Report on the brain of the monk seal (Monachus monachus, Hermann, 1779).

The Mediterranean monk seal (Monachus monachus, Hermann, 1779) is an endangered species of pinniped endemic to few areas of the Mediterranean Sea. Extensive hunting and poaching over the last two centuries have rendered it a rare sight, scattered mainly in the Aegean Sea and the western coast of North Africa. In a rare event, a female monk seal calf stranded and died in southern Italy (Brindisi, Puglia). During due necropsy, the brain was extracted and fixed. The present report is the first of a monk seal brain. The features reported are remarkably typical of a true seal brain, with some specific characteristics. The brain cortical circonvolutions, main fissures and the external parts are described, and an EQ was calculated. Overall, this carnivore adapted to aquatic life shares some aspects of its neuroanatomy and physiology with other seemingly distant aquatic mammals.

Serum Protein Concentration and Serum Protein Fractions in Bottlenose Dolphins (Tursiops truncatus) under Human Care Using Agarose Gel Electrophoresis.

Serum protein electrophoresis (SPE) is the most used and reliable method to determine the percentage of serum protein subfractions. The interpretation of the kinetics of total proteins and albumin and globulin fractions is receiving increased attention in wild animals, as well as in domestic animals, due to the possibility of identifying typical pathologic patterns. However, the interpretation of these data had to be performed in light of an appropriate method-and species- specific reference intervals (RIs). In marine mammals, as well as other non-domestic species, specific attention should also be given to the different environment (free ranging vs. human managed) and the associated different exposure to environmental stimuli. The aim of this report was to establish RIs for the serum protein fractions evaluated using agarose gel electrophoresis (AGE) in bottlenose dolphins under human care. Peripheral blood samples were collected from 40 bottlenose dolphins during standard veterinary procedures to evaluate their health status. Total protein concentration was determined using the biuret method while AGE was performed using an automated system. A pooled dolphin's serum sample was used to determine the intra-assay and inter-assay imprecision of AGE. The RIs were calculated using an Excel spreadsheet with the Reference Value Advisor set of macroinstructions. The intra and inter-assay imprecisions were 1.2% and 2.5%, respectively, for albumin; 2.9% and 5.7%, respectively, for α-globulins; 3.8% and 4.0%, respectively, for ß-globulins; and 3.4% and 4.8%, respectively, for γ-globulins. The total protein, albumin, α-globulin, ß-globulin, and γ-globulin concentrations were 65.5 ± 5.4 g/L, 45.5 ± 4.9 g/L, 8.0 ± 1.0 g/L, 5.0 ± 2.0 g/L, and 7.0 ± 2.0 g/L, respectively. We established the RIs for the total protein and serum protein fractions using AGE in bottlenose dolphins under human care.

Razorbills Alca torda in Italian Seas: A Massive Irruption of Historical Relevance and Role of Social Network Monitoring.

Reporting on uncommon wide animal movements could help in depicting potential carry-over effects at the population level, particularly in an era of rapid climate and environmental changes. The razorbill (Alca torda, Linnaeus 1758) is a regular passage migrant and winter visitor to Italian seas, but with sporadic presences usually involving small numbers of individuals. Irruptions have been occasionally documented, with the last records of an unusually large number dating back to 1982. However, in the past, irruptions have only been locally reported and poorly described. Here we report on an unprecedented massive irruption of hundreds of razorbills which occurred in the central Mediterranean Sea in November-December 2022. Using citizen science platforms and photos/videos shared on social networking sites (SNSs), we estimated the relative magnitude of the irruption and described the spatial distribution of birds at sea, as well as report cases of stranded individuals. We collected a total of 267 records, both from Italy and from neighboring countries. We also discuss the likely factors affecting razorbill irruption and stress the importance of open social platforms and data sharing to aid in the early detection and estimation of such events at a wide-scale, as well as for the monitoring of the mortality of the irrupted species.

General and species-specific recommendations for minimal requirements for the use of cephalopods in scientific research.

Here we list species-specific recommendations for housing, care and management of cephalopod molluscs employed for research purposes with the aim of contributing to the standardization of minimum requirements for establishments, care and accommodation of these animals in compliance with the principles stated in Directive 2010/63/EU. Maximizing their psychophysical welfare was our priority. General recommendations on water surface area, water depth and tank shape here reported represent the outcome of the combined action of the analysis of the available literature and an expertise-based consensus reached - under the aegis of the COST Action FA1301 - among researchers working with the most commonly used cephalopod species in Europe. Information on water supply and quality, environmental conditions, stocking density, feeding and handling are also provided. Through this work we wish to set the stage for a more fertile ground of evidence-based approaches on cephalopod laboratory maintenance, thus facilitating standardization and replicability of research outcomes across laboratories, at the same time maximizing the welfare of these animals.

Milk composition, fatty acids profile and fat globule size of bottlenose dolphin (Tursiops truncatus, Montagu 1821) milk at early lactation.

Research data on milk composition in cetaceans are scattered and fragmentary. This paper analyses the gross and mineral composition, the fatty acids profile and the fat globule size of bottlenose dolphin (Tursiops truncatus) colostrum and milk at early lactation. The milk samplings were carried out on three lactating female of bottlenose dolphins at the 1st, 4-5 and 20- 42 day post partum. High percentages of dry matter (51.88%), fat (26.08%) and protein (13.83%) were found in the colostrum on the first day, while there was a tendency for these components to decrease in the milk. The average diameter the milk fat globule (7.07 µm) of this species was assessed for the first time. The milk was rich in unsaturated fatty acids, which were more than twice compared to the saturated fatty acids (unsaturated / saturated ratio = 2.6). The main fatty acids of dolphin milk were C16: 0, C16:1 n-7, C18: 1n-9, C20: 1 n-11 C20: 5 and C22:6n-3. Several FA tended to increase from colostrum to milk: C12: 0 doubled, C20: 1 n-7 and C22:1 n-11 tripled, C20: 1 n-9 quintupled in later milk samples. Differently, other FA tended to decrease: C18: 3 n-3 was approximately 4 times lower and C24: 0 and C24: 1 halved in milk compared to colostrum. In conclusion, this paper contributes to the knowledge of the nutritional value of milk from bottlenose dolphins maintained under human care. Higher content of nutritional components such as fat and protein was found in the first day colostrum compared to milk. Moreover, the findings of this study highlight abundance of unsaturated fatty acids and large sized milk fat globules in dolphin milk.

Molecular and morphological characterization of Bolbosoma balaenae (Acanthocephala: Polymorphidae), a neglected intestinal parasite of the fin whale Balaenoptera physalus.

Post-mortem examination of a fin whale Balaenoptera physalus stranded in the Mediterranean Sea led to the finding of Bolbosoma balaenae for the first time in this basin. In this work, we describe new structural characteristics of this parasite using light microscopy and scanning electron microscopy approaches. Moreover, the molecular and phylogenetic data as inferred from both ribosomal RNA 18S-28S and the mitochondrial DNA cytochrome oxidase c subunit 1 (cox1) for adult specimens of B. balaenae are also reported for the first time. Details of the surface topography such as proboscis's hooks, trunked trunk spines of the prebulbar foretrunk, ultrastructure of proboscis's hooks and micropores of the tegument are shown. The 18S + 28S rRNA Bayesian tree (BI) as inferred from the phylogenetic analysis showed poorly resolved relationships among the species of Bolbosoma. In contrast, the combined 18S + 28S + mtDNA cox1 BI tree topology showed that the present sequences clustered with the species of Bolbosoma in a well-supported clade. The comparison of cox1 and 18S sequences revealed that the present specimens are conspecific with the cystacanths of B. balaenae previously collected in the euphausiid Nyctiphanes couchii from the North Eastern Atlantic Ocean. This study provided taxonomic, molecular and phylogenetic data that allow for a better characterization of this poor known parasite.

Physiological Parameters Monitored on Bottlenose Dolphin Neonates (Tursiops truncatus, Montagu 1821) over the First 30 Days of Life.

Bottlenose dolphins (Tursiops truncatus) have been bred under human care for more than 60 years. Calves up to 30 days of life have presented the highest mortality rate, although comparable data for free-ranging neonates within this age group is not available. Husbandry measures to increase survivability have therefore been constantly improved. This work shows the results of a structured veterinary program that established the procedures to collect relevant physiological parameters on 13 calves during their first 30 days of life. Standardized observation protocols facilitated statistical analysis of the respiratory pattern, nursing, morphometric measurements and bloodwork. These allow early detection of health issues. Healthy neonates had longer apnea duration, despite the shape and size of the facility. The nursing pattern showed that successful calves started nursing 3 to 18 h postpartum. Although with different individual patterns, a steady increase in suckling time occurs during the first 24 h of life. The healthy neonates grew 0.428 ± 0.102 kg per day and the complete blood count profile, glucose, iron, blood urea nitrogen (BUN), total protein, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Gamma-glutamyl transferase (GGT), creatinine and electrolytes values considered normal for healthy calves are provided. Furthermore, cholesterol, triglycerides, α-amylase, lipase, magnesium and cortisol are reported for the first time for such young calves. A list of indications for prompt intervention is included.

Cetacean Morbillivirus and Toxoplasma gondii Co-infection in Mediterranean Monk Seal Pup, Italy.

A Mediterranean monk seal (Monachus monachus) pup from the southern Adriatic coast of Italy showed cetacean morbillivirus (CeMV) and disseminated Toxoplasma gondii co-infection, which probably resulted from CeMV-induced immunosuppression. These findings are of concern for the conservation of this critically endangered species.

Correction: The use of Unmanned Aerial Vehicles (UAVs) to sample the blow microbiome of small cetaceans.

[This corrects the article DOI: 10.1371/journal.pone.0235537.].

The use of Unmanned Aerial Vehicles (UAVs) to sample the blow microbiome of small cetaceans.

Recent studies describe the use of UAVs in collecting blow samples from large whales to analyze the microbial and viral community in exhaled air. Unfortunately, attempts to collect blow from small cetaceans have not been successful due to their swimming and diving behavior. In order to overcome these limitations, in this study we investigated the application of a specific sampling tool attached to a UAV to analyze the blow from small cetaceans and their respiratory microbiome. Preliminary trials to set up the sampling tool were conducted on a group of 6 bottlenose dolphins (Tursiops truncatus) under human care, housed at Acquario di Genova, with approximately 1 meter distance between the blowing animal and the tool to obtain suitable samples. The same sampling kit, suspended via a 2 meter rope assembled on a waterproof UAV, flying 3 meters above the animals, was used to sample the blows of 5 wild bottlenose dolphins in the Gulf of Ambracia (Greece) and a sperm whale (Physeter macrocephalus) in the southern Tyrrhenian Sea (Italy), to investigate whether this experimental assembly also works for large whale sampling. In order to distinguish between blow-associated microbes and seawater microbes, we pooled 5 seawater samples from the same area where blow samples' collection were carried out. The the respiratory microbiota was assessed by using the V3-V4 region of the 16S rRNA gene via Illumina Amplicon Sequencing. The pooled water samples contained more bacterial taxa than the blow samples of both wild animals and the sequenced dolphin maintained under human care. The composition of the bacterial community differed between the water samples and between the blow samples of wild cetaceans and that under human care, but these differences may have been mediated by different microbial communities between seawater and aquarium water. The sperm whale's respiratory microbiome was more similar to the results obtained from wild bottlenose dolphins. Although the number of samples used in this study was limited and sampling and analyses were impaired by several limitations, the results are rather encouraging, as shown by the evident microbial differences between seawater and blow samples, confirmed also by the meta-analysis carried out comparing our results with those obtained in previous studies. Collecting exhaled air from small cetaceans using drones is a challenging process, both logistically and technically. The success in obtaining samples from small cetacean blow in this study in comparison to previous studies is likely due to the distance the sampling kit is suspended from the drone, which reduced the likelihood that the turbulence of the drone propeller interfered with successfully sampling blow, suggested as a factor leading to poor success in previous studies.