De novo assembly and functional annotation of blood transcriptome of loggerhead turtle, and in silico characterization of peroxiredoxins and thioredoxins

The aim of this study was to generate and analyze the atlas of the loggerhead turtle blood transcriptome by RNA-seq, as well as identify and characterize thioredoxin (Tnxs) and peroxiredoxin (Prdxs) antioxidant enzymes of the greatest interest in the control of peroxide levels and other biological functions. The transcriptome of loggerhead turtle was sequenced using the Illumina Hiseq 2000 platform and de novo assembly was performed using the Trinity pipeline. The assembly comprised 515,597 contigs with an N50 of 2,631 bp. Contigs were analyzed with CD-Hit obtaining 374,545 unigenes, of which 165,676 had ORFs encoding putative proteins longer than 100 amino acids. A total of 52,147 (31.5%) of these transcripts had significant homology matches in at least one of the five databases used. From the enrichment of GO terms, 180 proteins with antioxidant activity were identified, among these 28 Prdxs and 50 putative Tnxs. The putative proteins of loggerhead turtles encoded by the genes Prdx1, Prdx3, Prdx5, Prdx6, Txn and Txnip were predicted and characterized in silico. When comparing Prdxs and Txns of loggerhead turtle with homologous human proteins, they showed 18 (9%), 52 (18%) 94 (43%), 36 (16%), 35 (33%) and 74 (19%) amino acid mutations respectively. However, they showed high conservation in active sites and structural motifs (98%), with few specific modifications. Of these, Prdx1, Prdx3, Prdx5, Prdx6, Txn and Txnip presented 0, 25, 18, three, six and two deleterious changes. This study provides a high quality blood transcriptome and functional annotation of loggerhead sea turtles.

Nesting activity of the Loggerhead sea turtle, Caretta caretta, in Calabria: 2016-2020 reproductive seasons

The nesting activity of the loggerhead turtle along the coast of Calabria during five reproductive seasons (2016-2020) is presented. From May to August, survey methods were conducted using traditional observations on foot and utilizing additional innovative technologies, including electric fat bikes and drones. Monitoring was intensively focused on a key nesting area of approximately 40 km located on the southernmost Ionian coast of Reggio Calabria, and, for only the 2020 season, the area was extended an additional 40 km northeast. In the five nesting seasons, 419 female emergences were recorded, 192 of which were classified as nests. Overall, 65.1% (n = 125) of nests were found along the area regularly monitored in all nesting seasons. The maximum nesting activity was observed during July, and the mean nesting success was 45.8%. The average number of eggs for all seasons was 92.2 (n = 163; S.D.: ± 21.1; range: 39-160). The mean hatching and emergence success rates were 70.8% and 62.5%, respectively. The mean incubation period was 49 days (n = 122; S.D.: ± 5.4; range: 42-82). Considering previous data from the same area, our study shows an actual increase in nesting activity, probably due to the implementation of an effective monitoring method that includes the use of new technologies, which made it possible to expand the regularly monitored area, and a greater awareness campaign in recent years. Regarding the reproductive parameters, our data is similar to that reported for other Mediterranean nesting grounds.

Globally important refuge for the loggerhead sea turtle: Maio Island, Cabo Verde

One of the largest nesting colonies of the Vulnerable loggerhead sea turtle Caretta caretta is in Cabo Verde. Here we present the first comprehensive study of loggerhead turtle nesting on the island of Maio in Cabo Verde. During 2016–2019 we monitored 38 km of undeveloped sandy beaches that have minimal artificial lighting and where all nesting on Maio takes place. We counted 4,063 nests in 2016, 5,429 in 2017, 14,364 in 2018 and 7,937 in 2019. The estimated total number of females was 1,016, 1,357, 3,591 and 1,984 in each of these years, respectively. Our findings suggest there are more loggerhead turtles nesting in Cabo Verde than previously estimated, and that this could be the species’ largest nesting subpopulation (followed by Florida, USA and Oman). The inter-annual hatching success (the proportion of eggs producing hatchlings) was 29–38% for the whole island but varied between sites. Our study of 250 clutches showed that flooding affected 38–61% and predation by crabs 40–42%, with hatching success on different beaches in the range of 1–59%. Poaching of eggs was rare (< 2% of clutches), but dogs predated 68.4% of all clutches on the beach nearest the largest human settlement. We evaluated different nest management strategies at multiple sites and estimated productivity of hatchlings (the number of hatchlings that would reach the sea for each management strategy), finding that hatcheries are not always the best option for nest management. As the beaches on Maio are relatively undisturbed, and there is a high abundance and density of turtle nests, the island should be protected as a globally important site for the conservation of the loggerhead turtle, and of coastal biodiversity more broadly.

Testing a Novel Aggregated Methodology to Assess Hydrodynamic Impacts on a High-Resolution Marine Turtle Trajectory

We designed a novel aggregated methodology to infer the impact of ocean motions on the movements of satellite-tracked marine turtles adopting available oceanographic observations and validated products of a numerical oceanographic forecasting system. The method was tested on an 11-months trajectory of a juvenile loggerhead turtle (LT) wandering in the Tyrrhenian Sea (Mediterranean Sea) that was reconstructed with a high-resolution GPS tracking system. The application of ad-hoc designed metrics revealed that the turtle’s route shape, ground speed and periodicities of its explained variance mimic the inertial motions of the sea, showing that this methodology is able to reveal important details on the relation between turtle movements and oceanographic features. Inertial motions were also identified in the observed trajectory of a surface drifting buoy sampling the Tyrrhenian Sea in a common period. At each sampling point of the turtle trajectory, the sea current eddy kinetic energy (EKE) and a Sea Current Impact index were computed from a validated set of high-resolution ocean modeling products and their analysis showed the relevant effects of the highly variable local sea currents mechanical action. Specifically, the metric we adopted revealed that the turtle trajectory was favorably impacted by the encountered sea current advection for about 70% of its length. The presented oceanographic techniques in conjunction with high-resolution tracking system provide a practicable approach to study marine turtle movements, leading the way to discover further insights on turtle behavior in the ocean.

Transcriptional Analyses of Acute Exposure to Methylmercury on Erythrocytes of Loggerhead Sea Turtle

To understand changes in enzyme activity and gene expression as biomarkers of exposure to methylmercury, we exposed loggerhead turtle erythrocytes (RBCs) to concentrations of 0, 1, and 5 mg L−1 of MeHg and de novo transcriptome were assembled using RNA-seq. The analysis of differentially expressed genes (DEGs) indicated that 79 unique genes were dysregulated (39 upregulated and 44 downregulated genes). The results showed that MeHg altered gene expression patterns as a response to the cellular stress produced, reflected in cell cycle regulation, lysosomal activity, autophagy, calcium regulation, mitochondrial regulation, apoptosis, and regulation of transcription and translation. The analysis of DEGs showed a low response of the antioxidant machinery to MeHg, evidenced by the fact that genes of early response to oxidative stress were not dysregulated. The RBCs maintained a constitutive expression of proteins that represented a good part of the defense against reactive oxygen species (ROS) induced by MeHg.

Cucullanus carettae Baylis, 1923, in a loggerhead sea turtle (Caretta caretta) from the Adriatic sea: first detection and molecular characterization

Cucullanus carettae Baylis, 1923 (Nematoda: Cucullanidae) is found worldwide in loggerhead turtles (Caretta caretta). Regarding the Mediterranean, C. carettae has been identified in the Tyrrhenian and the Ionian Sea and a unique description of a Cucullanus sp. specimen in loggerheads from the Adriatic Sea has been reported in the literature so far. In the framework of a bio-monitoring project of the Abruzzo and Molise coasts, a parasitological survey was performed on stranded and by-caught sea turtles, at the Istituto Zooprofilattico of Abruzzo and Molise “G. Caporale.” During necropsy, the gastrointestinal system of 72 stranded loggerhead turtles was analyzed for the presence of endoparasites and fecal samples were collected for coprological examination. Adult C. carettae (n = 123) was found in the upper intestine of one loggerhead turtle, associated with chronic lymphoplasmocytic enteritis. Additionally, five stool samples (6.9%) were positive for Cucullanus sp. eggs. Molecular characterization of adult nematodes was carried out to study phylogenetic relationships among the Cucullanus species. To our knowledge, this is the first morphological and molecular identification of C. carettae in loggerhead turtles from the Adriatic Sea. Additional studies on the distribution of this parasite in the Mediterranean are encouraged.