Vermetid gastropods as key intermediate hosts for a lineage of marine turtle blood flukes (Digenea: Spirorchiidae), with evidence of transmission at a turtle rookery

Blood flukes of the family Spirorchiidae Stunkard, 1921 are significant pathogens of marine turtles, both in the wild and in captivity. Despite causing considerable disease and mortality, little is known about the life cycles of marine species, with just four reports globally. No complete life cycle has been elucidated for any named species of marine spirorchiid, but the group is reported to use vermetid and fissurellid gastropods, and terebelliform polychaetes as intermediate hosts. Here we report molecular evidence that nine related spirorchiid species infect vermetid gastropods as first intermediate hosts from four localities along the coast of Queensland, Australia. ITS2 rDNA and cox1 mtDNA sequence data generated from vermetid infections provides the first definitive identifications for the intermediate hosts for the four species of Hapalotrema Looss, 1899 and Learedius learedi Price, 1934. Additionally, we provide a new locality report for larval stages of Amphiorchis sp., and evidence of three additional unidentified spirorchiid species in Australian waters. Based on the wealth of infections from vermetids during this study, we conclude that the previous preliminary report of a fissurellid limpet as the intermediate host for L. learedi was likely mistaken. The nine species found infecting vermetids during this study form a strongly supported clade exclusive of species of the other two marine spirorchiid genera for which sequence data are available; Carettacola Manter & Larson, 1950 which falls sister to the vermetid-infecting clade + a small clade of freshwater spirorchiids, and Neospirorchis Price, 1934 which is distantly related to the vermetid-infecting clade. We provide further evidence that spirorchiid transmission can occur in closed system aquaria and show that spirorchiid transmission occurs at both an important turtle rookery (Heron Island, southern Great Barrier Reef, Australia) and foraging ground (Moreton Bay, Australia). We discuss the implications of our findings for the epidemiology of the disease, control in captivity, and the evolution of vermetid exploitation by the Spirorchiidae.     

Phylogeny and paraphyly among tetrapod blood flukes (Digenea: Schistosomatidae and Spirorchiidae)

The blood flukes of turtles (Digenea: Spirorchiidae) and the blood flukes of crocodilians, birds and mammals (Digenea: Schistosomatidae) have long been considered as closely related, but distinct evolutionary lineages. Recent morphological and molecular studies have considered these families as sister taxa within the Schistosomatoidea. Representatives of both families have similar furcocercous cercariae and similar two-host life cycles, but have different definitive hosts, distinct reproductive patterns and different morphologies. Sequences including approximately 1800 bases of the small subunit ribosomal DNA and 1200 bases of the large subunit ribosomal DNA were generated from representatives of eight spirorchiid genera. These sequences were aligned with pre-existing sequences of Schistosomatidae and other representatives of the Diplostomida and analysed for phylogenetic signal using maximum parsimony and Bayesian inference. These analyses revealed that the Spirorchiidae is paraphyletic and that the turtle blood flukes are basal to the highly derived schistosomatids. Three genera of spirorchiids from marine turtles form a sister group to the Schistosomatidae and five genera of spirorchiids from freshwater turtles occupy basal positions in the phylogeny of tetrapod blood flukes. Marine turtles are considered to be derived from freshwater turtles and the results of the current study indicate that the spirorchiid parasites of marine turtles are similarly derived from a freshwater ancestor. The close relationship of the marine spirorchiids to schistosomatids and the basal position of the marine transmitted Austrobilharzia and Ornithobilharzia in the schistosomatid clade suggests that schistosomatids arose after a marine turtle blood fluke ancestor successfully colonised birds.     

Development of an indirect immunofluorescence technique for the evaluation of generated antibody titers against Erysipelothrix rhusiopathiae in captive bottlenose dolphins (Tursiops truncatus)

Erysipelothrix rhusiopathiae is the causative agent of erysipelas, a disease of many mammalian and avian species, mainly swine and turkeys. In cetaceans, erysipelas is considered to be the most common infection in juvenile individuals, which have not been vaccinated. Moreover, the disease manifest in both forms, the dermatologic and the acute septicemic forms, has been reported in various species of dolphins and whales. It is difficult to diagnose erysipelas by currently available approaches. Moreover, it is mainly based on culture methods and also PCR methods, which are currently being developed. At the present stage, prophylactic approaches are based on antibiotic therapy and vaccination mostly with porcine erysipelas vaccines. In the present study, an Indirect Immuno Fluorescence method for the detection of dolphin antibodies levels against E. rhusiopathiae was developed and applied in two different groups of captive bottlenose dolphins (Tursiops truncatus) from Loro Parque (Tenerife, Canary Islands, Spain) and L’Oceanogràfic de Valencia (Valencia, Spain) in order to check the tittering levels of antibodies after application of porcine erysipelas vaccines in the studied dolphins.     

Helminth communities of loggerhead turtles (Caretta caretta) from Central and Western Mediterranean Sea: The importance of host's ontogeny

We investigated the factors providing structure to the helminth communities of 182 loggerhead sea turtles, Caretta caretta, collected in 6 localities from Central and Western Mediterranean. Fifteen helminth taxa (10 digeneans, 4 nematodes and 1 acanthocephalan) were identified, of which 12 were specialist to marine turtles; very low numbers of immature individuals of 3 species typical from fish or cetaceans were also found. These observations confirm the hypothesis that phylogenetic factors restrict community composition to helminth species specific to marine turtles. There were significant community dissimilarities between turtles from different localities, the overall pattern being compatible with the hypothesis that parasite communities reflect the ontogenetic shift that juvenile loggerheads undergo from oceanic to neritic habitats. The smallest turtles at the putative oceanic, pelagic-feeding stage harboured only the 2 digenean species that were regionally the most frequent, i.e. Enodiotrema megachondrus and Calycodes anthos; the largest turtles at the putative neritic, bottom-feeding stage harboured 11 helminth taxa, including 3 nematode species that were rare or absent in turtles that fed partially on pelagic prey. Mean species richness per host was low (range: 1.60–1.89) and did not differ between localities. Variance ratio tests indicated independent colonization of each helminth species. Both features are expected in ectothermic and vagrant hosts living in the marine environment.     

Detection and molecular characteristics of Rhytidodoides sp. (Digenea: Rhytidodidae) from the gall bladder of green sea turtles (Chelonia mydas) in the Ogasawara Islands,Japan

Trematodes of the genus Rhytidodoides are parasitic in marine turtles. Of the already known species, Rhytidodoides similis Price, 1939, occurs especially in the gall bladder. In this study, we surveyed 73 green sea turtles (Chelonia mydas) in the Ogasawara Islands, Japan, and detected Rhytidodoides sp. from the gall bladders of 18 turtles. A detailed morphological analysis revealed that the forebody of Rhytidodoides sp. differed slightly in shape from that of R. similis. There has been no information on DNA sequences of the family Rhytidodidae. A molecular phylogeny based on 28S rDNA sequences of Rhytidodoides sp. and related taxa suggested that the Rhytidodidae is sister to the other families of Echinostomatoidea. The intraspecific diversity of Rhytidodoides sp. was examined by using DNA sequences of mitochondrial cytochrome c oxidase subunit 1 gene (COI). The population genetic features of the COI haplotypes demonstrated that Rhytidodoides sp. is highly diverse in the Ogasawara Islands. The DNA sequences determined in this study will contribute to the species identification of congeners and the taxonomic reconsideration of the Echinostomatoidea.     

Fluke (Spirorchiidae) infections in sea turtles stranded on Taiwan: prevalence and pathology

The prevalence of spirorchiid fluke infections of marine turtles is high and may cause the death of the hosts throughout their ranges. Virtually nothing has been reported regarding the infective status of sea turtles stranded on Taiwan. Between 2007 and 2010, 30 green turtles (Chelonia mydas) and 2 loggerhead turtles ( Caretta caretta ), stranded and dead, were examined for spirorchiid flukes and their eggs. Twenty-four of the green turtles were juveniles, and the stranded loggerhead turtles were subadults. Adult spirorchiid flukes were found in 13 green turtles but not in the loggerheads. Four species of flukes were identified, namely, Leardius learedi , Hapalotrema postorchis , H. mehrai , and Carettacola hawaiiensis . The main infection sites were the major arteries and heart. Seventy percent of the green turtles harbored spirorchiid eggs, but no eggs were found in loggerheads. The largest eggs with bipolar spines, type I eggs, were found in every case. Although more than half of the stranded turtles were infected, parasite infections were not the main cause of death in the green turtles. Fishery by-catch is probably responsible for the mortality of these stranded turtles.     

Heavy Metal Accumulation in Four Species of Sea Turtles from the Baja California Peninsula, Mexico

Heavy metals were assessed in four species of sea turtles from the Baja California Peninsula, Mexico, representing the first report of heavy metal concentrations in tissues of post-yearling sea turtles from the Eastern Pacific. Concentrations of Cd measured in C. mydas kidney (653 μg/g dry wt) were the highest ever reported for any sea turtle species. Cd accumulated preferentially in kidney and the ratios of kidney to liver Cd in Baja California turtles were among the highest reported for sea turtles globally. Zn, Ni, and Mn concentrations were also significantly higher in kidney than other tissues, while Cu and Fe were greatest in liver, and all metals were lowest in muscle. With the exception of one value (69.9 μg/g in kidney of C. caretta), Pb was low in all tissues from Baja California. In comparisons across species, kidney of C. mydas had greater Zn and Ni concentrations as compared to other species, although there was no difference in liver metal levels among the species. Positive correlations were detected in the concentrations of Cd, Cu and Ni with the straight carapace length of C. caretta.     

The Head and Neck Anatomy of Sea Turtles (Cryptodira: Chelonioidea) and Skull Shape in Testudines

Background

Sea turtles (Chelonoidea) are a charismatic group of marine reptiles that occupy a range of important ecological roles. However, the diversity and evolution of their feeding anatomy remain incompletely known.

Methodology/Principal Findings

Using computed tomography and classical comparative anatomy we describe the cranial anatomy in two sea turtles, the loggerhead (Caretta caretta) and Kemp’s ridley (Lepidochelys kempii), for a better understanding of sea turtle functional anatomy and morphological variation. In both taxa the temporal region of the skull is enclosed by bone and the jaw joint structure and muscle arrangement indicate that palinal jaw movement is possible. The tongue is relatively small, and the hyoid apparatus is not as conspicuous as in some freshwater aquatic turtles. We find several similarities between the muscles of C. caretta and L. kempii, but comparison with other turtles suggests only one of these characters may be derived: connection of the m. adductor mandibulae internus into the Pars intramandibularis via the Zwischensehne. The large fleshy origin of the m. adductor mandibulae externus Pars superficialis from the jugal seems to be a characteristic feature of sea turtles.

Conclusions/Significance

In C. caretta and L. kempii the ability to suction feed does not seem to be as well developed as that found in some freshwater aquatic turtles. Instead both have skulls suited to forceful biting. This is consistent with the observation that both taxa tend to feed on relatively slow moving but sometimes armoured prey. The broad fleshy origin of the m. adductor mandibulae externus Pars superficialis may be linked to thecheek region being almost fully enclosed in bone but the relationship is complex.     

First satellite tracks of neonate sea turtles redefine the ‘lost years’ oceanic niche

Few at-sea behavioural data exist for oceanic-stage neonate sea turtles, a life-stage commonly referred to as the sea turtle ‘lost years’. Historically, the long-term tracking of small, fast-growing organisms in the open ocean was logistically or technologically impossible. Here, we provide the first long-term satellite tracks of neonate sea turtles. Loggerheads (Caretta caretta) were remotely tracked in the Atlantic Ocean using small solar-powered satellite transmitters. We show that oceanic-stage turtles (i) rarely travel in Continental Shelf waters, (ii) frequently depart the currents associated with the North Atlantic Subtropical Gyre, (iii) travel quickly when in Gyre currents, and (iv) select sea surface habitats that are likely to provide a thermal benefit or refuge to young sea turtles, supporting growth, foraging and survival. Our satellite tracks help define Atlantic loggerhead nursery grounds and early loggerhead habitat use, allowing us to re-examine sea turtle ‘lost years’ paradigms.     

Water temperature and internesting intervals for loggerhead (Caretta caretta) and green (Chelonia mydas) sea turtles

Temperature loggers were attached to the carapace of green turtles (Chelonia mydas) at Ascension Island and Cyprus and to loggerhead turtles (Caretta caretta) at Cyprus, in order to record the ambient temperature experienced by individuals during the internesting interval, i.e. the period between consecutive clutches being laid. Internesting intervals were relatively short (10–14 days) and mean ambient temperatures relatively warm (27–28°C), compared to previous observations for these species nesting in Japan, although a single internesting interval versus temperature relationship described all the data for these two species from the different areas. The implication is that water temperature has both a common and a profound effect on the length of the internesting interval for these two species: internesting intervals are shorter when the water is warmer.     

Variation in reproductive output of marine turtles

Most marine turtle species are non-annual breeders and show variation in both the number of eggs laid per clutch and the number of clutches laid in a season. Large levels of inter-annual variation in the number of nesting females have been well documented in green turtle nesting populations and may be linked to environmental conditions. Other species of marine turtle exhibit less variation in nesting numbers. This inter-specific difference is thought to be linked to trophic status. To examine whether individual reproductive output is more variable in the herbivorous green turtle (Chelonia mydas Linneaeus 1758) than the carnivorous loggerhead (Caretta caretta Linneaeus 1758), we examined the nesting of both species in Cyprus over nine seasons. Green turtles showed slower annual growth rates (0.11 cm year⁻¹ curved carapace length (CCL) and 0.27 cm year⁻¹ curved carapace width (CCW)) than loggerhead turtles (0.36 cm year⁻¹ CCL, 0.51 cm year⁻¹ CCW). CCL was highly correlated to mean clutch size in both green (R²=0.51) and loggerhead turtles (R²=0.61) and maximal clutch size of green turtles (R²=0.58). Larger females did not lay a greater number of clutches or have a shorter remigration interval than smaller females of either species. On average, the size of green turtle clutches increased and that of loggerhead turtles decreased as the season progressed. Individual green turtles, however, produced more eggs per clutch through the season to a maximum in the third or fourth clutch. In loggerhead turtles, clutches 1-4 were very similar in size but the fifth clutch was 38% smaller than the first. No individuals of either species were recorded laying more than five clutches. Green turtles may not be able to achieve their maximum reproductive output with respect to clutch size throughout the season, whereas only loggerhead turtles laying five clutches (n=5) appear to become resource depleted. Green turtles nesting in years when large numbers of nests were recorded laid a greater number of clutches than females nesting in years with lower levels of nesting.     

The life history of Sulcascaris sulcata (Nematoda: Ascaridoidea), a parasite of marine molluscs and turtles

Berry G. N. and Cannon L. R. G. 1981. The life history of Sulcascaris sulcata (Nematoda: Ascaridoidea), a parasite of marine molluscs and turtles. International Journal for Parasitoiogy11: 43–54. The morphology, development and hatching of Sulcascaris sulcata eggs are described. Two moults occurred in the egg. Third stage larvae spontaneously hatched and were found to develop in marine bivalves and gastropods. Larvae grew steadily and after three to four months, when about 5 mm long, they moulted to fourth stage larvae characteristic of natural infections in bivalves from commercial catches. Experimentally, when fed to laboratory-reared Caretta caretta, the fourth stage larvae first attached at the oesophago-gastric junction where they moulted to adults in 7–21 days. Subsequent growth to mature adults was obtained by at least 5 months after infection. It is suggested that under natural conditions the life history may take up to 2 years to complete. These findings are discussed in relation to the predatory mode of feeding and the breeding habits of C. caretta and the significance of a possible health hazard to man.     

Learedius learedi infection in black turtles (Chelonia mydas agassizii), Baja California Sur, Mexico

Black turtle (Chelonia mydas agassizii) carcasses, recovered as a result of incidental capture in Magdalena Bay, Mexico, revealed invasion by spirorchiid trematode eggs in liver, kidney, intestines, muscle, heart, pancreas, and duodenum. Seventy-five adult Learedius learedi Price, 1934, were recovered from the heart of 1 turtle. Most of the organs showed a mild or absent inflammatory response in histological sections, with the exception of a pancreatic-duodenal section that revealed severe lymphocyte and phagocyte infiltration associated with an infestation of more than 200 eggs. A linear formation of 35 eggs from the pancreas toward the intestinal lumen is described as resembling migration. This is among the first reports of a parasitic infection of L. learedi Price 1934, in C. m. agassizii in Mexico.     

First evidence of polychaete intermediate hosts for Neospirorchis spp. marine turtle blood flukes (Trematoda: Spirorchiidae)

Life cycles of spirorchiids that infect the vascular system of turtles are poorly understood. Few life cycles of these blood flukes have been elucidated and all intermediate hosts reported are gastropods (Mollusca), regardless of whether the definitive host is a freshwater or a marine turtle. During a recent survey of blood fluke larvae in polychaetes on the coast of South Carolina, USA, spirorchiid-like cercariae were found to infect the polychaetes Amphitrite ornata (Terebellidae) and Enoplobranchus sanguineus (Polycirridae). Cercariae were large, furcate, with a ventral acetabulum, but no eyespots were observed. Partial sequences of D1–D2 domains of the large ribosomal subunit, the internal transcribed spacer 2, and the mitochondrial cytochrome oxidase 1 genes allowed the identification of sporocysts and cercariae as belonging to two unidentified Neospirorchis species reported from the green turtle, Chelonia mydas, in Florida: Neospirorchis sp. (Neogen 13) in A. ornata and Neospirorchis sp. (Neogen 14) in E. sanguineus. Phylogenetic analysis suggests that infection of annelids by blood flukes evolved separately in aporocotylids and spirorchiids. Our results support the contention that the Spirorchiidae is not a valid family and suggest that Neospirorchis is a monophyletic clade within the paraphyletic Spirorchiidae. Since specificity of spirorchiids for their intermediate hosts is broader than it was thus far assumed, surveys of annelids in turtle habitats are necessary to further our understanding of the life history of these pathogenic parasites.     

New taxa described in Zoology in the Middle East

Abstract

The region between Mersin and ?skenderun was selected for studying marine turtles in the Turkish waters of the Mediterranean sea, as the most important nesting grounds of Green Turtles (Chelonia mydas) in the Mediterranean are situated there. In the 1995–96 fishing season, the 5 trawl boats taking part in the project reported that nets in the Eastern Mediterranean trapped 160 Green Turtles and 26 Loggerhead Turtles (Caretta caretta). In the following trawling season (1996/97), 306 Green Turtles, 116 Loggerhead Turtles (Caretta caretta) and 437 Nile Soft-shelled Turtles (Trionyx triunguis) were found to be trapped as a by-catch in the trawling nets. 87% of these turtles were captured by mid-trawling nets, the rest by bottom-trawling nets, mostly at depths of 11–30 m. 95% of all turtles were caught alive and healthy, and were usually released back into the sea immediately after capture by the fishermen. Training measures were given to local fishermen in order to raise their awareness of the threats to marine turtles.     

Accounting for Imperfect Detection Is Critical for Inferring Marine Turtle Nesting Population Trends

Assessments of population trends based on time-series counts of individuals are complicated by imperfect detection, which can lead to serious misinterpretations of data. Population trends of threatened marine turtles worldwide are usually based on counts of nests or nesting females. We analyze 39 years of nest-count, female-count, and capture-mark-recapture (CMR) data for nesting loggerhead turtles (Caretta caretta) on Wassaw Island, Georgia, USA. Annual counts of nests and females, not corrected for imperfect detection, yield significant, positive trends in abundance. However, multistate open robust design modeling of CMR data that accounts for changes in imperfect detection reveals that the annual abundance of nesting females has remained essentially constant over the 39-year period. The dichotomy could result from improvements in surveys or increased within-season nest-site fidelity in females, either of which would increase detection probability. For the first time in a marine turtle population, we compare results of population trend analyses that do and do not account for imperfect detection and demonstrate the potential for erroneous conclusions. Past assessments of marine turtle population trends based exclusively on count data should be interpreted with caution and re-evaluated when possible. These concerns apply equally to population assessments of all species with imperfect detection.     

Fibropapillomatosis of marine turtles

Cutaneous fibropapillomatosis in green sea turtles, Chelonia mydas (GTFP), was first reported over 50 years ago. In the last decade, GTFP has emerged as a significant worldwide epizootic with prevalences as high as 92% in some green turtle populations. Lesions similar to GTFP have been observed in other marine turtle species including olive ridleys, Lepidochelys olivacea, flatbacks, Natator depressus, and loggerheads, Caretta caretta, but disease in these species occurs at lower frequencies and is less well documented. The etiology of GTFP is unknown, and a variety of hypotheses concerning the possible etiology and pathogenesis of GTFP have been proposed and are discussed in this paper. Possible etiologies include viruses, metazoan parasites, ultraviolet radiation, and chemical carcinogens. Recent evidence from controlled transmission experiments implicates a filterable infectious agent as the primary etiology of GTFP. A herpesvirus has been identified in some lesions but has not been isolated and cultured; consequently, Koch's postulates have not yet been fulfilled for this agent. The epizootiology and pathogenesis of GTFP are poorly understood. Epizootiologic evidence, while limited to a few field studies, suggests that environmental conditions in certain near-shore marine habitats favor a high prevalence of disease expression. The possibility that immune system modulators play a role in the persistence and severity of this disease is discussed. Detailed investigations of the epizootiology of GTFP must await identification of the etiologic agent and development of specific diagnostic tests. In addition, until immune function tests can be developed and validated for free-ranging turtles, hypotheses about the role of immune system dysfunction in GTFP epizootics cannot be tested.