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Effective species identification is crucial for the conservation and management of marine mammals, particularly in regions such as the Mediterranean Sea, where several cetacean populations are endangered or vulnerable. In this study, we developed and validated a High-Resolution Melting (HRM) analysis protocol for the rapid, cost-effective, and reliable identification of the four representative marine cetacean species that occur in the Mediterranean Sea: the bottlenose dolphin (Tursiops truncatus), the striped dolphin (Stenella coeruleoalba), the sperm whale (Physeter macrocephalus), and the fin whale (Balaenoptera physalus). Species-specific primers targeting mitochondrial DNA regions (cytochrome b and D-loop) were designed to generate distinct melting profiles. The protocol was tested on both tissue and fecal samples, demonstrating high sensitivity, reproducibility, and discrimination power. The results confirmed the robustness of the method, with melting curve profiles clearly distinguishing the target species and achieving a success rate > 95% in identifying unknown samples. The use of HRM offers several advantages over traditional sequencing methods, including reduced cost, speed, portability, and suitability for degraded samples, such as those from the stranded individuals. This approach provides a valuable tool for non-invasive genetic surveys and real-time species monitoring, contributing to more effective conservation strategies for cetaceans and enforcement of regulations against illegal trade.

Ergasilus sieboldi are parasitic copepods belonging to the Ergasilidae family. They affect the gills of various freshwater and marine fish, causing mortality events worldwide. Like other copepods, E. sieboldi adult females are parasitic; while, the adult males die after fertilisation. Lesions caused by these parasites can create prerequisites for secondary infections by other pathogens, such as bacteria, viruses and fungi. This study aimed to describe the first case of E. sieboldi in largemouth bass (Micropterus salmoides) in Italy. Morphological analysis identified the genus of copepods collected from gills, distinguishing them from Neoergasilus, already described in Italy. The species was identified with molecular characterisation and phylogenetic analysis based on 28S rDNA sequencing. Histopathological analysis of the gills highlighted damage to the gill lamellae such as erosion, necrosis and desquamation typically caused by ergasilids.

Antimicrobial resistance affects all environments, endangering the health of numerous species, including wildlife. Increasing anthropic pressure promotes the acquisition and dissemination of antibiotic resistance by wild animals. Sea turtles, being particularly exposed, are considered sentinels and carriers of potential zoonotic pathogens and resistant strains. Therefore, this study examined the antibiotic resistance profiles of bacteria isolated from loggerhead sea turtles hospitalised in a rescue centre of Southern Italy over a 9-year period. Resistance to ceftazidime, doxycycline, enrofloxacin, flumequine, gentamicin, oxytetracycline and sulfamethoxazole-trimethoprim was evaluated for 138 strains isolated from the clinical samples or organs of 60 animals. Gram-negative families were the most isolated: Vibrionaceae were predominant, followed by Shewanellaceae, Pseudomonadaceae, Enterobacteriaceae and Morganellaceae. These last three families exhibited the highest proportion of resistance and multidrug-resistant strains. Among the three Gram-positive families isolated, Enterococcaceae were the most represented and resistant. The opportunistic behaviour of all the isolated species is particularly concerning for diseased sea turtles, especially considering their resistance to commonly utilised antibiotics. Actually, the multiple antibiotic resistance was higher when the sea turtles were previously treated. Taken together, these findings highlight the need to improve antimicrobial stewardship and monitor antibiotic resistance in wildlife, to preserve the health of endangered species, along with public and environmental health.

Caretta Caretta are endangered sea turtles leaving in the Mediterranean Sea. We carried out a study on 195 sea turtles stranded along the coast of Southern Italy, evaluating anthropogenic and opportunistic factors affecting animal health status and probably threatening the species. More than 60% of the animals showed body injuries, mainly caused by marine traffic and fishing. Ingestion of marine litter of various origins was observed in the digestive tract of 38.5% of the turtles investigated. Chelonid herpesvirus 5 was detected (for the first time in this basin) in 10.8% of the turtles, which were however free from fibropapillomatosis. Analyses on trace elements showed a time-dependent reduction in the mean concentrations of Cd and Hg, a decrease in the levels of Pb, and a time-dependent increase of As. Particularly, a site-dependent accumulation of As was detected in turtles from the coasts of Calabria. There is a significant correlation between the levels of As in the kidney of Campania turtles and their straight-line carapace length, indicating a size-dependent accumulation of this metal. Overall, besides dangerous debris items, high levels of trace elements may weaken the immune system of Caretta caretta , the most common turtle of the Mediterranean Sea, making it more likely vulnerable to viruses.

Toxoplasma gondii is an emerging cause of neurologic disease in cetaceans. In the present study, we combined molecular detection, microsatellite genotyping, and neuropathology in stranded striped dolphins along the coasts of Campania and Calabria (Southern Italy). During the years 2018–2023, 93 cetaceans were necropsied. Brain tissues were screened for T. gondii DNA by Real-Time PCR targeting the 529-bp repeat element; positive samples (Ct range 21–30) were genotyped using eight microsatellite markers (TUB2, W35, TgM-A, B18, B17, M33, IV.1 and XI.1) and compared with reference strains by hierarchical clustering. Histology and immunohistochemistry (IHC) for T. gondii were performed on available brains. T. gondii DNA in the brain was detected in 12/93 cetaceans; all positive cases were striped dolphins ( Stenella coeruleoalba ). Genotyping revealed two genotypes, with GII in 10/12 dolphins and GIII in 2/12. In 10 evaluable cases, lesions ranged from mild non-suppurative meningitis/meningoencephalitis to severe necrotizing encephalitis. IHC was positive in 7/10 positive cases; tissue cysts were observed in proximity to lesions in the more severely affected brains. These findings document the circulation of multiple T. gondii genotypes in pelagic dolphins from Southern Italy, supporting the land-to-sea transmission hypothesis and confirming the value of striped dolphins as sentinels of terrestrial pathogens from a One Health perspective.

The presence of cetaceans along the Campania coast has always been documented. Between 2016 and 2022, out of 65 cetaceans stranded along the Campania coast, 46 were studied for bacteriological, virological, parasitological, and histopathological investigations. The results highlighted that for 59% (n = 27) of the specimens, the cause of death was of natural origin, while for only 2% (n = 1) of animals, the origin of death was ascribed to anthropic causes. Unfortunately, for 39% (n = 18) of the cetaceans, it was impossible to determine the cause of death. All the cetaceans that died of natural causes showed viral, bacterial, and parasitic infections. The primary pathogens detected were Cetacean Morbillivirus (CeMV, 65.2%, n = 30/46), Toxoplasma gondii (10.9%, n = 5/46), and Brucella ceti (8.7%, n = 4/46). The animals showed typical lesions of the isolated pathogens, such as systemic infection, meningoencephalitis, and pneumonia. Moreover, even with a lower frequency, other relevant pathogens like Photobacterium damselae, Salmonella enteritidis, Erysipelothrix rhusiopathiae, and Chlamydia abortus were isolated. These data were useful to understand the spread and circulation of these pathogens, some zoonotic, in the coastal marine waters of the Campania region.

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.

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.

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.

Processing-using-DRAM (PUD) architectures impose a restrictive data layout and alignment for their operands, where source and destination operands (i) must reside in the same DRAM subarray (i.e., a group of DRAM rows sharing the same row buffer and row decoder) and (ii) are aligned to the boundaries of a DRAM row. However, standard memory allocation routines (i.e., malloc, posix_memalign, and huge pages-based memory allocation) fail to meet the data layout and alignment requirements for PUD architectures to operate successfully. To allow the memory allocation API to influence the OS memory allocator and ensure that memory objects are placed within specific DRAM subarrays, we propose a new lazy data allocation routine (in the kernel) for PUD memory objects called PUMA. The key idea of PUMA is to use the internal DRAM mapping information together with huge pages and then split huge pages into finer-grained allocation units that are (i) aligned to the page address and size and (ii) virtually contiguous. We implement PUMA as a kernel module using QEMU and emulate a RISC-V machine running Fedora 33 with v5.9.0 Linux Kernel. We emulate the implementation of a PUD system capable of executing row copy operations (as in RowClone) and Boolean AND/OR/NOT operations (as in Ambit). In our experiments, such an operation is performed in the host CPU if a given operation cannot be executed in our PUD substrate (due to data misalignment). PUMA significantly outperforms the baseline memory allocators for all evaluated microbenchmarks and allocation sizes.