Polystyrene Nanoplastics in Aquatic Microenvironments Affect Sperm Metabolism and Fertilization of Mytilus galloprovincialis (Lamark, 1819).

The continuous and unregulated discharge of wastes and pollutants into the aquatic environment has required constant monitoring of the risks incurred by aquatic ecosystems. Alarmism arises from plastic pollution as larger artifacts release nanoscale fragments that can contact free-living stages such as gametes, embryos, and larvae. Specifically, the interaction between spermatozoa, released in water in externally fertilizing species, and the surrounding microenvironment is essential for successful fertilization. Activation and kinematics of movement, proper maintenance of ionic balance, and chemotactism are processes highly sensitive to even minimal perturbations caused by pollutants such as polystyrene nanoplastics. Spermatozoa of Mytilus galloprovincialis (M. galloprovincialis), an excellent ecotoxicological model, undergo structural (plasma membrane ruptures, DNA damage) and metabolic (reduced motility, fertilizing capacity) damage upon exposure to 50 nm amino-modified polystyrene nanoplastics (nPS-NH2). Nanoplastics of larger diameter (100 nm) did not affect sperm parameters. The findings highlighted the negative impact that plastic pollution, related to nanoparticle diameter and concentration, could have on sperm quality and reproductive potential of organisms, altering the equilibrium of aquatic ecosystems.

Sublethal Effects of Polystyrene Nanoplastics on the Embryonic Development of Artemia salina (Linnaeus, 1758).

Currents, wave motion, solar radiation, and abrasion are mechanisms responsible for the degradation of large plastic artifacts and contribute to the dispersion of micro and nanoplastics into aquatic ecosystems, which are, currently, the most dangerous threats due to their invisibility and persistence. The present work evaluated the possible lethal and sublethal effects of amino-modified polystyrene nanoplastics (nPS-NH2) with diameters of 50 nm and 100 nm on Artemia salina (A. salina), an organism at the base of the trophic chain of the aquatic system, using a widely used model for the analysis of embryotoxicity from environmental pollutants. For this purpose, after evaluating the biodistribution of nanoplastics in the body of the tested animals, several endpoints such as anomalies, apoptosis, and ROS production were assessed. In addition, particular attention was dedicated to evaluating the correlation between toxicity and the particle size tested. The results reported that, despite the absence of a lethal impact, several sublethal effects involving gut and body size malformations, as well as the enhancement of apoptosis and oxidative stress in relation to an increase in tested concentration and a decrease in nanoparticle size.

Titanium Dioxide Nanoparticles: Effects on Development and Male Reproductive System.

Titanium dioxide nanoparticles (TiO2-NPs) are used intensively. Thanks to their extremely small size (1-100 nm), TiO2-NPs are more absorbable by living organisms; consequently, they can cross the circulatory system and then be distributed in various organs including the reproductive organs. We have evaluated the possible toxic effect of TiO2-NPs on embryonic development and the male reproductive system using Danio rerio as an organism model. TiO2-NPs (P25, Degussa) were tested at concentrations of 1 mg/L, 2 mg/L, and 4 mg/L. TiO2-NPs did not interfere with the embryonic development of Danio rerio, however, in the male gonads the TiO2-NPs caused an alteration of the morphological/structural organization. The immunofluorescence investigation showed positivity for biomarkers of oxidative stress and sex hormone binding globulin (SHBG), both confirmed by the results of qRT-PCR. In addition, an increased expression of the gene responsible for the conversion of testosterone to dihydrotestosterone was found. Since Leydig cells are mainly involved in this activity, an increase in gene activity can be explained by the ability of TiO2-NPs to act as endocrine disruptors, and, therefore, with androgenic activity.

Infection Due to Actinobacillus lignieresii in Cattle with Brain and Ocular Involvement: Histological and Microbiological Features.

The actinobacillosis is rare and to date the biological profile of the agent is not yet fully understood. The knowledge about the possible hosts of the pathogen is incomplete and is generally only associated with granulomatous lesions in cattle and sheep. The primary organs involved are the mouth, tongue and pharynx. Human infection is extremely rare. Actinobacillus lignieresii is the causative agent of a rare bovine granulomatous disease known as "wooden tongue". In this research, we describe a case of cerebral and ocular metastatic diffusion of granuloma due to infection with Actinobacillus lignieresii in cattle, probably resulting from primary oral localization. Diagnosis was made using histopathological assay which made it possible to highlight the typical lesion of actinobacillosis, and bacteriological assay that allowed to isolate the pathogen.

Effects of carnosine on the embryonic development and TiO2 nanoparticles-induced oxidative stress on Zebrafish.

Oxidative stress is due to an unbalance between pro-oxidants, such as reactive oxygen (ROS) and nitrogen (RNS) species, and antioxidants/antioxidant system. Under physiological conditions these species are involved in different cellular processes such as cellular homeostasis and immune response, while an excessive production of ROS/RNS has been linked to the development of various diseases such as cancer, diabetes, and Alzheimer's disease. In this context, the naturally occurring dipeptide carnosine has shown the ability to scavenge ROS, counteract lipid peroxidation, and inhibit proteins oxidation. Titanium dioxide nanoparticles (TiO2-NPs) have been widely used to produce cosmetics, in wastewater treatment, in food industry, and in healthcare product. As consequence, these NPs are often released into aquatic environments. The Danio rerio (commonly called zebrafish) embryos exposure to TiO2-NPs did not affect the hatching rate, but induced oxidative stress. According to this scenario, in the present study, we first investigated the effects of carnosine exposure and of a sub-toxic administration of TiO2-NPs on the development and survival of zebrafish embryos/larvae measured through the acute embryo toxicity test (FET-Test). Zebrafish larvae represent a useful model to study oxidative stress-linked disorders and to test antioxidant molecules, while carnosine was selected based on its well-known multimodal mechanism of action that includes a strong antioxidant activity. Once the basal effects of carnosine were assessed, we then evaluated its effects on TiO2-NPs-induced oxidative stress in zebrafish larvae, measured in terms of total ROS production (measured with 2,7-dichlorodihydrofluorescein diacetate probe) and protein expression by immunohistochemistry of two cellular stress markers, 70 kDa-heat shock protein (Hsp70) and metallothioneins (MTs). We demonstrated that carnosine did not alter the phenotypes of both embryos and larvae of zebrafish at different hours post fertilization. Carnosine was instead able to significantly decrease the enhancement of ROS levels in zebrafish larvae exposed to TiO2-NPs and its antioxidant effect was paralleled by the rescue of the protein expression levels of Hsp70 and MTs. Our results suggest a therapeutic potential of carnosine as a new pharmacological tool in the context of pathologies characterized by oxidative stress such as neurodegenerative disorders.

In Vitro Nano-Polystyrene Toxicity: Metabolic Dysfunctions and Cytoprotective Responses of Human Spermatozoa.

The ubiquitous spread of Polystyrene nanoplastics (PS-NPs) has rendered chronic human exposure an unavoidable phenomenon. The biodistribution of such particles leads to bioaccumulation in target organs including the testis, the site of sperm maturation. The purpose of this research has been to estimate the impact of PS-NPs (50 and 100 nm) on the metabolism of mature spermatozoa. The analysis of the semen parameters has revealed a higher toxicity of the smaller sized PS-NPs, which have negatively affected major organelles, leading to increased acrosomal damage, oxidative stress with the production of ROS, DNA fragmentation, and decreased mitochondrial activity. PS-NPs of 100 nm, on the other hand, have mainly affected the acrosome and induced a general state of stress. An attempt has also been made to highlight possible protective mechanisms such as the expression of HSP70s and their correlation among various parameters. The results have evinced a marked production of HSP70s in the samples exposed to the smaller PS-NPs, negatively correlated with the worsening in oxidative stress, DNA fragmentation, and mitochondrial anomalies. In conclusion, our results have confirmed the toxicity of PS-NPs on human spermatozoa but have also demonstrated the presence of mechanisms capable of counteracting at least in part these injuries.

Antioxidant activity of two Opuntia Mill. species fruit extracts on human sperm quality after a freeze-thaw cycle.

This study investigated the phenolic compounds and antioxidant capacity of fruit extracts of Opuntia dillenii (Ker Gawl.) Haw.(OD) and Opuntia ficus-indica (L.) Mill.(OFI), yellow (F1) and red (F2) varieties. In order to evaluate the antioxidant activity of these extracts on human sperm quality after thawing, the semen parameters (vitality, motility, acrosome reaction, oxidative stress, and DNA fragmentation) were analysed after 1 h of exposure. The results showed that OD has higher phenolic content and antioxidant power than OFI, and that they are higher in F2 than F1. Furthermore, regarding the activity of extracts on thawed sperm, the results showed a significant increase in motility in samples treated with OFI F1 and OD extracts, while an improvement in vitality and acrosome reaction and a reduction of DNA fragmentation were observed in all exposed samples compared to the control. Finally, a reduction of oxidative stress was observed in samples exposed to OFI F2 and OD than control.

Acute Toxicity of a Marine Emerging Pollutant (Promethazine Hydrochloride) on Artemia sp.

Recently, there has been a worrying increase in the pollution of the aquatic ecosystem caused by emerging contaminants (ECs) detected in wastewater effluent discharges. Although traces of ECs in waters have been found in low concentrations, it leads to negative effects for nontarget organisms. Antihistamines are a class of drugs largely used, whose metabolites are widespread in the aquatic ecosystem. The aim of the study was to evaluate the short-term effects of promethazine hydrochloride on nauplii of Artemia sp. A high percentage of mortality and morphological alterations were found. The results suggest a possible correlation between exposure to antihistamine and an acceleration of larval development.

Assessment of selected molecular factors and 17-ß estradiol dosage in response to Toxoplasma gondii infection in swine.

Background and Aim: Toxoplasma gondii is a global zoonotic parasite infecting virtually all warm-blooded species, although a species-specific variability is evident referring to symptoms frame. Both the success of T. gondii and the outcome of infection depend on a delicate balance between host cellular pathways and the evasion or modulation strategies elicited by the parasite. The hormonal and molecular mechanisms involved in this delicate host-parasite balance are still unclear, especially when considering intermediate host species other than mouse. This study aimed to assess any correlation between T. gondii infection and selected molecular and hormonal factors involved in responses to infection in susceptible species such as swine. Moreover, blood counts and hematochemical assays (glucose, total cholesterol, and triglycerides dosage) were performed to evaluate the overall health condition of animals. Materials and Methods: Toxoplasmosis was diagnosed by enzyme-linked immunosorbent assay for antibodies determination and real-time polymerase chain reaction (RT-PCR) for T. gondii DNA detection. Target genes coding for key factors of cell responses to T. gondii infection were selected, and their transcription was assessed in various tissues by quantitative RT-PCR. 17-ß estradiol concentrations were assessed by fluorimetric enzyme-linked immunoassay and the AIA-360 automated immunoassay analyzer. Blood count and hematochemical analyses were performed by a blood cell counter and a spectrophotometer, respectively. Results: The present research highlighted significant differences among infected and uninfected swine (control group) for both transcription profiles of some of the molecular factors considered and 17-ß estradiol concentrations. Referring to the assessed hematological and biochemical parameters, no statistically significant differences were observed in infected swine compared to the control group. Conclusion: Our results contribute to the enrichment of data available about the subject and could be useful for a deeper knowledge of the interaction between this parasite and its hosts. However, more aspects are still unclear, such as the effective response of downstream molecules from the same pathways to the variation of factors observed in this study either assessing how the same factors respond to Toxoplasma gondii infection in other host speciesand further analyses should be performed on other host species.

T-L technique for HoLEP: perioperative outcomes of a large single-centre series.

INTRODUCTION: The aim of this article was to describe, step-by-step, an original technique (T-L technique) in a single centre series of patients who underwent holmium laser enucleation of the prostate (HoLEP) for symptomatic benign prostatic hyperplasia and analyze perioperative outcomes. MATERIAL AND METHODS: We retrospectively analyzed data of 567 patients who underwent HoLEP. The T-L technique consists of a series of incisions used as landmarks, performed at the beginning of the procedure before enucleation. Two T-shape incisions are performed at the level of bladder neck (at the 5-7 and 12 o'clock positions); two L-shape incisions are performed at the level of verumontanum, bilaterally, to mark the apex and to limit the sphincter. Another T-shape incision is performed on the bladder neck at the 12 o'clock position posterior to the level of verumontanum. RESULTS: The median operative time (OT) was 80 minutes (IQR 64-105); 50 minutes (IQR 35-70) and 15 minutes (IQR 10-20) for enucleation and the morcellation phase, respectively. Conversion to transurethral resection of the prostate (TURP) was necessary in 3/567 (0.6%) patients. Intraoperative complications occurred in 3.4% of cases, capsule perforation occurred in 12/567 (2%) of cases, while bladder perforation during morcellation occurred in 8/567 (1.4%) of cases. Postoperative complications were observed in 20/567 (3.5%) of patients. Specifically, grade 1-2 occurred in 19/567 (3.3%) and grade 3 was recorded in 1/567 (0.2%). CONCLUSIONS: The T-L technique for HoLEP is safe and reproducible with a low rate of perioperative complications. The positioning of some landmarks before enucleation allows for the better orientation during enucleation and could be very useful in case of large prostates.

Artemia salina: A microcrustacean to assess engineered nanoparticles toxicity.

The focus of this work was to investigate the toxicity of different metal nanoparticles (gold nanoparticles [AuNPs], silver nanoparticles [AgNPs], titanium dioxide nanoparticles [TiO2 NPs]) on brine shrimp Artemia salina. We investigated if nanoparticles could have an influence on hatching of cysts and on mortality of larvae. Larvae (also called nauplii) and cysts were exposed to NPs for 24 hr in artificial seawater on microplates. At the end of the test, we assessed the endpoint (immobility/death) for the larvae by a stereomicroscope. Nauplii that appeared completely motionless, were counted as dead, and the percentages of mortality were calculated for each treatment. Instead for the cysts, the percentages of not-hatched nauplii for each concentration considered were calculated by counting the number of whole cysts. Currently, nanoparticles toxicity has been investigated in several research; in our study we highlighted the nontoxicity of TiO2 NPs on A. salina nauplii as shown by low percentages of immobilization and on cysts because TiO2 NPs do not affect their hatching. Despite AuNPs exerted toxic effects on hatching, they did not affect larvae development as well as TiO2 NPs. Otherwise, AgNPs induced mortality of the larvae and inhibited cysts hatching.

Edible Insects and Toxoplasma gondii: Is It Something We Need To Be Concerned About?

ABSTRACT: Novel foods, such as edible insects and food products on the basis of insects, could play an important role in both human and animal nutrition in the future. The identification of dangers associated with insect consumption is fundamental to guarantee consumer safety and adequate regulatory guidelines for operators of the food sector. Although former studies have focused on the microbiological contamination of fresh or processed edible insects, so far little information is available about the occurrence of foodborne parasites, such as Toxoplasma gondii, whose life cycles make them candidates for potential insect breeding substrate contamination. Hence, we investigated the presence of contaminating T. gondii in farmed edible insects to rule out this further hazard for consumers. Four species of insects most commonly used as food for human consumption were analyzed: mealworm; African migratory locust, house cricket, and silkworm. Samples included live specimens but also minimally (dehydrated) and highly processed edible insects. Traces of T. gondii DNA were detected in samples of dehydrated mealworm. These results highlight the need for implementing good farming and processing practices with particular care paid to safe storage and handling of feed and substrates used for edible insects to reduce the chance of T. gondii entering the human food chain.

Prognostic performance of magnetic resonance imaging-guided biopsy in defining prostate cancer anterior lesions.

PURPOSE: Diagnosis of anterior prostate cancer (PCa) can be quite challenging, often leading to delay in treatment. mpMRI-guided biopsy (GB) has been introduced aiming to increase the number of diagnoses of clinically significant PCa with fewer cores. The aim of our study is to compare pathological findings of prostate biopsy, In-bore or Fusion technique, with histopathological evaluation of radical prostatectomy. METHODS: We prospectively collected data from 90 consecutive patients who underwent either In-bore or Fusion biopsy following the detection of an index suspicious lesion at mpMRI in the anterior part of the prostatic gland. Bioptical pathological findings were compared with pathological findings reported after robot-assisted radical prostatectomy. RESULTS: Patients who underwent In-bore GB had a higher rate of previous negative prostate biopsies (19% vs 44%, p = 0.02). Median number of bioptic cores taken (13 vs 2) and number of positive cores (3 vs 2) were significantly superior in the Fusion group compared to the In-bore group (p < 0.001 and p = 0.002, respectively), whilst clinical International Society of Urological Pathology (ISUP) grade was homogeneous within groups. The concordance between anterior lesions detected at biopsy and those reported in the histopathological finding of radical prostatectomy was very high, without statistically significant difference between groups. CONCLUSION: Both Fusion and In-bore GB are accurate in detecting anterior PCa, with enhanced precision detecting clinically significant tumours, as evidenced by pathologic examinations which confirmed the presence of index anterior PCa in > 50% of patients overall. Additional sextant biopsy is still required, especially among biopsy-näive patients, to avoid missing clinically significant PCa.

Evaluation of dimethoate toxicity on fertilization and on embryonic development of Paracentrotus lividus (Lamarck, 1816).

Organophosphates are a large class of chemicals with anticholinesterase action insecticides. Dimethoate belongs to the class of organophosphates and it is used for agriculture purpose. Its main toxicological role in animals and humans is the inhibition of the activity of acetylcholinesterase. Although it is not considered genotoxic, carcinogenic and teratogen, there is evidence of increased pup mortality in developmental neurotoxicity studies. Since there is scant published literature about developmental toxicity, we investigated the adverse effects of dimethoate on fertilization and embryonic development in sea urchin (Paracentrotus lividus), a model organism widely used to assess the toxicity of contaminants on environmental matrices; so pesticide residues can be released into the environment, and could affect the health of organisms, including humans. Different solution of dimethoate (4 × 10-3, 4 × 10-4, 4 × 10-5, 4 × 10-6 and 4 × 10-7 g/10 ml) have been tested on spermatozoa of P. lividus to evaluate the fertilizing ability of them when we added egg cells untreated. We demonstrated that dimethoate does not interfere with fertilizing ability of spermatozoa but egg cells fertilized by treated spermatozoa showed alterations in the segmentation planes as asymmetric and/or asynchronous cell divisions.

Toxicity assessment of two-dimensional nanomaterials molybdenum disulfide in Gallus gallus domesticus.

Recently two-dimensional nanomaterials, such as graphene and molybdenum disulfide (MoS2), have received much attention as adsorbent materials for the effective removal of organic contaminants. MoS2 is attracting attention, not only for its chemical-physical properties, but also for its wide availability in nature as a constituent of molybdenite. The aim of this investigation was to assess the effects of different MoS2 concentrations (5 × 10-1, 5 × 10-2 and 5 × 10-3 mg/ml) on the embryonated eggs of Gallus gallus domesticus, according to Beck method. We evaluated the toxic effect of the MoS2 powder purchased at Sigma-Aldrich indicated as "received" and MoS2 powder treated via mechanical milling indicated as "ball mille". Subsequently, the embryos were sacrificed at different times of embryonic development (11th, 15th and 19th day after incubation) in order to evaluate their embryotoxic and teratogenic effects. The alterations of the embryonic development were studied by morphological and immunohistochemical analysis of the tissues. The results obtained have shown the toxicity of both powders of MoS2 with a high percentage of deaths and growth delays. Moreover, the immunohistochemical analysis performed on several tissue sections showed a strong positivity to the anti-metallothionein1 antibody only for the erythrocytes.

Morphological analysis reveals the presence of Hepatoxylon trichiuri (Holten, 1802) Bosc, 1811 (Cestoda: Sphyriocephalidae) in Lepidopus caudatus (Euphrasen, 1788) (Pisces: Trichiuridae) from the Mediterranean Sea.

In this article, we report for the first time the presence of the parasite Hepatoxylon trichiuri in specimens of Lepidopus caudatus from the Mediterranean Sea. A total of four plerocercoids were studied using microscopy techniques for both morphological and histological analysis. RESEARCH HIGHLIGHTS: We report for the first time for the Mediterranean Sea the presence of the parasite Hepatoxylon trichiuri in specimens of the commercial fish species Lepidopus caudatus. The plerocercoids of H. trichiuri not seems cause observable illness in the fish. However, further studies are needed in order to better assess the incidence and prevalence of the parasite in L. caudatus and other fish species of commercial interest.

Toxicity assessment of nano-TiO2 in Apis mellifera L., 1758: histological and immunohistochemical assays.

The aim of this study was to evaluate the toxicity of titanium dioxide nanoparticles (TiO2 NPs) by short-term toxicity tests in Apis mellifera, considered an excellent bioindicator organism mainly due to its sensitivity. Bees have been exposed to several concentrations of TiO2 NPs (1 × 10-3 , 1 × 10-4 , 1 × 10-5 , 1 × 10-6 mg/10 ml) for 10 days. Morphostructural and histological assays were done on gut and honey sac. The research of exposure biomarkers like metallothioneins 1 (MT1) and Heat Shock Protein 70 (HSP70) was performed to verify if a detoxification mechanism has been activated in the exposed animals. No histological alteration on the epithelium of the gut and honey sac were observed in exposed samples. A significant positivity for anti-MT1 antibody was observed only in the honey sac cells. A weak positivity for HSP70 was observed in both structures analyzed. In several studies have shown the non-toxicity of TiO2 NPs on other model organisms, in our study, titanium dioxide nanoparticles was proven to be highly toxic at the highest concentration tested (100% of lethality to 1 × 10-3 mg/10 ml) and moderately toxic at lower concentrations. Honey bees proved to be excellent models for study of NPs toxicity and for monitoring environment.

Morphostructural and immunohistochemical study for evaluation of nano-TiO2 toxicity in Armadillo officinalis Duméril, 1816 (Crustacea, Isopoda, Oniscidea).

Invertebrates are precious organisms in order to study environmental pollution. In particular, they appear to be suitable as a bioindicator species for pioneer ecotoxicity studies on new xenobiotics such as nanoparticles. In fact, they are able to absorb nanomaterials scattered in the environment in different ways and it's known the compartmentalization of nano-sized contaminants in selected tissues and intracellular organelles. Titanium dioxide represents the most used nanoparticulate, destined to become probably ubiquitous in the environment. Recently, some research has been published on the toxic potential of nano-TiO2 in several animal species. Among all invertebrates, Oniscidean Isopods are the only taxon of Crustaceans that has become completely terrestrial, known as excellent bioindicators and bioaccumulators. They have a digestive gland, the hepatopancreas, which is the location of election for the accumulation of pollutants. For this reason, they are considered efficient animal models to ecological studies. For this study, we collected Armadillo officinalis from Natural Oriented Reserve of "Vendicari" (Sicily, Italy), to evaluate the toxicity of titanium dioxide (TiO2 ) on their hepatopancreas, after a short period of exposure. We conducted morphostructural and immunohistochemistry assays. The results suggested a great capacity of the species of bioaccumulation of nanoparticles in the hepatopancreas, where a strong positivity to the metallothioneins was highlighted. Our study confirms that Oniscidean Isopods, in particular Armadillo officinalis, proved to be an appropriate indicator of pollution in terrestrial ecosystems from nanoparticles.

Is robotic approach useful to palliate advanced bladder cancer? A monocentric single surgeon experience.

INTRODUCTION: The aim of this study was to assess surgical and functional outcomes of 17 consecutive patients undergoing robot- assisted radical cystectomy (RARC) with palliative intent in a monocentric single surgeon series. MATERIAL AND METHODS: We collected data from 17 consecutive patients who underwent RARC with palliative intent performed by a single surgeon at our institution. Patients undergoing palliative RARC were those with advanced bladder cancer (BC) or advanced comorbidities. Clinical, surgical and functional outcomes were prospectively collected. Patients completed a specific questionnaire (Functional Assessment of Cancer Therapy-Bladder Cancer, FACT-BL) before and after surgery to assess the role of palliative RARC in terms of quality of life improvement. RESULTS: Median age at surgery was 78 years, with median Charlson Comorbidity Index (CCI) and age-adjusted CCI of 3 and 7, respectively. Clinical stage was T2, T3 or T4 in 7, 8 and 2 patients respectively, with 52.9% and 29.4% with cN+ and cM+ disease. Median estimated blood loss was 200 ml, with 1 patient requiring intra-operative blood transfusion. Median hospital stay was 7 days. A total of 3 and 2 patients were re-hospitalized during the first 30 and 30-90 post-operative days, respectively. One major Clavien grade complication was recorded.At median follow-up of 8 months, 9 and 2 patients succumbed due to tumor progression and other causes. Pre-operative and post-operative FACT-BL scores improved significantly in each domain. CONCLUSIONS: A RARC is a safe and feasible technique which could be offered as part of palliative care in patients with advanced BC or comorbidities. Precise guidelines for palliating BC patients should be better.

Biomarkers of Exposure to Chemical Contamination in the Commercial Fish Species Lepidopus caudatus (Euphrasen, 1788): A Particular Focus on Plastic Additives.

In recent years, the Mediterranean Sea has become an accumulation zone for waste generated by the 22 countries bordering its shores. Although the effects of plastic litter on the marine environment and on organisms have recently been studied in other areas, further information is needed for the Mediterranean Sea and, in particular, about plastics additives inputs and interactions with the biota and the trophic network, such as phthalates and bisphenol A. Plastic material production, use and disposal contribute also to the release of heavy metals into the environment, such as mercury (Hg), often used during the production of chlorine, the primary ingredient in PVC, lead (Pb) and cadmium (Cd), which are used as stabilizers in PVC and leach out of products during use and disposal. Our research aims to evaluate phthalates, bisphenol A and heavy metals contamination in Lepidopus caudatus (Pisces, Trichiuridae), which could be considered as a potential sentinel species. For the evaluation of toxicological effects, we evaluated the expression of vitellogenin and metallothioneins 1. In all samples analyzed, we have not found microplastics in the gastrointestinal tract but chemical analysis revealed the presence of high content of phthalates, and in particular high quantities of DIDP, DEHP, bis-benzylester phthalate, bis-butyl ester phthalate and mono-N-butyl ester phthalate in different organs. Instead, trace elements detected in tissue revealed a trend of concentrations generally higher in liver and intestine than gill and muscle tissues. Immunohistochemical analysis for anti-metallothionein 1 antibody showed a strong positivity of liver cells, both in females and males. Analysis for the anti-vitellogenin antibody showed in females a strong positivity both in the liver cells and in the gonads, in male specimens was found to be always negative except for a specimen, in which it was highlighted a positivity in some areas of the liver and of the gonad.