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.

An Epidemiological Update on Anisakis Nematode Larvae in Red Mullet (Mullus barbatus) from the Ligurian Sea.

Red mullet (Mullus barbatus) is a commercially relevant fish species, yet epidemiological data on anisakid nematode infestation in M. barbatus are scarce. To fill this gap, we report the occurrence of Anisakis larvae in red mullet in the Ligurian Sea (western Mediterranean). This survey was performed between 2018 and 2020 on fresh specimens of M. barbatus (n = 838) from two commercial fishing areas (Imperia, n = 190; Savona, n = 648) in the Ligurian Sea. Larvae morphologically identified as Anisakis spp. (n = 544) were characterized using PCR-RFLP as Anisakis pegreffii. The overall prevalence of A. pegreffii was 24.46%; the prevalence at each sampling site was 6.32% for Imperia and 29.78% for Savona. Furthermore, 3300 larvae of Hysterothylacium spp. were detected in the visceral organs of fish coinfected with A. pegreffii, showing that coinfection with two parasitic species is not rare. This study provides a timely update on the prevalence of ascaridoid nematodes in red mullet of the Ligurian Sea, an important commercial fishing area in the Mediterranean.

Content of Trace Elements and Human Health Risk Assessment via Consumption of Commercially Important Fishes from Montenegrin Coast.

Muscle tissues of Mullus barbatus and Merluccius merluccius were analyzed for the presence of selected trace elements (As, Hg, Cd, and Pb) to determine the value of the daily intake of trace elements from fish consumption and to assess the risk to human health. The mean concentrations of As in the muscle tissue of M. barbatus and M. merluccius for the entire period were 19.689 mg/kg wet weight (ww) and 8.356 mg/kg ww, Hg 0.497 mg/kg ww and 0.153 mg/kg ww, and Pb 0.031 mg/kg ww and 0.025 mg/kg ww, respectively. The concentrations of Cd in all fish sampled were below the detection limit (<0.02 mg/kg ww). The evaluation of the potential health risk assessments based on the target hazard quotient (THQ) and estimated daily intake (EDI) indicated that the intake of As in both fish species and Hg for M. barbatus could pose an appreciable risk to human health. The calculated hazard index (HI) was higher than 1 for both fish species. The continuous monitoring of trace elements' concentrations in fish is strongly recommended, as the results demonstrate potential health risks due to the presence of As and Hg.

Development of Multiplex PCR and Melt-Curve Analysis for the Molecular Identification of Four Species of the Mullidae Family, Available in the Market.

The authentication of food products and the verification of their identity are of major importance for consumers. Food fraud through mislabeling is an illegal practice consisting of the substitution of an expensive food product by a relatively cheaper one, misleading false labelling of their origin and adulteration in processed or frozen products. This issue is particularly of high importance concerning fish and seafood, which are easily adulterated primarily due to difficult morphological identification. Fish species of the Mullidae family are considered among the most high-valued seafood products traded in Greece and Eastern Mediterranean in general, in terms of the price and demand. Specifically, the red mullet (Mullus barbatus) and the striped red mullet (Mullus surmuletus) are both indigenous in the Aegean (FAO Division 37.3.1) and the Ionian (FAO Division 37.2.2) Seas, with high levels of consumers' preferences. However, they could be easily adulterated or misidentified by the invasive Aegean Sea Lessepsian migrator goldband goatfish (Upeneus moluccensis) as well as by the imported West African goatfish (Pseudupeneus prayensis). Keeping this in mind, we designed two novel, time-saving and easy-to-apply multiplex PCR assays and one multiple Melt-Curve analysis real-time PCR for the identification of these four species. These methodologies are based on species-specific primers targeting single nucleotide polymorphisms (SNPs) detected via sequencing analysis of the mitochondrial cytochrome C oxidase subunit I (CO1) and of the cytochrome b (CYTB) genes in newly collected individuals, with additional comparison with congeneric and conspecific haplotypes obtained from the GenBank database. Both methodologies, targeting CO1 or CYTB, utilize one common and four diagnostic primers, producing amplicons of different length that are easily and reliably separated on agarose gel electrophoresis, yielding a single clear band of diagnostic size for each species or a certain Melt-Curve profile. The applicability of this cost-effective and fast methodology was tested in 328 collected specimens, including 10 cooked samples obtained from restaurants. In the vast majority (327 out of the 328) of the specimens tested, one single band was produced, in agreement with the expected products with a single exception a M. barbatus sample that was identified as M. surmuletus, the identity of which was confirmed using sequencing, indicating erroneous morphological identification. The developed methodologies are expected to contribute to the detection of commercial fraud in fish authentication.

Microplastics in fish and sediments from the Montenegrin coast (Adriatic Sea): Similarities in accumulation.

The accumulation of microplastics (MPs) in the biotic and abiotic components of the marine environment poses a major threat to marine ecosystems worldwide. The objective of this study was to document, for the first time, differences in MP accumulation in the gastrointestinal tract of two commercially important fish species and to evaluate the possible correlation between MP accumulation in the biotic (fish) and abiotic (sediment) components of the marine environment of the Montenegrin coast (Adriatic Sea). Samples were collected from two areas of the Montenegrin coast, Boka Kotorska Bay and the coastal part of the open sea. The frequency of MP ingestion was 58.6 % for Mullus barbatus and 54 % for Merluccius merluccius, while the average number of ingested MPs was 2.9 ± 0.5 and 3.2 ± 1.0 items/individual, respectively. Average MP abundance in surface sediments from Boka Kotorska Bay and the coastal part of the open sea was 315 ± 45 and 435 ± 258 MPs/kg of dry sediment, respectively. Most MPs identified were filaments, followed by fragments and films, while the most abundant polymers found in fish and sediments samples were polypropylene and polyethylene. The present results indicate that MP pollution in the study area is reflected in the accumulation of MPs in the biotic (fish) and abiotic (sediment) components of the marine environment. Measures need to be taken to reduce the input of plastics/MPs into the marine environment.

Microplastic occurrence in the gastrointestinal tract and gill of bioindicator fish species in the northeastern Mediterranean.

Microplastic pollution is an extremely emerging problem and its potential threats to the aquatic organisms were investigated worldwide. In this study, four different commercial fish species (Mullus barbatus (Linnaeus, 1758), Mullus surmuletus (Linnaeus, 1758), Mugil cephalus (Linnaeus, 1758), Saurida undosquamis (Richardson, 1848)) were used as a bioindicator to assess the microplastic pollution in the northeastern Mediterranean. The frequency of occurrence in the gastrointestinal tract (GIT) and gill was varied between 66-100% and 68-90%, respectively. The highest microplastic abundance was detected at the GIT of M. cephalus sampled from Asi River estuary. The majority of extracted microplastics were fiber, black in color and less than 1 mm in size. Fourier Transform Infrared Spectroscopy (FTIR) indicated the most common polymer type as polyethylene. This study is the first study examining the microplastic existence in gill and results obtained in this study improve the knowledge about the relationship of microplastic ingestion in fish and environmental conditions in the Northeastern Mediterranean Sea.

Assessment of seasonal relationship between polycyclic aromatic hydrocarbon accumulation and expression patterns of oxidative stress-related genes in muscle tissues of red mullet (M. barbatus) from the Northern Adriatic Sea.

In this study, we examined the seasonal association between Polycyclic Aromatic Hydrocarbon (PAH) concentrations and mRNA expression profiles of some antioxidant genes (i.e. CAT, GST and SOD), as well as lipid peroxidation (LPO), in muscle of sexually inactive females of red mullet (Mullus barbatus). Fish were captured in a fishery area of the Northern Adriatic Sea during both winter and summer. We found significantly (p < 0.05) higher ∑HMW-PAHs concentrations in muscle of specimens caught during winter than summer. On the basis of sampling season, red mullets exhibited different gene expression profiles of antioxidant enzymes showing lower levels of both CAT and GST in winter than in summer. Accordingly, CAT was found to be negatively associated with ∑PAH concentrations, especially ∑LMW-PAH, in individuals collected during winter. Seasonal-related downregulation of some oxidative stress biomarker expression is suggestive of greater susceptibility of red mullets to PAHs during winter.

Interlaboratory comparison of microplastic extraction methods from marine biota tissues: A harmonization exercise of the Plastic Busters MPAs project.

In the framework of the Plastic Busters MPAs project, a harmonization exercise on two methods of microplastic extraction from biological samples i.e. 15% H2O2 digestion and 10% KOH digestion was carried out. The two methods were tested in four laboratories on fish gastrointestinal tracts and mussel tissues spiked with polyethylene, polypropylene and polyethylene terephthalate. The recovery percentage of microplastics for each method, species and polymer tested were overall similar among laboratories, and interlaboratory coefficient of variation was less than 11% for the majority of samples. Microplastic recovery rates for the two methods were similar for each sample tested, but overall mean interlaboratory recovery rate using KOH (96.67%) was higher than H2O2 (88.75%). Results validate the use of both methods for extracting microplastics from biota tissues. However, when comparing the two methods in terms of microplastic recovery rate, time consumed, technical difficulties and cost, digestion with 10% KOH is considered optimal.

Are anthropogenic fibres a real problem for red mullets (Mullus barbatus) from the NW Mediterranean?

Microfibres are among the most prevalent type of microplastics in marine environments. Man-made fibres derived from cellulose are distributed worldwide, but are often confused with synthetic plastic fibres and consequently neglected. All these fibres may adversely affect aquatic organisms, but their levels and potential effects in wild fish remain unknown. We analysed anthropogenic fibre (AF) ingestion in the red mullet (Mullus barbatus), at both temporal and geographical scales, to assess potential effects of these fibres on fish health condition. AFs were present in 50% of fish digestive tracts, with a mean of 1.48 AFs per individual (SD = 1.98). In Barcelona, an increase of 46% in AF ingestion was observed in 2018 compared to 2007. AF ingestion also increases by 20% when Barcelona is compared to a less urban area (the town of Blanes). Visual characterization of fibres by typologies-corroborated by Raman spectroscopy-allowed classification and identification of 88% of AFs as cellulosic (57%), and synthetic polymers (PET) (31%). In all sampling stations, the only histopathological alterations were cysts of unknown etiology, and the most abundant parasites were nematodes. None of these alterations, parasite load, or other fish health indicators (condition indices) indicate an effect of AF ingestion.

Gonadal macrophage aggregates in fish: A preliminary quantitative study in red mullet.

A histological study was conducted in red mullet, Mullus barbatus, collected from two sites characterized by different anthropogenic impacts. The aim of the study was to assess sex-, size-, season- and site-related variation in gonadal macrophage aggregate (MA) size, number, and relative area. Gonadal MAs were most abundant in males than in females. The number of MA was significantly higher in males from the most impacted site in October, with larger individuals showing more MA than smaller ones. MAs were always found in ripe testes, whereas they occurred only in regressing ovaries. These preliminary findings suggest that the presence of ovarian MA in red mullet is most likely related to ovary regression after spawning, whereas the presence of testicular MA is not necessarily associated to gonad regression, and may vary with season, size, and water quality.

Microplastics occurrence in edible fish species (Mullus barbatus and Merluccius merluccius) collected in three different geographical sub-areas of the Mediterranean Sea.

The gastrointestinal tracts of 229 demersal fish belonging to two species (Mullus barbatus, Merluccius merluccius) were examined for microplastic ingestion. Samples were collected in 3 different FAO Geographical Sub-Areas (GSA-9, GSA-17, GSA-19) of the Mediterranean Sea. Ingested microplastics were characterized using a stereo-microscope: observed, photographed, measured and categorized according to size class, shape and colour. Plastic fragments (ranging from 0.10 to 6.6 mm) were detected in 23.3% of the total investigated fish; a total of 65 plastic particles (66% constituted by fibers) were recorded. The percentage of plastic ingestion shows high variability between the two species and among the different sampling area. The highest frequency (48%) was found in European hake from GSA-19. These preliminary results represent a baseline for the implementation of the Marine Strategy Framework Directive descriptor 10 in Italy as well as an important step for detecting microplastics in bioindicator species from different GSAs.

Comparative Evaluation of Key Aroma-Active Compounds in Raw and Cooked Red Mullet (Mullus barbatus) by Aroma Extract Dilution Analysis.

Raw red mullet (Mullus barbatus) and its cooked samples, obtained from steam and oven cooking, were subjected to aroma and key odorant analysis for the first time using GC-MS-O. The extraction of the aroma compounds was carried out by the direct solvent extraction-solvent assisted flavor evaporation (DSE-SAFE) method. Principal component analysis (PCA) was used to determine the relations between cooking processes and fish aroma compounds. By applying the aroma extract dilution analysis (AEDA), 8 and 13 aroma-active compounds were detected in raw and cooked fish samples, respectively. The most prominent differences between raw and cooked fish samples were as follows: 3-hydroxybutan-2-one, 2,3-octadienone, (E,E)-2,4-heptadienal, linalool, γ-butyrolactone, 1-methylpyrrolidin-2-one, 2H-furan-5-one and pyrrolidin-2-one were only detected in cooked samples while hexanal and 2-phenoxyethanol in only raw fish samples. GC-MS-O results clearly indicated that cooking process results in the development of characteristics and pleasant aroma of red mullet samples due to the lipid oxidation. The most dominant aroma-active compound in the cooked fish samples was the 1-octen-3-ol which is responsible for the mushroom-like odor.

Assessing environmental quality status by integrating chemical and biological effect data: The Cartagena coastal zone as a case.

Cartagena coastal zone (W Mediterranean) was chosen for a practical case study to investigate the suitability of an integrated indicator framework for marine monitoring and assessment of chemicals and their effects, which was developed by ICES and OSPAR. Red mullet (Mullus barbatus) and the Mediterranean mussel (Mytilus galloprovincialis) were selected as target species. Concentrations of contaminants in sediment and biota, and contaminant-related biomarkers were analysed. To assess environmental quality in the Cartagena coastal zone with respect to chemical pollution, data were assessed using available assessment criteria, and then integrated for different environmental matrices. A qualitative scoring method was used to rank the overall assessments into selected categories and to evaluate the confidence level of the final integrated assessment. The ICES/OSPAR integrated assessment framework, originally designed for the North Atlantic, was found to be applicable for Mediterranean species and environmental matrices. Further development of assessment criteria of chemical and biological parameters in sediments and target species from the Mediterranean will, however, be required before this framework can be fully applied for determining Good Environmental Status (GES) of the Marine Strategy Framework Directive in these regions.

Assessment of biological effects of environmental pollution in Mersin Bay (Turkey, northeastern Mediterranean Sea) using Mullus barbatus and Liza ramada as target organisms.

The increasing emphasis on the assessment and monitoring of marine ecosystems has revealed the need to use appropriate biological indicators for these areas. Enzyme activities and histopathology are increasingly being used as indicators of environmental stress since they provide a definite biological end-point of pollutant exposure. As part of an ecotoxicological assessment of Mersin Bay, EROD enzyme activity and histopathological response in selected organs and tissues of two species of fish, Mullus barbatus (red mullet) and Liza ramada (thinlip grey mullet), captured from area were examined. Pollutant (Organochlorines (OC), alkylphenols (APs) and BPA) levels and biomarker responses in tissue samples were evaluated together for their potential to alter the metabolism and cellular aspects in liver and gonad. Elevated induction of EROD activity and histopathological alterations in contaminated samples from Mersin Bay was observed compared to reference site indicating the exposure to potential pollutants.

First isolation of Mycobacterium spp. in Mullus spp. in Turkey.

Ichthyozoonotic Mycobacterium spp. poses health risks both to fish and humans. In this study, the presence of ichthyozoonotic Mycobacterium spp. was investigated in red mullet (Mullus barbatus barbatus) and surmullet (Mullus surmuletus), widely caught species in the Mediterranean and the Aegean Sea. A total of 208 fish samples, provided from fishermen of Mersin province (Turkey) were studied. Using conventional methods, Mycobacterium spp. was isolated and identified at the genus level by PCR and at the species level by PCR-RFLP. Thirteen Mycobacterium spp. were detected in 13 (6.25%) fish samples. Four mycobacteria were identified as M. genavense, three as M. fortuitum, three as M. scrofulaceum, one as M. marinum, one as M. vaccae and one as M. aurum. No signs of mycobacteriosis were observed in fish samples. Findings of this study can contribute to future studies of onichthyozoonotic Mycobacterium spp. in seafood.

Presence of dentition in the premaxilla of juvenile Mullus barbatus and M. surmuletus.

The genus Mullus is usually described as lacking teeth in the premaxilla. Contrary to generic descriptions (based on adult specimens), the juveniles of M. barbatus and M. surmuletus have teeth in the premaxilla; however, the teeth are not visible in specimens >50 mm total length, because they are covered by lip tissue. The number of teeth decrease with juvenile growth and disappears after c. 100 mm total length is attained.