Hepatitis E Virus RNA Detection in Liver and Muscle Tissues Sampled from Home Slaughtered Domestic Pigs in Central Italy.

Hepatitis E virus is a worldwide emerging foodborne pathogen; raw or undercooked meats and liver pork products can cause infection through the orofecal route. In Central-Southern Italy, small traditional farming method, associated with the possibility of environmental sharing with wild species, can facilitate HEV diffusion and persistence. The aim of this study was to determine HEV genotype and subtype in Marche region from home slaughtered domestic pigs involved in small and traditional food chains. A total of 236 liver and muscle tissues and 6 pooled salami samples were screened. Laboratory workflow started with homogenization, followed by RNA extraction. Nested reverse transcription PCR and qRT-PCR were used to amplify specific parts of overlapping open reading frames belonging to the HEV genome. A total of 42/236 (17.79%) liver and 8/236 (3.39%) diaphragm specimens were positive; none of the pooled salami specimens showed positive HEV signal. The discovered HEV3c presented high nucleotide similarities with ones amplified from wild boar populations hunted in the same province. Extensive farming methods and environmental sharing with wild animal species support cross-infection infections, as observed in the present study. Although salami resulted negative for HEV RNA detection, the effects of food technologies on viral loads remain unclear. Therefore, further scientific investigations coupled with efficacious standardized laboratory procedures will be the next challenge.

Multidrug resistant Vibrio spp. identified from mussels farmed for human consumption in Central Italy.

AIMS: This study investigated phenotypic and genotypic antimicrobial resistance profiles of Vibrio strains identified from Mytilus galloprovincialis farmed for human consumption in the Adriatic Sea Central Italy. METHODS AND RESULTS: A total of 475 mussels (M. galloprovincialis) were involved in the present study, and culture-dependent microbiological methods permitted to identify a total of 50 Vibrio strains that were tested for antibiotic susceptibility followed by the genetic determinant detections. Antibiograms showed resistance against ampicillin (36.0%), amoxicillin-clavulanic acid (30.0%), gentamycin (14.0%), and imipenem (18.0%). Biomolecular assays amplified a total of 264 antibiotic resistance genes harbored by both susceptible and resistant Vibrio species. Among resistance genes, aacC2 (62.0%) and aadA (58.0%) for aminoglycosides, blaTEM (54.0%) for beta-lactams, qnrS (24.0%) for quinolones, tetD (66.0%) for tetracyclines, and vanB (60.0%) for glycopeptides were mainly amplified by PCR assays. CONCLUSIONS: Vibrio genus is involved in the antibiotic resistance phenomenon diffusion in the aquatic environments, as demonstrated by the harboring of many genetic determinants representing a kind of genetic "dark world".

Microplastics in Sepia officinalis caught on the central Adriatic coast: preliminary results.

Microplastics (µPs) represent an emerging problem for the marine environment given their wide bioavailability for all aquatic organisms, from zooplankton to top predators. This work aimed to evaluate a method of extracting microplastics from the gastrointestinal tract of 122 Sepia officinalis caught from the Adriatic Sea (along Abruzzo region coasts) to measure its quantity in a poorly investigated species. The extraction method used for gastrointestinal content was performed using 10% potassium hydroxide. In 98/122 (80.32-95% CI=73.27-87.37%) wild animals microplastics were detected with a mean concentration of 6.82±5.52 µPs/subject. Among the fragments, as supported by various authors, those of black color were the most represented ones; however, also blue fibers and transparent spheres were isolated. This study, in agreement with previous ones, poses further attention to the wide microplastic diffusion in the marine environment (surface, columns, sediments, and animals). The obtained results provide the basis for future investigations on this public health concern.

ARGs Detection in Listeria Monocytogenes Strains Isolated from the Atlantic Salmon (Salmo salar) Food Industry: A Retrospective Study.

Among bacterial foodborne pathogens, Listeria monocytogenes represents one of the most important public health concerns in seafood industries. This study was designed as a retrospective study which aimed to investigate the trend of antibiotic resistance genes (ARGs) circulation in L. monocytogenes isolates identified (in the last 15 years) from Atlantic salmon (Salmo salar) fresh and smoked fillets and environmental samples. For these purposes, biomolecular assays were performed on 120 L. monocytogenes strains collected in certain years and compared to the contemporary scientific literature. A total of 52.50% (95% CI: 43.57-61.43%) of these samples were resistant to at least one antibiotic class, and 20.83% (95% CI: 13.57-28.09%) were classified as multidrug resistant. Concerning ARGs circulation, tetracycline (tetC, tetD, tetK, tetL, tetS), aminoglycoside (aadA, strA, aacC2, aphA1, aphA2), macrolide (cmlA1, catI, catII), and oxazolidinone (cfr, optrA, poxtA) gene determinants were majorly amplified. This study highlights the consistent ARGs circulation from fresh and processed finfish products and environmental samples, discovering resistance to the so-called critical important antimicrobials (CIA) since 2007. The obtained ARGs circulation data highlight the consistent increase in their diffusion when compared to similar contemporary investigations. This scenario emerges as the result of decades of improper antimicrobial administration in human and veterinary medicine.

A Quality Index Method-based evaluation of sensory quality of red mullet (Mullus barbatus) and its shelf-life determination.

Every day the consumer must choose a food product rather than others based on its quality; for fishery products the main quality parameter is freshness. Implementation of the Quality Index Method (QIM) in the fish industry provides information on fish quality; therefore, it is important for effective quality and process management in the production of high-quality fish products. The present study aims to validate the shelflife study of fresh red mullet (Mullus barbatus) stored in ice previously presented by Özyurt in 2009 through Torry scheme in cooked filet and microbiological evaluation. Next, this revised scheme was applied to determine the shelf-life. It included seven descriptors and varies from 0 to 15 points. The use of a modified QIM scheme showed a good correlation between the quality index and days of storage time, with a R2 value of 0.9698. In fact, all the subjects examined reached the end of their commercial life, or the day of sensory rejection, in 9-10 days with a Quality Index of 13.83.

Hepatitis E Virus RNA Detection from Hunted Wild Boars in Central Italy: an Epidemiological Investigation.

Every year, foodborne pathogens, including the hepatitis E virus (HEV), cause thousands of infections in different continents. Final consumers become infected through the ingestion of contaminated animal origin foodstuffs. Generally, in industrialized countries, HEV genotype 3 is involved in sporadic outbreaks. Infections have been described, in Europe and Japan as consequence of pork products and contaminated wild boar's primary or processed products (liver and muscle tissues) consumption. In Central Italy, hunting activities are largely practiced. In these small and rural communities, game meat and liver are ingested by hunters' families or at local and traditional restaurants. Therefore, these food chains can be considered critical HEV reservoirs. In this study, 506 liver and diaphragm tissues were collected from hunted wild boars in the Southern Marche region (Central Italy) and were screened for HEV RNA detection. From the 10.87% of liver and 2.76% of muscle samples, HEV3 subtype c was discovered. The observed prevalence values resulted in line with previous investigations performed in other Central Italian regions, but higher than Northern ones (3.7% and 1.9% from liver tissue). Therefore, the obtained epidemiological data highlighted the wide occurrence of HEV RNA circulation in a low-investigated area. Basing on results, a One-health approach was adopted due to the sanitary relevance of this Public Health concern.

Antibiotic Resistance Profiles and ARG Detection from Isolated Bacteria in a Culture-Dependent Study at the Codfish Industry Level.

The antibiotic resistance phenomenon horizontally involves numerous bacteria cultured from fresh or processed seafood matrix microbiomes. In this study, the identified bacteria from food-producing processes and industrial environments were screened for phenotypic and genotypic resistance determinants. A total of 684 bacterial strains [537 from processed codfish (Gadus morhua and Gadus macrocephalus) products as salted and seasoned and soaked and 147 from environmental samples] were isolated. Antibiotic susceptibility tests showed resistance against tetracycline, oxacillin, and clindamycin in the Staphylococcus genus (both from food and environmental samples) and against beta-lactams (cefotaxime, carbapenems, etc.) and nitrofurans (nitrofurantoin) from E. coli and Salmonella enterica serovar. Enteritidis isolates. One-thousand and ten genetic determinants-tetracycline tetC (25.17%), tetK (21.06%), tetL (11.70%), clindamycin ermC (17.23%), ermB (7.60%), linezolid cfr (8.22%), optrA (3.62%), poxtA (2.05%), and oxacillin mecA (17.37%)-were amplified from Gram-positive resistant and phenotypically susceptible bacteria. Concerning Gram-negative bacteria, the beta-lactam-resistant genes (blaTEM, blaCIT, blaCTX-M, blaIMP, blaKPC, blaOXA-48-like) represented 57.30% of the amplified ARGs. This study found high antibiotic resistance genes in circulation in the fish food industry chain from the macro- to microenvironment. The obtained data confirmed the diffusion of the "antibiotic resistance phenomenon" and its repercussions on the One-health and food-producing systems.

Ultrasound Technology as Inactivation Method for Foodborne Pathogens: A Review.

An efficient microbiological decontamination protocol is required to guarantee safe food products for the final consumer to avoid foodborne illnesses. Ultrasound and non-thermal technology combinations represent innovative methods adopted by the food industry for food preservation and safety. Ultrasound power is commonly used with a frequency between 20 and 100 kHz to obtain an "exploit cavitation effect". Microbial inactivation via ultrasound derives from cell wall damage, the oxidation of intracellular amino acids and DNA changing material. As an inactivation method, it is evaluated alone and combined with other non-thermal technologies. The evidence shows that ultrasound is an important green technology that has a good decontamination effect and can improve the shelf-life of products. This review aims to describe the applicability of ultrasound in the food industry focusing on microbiological decontamination, reducing bacterial alterations caused by food spoilage strains and relative foodborne intoxication/infection.

Shelf Life and Safety of Vacuum Packed HPP-Treated Soaked Cod Fillets: Effects of Salt Content and Multilayer Plastic Film.

High microbiological quality standards, food safety, and environmental sustainability represent crucial topics in food production chains. For this reason, fish industries, which import salted and seasoned fish products from supplier countries, i.e., Norway, Denmark, USA (Alaska State), etc., have tried to reduce the salt content of each carton during transportation (reducing carbon emissions and the weight of major quantities of transported fish). In the present study, 360 differently processed fish fillet samples, belonging to the species Gadus macrocephalus caught in FAO zone 67, were microbiologically and chemically screened. This study aimed to provide original data concerning the applicability of sustainable solutions investigating the combined effects of salt content reduction combined with new recyclable multilayer plastic film packaging (vacuum skin packaging with two different oxygen transmission rate values). The microbiological results showed no substantial changes comparing the two differently salted products, highlighting their high hygienic characteristics which were also observed in their chemical analysis. The shelf life evolutions (comparing the two different studied plastic films) highlighted that, after 35 days from HPP treatments, bacterial loads gained high values, over 6 log cfu/g. This study highlights that, compared to the currently used plastic films, the results of the new and sustainable multilayer plastic films show that they can provide safe food matrices in combination with HPP technologies. Therefore, this preliminary investigation brings closer attention to alternative and environmentally sustainable production systems with their designs based on the multidisciplinary approach of food production systems.

Trichinella spp. detection in hunted wild boar (Sus scrofa) diaphragm biopsies in Central Italy.

Trichinellosis is a globally diffused foodborne parasitic disease caused by nematodes of the Trichinella complex. During evolution, guided by ecological interactions, natural selection, and biochemical "intelligence", these parasites developed admirable strategies to infect the host's organism. One of the most fascinating is represented by the nurse cell formation in muscular tissue (e.g., diaphragm, skeletal muscle, extrinsic ocular muscles, etc.). This strategy allowed the parasite to adapt and conquer the wider host species spectrum, including ungulates and humans. Consumption of undercooked meat from infected wild ungulates constitutes the most important source of infection for the human species. In this study, we show the prevalence of Trichinella spp. in hunted wild boars (Sus scrofa) in Central Italy. During the hunting season 2021/2022 in the province of Rieti, 554 wild boar diaphragm biopsies were collected for Trichinella spp. screening, in accordance with Regulation EU 1375/2015. An artificial digestion method was used for the detection of Trichinella spp. larval forms. The results revealed a positivity of 0.18% (1/554), and molecular biology identification demonstrated the presence of Trichinella britovi in the positive sample. This species is the most diffused in wild ungulate populations in Central Italy and the most frequently isolated in human patients with trichinellosis from this area, showing a close epidemiological relation between Homo sapiens and Sus scrofa for Trichinella spp. diffusion in an ecosystem. Epidemiological surveillance, in receptive animal species destined for human consumption and at any One Health level, represents the main "winning" strategy in the control of this worldwide, widespread foodborne parasitic disease.

Antibiotic Resistance in the Finfish Aquaculture Industry: A Review.

Significant challenges to worldwide sustainable food production continue to arise from environmental change and consistent population growth. In order to meet increasing demand, fish production industries are encouraged to maintain high growth densities and to rely on antibiotic intervention throughout all stages of development. The inappropriate administering of antibiotics over time introduces selective pressure, allowing the survival of resistant bacterial strains through adaptive pathways involving transferable nucleotide sequences (i.e., plasmids). This is one of the essential mechanisms of antibiotic resistance development in food production systems. This review article focuses on the main international regulations and governing the administering of antibiotics in finfish husbandry and summarizes recent data regarding the distribution of bacterial resistance in the finfish aquaculture food production chain. The second part of this review examines promising alternative approaches to finfish production, sustainable farming techniques, and vaccination that circumvents excessive antibiotic use, including new animal welfare measures. Then, we reflect on recent adaptations to increasingly interdisciplinary perspectives in the field and their greater alignment with the One Health initiative.

Intradiaphragmatic abscesses in a wild boar (Sus scrofa): Inspective implications based on anatomopathological evidences.

The intradiaphragmatic localization of an abscess is rarely described in humans and in other animal domestic and wild species, and can be caused by penetrative traumas (i.e., firearm injuries). Here we describe two intradiaphragmatic abscesses in a hunted adult male wild boar (Sus scrofa) pluck, associated with adhesion phenomena with the contiguous anatomical structures (pleural, phrenic, and glissonian serosas) and observed during the post mortem inspection, in accordance with the Reg. EU 627/2019. One of these lesions also presented a phreno-abdominal fistula. We found in cytopathological evaluation of the neoformations' content the presence of spheroidal bacterial soma, characterized by linearly concatenated "Streptococcus-like" aggregation pattern. Furthermore, microbiological assays revealed a polymicrobial pattern characterized by the presence of telluric microorganisms, some of which have a marked pyogenic action (Streptococcus suis type I, Sphingomonas paucimobilis, Carnobacterium divergens, and Lactobacillus sakei). Our results and collected evidence demonstrate the pathogenetic hypothesis of bacterial contamination secondary to penetrative trauma caused by a not-mortal projectile's wound, defining the inspective behaviour according to the cogent legislation. These lesions, in the reason of their potential relation to toxemia, bacteremia and septicemia phenomena, represent a sanitary risk that impose, from a normative point of view, the total condemnation of the carcass. These inspective implications, originating from the observation and interdisciplinary description of the anatomopathological and microbiological features of the lesions, are directly projected on the normative scenario, providing useful elements to guarantee the health of the consumer.

Hepatitis E Virus Detection in Hunted Wild Boar Liver and Muscle Tissues in Central Italy.

In different European countries, including Italy, hepatitis E virus (HEV) has been recognized as an emerging public health concern. Humans are infected through the orofecal route by the ingestion of contaminated uncooked or undercooked animal-origin foodstuffs. Wild boars (Sus scrofa) have gained a crucial role as viral reservoirs. HEV-3 is the most frequently identified genotype from hunted wild boar liver and muscle tissues. The Marche region, more specifically Ascoli Piceno province, is characterized by a rooted hunting tradition and related product consumption. In this research study, 312 liver and 296 muscle specimens were screened using biomolecular assays, and HEV RNA was detected from 5.45% and 1.35% of liver and muscle samples, respectively. Phylogenetic analysis revealed that positive animals were infected by genotype 3 subtype c. Based on the environmental pathogen characteristics, HEV has also evolved to guarantee its survival in a wild environment. Therefore, wild boars and ruminants have a key role in its persistence. Epidemiological data regarding HEV circulation have resulted as necessary, and biomolecular analysis represents an important means of monitoring and establishing preventive measures. A multidisciplinary approach could provide a wide perspective regarding HEV and infectious implications on human, animal, and environmental health.

Hepatitis E virus detection in hunted wild boar (Sus scrofa) livers in Central Italy.

Hepatitis E virus (HEV) is a zoonotic pathogen, responsible for numerous cases of infection in humans. Transmission occurs through the orofecal route, and ingestion of contaminated foods represents an important risk factor for final consumer's health. Wild animal species, in particular wild boar (Sus scrofa), are the main virus reservoirs; liver is the target organ, from which, through the hematic diffusion, HEV reaches different tissues and organs, as muscular one. The hygienic-sanitary critical issues connected with game meat food chain in general, and particularly wild boar, with special regards to any geographical area where this animal species can be directly in contact with humans, domestic ones (i.e., domestic pig), and other wild reservoirs (i.e., wild ruminants), finds favorable environmental conditions, have induced us to conduce the present scientific investigation. During the hunting season 2019/2020, a total of 156 wild boar livers were collected from provided plucks at slaughterhouse in Ascoli Piceno. Nested RT-PCR was used for the viral RNA detection. Results demonstrated a positivity of 5.12% (8/156), and the circulation in the screened area of genotype 3 subtype c, which is frequently identified in Central Italy. HEV sanitary relevance and the emerging role of any food chains in its transmission impose further detailed studies. The molecular screening of hunted wild boars' livers can provide important information about virus's circulation in wild animal populations in a specific area.

Hepatitis E Virus in the Food of Animal Origin: A Review.

Hepatitis E virus (HEV) is a cosmopolitan foodborne pathogen. The viral agent infects humans through the consumption of contaminated food (uncooked or undercooked). Most cases of infection are asymptomatic and for this reason, this pathology is considered underdiagnosed. Domestic and wild animals are considered natural reservoirs: that is, domestic pig, wild boar, sheep, goat, deer, rabbit, and so on. Therefore, various work categories are at risk: that is, veterinarians, farmers, hunters, slaughterhouse workers, and so on. In these last decades, researchers found a high percentage of positivity to the molecular viral detection in several food matrices included: ready-to-eat products, processed meat products, milk, and shellfish. This review aims to provide an international scenario regarding HEV ribonucleic acid (RNA) detection in several foodstuffs. From this investigative perspective, the study aims to highlight various gaps of the current knowledge about technologies treatments' impact on viral loads. The purpose was also to provide an innovative point of view "One Health"-based, pointing out the strategic role of environmental safety.