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In recent years, the consumption of fish products has surged in European countries, being an essential part of a healthy diet. Despite representing a small part of EU production, freshwater fisheries hold considerable significance for lake-dwelling populations and tourists seeking traditional dishes. This increased fish consumption has brought to light potential health risks associated with fish-borne zoonotic helminths (FBZHs), now acknowledged as global food-borne parasites. Fish-borne zoonotic helminths belong to various taxonomic groups, including nematodes (Anisakidae), trematodes (Opisthorchiidae and Heterophyidae), and cestodes (Diphyllobothriidae). More than 50 species of FBZH are known to cause human infections, derived from eating raw or undercooked aquatic foods containing viable parasites. Despite increased attention, FBZHs remain relatively neglected compared to other food-borne pathogens due to factors like chronic disease progression and under-diagnosis. This systematic review concentrates on the prevalence of six freshwater FBZHs (Clinostomum complanatum, Contracaecum rudolphii, Dibothriocephalus latus, Eustrongylides excisus, Opisthorchis felineus, and Pseudamphistomum truncatum) in Italy and neighbouring countries. The study explores the expansion of these parasites, analysing their biological and epidemiological aspects, and the factors that influence their proliferation, such as the increased cormorant population and the lake eutrophication phenomena. In summary, this research highlights the necessity for further research, the development of spatial databases, and the establishment of a unified European policy to effectively manage these multifaceted health concerns. It strongly advocates adopting a One-Health approach to address the growing incidence of parasitic zoonoses within the context of food safety in EU countries.

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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.

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The sub-Alpine lakes of Switzerland, Italy and France have long been reported as an endemic area of diphyllobothriosis, a parasitic zoonosis caused by Dibothriocephalus latus. With this study, we explored the hypothesis for a relationship between the prevalence of D. latus in Perca fluviatilis and the Escherichia coli load in lake water. To do this, we identified eleven sampling sites in three areas (north, centre and south) of Lake Iseo (north Italy) to determine E. coli load and the prevalence of D. latus in P. fluviatilis. Prevalence and 95% confidence interval (CI) of D. latus infestation ranged from 0% (95% CI: 0.71-0.0) in Sarnico (southern area) to 20% (95% CI: 33.0-11.2) in Pisogne (northern area). There were significant differences in prevalence between the sites (χ2  = 31.12; p-value = .0006) and in E. coli load (Kruskal-Wallis test; p-value = .0005). There was decreasing gradient of E. coli load and prevalence of D. latus infestation from north to south. A significant positive correlation (r = .881; p-value = .003) was found between E. coli load and prevalence of D. latus. Also, linear regression showed a significant relationship between E. coli load and prevalence of infestation (R2  = .775). Our findings offer an explanation for the link between E. coli load in water and D. latus prevalence. The potential factors in this link are the efficiency of the local wastewater treatment plant, the bathymetric profile of the lake and the life cycle of D. latus, which is mainly affected by light and water temperature.

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Fish mycobacteriosis is a widespread global problem caused by species of non-tuberculous mycobacteria (NTM). Mycobacterium marinum is one of the species most often involved in disease episodes of aquarium and farmed fish. Since there is currently no available effective therapy or vaccine, a prompt search for routes of entry is key to limiting the damage induced by the disease. Here we report a case of mycobacteriosis follow up in a European sea bass (Dicentrarchus labrax) farm located in Northern Italy, in which environmental samples and newly added fish batches were analyzed. Samples from fish present on the farm, sediment, and periphyton all resulted positive for M. marinum, whereas the new fish batches and the water samples resulted negative. The environmental resistance of NTM (alcohol-acid resistance, biofilm formation) and the lack of prophylactic and therapeutic strategies make these diseases difficult to manage. Prompt identification of biotic and abiotic reservoirs, combined with good zootechnical hygiene practices, are the most effective measures to control fish mycobacteriosis in intensive farms.

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The genus Pennella comprises hematophagous parasites of marine aquatic species, including cephalopods, marine mammals, and pelagic fish. Nine species have been officially included in the genus Pennella plus another six species inquirendae. They are most often found in the host's musculature, without penetrating internal organs. For the present study, 83 hearts from swordfish (Xiphias gladius) caught in the Mediterranean Sea were sampled and immediately fixed in formalin for histopathological analysis. In total, 10 (12.05%) hearts were found to be parasitized by copepods of the genus Pennella. Macroscopically, there was mild-to-severe fibrinous pericarditis with atrial wall thickening and multiple parasitic nodules. Histologically, the parasitic nodules were surrounded by an inflammatory-necrotizing reaction. Parasitic infestation by Pennella spp. is common in pelagic fish and in swordfish, in particular. Here, however, we report atypical cardiac localization. A future area of focus is the evaluation of cardiac Pennella spp. infestation by histopathology and genetic identification of the parasites.

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Bronopol and Detarox® AP are broad spectrum antimicrobial biocides of growing interest for the aquaculture sector. While their effectiveness against aquatic pathogens has been demonstrated, toxicity data on wild or farmed species are still lacking, as is information on their potential environmental risk for aquatic ecosystems. With this study, we assessed the acute and sublethal toxicity of Bronopol and Detarox® AP in the freshwater bivalve Sinanodonta woodiana and their theoretical risk for aquatic ecosystem. The 96-h median lethal concentration (LC50) was determined using the acute toxicity test, while for the sublethal toxicity test the bivalves were exposed to two concentrations for 14 days of Bronopol (2.5 and 50 mg/L) and Detarox® AP (1.11 and 22.26 mg/L) followed by a 14-day withdrawal period. Biocide-mediated oxidative processes were investigated via a panel of oxidative stress biomarkers (malondialdehyde, superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase). Theoretical environmental risk assessment of both biocides, with predicted concentration of no effect (PNEC), expected theoretical concentration (TEC) in the environment, and risk quotient (RQ) was performed. TEC was calculated using a model based on the size of the aquaculture facility and the receiving basin, the estimated quantity of biocide dissolved in water, and published data on biocide stability in water. Although the LC50 was higher for Bronopol (2440 mg/L) than for Detarox® AP (126 mg/L), fluctuations in oxidative stress biomarkers levels indicated that both biocides exert a slight oxidative pressure on S. woodiana. Theoretical environmental risk assessment suggested a muted risk with Detarox® AP and greater eco-sustainability compared to Bronopol.

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The Mycobacterium fortuitum group (MFG) consists of about 15 species of fast-growing nontuberculous mycobacteria (NTM). These globally distributed microorganisms can cause diseases in humans and animals, especially fish. The increase in the number of species belonging to MFG and the diagnostic techniques panel do not allow to clarify their real clinical significance. In this study, biomolecular techniques were adopted for species determination of 130 isolates derived from fish initially identified through biochemical tests as NTM belonging to MFG. Specifically, gene sequencing and phylogenetic analysis were used based on a fragment of the gene encoding the 65 KDa heat shock protein (hsp65). The analyzes made it possible to confirm that all the isolates belong to MFG, allowing to identify the strains at species level. Phylogenetic analysis substantially confirmed what was obtained by gene sequencing, except for six strains; this is probably due to the sequences present in NCBI database. Although the methodology used cannot represent a univocal identification system, this study has allowed us to evaluate its effectiveness as regards the species of MFG. Future studies will be necessary to apply these methods with other gene fragments and to clarify the real pathogenic significance of the individual species of this group of microorganisms.

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Carnobacteria are common bacteria in cold and temperate environments; they are also reported during fish mortality events. In a previous study, carnobacteria were isolated from the eyes of healthy wild salmonids from a high-mountain lake. To better understand these findings, salmonids were captured from three high-mountain lakes (Lower and Upper Balma Lake, Rouen Lake; northwest Italy) during August 2019 and subjected to bacteriological and histological examination. Although all were healthy, 8.7% (Lower Balma Lake), 24% (Upper Balma Lake), and 32.6% (Rouen Lake) were positive for carnobacteria colonization of the eyes. A Trojan-horse effect was hypothesized to explain carnobacteria isolation in the eye. This immune-escaping macrophage-mediated mechanism has been identified in other Gram-positive bacteria. Biochemical, molecular, and phylogenetic analysis were carried out on isolated bacteria (Carnobacterium maltaromaticum and C. divergens). Based on previous references for carnobacteria isolated from fish, C. maltaromaticum strains were tested for the pisA precursor gene of the bacteriocin piscicolin 126. Carnobacterium maltaromaticum strains were found to display genotypic heterogeneity and a low percentage of pisA positive amplification. Features of geomorphology, geographic isolation, and microbiota common to the three lakes are thought to be possibly related to our findings. Moreover, terrestrial insects collected from the lake shoreline and the stomach contents were screened for the presence of carnobacteria. The salmonids in these high-mountain environments feed mainly on terrestrial insects, which are considered possible vectors for carnobacteria that might catabolize the exoskeleton chitin. All insects tested negative for carnobacteria, but as a small number of samples were analyzed, their role as possible vectors of infection cannot be excluded. Further studies are needed to corroborate our research hypothesis.

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We appreciate the valuable comment of Pavlik et al. [...].

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Plastic pollution has become a pervasive environmental problem on a global scale, from the ocean depths to the aquatic ecosystems of the Tibetan Plateau. To date, data on plastic and microplastic occurrence in pristine ecosystems like high-mountain lakes are lacking. In this study, plastic (>5000 µm) and microplastic (10-5000 µm) levels were measured in snow at the end of the winter season (April 2020), and in water, sediment, and biological samples collected monthly (June-October 2019) during the ice-free season from the Dimon Lake, a high-mountain lake in the Carnic Alps, northeast Italy. Biological samples consisted of chironomids (Diptera, Chironomidae; n = 150) and stomach contents of Cottus gobio (n = 40). Analysis of the water, sediment, and biological samples revealed the absence of plastic and microplastics larger than 10 µm, whereas the snow samples contained microplastics of polyethylene terephthalate (PET) albeit at very low levels (0.11 ± 0.19 L-1). These results show that while the lake ecosystem could be considered unpolluted by microplastics, abundant snow precipitation in winter can trap microplastic particles that deposit on the ground. The very low levels of PET microparticles recorded in the snow samples suggest the need for further research to better understand the source of microplastic pollution in this environmental matrix.

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Integrated multi trophic aquaculture (IMTA) is well developed in seawater, while the only suitable organisms for bioremediation in freshwater are bivalves. The aim of this research has been to investigate a novel system, based on freshwater bivalves integrated with rainbow trout, for the bioremediation of inland aquaculture systems. The Unionid species selected for the experiments was Sinanodonta woodiana. Five mesocosm experiments were conducted to evaluate the efficiency of mussels in filtering the total bacteria and Aeromonas hydrophila, as well as their clearance efficiency on trout farm wastewater. The temperatures, dissolved oxygen, nitrates, and phosphates were monitored and were all within the physiological tolerance range of the species in all the experiments. The feasibility of bioremediation with S. woodiana in trout farming has thus been demonstrated, and among the tested rearing densities (3.75; 7.5; 15; 30 and 60 kg m-3) that of 7.5 kg m-3 was found to be optimal. The net reduction of the total bacterial concentration was as much as 72%, while that of A. hydrophila reached a level of 95-98%. No relevant effects of the mussels on particulate suspended material or sedimented material was observed, regardless of the temperature. The efficiency of freshwater bivalves in reducing the bacterial load, in particular toward A. hydrophila, indicates a bioremediation system with the possibility of interesting applications on inland fish farms, and as a biotechnological tool against the diffusion of antibiotic resistance in aquaculture.

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Dibothriocephalus latus (Linnaeus, 1758) (Cestoda: Diphyllobothriidea; syn. Diphyllobothrium latum), is a fish-borne zoonotic parasite responsible for diphyllobothriasis in humans. Although D. latus has long been studied, many aspects of its epidemiology and distribution remain unknown. The aim of this study was to investigate the prevalence, mean intensity of infestation, and mean abundance of plerocercoid larvae of D. latus in European perch (Perca fluviatilis) and its spatial distribution in three commercial fishing areas in Lake Iseo (Northern Italy). A total of 598 specimens of P. fluviatilis were caught in 2019. The total prevalence of D. latus was 6.5%. However, there were significant differences between areas (10.2% North; 7.3% Center; 1.5% South) (Chi-square test, p = 0.0018). The mean intensity of infestation ranged from 1 larva in southern area to 1.2 larvae in both the central and northern (Pisogne) areas. In addition, the mean abundance ranged from 0.02 in the southern area to 0.26 in the northern area (Pisogne). The total number of larvae (anterior dorsal-AD = 21; anterior ventral-AV = 1; posterior dorsal-PD = 15; posterior ventral-PV = 5) differed significantly between the four anatomical quadrants (Kruskal-Wallis test; p = 0.0001). The prevalence of D. latus plerocercoid larvae in European perch from Lake Iseo has long been investigated, but without an appropriate sampling design. With the present study, a broader analysis in spatial distribution has been added to the existing literature, revealing new information about D. latus distribution and occurrence in Lake Iseo, with new data that will be useful for health authorities and future studies.

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Mycobacterium pseudoshottsii, a slow-growing mycobacterium closely related to M. marinum, has been isolated only in wild fish in the United States and in Japanese fish farms to date. Here, we report cases of mortality in three farmed fish species (Dicentrarchus labrax, Sparus aurata, and Sciaenops ocellatus) caused by M. pseudoshottsii in Italy. Samples underwent necropsy, histology, and culture with pathogen identification based on PCR and sequencing of housekeeping genes (16S rRNA, hsp65, rpoB). Multifocal to coalescing granulomatous and necrotizing inflammation with acid-fast bacilli were observed in the parenchymatous organs, from which M. pseudoshottsii was isolated and identified. Phylogenetic analysis confirmed the results of gene sequencing and allowed subdivision of the isolates into three distinct groups. M. pseudoshottsii poses a potential threat for Mediterranean aquaculture. Its origin in the area under study needs to be clarified, as well as the threat to the farmed fish species.

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The genus Eustrongylides includes nematodes that infect fish species and fish-eating birds inhabiting freshwater ecosystems. Nematodes belonging to the genus Eustrongylides are potentially pathogenic for humans; infection occurs after the consumption of raw or undercooked fish. In the two-year period 2019-2020, a total of 292 fish belonging to eight species were examined for the occurrence of Eustrongylides spp. from Lake San Michele, a small subalpine lake in northwest Italy. The prevalence of infestation was 18.3% in Lepomis gibbosus, 16.7% in Micropterus salmoides, and 10% in Perca fluviatilis. The other five fish species (Ameiurus melas, Ictalurus punctatus, Squalius cephalus, Carassius carassius, and Scardinius erythrophthalmus) were all negative for parasite presence. There were no significant differences in prevalence between the three fish species (Fisher's exact test; p = 0.744). The mean intensity of infestation ranged from 1 (M. salmoides and P. fluviatilis) to 1.15 (L. gibbosus), and the mean abundance ranged from 0.1 (P. fluviatilis) to 0.28 (L. gibbosus). There were significant differences in the infestation site between the four muscle quadrants (anterior ventral, anterior dorsal, posterior ventral, and posterior dorsal) and the visceral cavity (Kruskal-Wallis test; p = 0.0008). The study findings advance our knowledge about the distribution and host range of this parasite in Italy.

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Crassostrea gigas is a sentinel species along the Italian coast. In mussels, the levels of oxidative stress biomarkers can be modulated by several environmental pollutants or pathogens and also fluctuate in response to reproductive stages and seasonal changes. In this study, adult Crassostrea gigas were sampled during summer and autumn from two lagoon and two offshore sites along the Adriatic coast of Italy in order to investigate the influence of seasonality on oxidative stress biomarkers. Trace elements load of Al, As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Se and Zn suggests low contamination for lagoon and offshore sites. Levels of total glutathione, superoxide dismutase, catalase, glutathione peroxidases, glutathione reductase and glutathione S-transferase were analyzed in digestive gland and gills of the Pacific oysters in June, July, September and October. OsHV-1 and Vibrio aestuarianus were detected in lagoon sites, but both pathogens did not affect the biomarkers levels in both tissues. Although several biological responses were found different among the four sites in the same month, principal component analysis revealed similar trend in biomarkers levels between sites during the whole sampling period. On the other hand, a different biochemical pattern through the months emerged, suggesting that the level of oxidative stress biomarkers in both tissues may be related to seasonal progress and biological cycle of oysters sampled from the two lagoons and offshore sites along the Italian coasts of the Mediterranean Sea.

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Although sturgeon production by aquaculture has increased worldwide, a major factor limiting its expansion are infectious diseases, although few data about viral diseases are available however. This review provides a rapid overview of viral agents detected and described to date. Following a general introduction on viral diseases are four sections arranged by virus classification: sturgeon nucleocytoplasmic large DNA viruses, herpesviruses, white sturgeon adenovirus 1, and other viruses. Molecular diagnosis is currently the best tool to detect viral diseases, since cell culture isolation is not yet applicable for the detection of most sturgeon viruses.

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Clinostomum complanatum, a digenean trematode of the Clinostomidae family, is a fish-borne zoonotic parasite responsible for Halzoun syndrome in humans and is transmitted through the consumption of raw or undercooked freshwater fish. Of the total of 112 specimens of European perch (Perca fluviatilis) sampled from a subalpine lake (Lake Endine) in North Italy in 2019, 21 (18.75%) tested positive for encysted metacercariae in the fillet. This study reports the first isolation of C. complanatum in P. fluviatilis and highlights the possible zoonotic risk for consumers, since P. fluviatilis is a food fish used in the traditional local cuisine.

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We report the first case of Acipenser iridovirus European (AcIV-E) infection in starry sturgeon (Acipenser stellatus) and in sterlet (A. ruthenus) reared in Northern Italy. During 2018, mortality began in A. stellatus and A. ruthenus specimens reared in co-habitation with Russian sturgeon positive for AcIV-E. Molecular analyses were done on the gills to amplify a fragment of the major capsid protein (MCP) gene using real-time PCR against AcIV-E. DNA of the positive samples was further sequenced and phylogenetic analyses were performed. The MCP gene sequences were highly similar to a virus previously identified in Italy (nucleotide identities between 99.38% and 99.69%). Phylogenetic analysis confirmed our hypothesis of passage of the virus from the infected Russian sturgeon. The detection of AcIV-E in new species of the Acipenseridae family may impact on sturgeon production, with relevant economic losses.

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High-mountain lakes are pristine ecosystems characterized by extreme environmental conditions. The atmospheric transport of pollutants from lowlands may add further stress to organisms inhabiting these environments. We investigated the environmental stress pressure on brook trout (Salvelinus fontinalis) from a high-mountain lake in the Cottian Alps (Piedmont, northwest Italy). To do this, males and females of brook trout were sampled from Balma Lake in summer (August) and autumn (October) 2017 in order to assess the influence of trace elements accumulation and environmental parameters (physicochemical parameters and nutrient characteristics of water) on oxidative stress biomarkers. Bioaccumulation of Al, As, Cd, Cr, Cu, Fe, Hg, Pb, Ni, Se, and Zn and metallothionein levels were measured in muscle tissue of males and females. Liver, gills, kidney, and spleen tissue samples were analyzed for superoxide dismutase, catalase, total glutathione peroxidase, selenium-dependent glutathione peroxidase, glutathione reductase, and glutathione S-transferase activity. Analysis of environmental parameters showed changes in biomarker levels with seasonal variations. Water temperature was significantly higher in summer than autumn (Wilcoxon test; p = .0078), while pH was significantly higher in autumn than in summer (Wilcoxon test; p = .0078). Sex-related differences in oxidative stress biomarkers in tissues were unremarkable, whereas seasonal variability of oxidative stress biomarkers was observed, with major differences occurred for liver in summer and for gills, kidney, spleen and muscle in autumn. Positive correlations between environmental parameters and biomarkers were noted. Major fluctuations in water temperature, pH, Cu, Pb and Hg produced changes in biomarker levels; however, increased food intake during the ice-free season was probably the main factor that influenced changes in oxidative stress biomarker levels in brook trout in this extreme ecosystem.

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