Renal oxalosis in free-ranging green turtles Chelonia mydas

Eighteen green turtles Chelonia mydas recovered from the Atlantic and Gulf coasts of Florida and Tortuguero National Park, Costa Rica, were diagnosed with renal oxalosis by histopathological examination. Affected sea turtles included 14 adults and 4 immature animals, which comprised 26% (18/69) of green turtle necropsy cases available for review. Calcium oxalate deposition ranged from small to moderate amounts and was associated with granuloma formation and destruction of renal tubules. All affected turtles died from traumatic events or health problems unrelated to renal oxalosis; however, 1 immature turtle had notable associated renal injury. Crystal composition was confirmed by infrared and scanning electron microscopy and energy dispersive X-ray analysis. The source of calcium oxalate is unknown and is presumed to be of dietary origin.     

Retrospective pathology survey of green turtles Chelonia mydas with fibropapillomatosis in the Hawaiian Islands, 1993--2003

We necropsied 255 stranded green turtles Chelonia mydas with fibropapillomatosis (FP) from the Hawaiian Islands, North Pacific, from August 1993 through May 2003. Of these, 214 (84 %) were euthanized due to advanced FP and the remainder were found dead in fresh condition. Turtles were assigned a standardized tumor severity score ranging from 1 (lightly tumored) to 3 (heavily tumored). Tumors were counted and measured and categorized as external, oral, or internal and tissues evaluated by light microscopy. Turtles in tumor score 2 and 3 categories predominated, and tumor score 3 turtles were significantly larger than the other 2 categories. More juveniles stranded than subadults or adults. Total cross-sectional area of tumors increased significantly with straight carapace length (SCL). Frequency distribution of total number of external tumors per turtle was significantly skewed to the right, and there were significantly more tumors at the front than rear of turtles. Eighty percent of turtles had oral tumors, and 51% of turtles with oral tumors had tumors in the glottis. Thirty-nine percent of turtles had internal tumors, most of them in the lung, kidney and heart. Fibromas predominated in lung, kidney and musculoskeletal system whereas myxofibromas were more common in intestines and spleen. Fibrosarcomas of low-grade malignancy were most frequent in the heart, and heart tumors had a predilection for the right atrium. Turtles with FP had significant additional complications including inflammation with vascular flukes, bacterial infections, poor body condition, and necrosis of salt gland. Turtles with oral tumors were more likely to have secondary complications such as pneumonia. Most turtles came from the island of Oahu (74%) followed by Maui (20 %), Hawaii, Molokai, and Lanai (<3 % each). On Oahu, significantly more turtles we necropsied stranded along the northwestern and northeastern shores.     

Immune status of free-ranging green turtles with fibropapillomatosis from Hawaii

Cell-mediated and humoral immune status of free-ranging green turtles (Chelonia mydas) in Hawaii (USA) with and without fibropapillornatosis (FP) were assessed. Tumored and non-tumored turtles from Kaneohe Bay (KB) on the island of Oahu and from FP-free areas on the west (Kona/Kohala) coast of the island of Hawaii were sampled from April 1998 through February 1999. Turtles on Oahu were grouped (0-3) for severity of tumors with 0 for absence of tumors, 1 for light, 2 for moderate, and 3 for most severe. Turtles were weighed, straight carapace length measured and the regression slope of weight to straight carapace length compared between groups (KB0, KB1, KB2, KB3, Kona). Blood was assayed for differential white blood cell count, hematocrit, in vitro peripheral blood mononuclear cell (PBMC) proliferation in the presence of concanavalin A (ConA) and phytohaemagglutinin (PHA), and protein electrophoresis. On Oahu, heterophil/lymphocyte ratio increased while eosinophil/monocyte ratio decreased with increasing tumors score. Peripheral blood mononuclear cell proliferation indices for ConA and PHA were significantly lower for turtles with tumor scores 2 and 3. Tumor score 3 turtles (KB3) had significantly lower hematocrit, total protein, alpha 1, alpha 2, and gamma globulins than the other four groups. No significant differences in immune status were seen between non-tumored (or KB1) turtles from Oahu and Hawaii. There was no significant difference between groups in regression slopes of body condition to carapace length. We conclude that turtles with severe FP are imunosuppressed. Furthermore, the lack of significant difference in immune status between non-tumored (and KB1) turtles from Oahu and Kona/Kohala indicates that immunosuppression may not be a prerequisite for development of FP.     

Dietary protein requirement for captive juvenile green turtles (Chelonia mydas)

Head-starting programs are extremely important for restoring the population of sea turtles in wild whereas husbandry conditions and feeding regimens of captive turtles are still limited. In the current study, the optimal dietary protein requirement for green turtle (Chelonia mydas) was investigated to support rearing in head-starting programs. Twenty-five-day-old turtles (44.5–46.2 g body weight, n = 45) were randomly distributed into 15 experimental plastic tanks, comprising three treatment replications of 3 turtles each. They were fed fishmeal-based feeds containing different levels of protein (30%, 35%, 40%, 45%, and 50%) for 8 weeks. At the end of feeding trial, growth performance (specific growth rate = 1.86% body weight/day) and feed utilization (protein efficiency ratio = 3.30 g gain/g protein) were highest in turtles fed with 40% protein in feed (p < .05). These nutritional responses were significantly supported by specific activities of fecal digestive enzymes, especially trypsin, chymotrypsin, amylase, and the amylase/trypsin ratio. Also, this dietary level improved the deposition of calcium and phosphorus in carapace, supporting a hard carapace and strong healthy bones. There were no negative effects in general health status of reared turtles, as indicated by hematological parameters. Based on a broken-line analysis between dietary protein levels and specific growth rate, the optimal protein level for green turtles was estimated as 40.6%. Findings from the current study support the use of artificial diets of specific protein levels to rear captive green turtle before release to natural habitats.     

Epizootiology of spirorchiid infection in green turtles (Chelonia mydas) in Hawaii

We describe the epizootiology of spirorchiid trematode infections in Hawaiian green turtles (Chelonia mydas) by quantifying tissue egg burdens in turtles submitted for necropsy and by assessing antibody response to crude adult worm and egg antigens among a variety of age groups. Hapalotrema sp. and Laeredius sp. predominated in turtles infected with spirorchiids. Tissue egg burdens decreased with increasing size and increased with deteriorating body condition of turtles. No relationship was found between tissue egg burdens and sex or fibropapillomatosis status. Tissue egg burdens increased in turtles from southeast to northwest in the main Hawaiian Islands (Hawaii to Kauai). Hatchling and captive-reared turtles had significantly lower levels of antibodies against crude worm and egg antigens. Based on tissue egg burdens and antibody status, we hypothesize that immature turtles become infected with spirorchiids shortly after recruiting into coastal foraging pastures from the pelagic environment, that infection levels decrease with age, and that spirorchiids detrimentally affect the body condition of sea turtles independent of tumor burden. The low intensity of infection in turtles with the endemic trematode Carettacola hawaiiensis suggests either that turtles are less susceptible to infection with this parasite or that the parasite is outcompeted by species of Hapalotrema and Laeredius. Given that the 2 latter species are found in the Pacific and other oceans, they are not likely endemic and were probably introduced into Hawaii through an undetermined route.     

Seasonality and numbers of green turtles Chelonia mydas nesting on the Pitcairn Islands

During 1991 and 1992 the first regular field observations of green turtles, Chelonia mydas , on the Pitcairn Islands (South Pacific) revealed that nesting occurs only on Henderson where about 10 females laid eggs (c. 1% of the French Polynesian population). Substrates on Pitcairn and Ducie Atoll are unsuitable for nesting and no signs of nesting were found on Oeno Atoll where the terrain appears suitable.     

Depot fatty acid composition in immature green turtles (Chelonia mydas) residing at two near-shore foraging areas in the Hawaiian Islands

The lipid content and fatty acid composition of depot fat were determined for 58 immature green turtles (Chelonia mydas) residing at two near-shore foraging areas, Ahu-O-Laka, located in Kaneohe Bay on Oahu, and Kiholo Bay located on the island of Hawaii. Benthic flora at Kiholo was limited to a single algal species but included algae and seagrass at Ahu-O-Laka. Turtle straight carapace length ranged from 38.6 to 59.2 cm, suggesting that the sample set included new recruits to up to 12-year residents. Fatty acid data were analyzed using principal components analysis (PCA). PC1 accounted for over 50% of the variance. Turtles were generally delineated along PC1 by the length of time on benthic foraging grounds, with high (>0.75) negative loadings for the fatty acids 22:6n-3, 7M7H, t16:1n-10, 15:0, and 17:0 associated with relatively new recruits (suggesting a pelagic dietary source for these fatty acids) and high positive loadings for 12:0 and 14:0 associated with long-term residents. PC2 separated turtles primarily by capture location, with high positive loadings for 18:2n-6 and 18:3n-3 [the primary seagrass polyunsaturated fatty acids] associated with the Ahu-O-Laka turtles. Fatty acid profiles of turtles from both locations differed substantially from those of their benthic diets, suggesting considerable modification of dietary fatty acids and de novo biosynthesis.     

Testing dispersal hypotheses in foraging green sea turtles (Chelonia mydas) of Brazil

Testing theories of dispersal is challenging in highly migratory species. In sea turtles, population size, geographic distance, natal homing, and ocean currents are hypothesized to affect dispersal. Little is known, however, about these mechanisms in sea turtles foraging along the South American coast. Green sea turtles feeding at Ubatuba (UB, n = 114) and Almofala (AF, n = 117), Brazil, were sequenced at the mitochondrial DNA (mtDNA) control region (486 bp) and genotyped at 7 microsatellite loci to test dispersal hypotheses. Fifteen mtDNA haplotypes were revealed, including a previously undescribed sequence, and the average observed heterozygosity (H(o)) was 76.4%. Overall short-term temporal differences were not detected, and differentiation was less pronounced in microsatellite than in mtDNA analyses. Mitochondrial results reveal significant differentiation between the Brazilian feeding grounds and most other Atlantic groups, whereas microsatellites uncover similarities to some of the geographically closest populations. Ubatuba and Almofala are mixed stocks, drawn primarily from Ascension, with lesser contributions from Surinam/Aves and Trindade. Costa Rica is also a significant source of individuals feeding at AF. The results are consistent with a model of juvenile natal homing impacted by other factors. Effective protection of turtles foraging along the extensive Brazilian coast may enhance breeding populations thousands of kilometers away.     

Basic oxidative stress metabolites in eastern Pacific green turtles (Chelonia mydas agassizii)

Analysis of hematological and biochemical parameters, including oxidative stress indicators, is an invaluable tool in wildlife health assessment, particularly for threatened or endangered species. This study was aimed at obtaining baseline information of oxidative stress indicators in eastern Pacific green turtles (Chelonia mydas agassizii) from a relatively undisturbed habitat at Bahía Magdalena, Baja California Sur, Mexico. Tissues were analyzed for superoxide radical (O(2)(*-) production, lipid peroxidation (measured as thiobarbituric acid reactive substances, TBARS), and antioxidant enzyme activity (superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST)). Overall levels for all variables were within ranges reported for other reptile species. Results suggest differences in oxidative metabolism among tissues (p< or =0.05). Liver, lung and muscle had the highest levels of O(2)(*-) production. Liver revealed the highest TBARS levels. Liver and muscle showed the highest SOD activity, while liver and kidney had the highest CAT and GST activities. These data provide baseline values of the oxidative stress indicators in tissues from eastern Pacific green turtles. Development of a biomarker system to assess the health of wildlife species, especially one that could detect early exposure to environmental pollutants or emerging diseases, would provide a useful tool in the long-term conservation of the species.     

Association of herpesvirus with fibropapillomatosis of the green turtle Chelonia mydas and the loggerhead turtle Caretta caretta in Florida.

Sea turtle fibropapillomatosis (FP) is a disease marked by proliferation of benign but debilitating cutaneous fibropapillomas and occasional visceral fibromas. Transmission experiments have implicated a chloroform-sensitive transforming agent present in filtered cell-free tumor homogenates in the etiology of FP. In this study, consensus primer PCR methodology was used to test the association of a chelonian herpesvirus with fibropapillomatosis. Fibropapilloma and skin samples were obtained from 17 green and 2 loggerhead turtles affected with FP stranded along the Florida coastline. Ninety-three cutaneous and visceral tumors from the 19 turtles, and 33 skin samples from 16 of the turtles, were tested. All turtles affected with FP had herpesvirus associated with their tumors as detected by PCR. Ninety-six percent (89/93) of the tumors, but only 9% (3/33) of the skin samples, from affected turtles contained detectable herpesvirus. The skin samples that contained herpesvirus were all within 2 cm of a fibropapilloma. Also, 1 of 11 scar tissue samples from sites where fibropapillomas had been removed 2 to 51 wk earlier from 5 green turtles contained detectable herpesvirus. None of 18 normal skin samples from 2 green and 2 loggerhead turtles stranded without FP contained herpesvirus. The data indicated that herpesvirus was detectable only within or close to tumors. To determine if the same virus infected both turtle species, partial nucleotide sequences of the herpesvirus DNA polymerase gene were determined from 6 loggerhead and 2 green turtle samples. The sequences predicted that herpesvirus of loggerhead turtles differed from those of green turtles by only 1 of 60 amino acids in the sequence examined, indicating that a chelonian herpesvirus exhibiting minor intratypic variation was the only herpesvirus present in tumors of both green and loggerhead turtles. The FP-associated herpesvirus resisted cultivation on chelonian cell lines which support the replication of other chelonian herpesviruses. These results lead to the conclusion that a chelonian herpesvirus is regularly associated with fibropapillomatosis and is not merely an incidental finding in affected turtles.     

Metabolic heating and the prediction of sex ratios for green turtles (Chelonia mydas)

We compared incubation temperatures in nests (n=32) of the green turtle (Chelonia mydas) on Ascension Island in relation to sand temperatures of control sites at nest depth. Intrabeach thermal variation was low, whereas interbeach thermal variation was high in both control and nest sites. A marked rise in temperature was recorded in nests from 30% to 40% of the way through the incubation period and attributed to metabolic heating. Over the entire incubation period, metabolic heating accounted for a mean rise in temperature of between 0.07 degrees and 2.86 degrees C within nests. During the middle third of incubation, when sex is thought to be determined, this rise in temperature ranged between 0.07 degrees and 2.61 degrees C. Metabolic heating was related to both the number of eggs laid and the total number of hatchlings/embryos produced in a clutch. For 32 clutches in which temperature was recorded, we estimate that metabolic heating accounted for a rise of up to 30% in the proportion of females produced within different clutches. Previous studies have dismissed any effect of metabolic heating on the sex ratio of marine turtle hatchlings. Our results imply that metabolic heating needs to be considered when estimating green turtle hatchling sex ratios.     

Caryospora cheloniae sp. n.: a coccidial pathogen of mariculture-reared green sea turtles (Chelonia mydas mydas).

Caryospora cheloniae sp. n. is described from mariculture-reared green sea turtles (Chelonia m. mydas). The sporulated oocyst has a thin, transparent, single-layered wall which often ruptures, leaving a naked sporulated sporocyst. Oocysts measured 33.8 to 40.1 micrometer by 11.0 to 14.6 micrometer (mean 37.4 by 12.8 micrometer). Greatest concentrations of developmental stages of C. cheloniae were found in the hindgut. Transverse binary fission was observed in dividing tissue stages. Pathologic alterations were most pronounced in the posterior third of the intestines (hindgut). The hindgut lumen was greatly dilated and filled with blood, oocysts and tissue debris. The hindgut wall was thinner than normal and the mucosal folds had sloughed into the intestinal lumen. Free blood escaped from the blood vessels of the tunica propria into the intestinal lumen. Epithelial hyperplasia was pronounced at the margins of denuded mucosal areas. Numerous inflammatory cells infiltrated the infected mucosal surface.     

Autopodial development in the sea turtles Chelonia mydas and Caretta caretta

The manus and pes were studied using whole-mount and histological preparations of ontogenetic series of Chelonia mydas and Caretta caretta. Patterns of connectivity and sequences of chondrification events are similar to those reported for other turtle species, with respect to both the primary axis and the digital arch. There is no evidence of anterior condensations in the region distal to the radius and the tibia, supporting the hypothesis that the radiale and tibiale are absent in turtles. The three middle metacarpals are the first elements to start ossification in the manus of C. mydas, while ossification has not started in the pes. In the hatchling of C. mydas, most carpals have started ossification, whereas tarsals are mostly still cartilaginous. In C. caretta, the first carpals to ossify are the ulnare and intermedium, followed by the pisiform. Among metatarsals, the fifth hooked metatarsal is the last one to start ossification. The fibulare and intermedium fuse early in chondrogenesis, later becoming the astragalocalcaneum. Ossification in the carpals of C. caretta starts while tarsals are still cartilaginous. The derived autopodial proportions in each autopodium of adults are laid out at the condensation stage, and features that were present in basal turtles are absent at all stages examined (developmental penetrance). In contrast to this, conservatism is expressed in the presence of similar patterns of connectivity during early chondrogenesis, and in the development of overall proportions of the manus versus pes. As in adult anatomy, the development of the autopodium of marine turtles is a mosaic of derived and plesiomorphic features.     

Feral cats (Felis catus) as predators of hatchling green turtles (Chelonia mydas)

In many parts of the world feral animals have been reported to have severe effects on marine turtle hatchling production. In this study, green turtle ( Chelonia mydas ) hatchlings were shown to be an important component of the diet of the feral cat ( Felis catus ) on Aldabra Atoll, Seychelles, and feral cat activity in the coastal areas was found to be concentrated on the beaches used most intensively by turtles for nesting. The impact of the feral cat's predation on green turtle recruitment could not be determined. However, despite cat predation, the size of the Aldabran green turtle nesting population has increased considerably since human exploitation ceased.     

Relating tumor score to hematology in green turtles with fibropapillomatosis in Hawaii

The relationship between hematologic status and severity of tumor affliction in green turtles (Chelonia mydas) with fibropapillomatosis (FP) was examined. During 1 wk periods in July 1997 and July 1998, we bled 108 free-ranging green turtles from Pala'au (Molokai, Hawaii, USA) where FP is endemic. Blood was analyzed for hematocrit, estimated total solids, total white blood cell (WBC) count and differential WBC count. Each turtle was assigned a subjective tumor score ranging from 0 (no visible external tumors) to 3 (heavily tumored) that indicated the severity of FP. There was a progressive increase in monocytes and a decrease in all other hematologic parameters except heterophils and total numbers of white blood cells as tumor score increased. These data indicate that tumor score can relate to physiologic status of green turtles afflicted with FP, and that tumor score is a useful field monitor of severity of FP in this species.     

Do female green turtles (Chelonia mydas) exhibit reproductive seasonality in a year-round nesting rookery?

In circumglobal populations of sea turtles, little nesting activity occurs during cold seasons when the sand temperature on a nesting beach is inappropriate for successful egg incubation. However, it is not known whether reproductive seasonality also occurs in year-round nesting rookeries where the sand temperature may be suitable throughout the year. Therefore, we examined the hypothesis that female turtles exhibit reproductive seasonality even in a year-round nesting population. To determine whether the time of nesting of individuals fluctuates between successive nesting seasons, a year-round nesting rookery of green turtles Chelonia mydas at Huyong Island, Thailand, was patrolled on foot every night for 3190 consecutive days, and nesting females were identified using microchips and metal tags. The date of first nesting within a season for an individual (nesting date) was considered the nesting season of each individual. We identified 94 females and 463 nests (including 47 unidentified nests) during the survey. Nesting dates were distributed throughout the year. Nineteen turtles remigrated to the nesting beach, and the nesting dates of the remigrated females occurred in the same seasons. This indicates that females have a fixed individual nesting season even in a year-round nesting rookery. The year-round nesting of green turtles may be attributed to a wide variation in the nesting seasons of individuals. Satellite tracking revealed that the mean travelling duration of post-nesting migrations in this population was 13.5±6.1 days. The proximity of locations for reproduction and feeding may also play an important role in sustaining year-round nesting in this population.     

Hydrodynamic stability in posthatchling loggerhead (Caretta caretta) and green (Chelonia mydas) sea turtles

Swimming animals may experience a wide range of destabilizing forces resulting from the movements of their propulsors. These forces often cause movements in directions other than the intended trajectory (i.e., recoil motions), potentially increasing locomotor costs. We quantified rectilinear swimming stability for posthatchling loggerhead (Caretta caretta) and green turtles (Chelonia mydas). Sea turtles predominantly swim via “aquatic flight”, which is characterized by synchronous dorsoventral flapping of their forelimbs. We tested four predictions about the effects of “aquatic flight” on stability: (1) it would produce little lateral recoil; (2) lateral recoil motions would be non-cyclic; (3) vertical recoil motions would be larger than lateral recoil motions; and (4) vertical recoil motions would be cyclic. Additionally, because posthatchling loggerheads possess dorsal keels on the shell that are absent in green turtles, we evaluated whether such keels might improve stability in swimming turtles. While our expectations for patterns of cyclicity in recoil motions (predictions 2 and 4) were met, our expectations for differences in their absolute and relative magnitudes (predictions 1 and 3) were not. We suggest that lateral recoil motions were greater than predicted due to slight asynchronies between the motions of the left and right foreflippers. Additionally, although minimum lateral recoil motions were smaller than minimum vertical recoil motions, maximum recoil motions were greater in the lateral direction, so that average recoil did not differ significantly between these directions. Finally, because loggerheads did not display higher levels of stability compared to green turtles, there is little evidence to support a stabilizing role for dorsal keels in loggerhead turtles.     

Detection of low plasma estradiol concentrations in nesting green turtles (Chelonia mydas) by HPLC/Ms-Ms

In previous studies on nesting green turtles under natural conditions from different geographical regions, 17-β-estradiol (E(2) ) was either undetectable or detected at very low levels. RIA and other related techniques were not sensitive enough to measure low E(2) values in the green turtles. In this study, a sensitive method was used in detecting low hormone concentrations: high performance liquid chromatography with tandem quadruple mass spectrometry (HPLC-MS/MS). Using this technique, estradiol for the first time was detected in nesting green turtles during the peak season (June-October) at Ras Al-Hadd Reserve, Oman. The E(2) values recorded from this study were the highest ever recorded from nesting green turtles in any geographical region, but the levels did not vary significantly throughout different phases of nesting. The presence of E(2) during nesting presumably plays a role in the physiology and behavior of this species. Ras Al-Hadd hosts one of the largest nesting populations of green turtles in the world, and an understanding of their nesting patterns may be of value in conservation and management programs for this endangered species.     

The influence of diet on fatty acids in the egg yolk of green sea turtles, Chelonia mydas

Fatty acid concentrations found in the yolk of green sea turtles reflect differences in the diet of the mothers. All of the 12 fatty acids measured in yolk samples were significantly different between eggs produced from the pellet and wild-type diets. However, the relative pattern of yolk fatty acids in the green turtle mirrored those of other reptiles. Yolk samples contained mostly (63–67%) 14:0. 16:0, 16:1n-7 and 18:1n-9. Yolks from captive animals on pellet diet contained an additional 17.64% of the total yolk lipid as 12:0 and 18:2n-6. Wild yolks contained an extra 11.41% of lipid as 18:0 and 18:1n-7. Selection of fatty acids for the yolk should balance the energetic and anabolic needs of the embryo. Eggs are provisioned based on maternal metabolism of available nutrients and subtle differences between natural foods and those available in captivity could affect the viability of future eggs.     

Digenetic trematode community in nesting green sea turtles (Chelonia mydas) from Tortuguero National Park, Costa Rica

The digenetic community of 40 green sea turtles (Chelonia mydas) was investigated at Tortuguero National Park, Costa Rica. In total, 24,270 trematodes belonging to 29 species and 6 families including Clinostomidae 1, Microscaphidiidae 5, Paramphistomidae 2, Pronocephalidae 15, Rhytidodidae 2, and Spirorchiidae 4 were recorded from chelonians examined. Turtles harbored a mean of 7.63 +/- 3.5 SD species. Only 3 species, i.e., Learedius learedi, Microscaphidium reticulare, and Pyelosomum cochlear, infected more than 50% of the hosts sampled. Learedius learedi was the most prevalent (97.5%) and the second most abundant species with a mean of 25.6 +/- 21.6 SD. Only 1 core species, M. reticulare, was recovered from the gastrointestinal tract; it was the most abundant parasite, with a mean intensity of 477 +/- 1,180 SD, and the second most prevalent (77.5%). Diversity values ranged from 0.10 to 2.10, with a mean of 1.00 +/- 0.43 SD for the total component community and from 0.10 to 1.84, with a mean of 0.79 +/- 0.41 SD, for the gastrointestinal component community. Species richness was the highest recorded from a sea turtle species. All digenetic species were recorded from Costa Rica for the first time. This represents the first report on the helminth community of the green turtle.