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Effects of organochlorine contaminants on loggerhead sea turtle immunity: comparison of a correlative field study and in vitro exposure experiments.

Several laboratory and field studies indicate that organochlorine contaminants (OCs), such as polychlorinated biphenyls (PCBs) and pesticides, modulate immune responses in rodents, wildlife, and humans. In the present study we examined the effects of OCs on immunity in free-ranging loggerhead sea turtles (Caretta caretta). Mitogen-induced lymphocyte proliferation responses, lysozyme activity, and OC concentrations were measured from blood samples. Mitogens chosen in the lymphocyte proliferation assay were phytohemagglutinin (PHA) and concanavalin A (ConA) for T-lymphocyte stimulation, and lipopolysaccharide (LPS) and phorbol 12,13-dibutyrate (PDB) for B-lymphocyte stimulation. Lysozyme activity was significantly and negatively correlated with whole-blood concentrations of 4,4 -dichlorodiphenyldichloroethylene (4,4 -DDE) and the sum of chlordanes. Lymphocyte proliferation responses stimulated by PHA, LPS, and PDB were significantly and positively correlated with concentrations of the sum of PCBs measured in whole blood. LPS- and PDB-induced proliferation were also significantly and positively correlated with 4,4 -DDE blood concentrations. These correlative observations in free-ranging turtles suggest that current, chronic exposure to OCs may suppress innate immunity and enhance certain lymphocyte functions of loggerhead sea turtles. To further test this hypothesis, lymphocyte proliferation was measured after in vitro exposure of peripheral blood leukocytes from 16 turtles to Aroclor 1254 (0-13.5 microg/mL) or 4,4 -DDE (0-13.4 microg/mL). Both contaminants increased PHA- and PDB-induced proliferation at concentrations below those that affected cell viability. Moreover, the concentrations that enhanced PDB-induced proliferation in vitro were similar to concentrations measured in turtles with the highest proliferative responses. The similarities between the in vitro experiments and the correlative field study suggest that OC exposure modulates immunity in loggerhead turtles.

Animals↗

Effects of acute fresh water exposure on water flux rates and osmotic responses in Kemp's ridley sea turtles (Lepidochelys kempi).

Water flux rates and osmotic responses of Kemp's Ridley sea turtles (Lepidochelys kempi) acutely exposed to fresh water were quantified. Salt-water adapted turtles were exposed to fresh water for 4 d before being returned to salt water. During the initial salt water phase, absolute and relative water flux rates were 1.2+/-0.1 l d(-1) and 123.0+/-6.8 ml kg(-1) d(-1), respectively. When turtles were exposed to fresh water, rates increased by approximately 30%. Upon return to salt water, rates decreased to original levels. Plasma osmolality, Na(+), K(+), and Cl(-) decreased during exposure to fresh water, and subsequently increased during the return to salt water. The Na(+):K(+) ratio was elevated during the fresh water phase and subsequently decreased upon return to salt water. Aldosterone and corticosterone were not altered during exposure to fresh water. Elevated water flux rates during fresh water exposure reflected an increase in water consumption, resulting in a decrease in ionic and osmotic concentrations. The lack of a change in adrenocorticoids to acute fresh water exposure suggests that adrenal responsiveness to an hypo-osmotic environment may be delayed in marine turtles when compared to marine mammals.

Adaptation, Physiological↗

Global phylogeography of the ridley sea turtles (Lepidochelys spp.) as inferred from mitochondrial DNA sequences.

The Kemp's ridley sea turtle (Lepidochelys kempi) is restricted to the warm temperate zone of the North Atlantic Ocean, whereas the olive ridley turtle (L. olivacea) is globally distributed in warm-temperate and tropical seas, including nesting colonies in the North Atlantic that nearly overlap the range of L. kempi. To explain this lopsided distribution, Pritchard (1969) proposed a scenario in which an ancestral taxon was divided into Atlantic and Pacific forms (L. kempi and L. olivacea, respectively) by the Central American land bridge. According to this model, the olive ridley subsequently occupied the Pacific and Indian Oceans and recently colonized the Atlantic Ocean via southern Africa. To assess this biogeographic model, a 470 bp sequence of the mtDNA control region was compared among 89 ridley turtles, including the sole L. kempi nesting population and 7 nesting locations across the range of L. olivacea. These data confirm a fundamental partition between L. olivacea and L. kempi (p = 0.052-0.069), shallow separations within L. olivacea (p = 0.002-0.031), and strong geographic partitioning of mtDNA lineages. The most divergent L. olivacea haplotype is observed in the Indo-West Pacific region, as are the central haplotypes in a parsimony network, implicating this region as the source of the most recent radiation of olive ridley lineage. The most common olive ridley haplotype in Atlantic samples is distinguished from an Indo-West Pacific haplotype by a single nucleotide substitution, and East Pacific samples are distinguished from the same haplotype by two nucleotide substitutions. These shallow separations are consistent with the recent invasion of the Atlantic postulated by Pritchard (1969), and indicate that the East Pacific nesting colonies were also recently colonized from the Indo-West Pacific region. Molecular clock estimates place these invasions within the last 300,000 years.

Animals↗

Hatchling sea turtles use surface waves to establish a magnetic compass direction

Hatchling sea turtles emerge from underground nests, crawl to the ocean, and swim away from the land. In shallow water near shore, hatchlings maintain offshore headings by swimming into oceanic waves; in deeper water, however, turtles appear to rely on different mechanisms to maintain their courses. To determine whether loggerhead hatchlings, Caretta caretta L., are able to transfer a course initiated on the basis of waves to a course maintained by a magnetic compass, we studied the orientation behaviour of turtles that had been exposed to waves for either 15 or 30 min before being tested in still water. Hatchlings that swam into waves for 15 min failed to continue swimming on similar courses when the waves were discontinued, but turtles that swam into waves for 30 min maintained similar mean headings after the waves stopped. Inverting the vertical component of the magnetic field during the test period reversed the direction of orientation of this latter group of turtles. Thus, hatchlings can transfer a heading induced by waves to a magnetic compass, and thereby continue to migrate away from land after contact with the coast is lost. Migratory orientation in turtles resembles that of birds in that both rely on multiple cues and an ability to transfer information between various cues and compasses at appropriate times during the journey.Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗

Plasma corticosterone concentrations associated with acute captivity stress in wild loggerhead sea turtles (Caretta caretta).

Plasma corticosterone concentrations were measured in wild loggerhead sea turtles (Caretta caretta) in response to acute captivity (capture, serial bleeding, and restraint up to 6 hr). In general, concentrations of corticosterone dramatically increased 1 hr after capture, peaked at 3 hr, and decreased by 6 hr. Initial corticosterone concentrations were significantly lower in animals captured by tangle net than in those captured by trawl and were thought to more closely represent baseline levels. Significant effects of season and size class on corticosterone concentrations were found for turtles captured by trawl. Corticosterone concentrations of small turtles captured in summer were higher than those of large turtles captured in the same season and of all turtles captured during winter. In winter, corticosterone concentrations for small turtles were higher than those for large turtles at 3 hr after capture. Large turtles captured during winter experienced the slowest rate of increase in plasma corticosterone and a decline at 3 hr after capture. Although cloacal temperatures were significantly higher in summer samples, corticosterone concentrations of large turtles did not differ between seasons until 1 hr after capture. In addition, several large turtles during summer did not experience an increase in corticosterone concentrations 1 hr after capture. It is possible that the lower corticosterone response of large turtles captured during summer may be associated with reproductive condition.

Animals↗

Studies on cardiovascular fluke (Digenea: Spirorchiidae) infections in sea turtles from the Great Barrier Reef, Queensland, Australia.

Twenty-seven sea turtles (23 Chelonia mydas and four Eretmochelys imbricata) from northeast Queensland were found to be infected with cardiovascular flukes and/or their eggs. Five had originated from turtle farms in the Torres Strait, five from an oceanarium on Magnetic Island (146 degrees 56'E, 19 degrees S) and the remainder from coral reefs in the Torres Strait or near Townsville. The incidence of flukes and/or eggs in the groups was 4.8 per cent (5 of 104), 33.3 per cent (5 of 15) and 72.2 per cent (17 of 22), respectively. Affected animals ranged in size from 18 to 108 cm (curved carapace length) and weighed between 0.5 and 77 kg. The average number of flukes per host was 47. Flukes were recovered from the three chambers of the heart and major vessels (right aortic arch and brachiocephalic artery), where they were attached to the walls or free in the lumen. They were subsequently identified as Haplotrema spp. and/or Learedius spp. In 59.2 per cent (16 of 27) of turtles, flukes were not found, although their eggs were detected microscopically. Gross pathological changes associated with the presence of flukes included thickening and hardening of arterial walls (four turtles), thrombus formation (three), chronic pneumonia (two) and an excess of pericardial or peritoneal fluid (four). Microscopically, the essential changes was that of chronic inflammation, as evidenced by the proliferation of epithelial cells, reticulo-endothelial cells and fibroblasts in areas accessible to flukes and/or eggs. Multiple diffuse egg granulomas were a prominent feature of most organs, the spleen and lungs being predilection sites. Proliferative changes had occurred in the endocardium and in the endothelium of vessels supplying the spleen, stomach, intestine and pancreas (18 turtles). The walls of major arteries, lungs, liver, brain, crop and stomach were also acutely inflamed (eight turtles). Haemorrhage was recorded in the lungs and/or brain of eight turtles with heavy fluke infestations. Other vascular changes, viz. congestion, oedema and hypertrophy of arterial/arteriolar walls, resulted from the inhibition of blood flow by parasitic emboli.

Animals↗

Oxygen affinity of sea turtle blood.

The oxygen binding properties of whole blood in adult sea turtles (green, Chelonia mydas; loggerhead, Caretta caretta) were measured as a function of pH, and carbon dioxide partial pressure using the 'Hemoscan Oxygen Dissociation Analyser' with modifications that improved the performance of the method. Under approximately physiological conditions (25.7 degrees C, PCO2 = 37 Torr; pH 7.45) there is a marked affinity difference between the two species (green, P50 = 29 Torr; loggerhead, 47 Torr). It is suggested that the large affinity difference between blood of green and loggerhead turtles may be related to differences in depths to which they typically dive. The Hill coefficient at 50% saturation is 2.4 in the green under these conditions, and 2.7 in loggerhead. Hill plots of both species curve upward and have significant regions with n approaching 1 at low saturation with the effect that the lower end of the binding curve is more hyperbolic than sigmoidal. The fixed acid and CO2 Bohr effects in both species are strongly saturation dependent, being very low at low saturation and increasing markedly at higher saturation. The fixed acid Bohr effects are similar in the two species but the green is more sensitive to CO2, at constant pH, and so has a larger CO2 Bohr effect. The hyperbolic shape of the lower end of the binding curves, and low pH sensitivity in this region, would tend to favor oxygen loading during a dive, when lung PO2 and blood pH are falling. These properties might therefore favor more complete utilization of the lung oxygen store. We suggest that diving vertebrates can usefully be divided into two groups, those that use the lung as the main oxygen stores and those that use tissue (blood and myoglobin) stores.

Animals↗

Chemosensory orientation behavior in juvenile sea turtles.

It has been widely believed for several decades that hatchling sea turtles imprint to chemical cues characteristic of their natal beach and use this information as part of a repertoire of mechanisms enabling their return to the same beach for mating and nesting. This has proven very difficult to test. Although the imprinting theory is conceptually simple, functionally it is quite complex. This involves not only chemical imprinting of nestlings but growth and migration to habitats where the adults are found, long-term memory of their earlier chemical exposure, reproductive maturation, and homing. A few studies have been conducted to examine these elements of the imprinting theory. Experiments involving the exposure of embryos and hatchlings to chemicals suggest that juvenile turtles 'imprint' to the chemical environment of their nest. This can be termed chemical imprinting. Loggerhead turtles, Caretta caretta, and ridley turtles, Lepidochelys kempi, appear to be attracted to chemicals (morpholine and natural seawater, respectively) to which they were exposed as embryos. The strongest support for chemical imprinting is that six-month-old green turtles, Chelonia mydas, exposed to either morpholine or 2-phenylethanol in the nest and for a period of time after hatching, respond similarly to the chemical to which they were exposed as nestlings. Although chemical imprinting does not 'prove' the imprinting theory of turtle homing, it is a necessary component of the theory not previously examined.

Animals↗

Salt and water regulation by the leatherback sea turtle Dermochelys coriacea.

We measured the salt and water balance of hatchling leatherback sea turtles, Dermochelys coriacea, during their first few days of life to investigate how they maintain homeostasis under the osmoregulatory challenge of a highly desiccating terrestrial environment and then a hyperosmotic marine environment. Hatchlings desiccated rapidly when denied access to sea water, with their hematocrit increasing significantly from 30.32+/-0.54 % to 38.51+/-1.35 % and plasma Na(+) concentration increasing significantly from 138.2+/-3.3 to 166.2+/-11.2 mmol l(-1) in 12 h. When hatchlings were subsequently put into sea water, hematocrit decreased and plasma Na(+) concentration was unchanged but both were significantly elevated above pretreatment values. In other hatchlings kept in sea water for 48 h, body mass and plasma Na(+) concentration increased significantly, but hematocrit did not increase. These data show that hatchlings were able to osmoregulate effectively and gain mass by drinking sea water. We stimulated hatchlings to secrete salt from the salt glands by injecting a salt load of 27 mmol kg(-1). The time taken for secretion to begin in newly hatched turtles was longer than that in 4-day-old hatchlings, but the secretory response was identical at 4.15+/-0.40 and 4.13+/-0.59 mmol Na(+) kg(-1) h(-1) respectively. Adrenaline and methacholine were both potent inhibitors of salt gland secretion in a dose-dependent manner, although methacholine administered simultaneously with a subthreshold salt load elicited a transient secretory response. The results showed that hatchling leatherbacks are able to tolerate significant changes in internal composition and efficiently use their salt glands to establish internal ionic and water balance when in sea water.

Animals↗

Seasonal reproductive cycle of the Kemp's ridley sea turtle (Lepidochelys kempi).

The seasonal reproductive cycle of the Kemp's ridley sea turtle (Lepidochelys kempi) was studied under seminatural conditions at the Cayman Turtle Farm, Grand Cayman, British West Indies, from June 1987 to July 1988. Male L. kempi displayed a prenuptial rise in serum testosterone 4 to 5 months prior to the mating period (March). Male testosterone then declined sharply during the mating period. Female L. kempi also displayed a prenuptial rise in serum testosterone, estradiol, and total calcium 4 to 6 months prior to the mating period (March). Female testosterone and estradiol declined during the nesting period (April to July) immediately following the mating period (March). Elevated levels in female estradiol and total calcium corresponded with the period of vitellogenesis as determined from gel electrophoresis and ultrasonography. Serum thyroxine also fluctuated seasonally with elevated levels observed in females associated with the period of vitellogenesis. L. kempi displayed a distinct seasonal reproductive cycle in captivity. Nesting in the captive study group corresponded with nesting in the wild population at Rancho Nuevo, Mexico (April to July). Female endocrine cycles during the nesting period were similar to those observed in the wild population.

Animals↗

Arsenic accumulation in three species of sea turtles.

Arsenic in the liver, kidney and muscle of three species of sea turtles, e.g., green turtles (Chelonia mydas), loggerhead turtles (Caretta caretta) and hawksbill turtles (Eretmochelys imbricata), were determined using HG-AAS, followed by arsenic speciation analysis using HPLC-ICP-MS. The order of arsenic concentration in tissues was muscle > kidney > liver. Unexpectedly, the arsenic concentrations in the hawksbill turtles feeding mainly on sponges were higher than the two other turtles primarily eating algae and mollusk which accumulate a large amount of arsenic. Especially, the muscles of the hawksbill turtles contained remarkably high arsenic concentrations averaging 153 mg kg(-1) dry weight with the range of 23.1-205 mg kg(-1) (n = 4), even in comparison with the data from other organisms. The arsenic concentrations in the tissues of the green turtles were significantly decreased with standard carapace length as an indicator of growth. In arsenic compounds, arsenobetaine was mostly detected in the tissues of all the turtles. Besides arsenobetaine, a small amount of dimethylarsinic acid was also observed in the hawksbill turtles.

Animals↗

Organochlorine contaminants in sea turtles from the Eastern Pacific.

We measured organochlorine residues in three species of sea turtles from the Baja California peninsula, Mexico. Seventeen of 21 organochlorine pesticides analyzed were detected, with heptachlor epoxide and gamma-hexachlorocyclohexane the most prevalent (14 (40%) and 11 (31%) of the 35 tissue samples, respectively). PCBs were detected in all but one of the 9 turtles studied, with congener 18 the most commonly detected (8 (23%) of the samples). The dioxin-like congeners 118 and 180 were detected in 4 (11%) and 3 (9%) of the samples, respectively. Percent contribution of congeners was negatively correlated to their octanol-water partition coefficients, with kidney and muscle containing more lower-chlorinated congeners than liver or adipose samples. Levels of organochlorines detected in the present study were low, potentially attributable to the feeding habits of the predominant species collected in this study (herbivorous) and/or the samples obtained in an unindustrialized region.

Adipose Tissue↗

The sea-finding behavior of hatchling olive ridley sea turtles, Lepidochelys olivacea, at the beach of San Miguel (Costa Rica).

Newly hatched olive ridley sea turtles (Lepidochelys olivacea) were tested for their directional preferences in a sand-filled circular arena in total darkness. Hatchlings that had crawled about 5 m on the beach, toward the sea preferred the southwesterly direction that would have brought them to the water line, whereas hatchlings that had been denied this experience headed eastward, a direction of unclear origin. These data suggest that a short crawl across the natural beach can set the direction in which the young turtles subsequently move. The crawling experience was sufficient to acquire the compass course that they later follow, probably with the help of a magnetic compass, not only in the water, but already while still on land.

Animals↗

Characterization of gamma-glutamyl transpeptidase from the Rathke's gland secretions of Kemp's ridley sea turtles (Lepidochelys kempi).

The secretion produced by Rathke's glands of Kemp's ridley sea turtles (Lepidochelys kempi) contains the enzyme gamma-glutamyl transpeptidase. The approximately 200 kDa enzyme contains two different subunits, alpha (54 kDa) and beta (21 kDa), in an unknown stoichiometry. The enzyme transfers gamma-Glu from a number of different donors, such as glutamine, glutathione, S-Me-glutathione, N epsilon(gamma-Glu)-Lys, gamma-Glu-Ala, and other gamma-glutamyl amino acids, either to water or to a variety of acceptor substrates. It appears that a free alpha-amino group is the preferred acceptor. The enzyme is not inhibited by typical sulfhydryl reagents such as N-ethyl-maleimide, p-(chloro)mercuri-benzoate or 5,5'-dithio-bis-(2-nitrobenzoate) or by the active Ser reagent tosyl fluoride. Maleate stimulates the activity of the enzyme, and in the presence of 100 mM maleate 2 mM tosyl fluoride becomes an inactivator of the enzyme. The catalytic and molecular properties of the turtle gamma-glutamyl transpeptidase are similar to those established for mammalian gamma-glutamyl transpeptidase. Neither the physiological role of the enzyme nor the biological function of the secretion in which it occurs is understood at this time.

Amino Acid Sequence↗

Ontogeny of energetics in leatherback (Dermochelys coriacea) and olive ridley (Lepidochelys olivacea) sea turtle hatchlings.

Changes in activity related oxygen consumption were measured in leatherback and olive ridley sea turtle hatchlings over their first month after emergence from the nest. Leatherbacks emerged with 75-90 KJ of energy in the residual yolk for growth and activity whereas olive ridleys emerged with 45 KJ. In leatherbacks (n=8), resting mass-specific oxygen consumption rates decreased by 53% over the first post-hatching month (0.34+0.03 mL O(2) h(-1) g(-1) to 0.16+0.01 mL O(2) h(-1) g(-1), respectively), while for ridleys (n=8) the fall was 35% (0.20+0.03 mL O(2) h(-1) g(-1) to 0.13+0.01 mL O(2) h(-1) g(-1), respectively). Olive ridley factorial aerobic scope doubled (1.93+0.30 to 3.97+0.51) over the first month but there was no significant increase in leatherback factorial aerobic scope (1.39+0.21 to 1.60+0.13). Leatherback hatchlings gained on average 20% initial body mass (7.68+1.66 g) over the first week, with 70 to 80% of this increase due to water accumulation. Olive ridleys gained 14% (1.83+0.16 g) in initial mass over the first week of age. We propose that the differences in aerobic scope and energy reserves are related to differences in early life ecological stratagems of these species.

Aerobiosis↗

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.

Animals↗

Hearing in the giant sea turtle, Chelonia mydas.

Measurement were made of the cochlear potentials in three specimens of the giant sea turtle, Chelonia mydas, in response to both aerial and mechanical stimulation of the ear. The results show maximum sensitivity in the region of 300 to 400 Hz, with a rather rapid decline in sensitivity for lower and especially for higher tones. The upper limit for the observation of cochlear potentials without injury is 2000 Hz, and a practical limit of usefulness of this ear is probably about 1000 Hz.

Animals↗

Intraocular pressure of juvenile loggerhead sea turtles (Caretta caretta) held in different positions.

The intraocular pressure of 12 apparently healthy juvenile loggerhead sea turtles (Caretta caretta) was determined by applanation tonometry while the turtles were held in dorsoventral, ventrodorsal, and head-down suspended positions. The median intraocular pressures were 5 mmHg (range 4 to 9 mmHg) in the dorsoventral position, 7 mmHg (range 5 to 12 mmHg) in the ventrodorsal position, and 23 mmHg (range 17 to 33 mmHg) in the suspended position.

Animals↗