Food Safety Assessment and Nutraceutical Outcomes of Dairy By-Products: Ovine Milk Whey as Wound Repair Enhancer on Injured Human Primary Gingival Fibroblasts.

The valorization of milk whey appears to be a promising strategy for managing by-products from dairy food industries, which incur demanding economic costs for treatment and/or disposal. Thanks to its numerous bioactive components, whey is expected to be increasingly incorporated into foods in the future. We investigated the safety of ovine milk whey through in vitro experiments on human primary gingival fibroblast (HGF-1) proliferation and wound healing. Fibroblasts play a crucial role in the repair processes from the late inflammatory phase until the final stages. Cells treated with varying concentrations of ovine whey (0.01%, 0.1%, 1%, and 10%) were able to close wounds more rapidly than vehicle-treated cells. Time- and dose-dependent responses were observed in cell populations exposed to ovine whey. Specifically, wounds treated with 0.1% and 10% milk whey showed better migratory capabilities compared to those treated with 0.01% and 1% milk whey after 24 and 48 h. In addition, ovine milk whey stimulates extracellular matrix deposition, as evidenced by the increasing levels of CD44 antigen density evaluated through FACS analysis, as well as COL1A1 expression measured both via RT-qPCR and immunofluorescence. This phenomenon was particularly evident at concentrations of 0.01% and 10%. Ensuring quality and safety has become a major concern for health authorities in the food industry. Our findings suggest that ovine milk whey is safe and possesses regenerative properties. It facilitates tissue re-establishment following exposure to environmental stress, particularly accelerating gingival wound closure.

Effect of a new sustainable cooling system used during firming and brining on the microbiological, chemical, and sensory characteristics of buffalo mozzarella cheese.

The cooling applied during the firming and brining processes represents an important production step in mozzarella cheese-making. The temperature fluctuations of the cooling water can negatively affect the hygiene, composition, and quality of mozzarella. Some sustainable cooling systems can minimize this problem by using hot process fluids as heat sources to generate refrigerated energy. This study aimed to evaluate the effects of a new cooling system equipped with a water-ammonia absorption chiller (MA) on the characteristics of buffalo mozzarella through a comparative study with products cooled with a traditional ice water chiller (MT). The buffalo mozzarella cheese manufacture was monitored, and the samples were analyzed for chemical, nutritional, microbiological, and sensory characteristics. The MT samples showed an overall weight loss of 7.4% compared to an average of 2.8% for the MA samples. The MT samples were characterized by greater sapidity than the MA ones, which instead showed a higher moisture content that increased juiciness. The microbiological analysis showed a lower concentration of mesophilic bacterial load in the MA samples than in the MT ones [difference of 1 Log (CFU/g)], which is probably due to the low and constant temperatures that reduced the permanence time of the mozzarella in the vats (firming and brining). This study represents a preliminary positive evaluation of the use of this sustainable cooling system for mozzarella cheese, which is useful for dairy plants with an annual cheese production volume sufficient to justify the operating cost of the plant and the annual energy cost.

The Sous Vide Cooking of Mediterranean Mussel (Mytilus galloprovincialis): Safety and Quality Assessment.

This study involves an investigation of the effects of various cooking temperatures, freeze-thaw processes, and food preservatives on the quality and shelf-life of sous vide Mediterranean mussels. Cooking temperatures of 80 °C or above significantly improved the microbiological quality, with bacterial counts remaining within the acceptability range for human consumption even after 21 days of refrigerated storage. Fast freezing followed by slow thawing preserved the highest moisture content, potentially improving texture. Sensory analysis revealed that refrigerated sous vide mussels maintained a comparable taste to freshly cooked samples. Frozen samples reheated via microwaving exhibited more intense flavour than pan-reheated or fresh mussels. Food additives, including citric acid, potassium benzoate, and potassium sorbate, alone or in combination with grape seed oil, significantly reduced total volatile basic nitrogen and thiobarbituric acid-reactive substances during 28 days of storage, indicating decreased spoilage and lipid oxidation. Mussels with a combination of these additives registered a nitrogen content as low as 22 mg of N/100g after 28 days, well below the limit of acceptability (<35 mg of N/100g). Food additives also inhibited bacterial growth, with mesophilic bacteria count below 3.35 Log CFU/g after 28 days, compared with 5.37 Log CFU/g in control samples. This study provides valuable insights for developing optimal cooking and preservation methods for sous vide cooked seafood, underscoring the need for further research on optimal cooking and freeze-thaw protocols for various seafood types.

Utilization of Dairy By-Products as a Source of Functional and Health Compounds-The Role of Ovine Colostrum and Milk Whey on Chronic Myeloid Leukemia Cells.

Nowadays, the search for food products that promote consumers' health has gained interest, and dairy by-products, due to their biological quality, could have a prominent position among products with health benefits. However, little is known about their activity on cancer cells. This study aimed to provide evidence about the effect of ovine colostrum and milk whey on K562 cells, a model of the human chronic myeloid leukemia cell line. The exposure of K562 cells to a single administration of sheep by-products at different concentrations for three days and three treatments for three days was carried out. Using a flow cytometric approach, we found that CD235a expression remained stable in the cells exposed to ovine whey (milk and colostrum) at concentrations ranging from 1 ng/mL to 100 µg/mL, after three days from one or three administrations, respectively. A significant reduction in fluorescent cells was observed in the populations exposed to 1 mg/mL of both milk and colostrum at the same time points. In these conditions, the size and granularity of the leukemic cells also changed, with a substantial reduction in the number of actively dividing cells in the S phase of the cell cycle. This phenomenon was highlighted by the Annexin V/PI cytofluorimetric test, which is able to provide quantitative results regarding the population of cells in early or late apoptosis or necrotic cells after exposure to a single dose or three doses of colostrum or sheep whey for three days, respectively. This report showed that both colostrum and milk whey were able to modify the phenotypic profile and cell cycle of the K562 cell line, inducing apoptosis at the highest concentration.

Role of Feeding and Novel Ripening System to Enhance the Quality and Production Sustainability of Curd Buffalo Cheeses.

The buffalo dairy sector is extending its boundaries to include new buffalo cheese productions beyond mozzarella, overcoming some barriers that make cheeses expensive and unsustainable. This study aimed to evaluate the effects of both the inclusion of green feed in the diet of Italian Mediterranean buffaloes and an innovative ripening system on buffalo cheese quality, providing solutions capable of guaranteeing the production of nutritionally competitive and sustainable products. For this purpose, chemical, rheological, and microbiological analyses were carried out on cheeses. Buffaloes were fed with or without the inclusion of green forage. Their milk was used to produce dry ricotta and semi-hard cheeses, ripened according to both respective traditional (MT) and innovative methods (MI); these are based on automatic adjustments of climatic recipe guided by the continuous control of pH. Green feed enhances the nutritional profile of the final products (high content of MUFAs and PUFAs). As far as the ripening method is concerned, to our knowledge, this is the first study that tests aging chambers, commonly used for meat, for the maturing of buffalo cheeses. Results pointed out the MI validity also in this field of application, as it shortens the ripening period without negatively compromising any of desirable physicochemical properties and the safety and hygiene of the final products. Conclusively, this research highlights the benefits of diets rich in green forage on productions and provides support for the ripening optimization of buffalo semi-hard cheeses.

Trichinella britovi in wild boar meat from Italy, 2015-2021: A citizen science approach to surveillance.

As a result of the increase of game meat intended for human consumption through Europe, a plethora of food-borne diseases, including trichinellosis, may occur in consumers, posing a relevant public health threat. Thus, this study aims to a citizen science approach to monitor the occurrence of Trichinella spp. in wild boar meat intended for human consumption, evaluating the risk of infection for consumers. Following the European Regulation 2015/1375 (laying down specific rules on official controls for Trichinella in meat), from 2015 to 2021, hunters (n = 478) were involved to collect diaphragm pillar samples of wild boars from mainland southern Italy, which were tested for Trichinella spp. L1 larvae via HCl-pepsin digestion and Multiplex PCR. Overall, 139,160 animals were collected (average of 19,880 per year), being 14 (i.e., 0.01%) tested positive to Trichinella britovi by the combined biochemical and molecular approach. An average larval burden of 28.4 L1 per gram of meat was found (minimum 3.2 - maximum 132.6). A statistically significant difference was found in the prevalence according to hunting seasons (p < 0.01, with higher values in 2016 and 2021) and regions of the study area (p < 0.01). No statistically significant decrease in the prevalence of T. britovi throughout the study period was found (p = 0.51), except in Apulia region (p < 0.01). These findings revealed a stable prevalence of T. britovi in wild boar meat intended for human consumption, suggesting a risk of infection for consumers, especially hunters and local markets users. Citizen science surveillance models could be promoted to improve trichinellosis control and prevention in a One Health perspective.

The Antimicrobial Peptide 1018-K6 Interacts Distinctly with Eukaryotic and Bacterial Membranes, the Basis of Its Specificity and Bactericidal Activity.

Since penicillin was discovered, antibiotics have been critical in the fight against infections. However, antibiotic misuse has led to drug resistance, which now constitutes a serious health problem. In this context, antimicrobial peptides (AMPs) constitute a natural group of short proteins, varying in structure and length, that act against certain types of bacterial pathogens. The antimicrobial peptide 1018-K6 (VRLIVKVRIWRR- NH2) has significant bactericidal and antibiofilm activity against Listeria monocytogenes isolates, and against different strains and serotypes of Salmonella. Here, the mechanism of action of 1018-K6 was explored further to understand the peptide-membrane interactions relevant to its activity, and to define their determinants. We combined studies with model synthetic membranes (liposomes) and model biological membranes, assessing the absorption maximum and the quenching of 1018-K6 fluorescence in aqueous and lipid environments, the self-quenching of carboxyfluorescein, as well as performing lipid sedimentation assays. The data obtained reflect the differential interactions of the 1018-K6 peptide with eukaryotic and prokaryotic membranes, and the specific interactions and mechanisms of action in the three prokaryotic species studied: Salmonella Typhimurium2GN, Escherichia coli3GN, and Staphylococcus aureus3GP. The AMP 1018-K6 is a candidate to prevent (food preservation) or treat (antibiotic use) infections caused by certain pathogenic bacteria, especially some that are resistant to current antibiotics.

Effect of novel active packaging containing antimicrobial peptide on the shelf-life of fish burgers (Coryphaena hippurus) during refrigerated storage.

Fresh fishery products are highly perishable foods mainly due to their high-water content and high level of pH which act as promoters of spoilage processes. In these matrices, the deterioration phenomena are the result of the action of oxidative, and enzymatic processes due in part to the presence of specific microorganisms. Indeed, the microbial communities responsible for spoilage are a small fraction of the flora detectable in the fish and are known as specific spoilage organisms (SSOs). In the last decades, the scientific community has worked to achieve the ambitious goal of reducing the impact of microbial deterioration on food losses through innovative solutions, including antimicrobial packaging. The goal of this study was to evaluate the efficacy of an active polypropylene (PP)- based packaging functionalized with the antimicrobial peptide 1018K6 to extend the shelf life of dolphinfish burgers (Coryphaena hippurus) by evaluating its effect on sensorial and microbiological profile. The microbiological results showed an evident antimicrobial activity of the active packaging against hygiene indicator microorganisms and SSOs, recording a reduction of about 1 Log (CFU/g) of their concentrations compared to those of the control groups. Furthermore, a significant influence of functionalized packaging on the organoleptic characteristics was noted, accentuating the differences in freshness between the two experimental groups. This work confirmed the hypothesis of considering antimicrobial packaging as a potential tool capable of slowing down surface microbial replication and, therefore, extending the shelf-life and improving the health and hygiene aspect of fresh fish products.

Recombinant Expression of Archaeal Superoxide Dismutases in Plant Cell Cultures: A Sustainable Solution with Potential Application in the Food Industry.

Superoxide dismutase (SOD) is a fundamental antioxidant enzyme that neutralises superoxide ions, one of the main reactive oxygen species (ROS). Extremophile organisms possess enzymes that offer high stability and catalytic performances under a wide range of conditions, thus representing an exceptional source of biocatalysts useful for industrial processes. In this study, SODs from the thermo-halophilic Aeropyrum pernix (SODAp) and the thermo-acidophilic Saccharolobus solfataricus (SODSs) were heterologously expressed in transgenic tomato cell cultures. Cell extracts enriched with SODAp and SODSs showed a remarkable resistance to salt and low pHs, respectively, together with optimal activity at high temperatures. Moreover, the treatment of tuna fillets with SODAp-extracts induced an extension of the shelf-life of this product without resorting to the use of illicit substances. The results suggested that the recombinant plant extracts enriched with the extremozymes could find potential applications as dietary supplements in the nutrition sector or as additives in the food preservation area, representing a more natural and appealing alternative to chemical preservatives for the market.

An Active Peptide-Based Packaging System to Improve the Freshness and Safety of Fish Products: A Case Study.

Fresh fish are highly perishable, owing mainly to their moisture content, high amount of free amino acids and polyunsaturated fatty acids. Microorganisms and chemical reactions cause the spoilage, leading to loss in quality, human health risks and a market value reduction. Therefore, the fishing industry has always been willing to explore new technologies to increase quality and safety of fish products through a decrease of the microbiological and biochemical damage. In this context, antimicrobial active packaging is one such promising solution to meet consumer demands. The main objective of this study was to evaluate the effects of an active polypropylene-based packaging functionalized with the antimicrobial peptide 1018K6 on microbial growth, physicochemical properties and the sensory attributes of raw salmon fillets. The results showed that application of 1018K6-polypropylene strongly inhibited the microbial growth of both pathogenic and specific spoilage organisms (SSOs) on fish fillets after 7 days. Moreover, salmon also kept its freshness as per volatile chemical spoilage indices (CSIs) during storage. Similar results were obtained on hamburgers of Sarda sarda performing the same analyses. This work provides further evidence that 1018K6-polymers have good potential as antimicrobial packaging for application in the food market to enhance quality and preserve the sensorial properties of fish products.

Surveillance of Human Cases of Salmonellosis, Campylobacteriosis, Listeriosis, and Hepatitis A in Campania (Southern Italy): Seven-Year Monitoring (2013-2019).

Foodborne infections cause illness and death every year worldwide. The aim of this study was to describe trends in 2013-2019 in the occurrence of human cases of salmonellosis, campylobacteriosis, listeriosis, and hepatitis A in the Campania region. Human case data were provided by the National Surveillance System of disease and were grouped by year, province, age group, and sex. Moreover, the number of people hospitalized was recorded. In the Campania region, the total number of confirmed human cases for the diseases investigated was 1924, with Hepatitis A and the Salmonellosis as the first most reported (1009 and 825 cases, respectively). The incidence rates of gastroenteritis under study were lower than those in Italy and European Union in the same period, with the exception of Hepatitis A whose incidence was higher than that recorded in Italy. Data on hospitalizations pointed out the onset of severe forms of infection also for listeriosis and campylobacteriosis, whose incidence was very low (27 and 63 cases, respectively). Unfortunately, no information on the foods implicated is available. Although probably underestimated, gastroenteritis due to foodborne agents still represents a burden in Campania, and continuous monitoring and implementation of the currently available regional surveillance system is required.

Effects of Different Levels of Inclusion of Apulo-Calabrese Pig Meat on Microbiological, Physicochemical and Rheological Parameters of Salami during Ripening.

This study focused on the characterization of salami produced with meat from different pig breeds. The aim consisted in evaluating the added value of the inclusion of Apulo-Calabrese meat in the production of salami, which was characterized by production until the end of maturation (1, 30, 60, and 120 days). The experimental design involved three types of salami, two of which were produced by partial inclusion of 50 and 75% of the Italian breed pork meat (S50 and S75, respectively). Physicochemical (pH, aw, fatty acid analysis, and malondialdehyde concentration), rheological parameters (texture analyses and color measurement), and bacterial biodiversity were evaluated. Results showed that the partial inclusion of Apulo-Calabrese meat influences the fatty acid profile of final products, which were characterized by a higher percentage of monounsaturated fatty acids compared to traditional salami; however, due to the high content of unsaturated fatty acids, S50 and S75 showed higher values of secondary lipid oxidation up to the 120th day. The linoleic and palmitic acid content significantly affected hardness and brightness. Overall, the ripening process was able to control the microbiological profile and the S50 formulation appeared as a suitable choice that could satisfy consumers for nutritional expectations and sensory profiles.

The complete mitochondrial genome of the white seabream Diplodus sargus (Perciformes: Sparidae) from the Tyrrhenian sea.

The white seabream Diplodus sargus (Linnaeus, 1758) is a species of interest for commercial fisheries throughout its range of distribution and it is also reared using aquaculture techniques. Herein, we present the first complete sequence and annotation of the mitochondrial genome of this species. The D. sargus mitogenome is 16,515 base pairs in length and contains 13 protein-coding genes, 2 rRNA, 22 tRNA, and 2 non-coding regions (D-loop and L-origin). The overall nucleotide composition is: 27.3% A, 28.9% C, 26.8% T, and 17.0% G. Maximum likelihood analyses placed D. sargus as a sister species of Diplodus puntazzo. This study provides valuable information for further studying identification methods and evolutionary relationships of Sparidae species.

Monitoring Cyanobacterial Blooms during the COVID-19 Pandemic in Campania, Italy: The Case of Lake Avernus.

Cyanobacteria are ubiquitous photosynthetic microorganisms considered as important contributors to the formation of Earth's atmosphere and to the process of nitrogen fixation. However, they are also frequently associated with toxic blooms, named cyanobacterial harmful algal blooms (cyanoHABs). This paper reports on an unusual out-of-season cyanoHAB and its dynamics during the COVID-19 pandemic, in Lake Avernus, South Italy. Fast detection strategy (FDS) was used to assess this phenomenon, through the integration of satellite imagery and biomolecular investigation of the environmental samples. Data obtained unveiled a widespread Microcystis sp. bloom in February 2020 (i.e., winter season in Italy), which completely disappeared at the end of the following COVID-19 lockdown, when almost all urban activities were suspended. Due to potential harmfulness of cyanoHABs, crude extracts from the "winter bloom" were evaluated for their cytotoxicity in two different human cell lines, namely normal dermal fibroblasts (NHDF) and breast adenocarcinoma cells (MCF-7). The chloroform extract was shown to exert the highest cytotoxic activity, which has been correlated to the presence of cyanotoxins, i.e., microcystins, micropeptins, anabaenopeptins, and aeruginopeptins, detected by molecular networking analysis of liquid chromatography tandem mass spectrometry (LC-MS/MS) data.

A Study on the Antimicrobial and Antibiofilm Peptide 1018-K6 as Potential Alternative to Antibiotics against Food-Pathogen Salmonella enterica.

Antimicrobial resistance has become one of the major global public health concerns, and it is indispensable to search for alternatives to conventional antibiotics. Recently, antimicrobial peptides have received great attention because of their broad-spectrum antimicrobial activity at relatively low concentrations, even against pathogens such as Salmonella enterica, which is responsible for most food-borne illnesses. This work aimed at evaluating the antimicrobial and antibiofilm activity of the innate defense peptide, named 1018-K6, against S. enterica. A total of 42 strains, belonging to three different subspecies and 32 serotypes, were included in this study. The antibiotic resistance profile of all the strains and the cytotoxic effects of 1018-K6 on mammalian fibroblast cells were also investigated. Results revealed that MIC (minimum inhibitory concentrations) and MBC (minimum bactericidal concentrations) values were in the ranges of 8-64 µg/mL and 16-128 µg/mL, respectively, although most strains (97%) showed MICs between 16 and 32 µg/mL. Moreover, sub-inhibitory concentrations of 1018-K6 strongly reduced the biofilm formation in several S. enterica strains, whatever the initial inoculum size. Our results demonstrated that 1018-K6 is able to control and manage S. enterica growth with a large potential for applications in the fields of active packaging and water disinfectants.

The Influence of Broilers' Body Weight on the Efficiency of Electrical Stunning and Meat Quality under Field Conditions.

Water-bath stunning represents the most-applied stunning system in poultry slaughtering, but within the European Union, specific indications on electric parameters that should be used, such as voltage, are missing. The objective of this study was to evaluate the efficiency of two commercially available types of electrical equipment (A and B) on broilers with different live body weights and the influence of the tested parameters on meat quality. Experimental trials in a European Union-approved slaughterhouse were carried out using two different stunners. 6600 broilers, divided into three weight groups, were stunned applying different protocols based on the same current frequencies and intensity but different voltages. The state of unconsciousness (presence of corneal reflex and wings flapping) and post-mortem defects (pectoral hemorrhages and dark meat) were evaluated by blinded trained operators. The presence of corneal reflex and petechiae were the most reported consciousness signs and post-mortem injuries, respectively. Different weights played an important role within stunner A, registering statistical differences (p < 0.01) among groups. Considering injuries, an inverse relationship between body weight and lesions was found. The results highlighted the effectiveness of both stunning systems applying the best combination of electrical parameters considering the weight of the animal and ensuring its well-being.

Effect of beetroot (Beta vulgaris) extract on Black Angus burgers shelf life.

Beef burgers are meat preparations with easy perishability. To ensure a longer shelflife, the Regulation EU 1129/11 allows the use of some additives. However, healthconscious consumers prefer products which do not contain synthetic substances. Aim of the present study was to evaluate the effect of Red Beetroot (Beta vulgaris) integration on Black Angus made burgers shelf life. Red beet was prepared as powder and added to meat mixture as the same or in water solution. The study was split into 2 trials to assess the extract activity also in burgers vacuum-packaged stored. Burgers were analysed (up to 9 days at 4°C) in terms of sensory properties, microbiological profile, pH, aw and lipid oxidation (TBARS). At the end of storage, treated samples showed the highest values of redness and the lowest content of malondialdehyde, probably due to antioxidant properties of red beet towards myoglobin and lipid oxidation processes. Moreover, results highlighted that Red Beetroot activities were dose-dependent and intensified if dissolved in water. The aw values did not appear to be conditioned by extract integrations, unlike the pH that was lower in treated samples than control ones. Microbiological analyses identified beetroot as a potential antimicrobial substance, especially in high concentration. In conclusion, Beta vulgaris extract could be proposed as natural compound exploitable in beef burgers to preserve qualities and extend their shelf-life.

Green feed increases antioxidant and antineoplastic activity of buffalo milk: A globally significant livestock.

The effect of green feed on health-promoting biomolecules in milk was examined in dairy buffaloes. Buffaloes received a total mixed ration (TMR) (Control, C; n = 40) or TMR + alfalfa green feed (30% of diet) (Treated, T; n = 40). Biomolecules and functional activity were measured in milk obtained twice-monthly. Treated buffaloes had higher milk l-carnitine, acetyl-l-carnitine, propionyl-l-carnitine and δ-valerobetaine (P < 0.01). They also had higher antioxidant activity (P < 0.01). Compared with C buffaloes, milk of T buffaloes improved the viability of endothelial cells exposed to high-glucose (P < 0.01), and reduced intracellular lipid peroxidation, reactive oxygen species (ROS), and cytokine release (P < 0.01). Milk of T buffaloes inhibited with greater potency the viability of human HCT116 and Cal 27 cancer cells (P < 0.001). The findings show that including green feed in the diet of dairy buffaloes enhances health-promoting biomolecules and the antioxidant and antineoplastic properties of milk.

Anisakis spp. Larvae in Deboned, in-Oil Fillets Made of Anchovies (Engraulis encrasicolus) and Sardines (Sardina pilchardus) Sold in EU Retailers.

Sardina pilchardus and Engraulis encrasicolus are considered the principal target species for commercial fishing in Europe and are widely consumed as semipreserved products. Although they are considered shelf-stable products, if treatment is not correctly applied, their consumption may represent a public health risk in regard to anisakiasis and allergic reactions. Little is known about the prevalence of Anisakis spp. in ripened products. This study aimed to evaluate the presence of Anisakis spp. larvae in deboned, in-oil anchovy and sardine fillets marketed in the EU to assess the influence of processing techniques on the prevalence of larvae. Ninety semipreserved anchovy and sardine products deriving from the Mediterranean Sea or Atlantic Ocean were collected from different EU retailers and examined using chloropeptic digestion to evaluate the presence of larvae and identify them. Thirty nonviable Anisakid larvae-A. pegreffii (30%) and A. simplex (70%)-were found. The frequency of larvae was higher in anchovies (28.8%). The low frequency of parasites found proved that processing technologies can influence the presence of larvae in final products, but it is important that visual inspection is performed only by trained people. The sources of raw materials should be considered in the production flow chart.

A Rapid Method for the Identification of Fresh and Processed Pagellus erythrinus Species against Frauds.

The commercialization of porgies or seabreams of the family Sparidae has greatly increased in the last decade, and some valuable species have become subject to seafood substitution. DNA regions currently used for fish species identification in fresh and processed products belong to the mitochondrial (mt) genes cytochrome b (Cytb), cytochrome c oxidase I (COI), 16S and 12S. However, these markers amplify for fragments with lower divergence within and between some species, failing to provide informative barcodes. We adopted comparative mitogenomics, through the analysis of complete mtDNA sequences, as a compatible approach toward studying new barcoding markers. The intent is to develop a specific and rapid assay for the identification of the common pandora Pagellus erythrinus, a sparid species frequently subject to fraudulent replacement. The genetic diversity analysis (Hamming distance, p-genetic distance, gene-by-gene sequence variability) between 16 sparid mtDNA genomes highlighted the discriminating potential of a 291 bp NAD2 gene fragment. A pair of species-specific primers were successfully designed and tested by end-point and real-time PCR, achieving amplification only in P. erythrinus among several fish species. The use of the NAD2 barcoding marker provides a rapid presence/absence method for the identification of P. erythrinus.