Phthalate esters (PAEs) concentration pattern reflects dietary habitats (δ13C) in blood of Mediterranean loggerhead turtles (Caretta caretta).

Phthalic acid esters (PAEs) are classified as endocrine disruptors, but it remains unclear if they can enter the marine food-web and result in severe health effects for organisms. Loggerhead turtles (Caretta caretta) can be chronically exposed to PAEs by ingesting plastic debris, but no information is available about PAEs levels in blood, and how these concentrations are related to diet during different life stages. This paper investigated, for the first time, six PAEs in blood of 18 wild-caught Mediterranean loggerhead turtles throughout solid-phase extraction coupled with gas chromatography-ion trap/mass spectrometry. Stable isotope analyses of carbon and nitrogen were also performed to assess the resource use pattern of loggerhead turtles. DEHP (12-63 ng mL-1) and DBP (6-57 ng mL-1) were the most frequently represented PAEs, followed by DiBP, DMP, DEP and DOP. The total PAEs concentration was highest in three turtles (124-260 ng mL-1) whereas three other turtles had concentrations below the detection limit. PAEs were clustered in three groups according to concentration in all samples: DEHP in the first group, DBP, DEP, and DiBP in the second group, and DOP and DMP in the third group. The total phthalates concentration did not differ between large-sized (96.3 ± 86.0 ng mL-1) and small-sized (67.1 ± 34.2 ng mL-1) turtles (p < 0.001). However, DMP and DEP were found only in large-sized turtles and DiBP and DBP had higher concentrations in large-sized turtles. On the other hand, DEHP and DOP were found in both small- and large-sized turtles with similar concentrations, i.e. ~ 21.0/32.0 ng mL-1 and ~ 7.1/9.9 ng mL-1, respectively. Winsored robust models indicated that δ13C is a good predictor for DBP and DiBP concentrations (significant Akaike Information criterion weight, AICwt). Our results indicate that blood is a good matrix to evaluate acute exposure to PAEs in marine turtles. Moreover, this approach is here suggested as a useful tool to explain the internal dose of PAEs in term of dietary habits (δ13C), suggesting that all marine species at high trophic levels may be particularly exposed to PAEs, despite their different dietary habitats and levels of exposure.

COMPARISON OF INTRAOCULAR PRESSURE MEASUREMENT USING REBOUND AND APPLANATION TONOMETRY IN LOGGERHEAD (CARETTA CARETTA) SEA TURTLES.

This study aimed to evaluate intraocular pressure (IOP) estimates in healthy eyes of Caretta caretta using rebound tonometry in comparison with applanation tonometry. Twenty-three healthy C. caretta (housed at the Marine Turtle Research Center) without preexisting ophthalmic disease were enrolled in the study. IOP measurements were obtained by the same ophthalmologist, with the turtle in ventral recumbency between 2:30 p.m. and 4:30 p.m., using a rebound tonometer (RT; TonoVet) in dog calibration mode, and after topical anesthesia, an applanation tonometer (AT; Tono-Pen) in both eyes. The average of three readings per instrument was used for analysis. The agreement between the two tonometers was assessed by Bland-Altman analysis and intraclass correlation coefficient (ICC). Moreover, differences in IOP between the two tonometers were analyzed using the Mann-Whitney test. Moderate agreement was found between the two tonometers (ICC, 0.663; 95% confidence interval, 0.206-0.857). The median, Q1, and Q3 IOP obtained with AT (6.2, 4.7, and 9.1 mm Hg) were significantly lower (P = 0.001) than that obtained with RT (9.7, 8.3, and 11.6 mm Hg). It was not possible to obtain an instrument automatically generated mean of four values with AT because of retraction of the globe by the animals, and IOP measurement was unsuccessful in 7 eyes. In conclusion, IOP readings from the RT were statistically higher than those from the AT. RT proved to be more feasible because of the light, short-lasting contact with the cornea.

Gastrointestinal investigation of parasites and Enterobacteriaceae in loggerhead sea turtles from Italian coasts.

BACKGROUND: Caretta caretta is the most abundant sea turtle species in the Mediterranean, and studies on this species have vastly expanded during recent years, including those investigating gut bacterial and parasitic communities. Members of these communities have been reported with variable prevalence and pathogenicity, mainly depending on their host and environment (e.g. lifespan, distribution, habitat, diet, health status and stressors). Indeed, many species commonly inhabiting the sea turtle gastrointestinal tract exhibit an opportunistic behaviour. This study aimed to provide baseline data on enterobacterial and parasitic composition, through bacteriological culture-based methods and the FLOTAC parasitological technique, in cloacal and faecal samples of 30 live Caretta caretta, examined upon their arrival at the Marine Turtle Research Centre (Portici, Italy). RESULTS: Enterobacteriaceae were isolated in 18/23 cloacal samples (78.3%), with Citrobacter and Morganella as the most common genera, followed by Proteus, Enterobacter, Providencia, and Hafnia. Parasitic elements were detected in 11/30 faecal samples (36.7%), with Enodiotrema, Rhytidodes, and Eimeria as most common genera, followed by Pachypsolus and Cymatocarpus. Additionally, Angiodyctium is reported for the first time in this host. The majority (47.8%) of sea turtles hosted exclusively Enterobacteriaceae, whereas 30.4% hosted both parasites and Enterobacteriaceae; the remaining 21.8% hosted neither of the agents. CONCLUSIONS: Bacteria and parasites evaluated in the present study are common in Mediterranean loggerhead sea turtles, with slight differences between the western and eastern basin. Although naturally present in the gastrointestinal system of free-living sea turtles, their relationship with these hosts might range from mutualism to parasitism. Indeed, members of the gut community might express their pathogenic potential in immune-compromised animals, such as those in rehabilitation facilities. Therefore, it is advisable to include in the standard work-up of rescued sea turtles a screening procedure for such opportunistic agents, in order to better evaluate the animal's health status and achieve timely intervention with appropriate treatment, thus improving rehabilitation. Furthermore, data collected from free-living sea turtles represent a starting point for investigating wild populations. However, further studies are needed to clarify the differences between sea turtle's normal gut microbiome and pathobiome.

Aeromonas induced polyostotic osteomyelitis in a juvenile loggerhead sea turtle Caretta caretta.

Bacterial bone infections have been occasionally reported in wild sea turtles. This study reports on a sub-adult Caretta caretta affected by Aeromonas hydrophila osteomyelitis, with extensive involvement of cranial and caudal flippers. The turtle was severely debilitated, had limited flipper mobility and showed signs of pain in reaction to manipulation. Radiographs and computed tomography revealed multiple lytic bone lesions. Since an infectious polyostotic osteomyelitis was suspected, the turtle was subjected to echo-assisted fine needle aspiration to characterize the etiology of the disease. Bacterial cultures and antibiotic susceptibility testing led to the isolation of Aeromonas hydrophila responsive to amikacin and doxycycline. Therefore, the turtle was treated with these antibiotics and monitored through repeat bacterial cultures and diagnostic imaging. The turtle was released 17 mo after admission, upon resolution of clinical signs. The documentation of this case provides a treatment approach that may improve the outcome of Aeromonas-associated osteomyelitis, especially in endangered wildlife species.

Occurrence and pattern of legacy and emerging per- and Poly-FluoroAlkyl substances (PFAS) in eggs of loggerhead turtle Caretta caretta from western Mediterranean.

Per-and Poly-FluoroAlkyl Substances (PFAS) are a class of persistent, toxic, and mobile and chemicals both from industrial sources and from the use and disposal of Consumers products containing PFAS, whose concentration in marine food webs could pose a toxicological risk for biota and humans. In 2021, unhatched eggs were sampled from 41 loggerhead turtle Caretta caretta nests from the Italian shores of the Campania Region (Southern Italy). Whole eggs were analysed for the presence of 66 legacy and emerging PFAS with Liquid Chromatography coupled to Hybrid High Resolution Mass Spectrometry. A median Σ66 Per- and Poly-FluoroAlkyl Substances value of 3.34 ng/g egg fresh weight was found; perfluoroctane sulfonate (PFOS) represented the most contributing congener (47%), followed by perfluoro-n-undecanoic acid, perfluoro-n-tridecanoic acid, perfluoro-n-decanoic acid, perfluoro-n-decanoic acid, and perfluoro-n-tetradecanoic acid, respectively. Such compounds showed a log-norm distribution, suggesting found concentrations could represent the baseline levels in the considered sampling area. Emerging ChloroPolyFluoroPolyEthers Carboxylic Acids (ClPFECAs) were found in 20 out of 41 samples in the range 0.01-1.59 ng/g. Four samples had 20-100 fold higher concentration compared to that of other samples, suggesting the presence of hot spot areas possibly related to presence of fluoropolymer-based marine litter turtles may ingest. The analysis of two paired eggs/liver samples recovered from stranded animals revealed PFAS concentration in the same order of magnitude, supporting the role of vitellogenin in their selective transfer to yolk. Significant (P = 0.0155) Kendall negative correlation coefficient of -0.2705 among PFOS content in eggs and the recorded hatching success prompts for further investigation on associated exposure assessment and related eco-toxicity risk. This work reports for the first time PFAS presence in georeferenced loggerhead turtle eggs of the Mediterranean Sea and results represent a starting point to study PFAS time-trends in this vulnerable species.

Are we even close? Five years marine litter ingestion monitoring in loggerhead turtles along Italian coast reveals how far we are from the Good Environmental Status.

The loggerhead sea turtle Caretta caretta has been chosen as bioindicator to monitor the amount of litter ingested by marine animals within the European Marine Strategy Framework Directive and the Barcelona Regional Sea Convention. European Member States and Contracting Parties are committed to achieve the Good Environmental Status (GES), which is reached when the quantity of ingested litter does not adversely affect the health of the species concerned. Although the monitoring strategy has been outlined for more than a decade, to date no threshold values have been adopted to verify GES achievement. After five years of extensive monitoring along the Italian coasts, this study evaluates the suitability of five different GES scenarios and proposes a new threshold value (i.e., "there should be less than 33% of sea turtles having more than 0.05 g of ingested plastic in the GI") for its implementation in the European seas and the Mediterranean basin.

Increase of nesting habitat suitability for green turtles in a warming Mediterranean Sea.

Climate change is reshaping global ecosystems at an unprecedented rate, with major impacts on biodiversity. Therefore, understanding how organisms can withstand change is key to identify priority conservation objectives. Marine ectotherms are being extremely impacted because their biology and phenology are directly related to temperature. Among these species, sea turtles are particularly problematic because they roam over both marine and terrestrial habitats throughout their life cycles. Focusing on green turtles (Chelonia mydas) in the Mediterranean Sea, we investigated the future potential changes of nesting grounds through time, assuming that marine turtles would shift their nesting locations. We modeled the current distribution of nesting grounds including both terrestrial and marine variables, and we projected the potential nesting distribution across the Mediterranean basin under alternative future greenhouse gas emission scenario (2000-2100). Our models show an increase in nesting probability in the western Mediterranean Sea, irrespective of the climate scenario we consider. Contrary to what is found in most global change studies, the worse the climate change scenario, the more suitable areas for green turtles will potentially increase. The most important predictors were anthropogenic variables, which negatively affect nesting probability, and sea surface temperature, positively linked to nesting probability, up to a maximum of 24-25 °C. The importance of the western Mediterranean beaches as potential nesting areas for sea turtles in the near future clearly call for a proactive conservation and management effort, focusing on monitoring actions (to document the potential range expansion) and threat detection.

Electroretinography, Ocular Ultrasonography, and Phacoemulsification of Bilateral Cataracts in Two Juvenile Loggerhead Sea Turtles (Caretta caretta) of the Mediterranean Region.

Bilateral cataracts were diagnosed in two rescued juvenile, immature loggerhead sea turtles (Caretta caretta), weighing 1.65 and 1.7 kg. Both animals showed vision impairment and difficulty in feeding without assistance. In fact, they did not notice the presence of the food in the tank unless it was brought close to touching the mouth. Ocular ultrasonography and electroretinography showed no lesions of the vitreal body and retinal layer, therefore, both animals were candidates for bilateral cataract surgery. Topical administration of tropicamide + phenylephrine alternating with rocuronium resulted in only minimal mydriasis. Administration of intracameral rocuronium did not improve mydriasis. Phacoemulsification using a one-handed technique was performed bilaterally with a phacoemulsification device (Sovereign, AMO (Abbott Medical Optics®). After surgery, the systemic anti-inflammatory drug (dexamethasone 0.2 mg/kg, IM daily for one week) and antibiotics (enrofloxacin 10 mg/kg IM q 72 h, for 4 weeks; ceftazidime 20 mg/kg IM q 72 h for 3 weeks) were administered. Topical ofloxacin, flurbiprofen and tobramycin/dexamethasone were instilled TID for 4 weeks. Both turtles regained vision in both eyes. Results at a 10-month follow-up were satisfactory. This is the first report of cataracts in turtles rescued in the Mediterranean Sea and the first description of surgical treatment of cataracts in loggerhead turtles so young.

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.

Dietary Preferences of Loggerhead Sea Turtles (Caretta caretta) in Two Mediterranean Feeding Grounds: Does Prey Selection Change with Habitat Use throughout Their Life Cycle?

According to their life stage, the loggerhead sea turtle (Caretta caretta) is found in a wide range of habitats, from neritic to more oceanic areas. Their feeding habits are expected to change as they develop, along with habitat use. Juvenile sea turtles are hypothesized to feed on pelagic species in oceanic areas, shifting to more benthic prey during the subadult and adult stages. We analyzed the gastrointestinal content from 150 loggerhead sea turtles stranded and/or bycaught along the Adriatic coast of the Abruzzo and Molise regions (n = 89) and the Tyrrhenian coast of the Lazio and Campania regions (n = 61) from 2018 to 2021. Food items were identified to the lowest taxonomic level, and the frequency of occurrence was calculated for each taxon and most recurrent species to assess changes in prey selection during the development. The marine litter was categorized, and the frequency of occurrence was calculated for the ingestion of litter. The most recurrent taxonomic prey group recorded in the Adriatic sample was Arthropoda (94%), followed by Mollusca (63%) and Chordata (34%). In the Tyrrhenian sample, loggerhead sea turtles fed mostly on Mollusca (84%), Arthropoda (38%), and Chordata (26%). Surprisingly, the Adriatic-Tyrrhenian sample groups showed similar feeding behavior between juveniles, subadults, and adults. A similar correlation has been observed concerning the ingestion of litter. Moreover, this study confirms the opportunistic feeding behavior of loggerhead sea turtles and their high adaptability.

Unveiling the egg microbiota of the loggerhead sea turtle Caretta caretta in nesting beaches of the Mediterranean Sea.

Microbes have central roles in the development and health of animals, being the introduction of specific microbial species a potential conservation strategy to protect animals from emerging diseases. Thus, insight into the microbiota of the species and their habitats is essential. In this manuscript, we report for the first time the bacterial composition of all the components (eggshells of hatched and unhatched eggs, internal content of unhatched eggs, intestinal content of hatchling and pipping sea turtles, and sand) of three nesting beaches of Caretta caretta along the Italian coasts of the Mediterranean Sea. The analysis of 26 amplicon samples was carried out using next-generation sequencing analysis, targeting V3-V4 regions of the bacterial 16S rRNA gene. Samples featured mainly Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes, whose percentages depended on the sample type. Our results showed that, although from different sampling sites, the internal content of the unhatched eggs, intestinal content of hatchling and pipping sea turtles share the microbiota, which was yet different from that of eggshells and sand of the same nesting beach. This study suggests the maternal and environmental influence alongside a protective role of eggshells in shaping the egg microbiota of Caretta caretta sea turtles.

Detection of Chlamydial DNA from Mediterranean Loggerhead Sea Turtles in Southern Italy.

Chlamydiae are obligate intracellular bacteria that include pathogens of human and veterinary importance. Several reptiles were reported to host chlamydial agents, but pathogenicity in these animals still needs clarification. Given that only one report of chlamydiosis was described in sea turtles, and that chlamydiae might also be detected in hosts without clinical signs, the current study examined asymptomatic Mediterranean loggerhead sea turtles for the presence of chlamydial DNA. Twenty loggerhead sea turtles, rehabilitated at the Marine Turtle Research Centre (Portici, Italy), were examined collecting ocular-conjunctival, oropharyngeal and nasal swabs. Samples were processed through quantitative and conventional PCR analyses to identify Chlamydiales and Chlamydiaceae, with particular attention to C. pecorum, C. pneumoniae, C. psittaci, and C. trachomatis. Although it was not possible to determine the species of chlamydiae involved, the detection of chlamydial DNA from the collected samples suggests that these microorganisms might act as opportunistic pathogens, and underlines the role of sea turtles as potential carriers. This study highlights the presence of chlamydial agents in sea turtles, and encourages further research to fully characterize these microorganisms, in order to improve the management of the health and conservation of these endangered species, and prevent potential zoonotic implications.

Symblepharon, Ankyloblepharon, and Salt Gland Dysfunction in a Loggerhead Sea Turtle (Caretta caretta).

Adhesions involving the bulbar and the palpebral conjunctiva (Symblepharon) may interfere with tear drainage, cause chronic conjunctivitis, and reduce ocular motility. This condition may be associated with adhesion of the edges of the upper and lower eyelids (ankyloblepharon). The present case describes bilateral symblepharon, ankyloblepharon and salt gland dysfunction in a juvenile Caretta caretta. The loggerhead presented both eyelids swollen, ulcerated, and not separable when rescued. Eye examination was not possible, but ultrasonography showed right bulbar integrity, while the left eye was smaller, with a thicker cornea that had lost its normal doubled lined structure. Surgical dissection of the fibrous adhesions between the palpebral and bulbar conjunctiva, cornea, and third eyelid was performed, and large dacryoliths were removed. The microscopic findings were consistent with chronic keratoconjunctivitis. Ultrastructurally, no virus-like particles were observed. In addition, tissue samples were negative for herpesvirus by qualitative PCR. The eyelids of both eyes and the corneal epithelium of the right eye healed; moreover, the vision was restored in the right eye. There were no recurrences after 12 months of follow-up, and the turtle was released 16 months after the end of treatments on the southern Tyrrhenian coast in the western Mediterranean Sea. To the authors' knowledge, this is the first report of symblepharon with ankyloblepharon and salt gland dysfunction in Caretta caretta turtle. Ocular ultrasonography was helpful in the preliminary diagnostic work-up.

Polymer composition assessment suggests prevalence of single-use plastics among items ingested by loggerhead sea turtles in the western mediterranean sub-region.

The ingestion of plastic is becoming a major concern for various species and particularly for marine turtles across the globe. The loggerhead sea turtle (Caretta caretta) was recently chosen by the European Commission as a bio-indicator for plastic pollution within the Mediterranean basin. We further investigated which items this key species is more prone to ingest, following the standardised Marine Strategy Framework Directive protocols. Moreover, we integrated to this protocol the Fourier Transform Infrared Spectroscopy, which allowed us to determine the polymer type of each item. We analysed samples from 226 sea turtles from 2008 to 2017 in two areas of the western Mediterranean sub-region (sensu MSFD). In the Lazio area we found a frequency of occurrence of plastic ingestion of 78.33%, while in Sardinia 41.79%. The analysis of the litter categories, among all individuals, highlights a prevalence of user-sheet (Use-She; 69.13%) and user-fragment plastics (Use-Fra; 20.84%). In addition, the polymer analysis showed a dominance of polyethylene (65.98%) and polypropylene (26.23%). As a result, by looking at other works that have investigated polymer types and items sources, we are able to infer that 77.25% of the objects ingested by the C. caretta individuals are attributable to disposable daily-life objects managed in an improper way. Therefore, C. caretta apart from being an efficient bio-indicator for plastic pollution, highlighting spatial and temporal concentration differences, it could also be used to verify the effectiveness of the Single-use Plastic Directive (EU 2019/904).

Movement patterns of large juvenile loggerhead turtles in the Mediterranean Sea: Ontogenetic space use in a small ocean basin.

Mechanisms that determine how, where, and when ontogenetic habitat shifts occur are mostly unknown in wild populations. Differences in size and environmental characteristics of ontogenetic habitats can lead to differences in movement patterns, behavior, habitat use, and spatial distributions across individuals of the same species. Knowledge of juvenile loggerhead turtles' dispersal, movements, and habitat use is largely unknown, especially in the Mediterranean Sea. Satellite relay data loggers were used to monitor movements, diving behavior, and water temperature of eleven large juvenile loggerhead turtles (Caretta caretta) deliberately caught in an oceanic habitat in the Mediterranean Sea. Hidden Markov models were used over 4,430 spatial locations to quantify the different activities performed by each individual: transit, low-, and high-intensity diving. Model results were then analyzed in relation to water temperature, bathymetry, and distance to the coast. The hidden Markov model differentiated between bouts of area-restricted search as low- and high-intensity diving, and transit movements. The turtles foraged in deep oceanic waters within 60 km from the coast as well as above 140 km from the coast. They used an average area of 194,802 km2, where most individuals used the deepest part of the Southern Tyrrhenian Sea with the highest seamounts, while only two switched to neritic foraging showing plasticity in foraging strategies among turtles of similar age classes. The foraging distribution of large juvenile loggerhead turtles, including some which were of the minimum size of adults, in the Tyrrhenian Sea is mainly concentrated in a relatively small oceanic area with predictable mesoscale oceanographic features, despite the proximity of suitable neritic foraging habitats. Our study highlights the importance of collecting high-resolution data about species distribution and behavior across different spatio-temporal scales and life stages for implementing conservation and dynamic ocean management actions.

Humerus osteology, myology, and finite element structure analysis of Cheloniidae.

Adaptation of osteology and myology lead to the formation of hydrofoil foreflippers in Cheloniidae (all recent sea turtles except Dermochelys coriacea) which are used mainly for underwater flight. Recent research shows the biomechanical advantages of a complex system of agonistic and antagonistic tension chords that reduce bending stress in bones. Finite element structure analysis (FESA) of a cheloniid humerus is used to provide a better understanding of morphology and microanatomy and to link these with the main flipper function, underwater flight. Dissection of a Caretta caretta gave insights into lines of action, that is, the course that a muscle takes between its origin and insertion, of foreflipper musculature. Lines of action were determined by spanning physical threads on a skeleton of Chelonia mydas. The right humerus of this skeleton was micro-CT scanned. Based on the scans, a finite element (FE) model was built and muscle force vectors were entered. Muscle forces were iteratively approximated until a uniform compressive stress distribution was attained. Two load cases, downstroke and upstroke, were computed. We found that muscle wrappings (m. coracobrachialis magnus and brevis, several extensors, humeral head of m. triceps) are crucial in addition to axial loading to obtain homogenous compressive loading in all bone cross-sections. Detailed knowledge on muscle disposition leads to compressive stress distribution in the FE model which corresponds with the bone microstructure. The FE analysis of the cheloniid humerus shows that bone may be loaded mainly by compression if the bending moments are minimized.

Detection and partial characterization of herpesviruses from Egyptian tortoises (Testudo kleinmanni) imported into Italy from Libya.

A group of approximately 370 Egyptian tortoises (Testudo kleinmanni) and 36 spur-thighed tortoises (Testudo graeca) were illegally imported into Italy from Libya. Within 6 mo of their entry into Italy, all but 40 of the Egyptian tortoises had died with signs of severe stomatitis. Herpesviruses were detected from the tongues of seven Egyptian tortoises by polymerase chain reaction (PCR) and virus isolation. Sequencing of a portion of the UL39 homologue of the herpesviruses from three different tortoises demonstrated that the viruses were identical to one another and identical to a herpesvirus isolated from a Hermann's tortoise (Testudo hermanni) in Germany. This is the first description of the detection of a herpesvirus from diseased Egyptian tortoises. That these animals were imported into Europe from Libya provides circumstantial evidence for the presence of herpesviruses among tortoises in northern Africa.

Loggerhead sea turtles as sentinels in the western Mediterranean: antibiotic resistance and environment-related modifications of Gram-negative bacteria.

Sea turtles possess relevant characteristics to serve as sentinel species for monitoring the health of marine ecosystems, which is currently threatened. This study examined 35 loggerhead turtles from the western Mediterranean, focusing on the oral and cloacal prevalence of aerobic Gram-negative bacteria, their antibiotic resistance and the influence of several variables linked both to the animal and the environment (i.e. estimated life stage; area, season and cause of recovery; plastic ingestion). Conventional bacteriology methods led to the isolation of bacterial families commonly regarded as opportunistic pathogens (i.e. Aeromonadaceae; Enterobacteriaceae; Pseudomonadaceae; Shewanellaceae; Vibrionaceae), but pointing out sea turtles as carriers of potential zoonotic agents. The high rates of antibiotic resistance, here detected, raise important concerns on the dissemination of this phenomenon in marine environments. Moreover, several of the examined variables showed a significant influence on the prevalence of bacterial families, strengthening the role of sea turtles as mirrors of their ecosystems.

Parallel pattern of differentiation at a genomic island shared between clinal and mosaic hybrid zones in a complex of cryptic seahorse lineages.

Diverging semi-isolated lineages either meet in narrow clinal hybrid zones, or have a mosaic distribution associated with environmental variation. Intrinsic reproductive isolation is often emphasized in the former and local adaptation in the latter, although both reduce gene flow between groups. Rarely are these two patterns of spatial distribution reported in the same study system. Here, we report that the long-snouted seahorse Hippocampus guttulatus is subdivided into discrete panmictic entities by both types of hybrid zones. Along the European Atlantic coasts, a northern and a southern lineage meet in the southwest of France where they coexist in sympatry-i.e., in the same geographical zone-with little hybridization. In the Mediterranean Sea, two lineages have a mosaic distribution, associated with lagoon-like and marine habitats. A fifth lineage was identified in the Black Sea. Genetic homogeneity over large spatial scales contrasts with isolation maintained in sympatry or close parapatry at a fine scale. A high variation in locus-specific introgression rates provides additional evidence that partial reproductive isolation must be maintaining the divergence. We find that fixed differences between lagoon and marine populations in the Mediterranean Sea belong to the most differentiated SNPs between the two Atlantic lineages, against the genome-wide pattern of structure that mostly follow geography. These parallel outlier SNPs cluster on a single chromosome-wide island of differentiation. Since Atlantic lineages do not map to lagoon-sea habitat variation, genetic parallelism at the genomic island suggests a shared genetic barrier contributes to reproductive isolation in contrasting contexts-i.e., spatial versus ecological. We discuss how a genomic hotspot of parallel differentiation could have evolved and become associated both with space and with a patchy environment in a single study system.

Author Correction: Sporadic nesting reveals long distance colonisation in the philopatric loggerhead sea turtle (Caretta caretta).

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.