Comparative study of polycyclic aromatic hydrocarbons (PAHs) in salt gland and liver of loggerhead turtle Caretta caretta (Linnaeus, Cheloniidae) stranded along the Mediterranean coast, Southern Italy.

The levels of polycyclic aromatic hydrocarbons, PAHs, were determined in the liver and salt gland of 19 loggerhead turtles (Caretta caretta Linnaeus, Cheloniidae) stranded along the coasts of the south Tyrrhenian Sea, Italy, from 2019 to 2021. The 16 EPA's priority PAHs were determined by gas chromatography coupled with mass spectrometry (GC-MS). The average values of ΣPAHs in liver, 139 ± 55.0, were exceptionally high and up to one hundred times those of literature and appeared even more worrying in salt gland, 320 ± 97.6 ng/g, w.w. Naphthalene was the predominant contributor to PAHs richness and accounted for 90.0% and 93.7% of ΣPAHs in the two matrices, highlighting the petrogenic source of exposure. An overall higher bioaccumulation of NAP, more than two-fold, was detected in salt gland and especially in female and adults. Data of PAHs richness highlighted a potential risk of neoplastic disease development and that anthropogenic activities may seriously impair healthy state conditions of C. caretta. populations in south Tyrrhenian Sea.

Epidemiology of Sulcascaris sulcata (Nematoda: Anisakidae) ulcerous gastritis in the Mediterranean loggerhead sea turtle (Caretta caretta).

Sulcascaris sulcata Rudolphi 1819 is a gastric nematode parasite of sea turtles. Here, we report the occurrence and describe for the first time the pathological changes caused by S. sulcata in the Mediterranean loggerhead sea turtle (Caretta caretta) stranded along the Tyrrhenian coast and northern Adriatic coast of Italy. Prevalence of infection was significantly higher in loggerhead sea turtles from the Adriatic Sea. Both prevalence and abundance of infection showed an increasing trend along with host age classes from both geographical localities. Nevertheless, while many small loggerhead sea turtles were found infected from the Adriatic Sea, only bigger individuals were infected from the Tyrrhenian Sea. The most common gross pathological change was a mucous gastritis with focal to multifocal raised ulcerous lesions roundish to irregular in shape ranging from 1 to over 20 cm in length, and cream-yellowish to greenish in color. The severity grade of gastritis increased with higher number of S. sulcata individuals. Microscopic pathological changes ranged from atrophic gastritis with heterophilic infiltration in the lamina propria to the destruction of the mucosal and sub-mucosal surfaces and necrosis. Results here obtained demonstrate that S. sulcata may cause ulcerous gastritis in both samples of loggerhead sea turtles studied from the Mediterranean Sea. Observed differences in S. sulcata infection among the different host age classes and between the two studied basins are likely linked to the differences of regional habitat and intermediate prey host availability.

Correction to: Epidemiology of Sulcascaris sulcata (Nematoda: Anisakidae) ulcerous gastritis in the Mediterranean loggerhead sea turtle (Caretta caretta).

The original version of this article contained a mistake in the value of Bar in figure 3 caption. It should be Bar = 200 µm instead of Bar = 500 µm.

Grillotia (Cestoda: Trypanorhyncha) plerocerci in an anglerfish (Lophius piscatorius) from the Tyrrhenian Sea.

Trypanorhynch cestodes are common parasites of marine fish with complicated life cycles which have been suggested as model taxa to study the evolution of marine helminth parasites and their life cycles. Among the Trypanorhyncha, the genus Grillotia includes 18 valid species, of which only four have been found in Mediterranean fish hosts. Morphological, histopathological, and molecular data are presented on a massive Grillotia plerocercus infection in an anglerfish (Lophius piscatorius) from the Tyrrhenian Sea. BLAST analysis of the 28S rDNA sequences revealed 99% similarity between specimens here found and a G. (Bathygrillotia) rowei sequence available in GenBank with a total of six nucleotide site differences. A morphological study suggested that the Grillotia sp. here reported did not match important characters to those previously reported from the Mediterranean Sea. Taking in account these differences, we prefer to place these specimens within Grillotia sensu lato until more material is available for study including sequences from adult specimens of Grillotia spp. from the Mediterranean Sea.

Genomic Characterization of Listeria monocytogenes and Other Listeria Species Isolated from Sea Turtles.

Listeria monocytogenes is a ubiquitous pathogen found both in the environment and food. It can cause listeriosis in a wide range of animals as well as in humans. Investigations on presence, spread and virulence are still limited to terrestrial and human environments. Embracing the One Health Approach, investigating the presence and spread of L. monocytogenes in marine ecosystems and among wildlife, would provide us with useful information for human health. This study investigated the presence of L. monocytogenes and Listeria spp. in two species of sea turtles common in the Mediterranean Sea (Caretta caretta and Chelonia mydas). A total of one hundred and sixty-four carcasses of sea turtles (C. caretta n = 161 and C. mydas n = 3) stranded along the Abruzzo, Molise, Campania, and Calabria coasts, were collected. Brain and fecal samples were taken, enriched, and cultured for the detection of Listeria spp. From the specimens collected, strains of L. monocytogenes (brain n = 1, brain and feces n = 1, multiorgan n = 1 and feces n = 1), L. innocua (feces n = 1 and brain n = 1), and L. ivanovii (brain n = 1) were isolated. Typical colonies were isolated for Whole Genome Sequencing (WGS). Virulence genes, disinfectants/metal resistance, and antimicrobial resistance were also investigated. L. monocytogenes, L. innocua, and L. ivanovii were detected in C. caretta, whilst only L. monocytogenes and L. innocua in C. mydas. Notable among the results is the lack of significant differences in gene distribution between human and sea turtle strains. Furthermore, potentially pathogenic strains of L. monocytogenes were found in sea turtles.

First description of Rhodococcus equi infection in common bottlenose dolphin (Tursiops truncatus).

Rhodococcus equi is a terrestrial bacterium and a common pathogen in foals (Equus caballus), in which causes pneumonia. This report describes for the first time the infection caused by R. equi in a common bottlenose dolphin (Tursiops truncatus) stranded in the Calabrian coast, Italy. The post mortem examination of the animal revealed lesions in lung and colon. The animal was also positive to dolphin morbillivirus. The histological study showed lesions attributable to R. equi infection, such as pyogranulomatous bacterial pneumonia and chronic granulomatous colitis. Whole genome sequencing of the isolated strain confirmed its identification as R. equi.

Trace Elements and Persistent Organic Pollutants in Unhatched Loggerhead Turtle Eggs from an Emerging Nesting Site along the Southwestern Coasts of Italy, Western Mediterranean Sea.

Marine pollution is one of the major threats affecting loggerhead turtles, which due to their long life span, highly migratory behavior, and carnivorous diet, may be exposed to elevated levels of toxic elements throughout their life. The transfer of chemicals from mothers to their offspring is of particular conservation concern because it may affect embryonic development and hatching success. In this study, the concentrations of 16 toxic and potentially toxic trace elements, 6 indicator polychlorinated biphenyls (PCBs), and organochlorine pesticide residues (OCPs) were determined in 138 eggs from 46 loggerhead turtle nests laid during the 2021 nesting season in Campania, Italy, western Mediterranean Sea. The possible impact of pollutant levels on hatching success and early embryonic death was also investigated. Trace element analysis was performed using an ICP-MS, except for mercury, which was determined using a Direct Mercury Analyzer® (DMA). PCBs and OCPs were analyzed with high-resolution gas chromatography coupled with high-resolution mass spectrometry (HRGC-HRMS) and gas chromatography tandem quadrupole mass spectrometry GC-MS /MS, respectively. The concentrations of essential elements in the eggs were higher than those of non-essential elements. In addition, the highly chlorinated PCBs (153, 138, and 180) contributed the most to the total PCBs, while OCPs were not detected. No correlations were found between contaminant concentrations and reproductive parameters (hatching success and no obvious embryos). The results obtained suggest that the levels of contaminants found in the eggs do not affect the reproductive success of the species in the study area.

Impact of polystyrene microplastic exposure on gilthead seabream (Sparus aurata Linnaeus, 1758): Differential inflammatory and immune response between anterior and posterior intestine.

Plastics are the most widely discharged waste into the aquatic ecosystems, where they break down into microplastics (MPs) and nanoplastics (NPs). MPs are ingested by several marine organisms, including benthic and pelagic fish species, contributing to organ damage and bioaccumulation. This study aimed to assess the effects of MPs ingestion on gut innate immunity and barrier integrity in gilthead seabreams (Sparus aurataLinnaeus, 1758) fed for 21 days with a diet enriched with polystyrene (PS-MPs; 1-20 µm; 0, 25 or 250 mg /kg b.w./die). Physiological fish growth and health status were not impacted by PS-MPs treatments at the end of experimental period. Inflammation and immune alterations were revealed by molecular analyses in both anterior (AI) and posterior intestine (PI) and were confirmed by histological evaluation. PS-MPs triggered TLR-Myd88 signaling pathway with following impairment of cytokines release. Specifically, PS-MPs increased pro-inflammatory cytokines gene expression (i.e., IL-1ß, IL-6 and COX-2) and decreased anti-inflammatory ones (i.e., IL-10). Moreover, PS-MPs also induced an increase in other immune-associated genes, such as Lys, CSF1R and ALP. TLR-Myd88 signaling pathway may also lead to the mitogen-activated protein kinases (MAPK) signaling pathway activation. Here, MAPK (i.e., p38 and ERK) were activated by PS-MPs in PI, following the disruption of intestinal epithelial integrity, as evidenced by reduced gene expression of tight junctions (i.e. ZO-1, Cldn15, Occludin, and Tricellulin), integrins (i.e., Itgb6) and mucins (i.e., Muc2-like and Muc13-like). Thus, all the obtained results suggest that the subchronic oral exposure to PS-MPs induces inflammatory and immune alterations as well as an impact on intestinal functional integrity in gilthead seabream, with a more evident effect in PI.

Levels of non-dioxin-like PCBs (NDL-PCBs) in liver of loggerhead turtles (Caretta caretta) from the Tyrrhenian Sea (Southern Italy).

The levels of six non-dioxin-like polychlorinated biphenyls (PCBs 28, 52, 101, 138, 153, and 180) were determined in the liver of 84 loggerhead turtles (Caretta caretta) stranded along the coasts of the Tyrrhenian Sea in Campania Region (Southern Italy), from 2017 to 2021. The average value of the sum (∑6PCBIND) was 28.0 ± 52.2 ng/g (w.w.). The hexachlorobiphenyls PCB 153 and PCB 138 and the heptachlorobiphenyl PCB 180 were the main contributors to the ∑6PCBIND. A weak positive correlation was found between CCL and highly chlorinated PCBs, with adult females having lower PCB concentrations than juvenile females and adult males. This study provides more comprehensive information on the levels of NDL-PCB in Mediterranean loggerhead turtles and sets the basis for assessing anthropogenic impacts on this species.

Polychlorinated organic pollutants (PCDD/Fs and DL-PCBs) in loggerhead (Caretta caretta) and green (Chelonia mydas) turtles from Central-Southern Tyrrhenian Sea.

This study assesses for the first time the levels of PCDD/Fs and DL-PCBs in sea turtles coming from Tyrrhenian Sea. The concentrations measured in liver of the 24 specimens analysed were 6.90 vs 5.65 pg g-1 wet weight (ww) for PCDD/Fs and 10.95 vs 0.79 ng g-1 ww for DL-PCBs in Caretta caretta and Chelonia mydas, respectively. The DL-PCB levels resulted very higher in Caretta caretta than Chelonia mydas probably due to the different eating habits between the two species investigated. Furthermore, the highest levels of DL-PCBs were determined in livers of the adult Caretta caretta turtles of male sex. Positive correlations were found out between PCB-81 and the body mass (BM) of turtles (r2 = 0.6561; p = 0.001) and between PCB-81 and the curved carapace length (CCL) (r2 = 0.6250; p = 0.006) suggesting that the body burden of contaminants is related to the body size. The mean TEQ values, as a matter of risk assessment for turtles, were 3.64 vs 1.62 pg TEQ g-1 ww for PCDD/Fs and 8.72 vs 2.16 pg TEQ g-1 ww for DL-PCBs in Caretta caretta and Chelonia mydas, respectively. The results reported in this study increase the data available about the consequences of the Mediterranean Sea contamination by organochlorine pollutants and highlight an evident PCDD/F and PCB bioaccumulation in sea turtle tissues that threatens the survival of these marine organisms.

Host biological factors and geographic locality influence predictors of parasite communities in sympatric sparid fishes off the southern Italian coast.

Host biological factors and habitat influence the faunal assemblages and biodiversity worldwide, including parasite communities of vertebrate and invertebrate hosts. The ecological relationship between hosts and parasites can be mediated by interaction of host's biological factors, as their physiological condition, diet and size, with the environmental components, somehow influencing the features of parasite infection in host populations. Here, we used boosted regression tree models to study the parasite communities of two sympatric sparid fishes, the salema Sarpa salpa and the white seabream Diplodus sargus, to investigate the role of specific host's traits in two contiguous coastal areas along the southern-western Tyrrhenian coast of Italy characterized by different degree of deterioration. Results showed that overall and across all parasite groups (ecto-, endo- and ecto- plus endo-parasites), sampling localities were the most important predictors of abundance, species richness, and diversity for salema. Moreover, seasonality was the main predictor of endo-parasite abundance, while size-related factors explained most of the variation in species richness and diversity. In the white seabream, size-related factors and reproductive cycle-related factors were the most important predictors for the overall parasite abundance and parasite richness, respectively. Our findings suggest that the parasite community of salema and white seabream responded differently to specific biological factors, highlighting how the environmental conditions under which they live may exert a strong influence on the parasite communities of each host fish.

Non-essential toxic element (Cd, As, Hg and Pb) levels in muscle, liver and kidney of loggerhead sea turtles (Caretta caretta) stranded along the southwestern coasts of Tyrrhenian sea.

Non-essential toxic metals are environmental pollutants that contaminate the marine ecosystem due to their extensive use and long-range transport (by rivers and air). Their presence into the environment is often linked to the human activity, with an expected bioaccumulation in the food chain. Within the marine animals, sea turtles may be considered as potential sentinel species for environmental assessment because of their long lifetime, habitat use and migratory nature. In this study, non-essential toxic metals in the coastal of Tyrrhenian Sea were monitored, during the 2017-2018 period, using the Mediterranean loggerhead sea turtle (Caretta caretta) as indicator species. The levels of Cadmium (Cd), Arsenic (As), Mercury (Hg) and Lead, (Pb) were determined in 30 turtles stranded in different locations along the coasts of Campania region of Southern Italy. After morphometric analysis, the non- essential toxic metal accumulation in loggerhead sea turtle muscle and organs have been measured analysing the total concentrations of metals in samples of liver, kidney, and muscle by Atomic Absorption Spectrophotometry (AAS) after microwave assisted digestion with nitric acid and hydrogen peroxide. The pattern of contaminants revealed a similar distribution for Pb and total mercury, that were not significantly high in all the analysed matrices. On the contrary, elevated concentrations of total As were found out in all matrices, mainly in muscle (25.7 mg/kg w.w.), and of Cd in liver (1.23 mg/kg w.w.) and kidney (6.82 mg/kg w.w.). These studies allow us to estimate the exposure degree to which these species are subjected.

Epidemiology of blood flukes (Digenea: Spirorchiidae) in sea turtles from Tyrrhenian and Adriatic Seas, off Italy.

BACKGROUND: The Spirorchiidae is a family of blood flukes parasitizing turtles. Spirorchiids may cause a wide range of inflammatory reactions in the vascular system of their host being frequently implicated with stranding and death of sea turtles worldwide. Recent studies revealed the presence of two spirorchiid species in the Mediterranean basin. Our study presents comparative epidemiological data of spirorchiid infections in loggerhead turtles (Caretta caretta) stranded during an eight-year period from Adriatic and Tyrrhenian Seas, and the first report of Neospirorchis Neogen-11 in a green turtle (Chelonia mydas). METHODS: We screened a total of 319 carcasses of loggerhead turtles stranded from January 2011 to December 2018 along the Tyrrhenian coast (n = 111) and the north-western Adriatic coast (n = 208) of Italy using traditional (copromicroscopy and histopathology) and molecular assays. Three green turtles from the Tyrrhenian coast were also included in the study. RESULTS: A total of 56 (17.5%) loggerhead turtles and one green turtle (33.3%) were found to be infected with spirorchiid flukes. Amplification, sequencing of the ITS2 region of the ribosomal RNA gene cluster and BLAST analysis confirmed the presence of Hapalotrema mistroides and Neospirorchis Neogen-11 in 51 (16.0%) and 24 (7.5%) loggerhead turtles, respectively, and Neospirorchis Neogen-11 in an infected green turtle. Differences in prevalence of infection between the two sampling areas were found. CONCLUSIONS: The risk of spirorchiid infection in the Tyrrhenian Sea is lower than in the Adriatic Sea and in general the risk of infection in the Mediterranean is lower than in other geographical locations. Differences in the prevalence of infection between the two sampling areas were related to the differences of regional habitats supporting different abundance of spirorchiid intermediate hosts. A systematic monitoring to evaluate the progress of the infection is recommended, as well as studies on the occurrence and distribution of spirorchiid species from other Mediterranean areas.

Dolphin Morbillivirus in a Cuvier's Beaked Whale (Ziphius cavirostris), Italy.

Dolphin morbillivirus (DMV) has caused several mortality events in Mediterranean striped (Stenella coeruleoalba) and bottlenose (Tursiops truncatus) dolphins populations since 19; in the last 5 years, the virus was reported to infect new hosts in this basin, such as fin whales (Balaenoptera physalus), sperm whales (Physeter macrocephalus), and even a harbor seal (Phoca vitulina). Very recently, a calf Cuvier's beaked whale (Ziphius cavirostris) calf stranded on the Southern Italian coastline with mild pathological findings suggestive of morbilliviral infection, received the first confirmation of DMV infection in this species by biomolecular evidences on lung tissue. This new cross-species infection report, along with 19% of the cetaceans specimens examined by the Italian Stranding Network being found positive to DMV, support the hypothesis of an endemic circulation of this virus among Mediterranean cetaceans.