una fattrice ed il suo re- do giacevano in decubito laterale, avevano fre- quenza respiratoria e cardiaca aumentate, mu- cose pallide e occasionali manifestazioni convul- sive. L’esame ematologico ed il profilo biochimi- co evidenziavano anemia, ipoalbuminemia, ipo- creatininemia, iperuremia, disequilibrio elettroli- tico ed assenza di emoparassiti. Nonostante una fluidoterapia disintossicante (80 ml/kg/die) con glucosio ed epatoprotettori non si evidenziava- no miglioramenti visibili e, dopo 48 ore, si ese- guiva l’eutanasia. All’esame necroscopico si os- servavano: edema polmonare, enterite catarrale con diffuse petecchie emorragiche, presenza di essudato in peritoneo, fegato ingrossato, induri- to e con piccole aree di necrosi, rene iperemico e allargamento della pelvi. Attraverso l’esame chimico dei campioni epati- ci e renali è stato possibile rilevare la presenza del metabolita dell’erbicida paraquat (N,N’-di- metill-4,4’-bipiridinio dicloruro) in concentra- zioni di 0,23 mg/l ed al successivo controllo del pascolo si rilevava una vasta area trattata. Il pa- raquat è un composto inodore che non altera subito le caratteristiche organolettiche della pianta, altamente tossico per gli equini. Il resto della mandria è stato tenuto a regime stallino e trattato con disintossicanti (colina, metionina, trimetilglicina e silimarina) e antios- sidanti (vitamina C ed E) per via orale. Al follow up a 15 giorni si evidenziava un quadro clinico normale degli animali e a 2 mesi la loro com- pleta guarigione e il raggiungimento di un Body Condition Score (BCS) pari a 3. Questo articolo è una segnalazione di un caso clinico di intossicazione da paraquat, la cui uni- ca via di prevenzione sembra essere il control- lo della materia prima: ovvero dei pascoli.

Beak and Feather Disease Virus (BFDV) is a member of the genus Circovirus and causes the Psittacine Beak and Feather Disease (PBFD) in Psittaciformes. PBFD is a severe disease generally characterized by immunodeficiency and beak and feather disorders. Although Circovirus spp. have been detected in several non-psittacine specie, little is known about the symptoms and the disease associated to this infection in birds other than Psittaciformes. Nevertheless, PBFD has never been described in non-psittacine specie. In this study, we reported an outbreak of Circovirus infection in a flock of Gouldian finches where beak and feather disorders were observed. Sequence analyses on the rep gene of the virus highlighted a strong similarity at nucleotide and amino acid level with the corresponding regions of BFDV

The aim of the study was to evaluate the occurrence of Arcobacter spp. in 20 samples of Mytilus galloprovincialis purchased at fish markets in Apulia region. The detection of Arcobacter spp. was performed, after selective enrichment, on modified charcoal cefoperazone deoxycholate (mCCD) agar supplemented with Cefoperazone, Amphotericin B and Teicoplanin (CAT). In 6 out of the 20 tested samples the presence of Arcobacter spp. was found and confirmed by genus-based polymerase chain reaction. All the isolates were identified as belonging to the species Arcobacter butzleri using 16S rDNA sequencing and BLAST online. The results represent the first report in Italy of A. butzleri detection in marketed Mytilus galloprovincialis. The survey underlines the epidemiological importance of A. butzleri as an emerging pathogen, and highlights that mussels should be considered as a potential cause of foodborne disease outbreak.

Ochratoxins are fungal secondary metabolites that may contaminate various foods and beverages. The intake of ochratoxins by humans may result in typical syndromes (nefrotoxicity, carcinogenity, teratogenicity and immunotoxicity) and has been associated with Balkan Endemic Nephropathy (BEN). In this study we evaluated the effects of accumulation of ochratoxin A throughout the chain production of eggs, by investigating the dynamics of OA accumulation in eggs placed by laying hens experimentally exposed to OA. It was demonstrated that after exposure at the concentrations admitted by the current European legislation (100 lg/kg) and at concentrations 20-folds as much the European Legislation limit (2000 lg/kg), OA was not detectable in the eggs, although a number of eggs were found to have altered structure or conformation and/or pathological lesions. Monitoring fungal contamination and toxins in animal feeds is necessary to guarantee animal health and to prevent the risk of decreased productions in livestock animals.

The plasmatic cortisol levels of 60 eight-month-old calves (Charolais breed) were measured as stress indicators resulting from two types of slaughter: traditional and religious rite. The plasmatic cortisol levels were evaluated during three different stages of their productive life: during growth, after transport and during slaughter. The thirty calves slaughtered after stunning showed plasmatic cortisol values of 4.85±3.2; 36.36±12.2 and 45.08±14.1 nmol/L, during growth, in the slaughterhouse stables and during exsanguination, respectively. Conversely, the average values found in the thirty calves subjected to ritual slaughter were 2.96±1.2; 31.65±25.4 and 68.70±30.6 nmol/L. The results of the study showed that animal welfare should be improved in both forms of slaughter.

The extensive use of pesticides in agriculture plays an important role in bees die-off and allows the presence of residues in hive products, particularly in honey. An accurate and reliable analytical method, based on QuEChERS extractive technique, has been developed for the quantitative determination by high-performance liquid chromatography UV-visible detector of 5 pesticides (Deltamethrin, Dimethoate, Imidacloprid, Acetamiprid, Chlorfenvinphos) in honey. The method, according to Commission Directive 2002/63/EC and Regulation 882/2004/EC, provided excellent results with respect to linearity (correlation coefficient up to 0.993), limits of detection and quantification (0.005 and 0.01 μg/mL for Dimethoate, Deltamethrin and Chlorfenvinphos; 0.02 and 0.05 μg/mL for Acetamiprid and Imidacloprid), recovery values (86.4 to 96.3%), precision and relative expanded uncertainty of a measurement, demonstrating the conformity of the this method with the European directives. The proposed method was applied to 23 samples of Apulian honey. None of the investigated pesticides was detected in these samples

The Italian Ministry of Agricultural, Food and Forestry Policies (MiPAAF) Decree dated 31 January 2008, which reports the Italian name for fish species of commercial interest, establishes that baccalà can be obtained exclusively from G. macrocephalus (Pacific cod) and G. morhua (Atlantic cod). This paper describes the COI-based DNA identification system to verify the substitution or misbranding of gadoid fish species and, consequently, its concordance with the labels on salted cod fillets shown as baccalà and on battered cod chunks labelled as bocconcini di baccalà. The analysis of interpretable sequences revealed that 55/65 dried salted cod fillet samples were detected as belonging to the family Gadidae, while 10/65 samples appeared to belong to the Lotidae family, while among battered cod chunks labelled as bocconcini di baccalà, the post-sequencing data analysis shows that the labels were completely wrong, with 28/40 samples from Pollachius virens and 12/40 samples from Brosme brosme. The substitution rate for products labelled on the market as baccalà in this study raises significant issues relating to food safety and consumer protection.

Sarcocystis spp. are protozoa belonging to the Phylum Apicomplexa, order Eucoccida, family Sarcocystis that cause sarcocystosis in humans, other primates and in many animal species. In this study the authors report the results of a survey on the presence of these parasites, in 50 horses slaughtered in the province of Bari. The inspection visit, by a veterinarian inspector, resulted positive for tne examined animals. On the contrary, the microscopic investigations revealed the presence of sarcocystis in 5 of 50 examined and permitted to classify the parasite as belonging to the species equicanis.

Human astroviruses (HAstVs) are important enteric pathogens and can be classified genetically and antigenically into eight types. During surveillance of HAstVs in Italy, type-4 HAstVs were detected only sporadically and found to cluster into two distinct genetic groups. Upon sequence analysis of the 3' end of the polymerase gene (ORF1b) and of the full-length ORF2, the 2008 type-4 HAstV strains were characterised as a novel ORF2 genetic lineage, designated as 4c. The 2008 type-4 HAstVs also shared the ORF1b gene with similar HAstV-4c strains detected globally, thus displaying a conserved ORF1b/ORF2 asset. By interrogation of the databases, this novel lineage 4c accounted for 60.8% of the type-4 strains identified worldwide and the vast majority of recent type-4 HAstVs. The 2002 type-4 HAstVs displayed a type-4b ORF2, whereas in the ORF1b they resembled type-1 HAstVs. This inconsistency suggests a possible recombinant origin, with the RNA switch taking place upstream the ORF1b/ORF2 junction region. Also, recombination likely played a role in the diversification of the ORF2 of the three type-4 lineages. Multi-target analysis is required for appropriate characterisation and identification of recombinant HAstVs.

Considering that the authentication of food contents is one of the most important issues for the food quality sector, and given the increasing demand for transparency in the meat industry followed the horsemeat scandal in Europe, this study investigates processed-meat products from Italian markets and supermarkets using the mitochondrial cytochrome b gene qualitative PCR identification system in order to verify any species substitution or mislabeling. The results revealed a high substitution rate among the meat products, highlighting a mislabeling rate of 57 %, and consequently, considerable discordance with the indications on the labels, which raises significant food-safety and consumer-protection concerns.

Considering that powdered infant milk formula effectively supports the growth of numerous pathogens, this study investigates the prevalence of potentially pathogenic Bacillus cereus in dried milk products by evaluating the occurrence of B. cereus and the presence of virulenceassociated genes. The approach consisted of enriching, isolating and biochemical identifying isolates, followed by PCR assays aimed at the hbl (C, D, A, B), nhe (A, B, C) and cytK enterotoxin genes coding HBL complex, NHE complex and cytotoxin K, respectively. Among cytKpositive strains, the discrimination of two different forms for cytotoxin K, cytK-1 and cytK-2 was performed. Bacillus cereus was detected in powdered infant milk formula samples. All the strains harbored at least one gene of the cytK, HBL and NHE enterotoxins. Because of an increasing trend in invasive infections by B. cereus in infants and immunocompromised children, our PCR findings highlight the need to implement an adequate control plan in order to guarantee the health of potentially fragile consumers. From a hygiene point of view, intensive and continuous monitoring of potentially pathogenic B. cereus may be crucial for powdered infant milk formula safety and even recommended in order to assess the infant health risk, as proposed by Commission Regulation (EC) no. 1441/2007 on microbiological criteria for foodstuffs. Furthermore, the detection in this study of B. licheniformis, B. subtilis and B. mycoides strains raises significant health issues regarding Bacillus spp. in powdered infant milk formula.

Ochratoxins are fungal secondary metabolites that may contaminate a broad variety of foodstuffs, such as grains, vegetables, coffee, dried fruits, beer, wine and meats. Ochratoxins are nephrotoxins, carcinogens, teratogens and immunotoxins in rats and are also likely to be in humans. In 2009/2010, a survey of the presence of Ochratoxin A (OTA) in regularly hunted wild boars in the Calabria region of southern Italy detected OTA in 23 animals in the kidney, urinary bladder, liver and muscles: 1.1 ± 1.15, 0.6 ± 0.58, 0.5 ± 0.54 and 0.3 ± 0.26 μg/kg, respectively. Twelve tissue samples showed levels of OTA higher than the guideline level (1 μg/kg) established by the Italian Ministry of Health. In five wild boars, gross-microscopic lesions were described for the organs displaying the highest concentrations of OTA determined by HPLC-FLD analysis, i.e., the kidney, liver and urinary bladder.

Protothecosis is a potential zoonotic disease associated with bovine mastitis which can be transmitted to humans through contaminated milk. Considering the increasing prevalence of bovine mastitis due to Prototheca species, individual cow milk samples were analyzed using microbiological examination and biomolecular assay. Aspects related to health requirements for milk production, clinical and histological bovine mastitis were also described. The results showed 24/257 (9.3%) culture-positive samples and 42/257 (16.3%) PCR-positive samples. Moreover in 5 cows with somatic cell count over 106/mL presented histological features of mastitis. This study reveals that the presence of Prototheca species in dairy herds was related to the hygienic conditions of the milking equipment, showing an emerging public health issue.

Ochratoxins are fungal secondary metabo- lites that may contaminate a broad variety of foodstuffs, such as grains, vegetables, coffee, dried fruits, beer, wine and meats. Ochratoxins are considered powerful nephrotoxins, car- cinogens, teratogens in rats and likely in humans. In 2011, during a programme aimed to survey the presence of ochratoxin A in 35 regularly slaughtered wild boars in Calabria region (Southern Italy), ochratoxin A (OTA) was detected in 35 kidneys, 33 urinary blad- ders, 33 livers and 32 muscles of 35 animals at the following levels: 1.05 ppb (0.1-3.9 ppb), 0.5 ppb [not detected (ND)-2.6 ppb], 0.4 ppb (ND- 2 ppb), 0.2 ppb (ND-0.5 ppb), respectively. A total of 12 samples of kidney, 4 samples of liver, and 4 samples of urinary bladder showed levels of OTA higher than the level (1 ppb) estab- lished by the guidelines of the Italian Ministry of Health circular No. 10.

Given that changes in consumer food behaviours have led to an increase in the demand for pre-cut ready-to-eat (RTE) vegetables, and that few data are currently available on the occurrence of Arcobacter spp. in such foods, the aim of the present study was to assess the occurrence of Arcobacter spp. that carry virulence-associated genes on pre-cut RTE vegetables, using cultural and molecular methods. Arcobacter was detected using biomolecular iden- tification methods in 44/160 (27.5%) of the samples, of which 40/44 (90.9%) isolates corresponded to A. butzleri and 4/44 (9.1%) to A. cryaerophilus. Studying the incidence of 9 virulence-associated genes revealed the wide- spread distribution of these genes among the Arcobacter isolates tested. The results obtained in our research provided plenty of information on the health risks associated with the direct consumption of raw vegetables, and highlight the need to implement further studies at each level of the produc- tion chain, in order to obtain further information to help protect human health.

Fish authentication is a major concern not only for the prevention of commercial fraud, but also for the assessment of safety risks deriving from the undeclared introduction of potentially dangerous toxic or allergenic substances or environmentally damaging fish where endangered species are involved. Moreover, food authentication might affect the diet of certain groups of consumers, such as followers of religious practices. Considering the authentication of fish products is one of the key issues in food safety, quality and sustainability, the aim of this work was to investigate the prevalence of mislabelling in sole (Solea solea), plaice (Pleuronectes platessa), Atlantic salmon (Salmo salar), and hake (Merluccius merluccius) fillets from markets and supermarkets located in Apulia (Southern Italy) using DNA barcoding. The results of the molecular investigations reveal that 42/98 (42.8%) fillet samples were not correctly labelled. In particular, 12/27 (44.4%) fillets of sole (Solea solea) were identified as belonging to Solea senegalensis. In addition, 13/28 (46.4%) plaice (Pleuronectes platessa) samples were identified as Pangasius hypophtalmus. All Atlantic salmon (Salmo salar) samples were correctly labelled. Post-sequencing data analysis revealed that 17/30 (56.6%) hake fillets (Merluccius merluccius) were not correctly labelled, of which 8/30 samples identified as Merluccius hubbsi, 5/30 samples as Merluccius products and 4/30 as Merluccius capensis. The study reveals a high occurrence of species mislabelling in the prepared fish fillet products, further evidence of the need for increased traceability and assessment of the authenticity of food products.

Considering that the authentication of food contents is one of the most important issues for the food quality sector, and given the increasing demand for transparency in the meat industry followed the horsemeat scandal in Europe, this study investigates processed-meat products from Italian markets and supermarkets using the mitochondrial cytochrome b gene qualitative PCR identification system in order to verify any species substitution or mislabeling. The results revealed a high substitution rate among the meat products, highlighting a mislabeling rate of 57 %, and consequently, considerable discordance with the indications on the labels, which raises significant food-safety and consumer-protection concerns.

Considering that several recent cases of human gastroenteritis have been associated with species from the Arcobacter genus, and that few data are currently available about the occurrence of this genus in Italian shellfish, the aim of the present study was to evaluate the occurrence of Arcobacter spp. and the presence of virulence-associated genes. The approach consisted of cultural and biomolecular (multiplex- PCR and 16S-RFLP) methods identifying isolates, followed by PCR assays aimed at the cadF, ciaB, cjl349, irgA, hecA putative virulence genes. Arcobacter spp. was detected in 16/70 (22.8%) shellfish samples. Specifically, Arcobacter spp. was highlighted in 10/42 (23.8%) mussel and in 6/28 (21.4%) clam samples. Subsequently, biomolecular assays revealed Arcobacter butzleri in 12/16 (75%) and Arcobacter cryaerophilus 1B in 4/16 (25%) isolates. PCRs aimed at the five putative virulence genes demonstrated widespread distribution of these genes among Arcobacter isolates and some differences from the results published by other authors. Our research provides more information regarding the health risks associated with the consumption of raw bivalve molluscs and underlines the need to implement an adequate control plan by performing intensive and continuous monitoring in order to guarantee human health

Hepatitis A virus (HAV) was detected in a batch of imported non-packaged frozen redcurrants purchased in a Bari grocery. Sequence and phylogenetic analysis showed the HAV strain clustered tightly with the HAV strain from the 2013 Italian epidemic, providing additional evidence that frozen redcurrants were the main vehicle of the HAV outbreak.

Given the increase in the international trade of packaged frozen fishery products, this study used DNA barcoding to investigate the breaded hake and plaice species, sold in Italian markets. The results of this study generally matched the ingredient list on the food label. Only 6 of the 120 samples were non-compliant. Specifically, breaded merluccius samples match the species shown in the list of ingredients on the label. Of the ‘‘breaded plaice’’ samples, 4/14 contained Lepidopsetta polyxystra and 2/14 Merluccius gayi, thus failing to match the ingredient list on the label. Considering the European legislation indicates that the label must not mislead consumers, but international trade and the use of similar terms for different products makes it complicated when a product from one country is introduced into another in which the niche already exists, clear labeling is strongly recommended in order to ensure that consumers can make conscious choices.

Beak and Feather Disease Virus (BFDV) is a member of the genus Circovirus and causes the Psittacine Beak and Feather Disease (PBFD) in Psittaciformes. PBFD is a severe disease generally characterized by immunodeficiency and beak and feather disorders. Although Circovirus spp. has been detected in several nonpsittacine species, little is known about the symptoms and the disease associated with this infection in birds other than Psittaciformes. In this study, we reported the identification of Circovirus infection in a flock of Gouldian finches showing beak and feather disorders. Sequence analyses on the rep gene of the virus highlighted a strong similarity at nucleotide and amino acid level with the corresponding regions of BFDV from psittacine species. By contrast, it was more distant to circoviruses identified in finch and canary

Silter cheese is a traditional hard cheese, produced in Valcamonica, Brescia, Italy. A total of 426 lactic strains isolated from Silter were analyzed to determine their probiotic characteristics. 274 out of 426 strains were found to produce bacteriocins against at least one of eight different pathogens (Salmonella enterica, Listeria monocytogenes, Salmonella derby, Salmonella thyphimurium, Salmonella napoli, Staphylococcus aureus, E. coli O157:H7, Salmonella enteritidis). In addition, 211 of 274 bactericin-producer strains adhered to Caco-2 cells and were characterized by RiboPrinter, revealing predominance of Enterococcus faecalis (26%) and Enterococcus durans-faecium (22%). These findings suggest that Silter may qualify as an important source of feasible probiotic strains.

Considering that seafood mislabeling has been widely reported throughout the world and that the authentication of food components is one of the key issues in food safety, quality and sustainability, the aim of this study was to use DNA barcoding to investigate the prevalence of mislabeling among fish fillet products from markets and supermarkets located in Apulia (SE Italy). The study reveals a high degree of species mislabeling in fish fillet products. In particular, this study shows that the labels of only 32/200 fish fillet samples provided comprehensive information relating to commercial designation, scientific name, geographical area, production method and whether previously frozen. The labeling of other samples was not compliant with European legislation. Indeed, the scientific name, which must also be indicated from 1st January 2012, according to Article 68 of EU Commission Implementing Regulation No. 404/2011, was missing in 157/168 samples, the geographical area was missing in 152/168, while the commercial designation and the production method were reported in all samples. Furthermore, results from molecular investigations reveal a high occurrence of incorrect species declaration in fish fillet products. The commercial and/or scientific name declared failed to match the species identified in 164/200 (82%) samples. The study also highlighted that threatened, Vulnerable (VU), Endangered (EN) and Critically Endangered (CR) species considered to be facing a high risk of extinction has been used in the place of commercial species. This study thus provides further evidence of the need for increased traceability and assessment of food product authenticity. Additionally, traceability may improve the management of hazards related to fish safety, as well as guaranteeing product authenticity, providing reliable information to customers, enhancing supply-side management and improving product quality and sustainability.

Monitoring animal welfare at a slaughterhouse is somehow different from the situation on the farm. The throughput numbers can be very high, and there is no possibility to know the individual history, background or habits of each animal. The animals are unfamiliar to the staff and viceversa. Furthermore, most animals will be unknown to each other even if they may be delivered in groups or batches, and it is not rare to see mixing of completely unfamiliar animals. The slaughter plants’ environment is of course completely unknown to the animals, with large areas not always designed to minimize noise or visual distraction for animals. Sub-optimal handling of animals can unfortunately be seen at many slaughterhouses, where animals are coerced roughly, using sticks or prods and loud voices, forcing the animals to move fast in the desired direction. Vets need to be aware of these factors because they can cause excessive prodding for animals, which may become balking, excited and stressed. A survey of 10 italian slaughter plants revealed that: 5 plants had slick floors that would cause animals to slip and fall, 9 plants had high pitched motor noise or hissing air that made animals balking; air drafts blowing down the chutes, which will often impede animal movement, were a problem in 9 of the observed plants. Simple modifications of lighting and elimination of air drafts and hissing will often greatly improve animal movement.

Ochratoxins are fungal secondary metabolites that may contaminate various foods and beverages. The intake of ochratoxins by humans may result in typical syndromes (nefrotoxicity, carcinogenity, teratogenicity and immunotoxicity) and has been associated with Balkan Endemic Nephropathy. In this study the authors describe a simple and highly specific method for the determination of Ochratoxin A in infant milk products and human breast milk. The method involves: the extraction of Ochratoxin A, clean-up with immunoaffinity columns having specific antibodies and quantification using high performance liquid chromatography with fluorescence detection. The minimum detectable concentration values of OTA obtained were appropriate for the analytical findings, designed to detect OTA in low concentrations.